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  • 1985-1989  (519)
  • 1986  (519)
  • Biochemistry and Biotechnology  (519)
  • 1
    Electronic Resource
    Electronic Resource
    Masthead (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010101
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  • 2
    Electronic Resource
    Electronic Resource
    Editorial (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 1-1 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010102
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  • 3
    Electronic Resource
    Electronic Resource
    Comment: “Dry” enzymes (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 2-3 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010103
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  • 4
    Electronic Resource
    Electronic Resource
    Proteins and peptides in water-restricted environments (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 4-15 
    Details
    ISSN: 0887-3585
    Keywords: membrane biomimetic system ; reverse micelles ; interfacial water ; myelin proteins ; solid enzyme activity ; organic solvents ; biotechnology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010104
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  • 5
    Electronic Resource
    Electronic Resource
    The design, synthesis, and crystallization of an alpha-helical peptide (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 16-22 
    Details
    ISSN: 0887-3585
    Keywords: protein design ; alpha-helical bundle ; x-ray crystallography ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Twelve- and sixteen-residue peptides have been designed to form tetrameric alphahelical bundles. Both peptides are capable of folding into amphiphilic alpha-helices, with leucyl residues along one face and glutamyl and lysyl residues along the opposite face. Four such amphiphilic alpha-helices are capable of forming a noncovalently bonded tetramer. Neighboring helices run in antiparallel directions in the design, so that the complex has 222 symmetry. In the designed tetramer, the leucyl side chains interdigitate in the center in a hydrophobic interaction, and charged side chains are exposed to the solvent. The designed 12-mer(ALPHA-1) has been synthesized, and it forms helical aggregates in aqueous solution as judged by circular dichroic spectroscopy. It has also been crystallized and characterized by x-ray diffraction. The crystal symmetry is compatible with (but does not prove) the design. The design can be extended to a four-alpha-helical bundle formed from a single polypeptide by adding three peptide linkers.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010105
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  • 6
    Electronic Resource
    Electronic Resource
    The design and production of semisynthetic ribonucleases with increased thermostability by incorporation of S-peptide analogues with enhanced helical stability (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 23-33 
    Details
    ISSN: 0887-3585
    Keywords: peptide helix ; protein stability ; framework model of folding ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Recent work has shown that, with synthetic analogues of C-peptide (residues 1-13 of ribonuclease A), the stability of the peptide helix in H2O depends strongly on the charge on the N-terminal residue. We have asked whether, in semisynthetic ribonuclease S reconstituted from S-protein plus an analogue of S-peptide (1-15), the stability of the peptide helix is correlated with the Tm of the reconstituted ribonuclease S. Six peptides have been made, which contain Glu9 → Leu, a blocked α-COO- group (—CONH2), and either Gln11 or Glu11. The N-terminal residue has been varied; its charge varies from +2 (Lys) to -1 (succinyl-Ala). We have measured the stability of the peptide helix, the affinity of the peptide for S-protein (by C.D. titration), and the thermal stability of the reconstituted ribonuclease S.All six peptide analogues show strongly enhanced helix formation compared to either S-peptide (1-15) or (1-19), and the helix content increases as the charge on the N-terminal residue changes from +2 to -1. All six peptides show increased affinity for S-protein compared to S-peptide (1-19), and all six reconstituted ribonucleases S show an increase in Tm compared to the protein with S-peptide (1-19). The Tm increases as the charge on residue 1 changes from +2 to -1. The largest increment in Tm is 6°.The results suggest that the stability of a protein can be increased by enhancing the stability of its secondary structure.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010106
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  • 7
    Electronic Resource
    Electronic Resource
    Stabilization of λ repressor against thermal denaturation by site-directed Gly→Ala changes in α-helix 3 (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 43-46 
    Details
    ISSN: 0887-3585
    Keywords: protein stability ; helix-coil ; mutant ; calorimetry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Oligonucleotide-directed mutagenesis has been used to replace α-helical glycines in the N-terminal domain of λ repressor with alanines. Since alanine is a significantly better helix-forming residue than glycine, these changes were predicted to have a stabilizing effect. We show that the Gly46→Ala substitution, the Gly48→Ala substitution, and the double substitution increase the melting temperature of the N-terminal domain by 3-6°.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010108
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  • 8
    Electronic Resource
    Electronic Resource
    Protein folding kinetics by combined use of rapid mixing techniques and NMR observation of individual amide protons (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 34-42 
    Details
    ISSN: 0887-3585
    Keywords: hydrogen exchange ; BPTI ; folding pathway ; protein dynamics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A method to be used for experimental studies of protein folding introduced by Schmid and Baldwin (J. Mol. Biol. 135: 199-215, 1979), which is based on the competition between amide hydrogen exchange and protein refolding, was extended by using rapid mixing techniques and 1H NMR to provide site-resolved kinetic information on the early phases of protein structure acquisition. In this method, a protonated solution of the unfolded protein is rapidly mixed with a deuterated buffer solution at conditions assuring protein refolding in the mixture. This simultaneously initates the exchange of unprotected amide protons with solvent deuterium and the refolding of protein segments which can protect amide groups from further exchange. After variable reaction times the amide proton exchange is quenched while folding to the native form continues to completion. By using 1H NMR, the extent of exchange at individual amide sites is then measured in the refolded protein. Competition experiments at variable reaction times or variable pH indicate the time at which each amide group is protected in the refolding process. This technique was applied to the basic pancreatic trypsin inhibitor, for which sequence-specific assignments of the amide proton NMR lines had previously been obtained. For eight individual amide protons located in the β-sheet and the C-terminal α-helix of this protein, apparent refolding rates in the range from 15s-1 to 60 s-1 were observed. These rates are on the time scale of the fast folding phase observed with optical probes.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010107
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  • 9
    Electronic Resource
    Electronic Resource
    Focusing of electric fields in the active site of Cu-Zn superoxide dismutase: Effects of ionic strength and amino-acid modification (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 47-59 
    Details
    ISSN: 0887-3585
    Keywords: protein electrostatics ; substrate diffusion ; Poisson-Boltzmann ; electrostatic potentials ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In this paper we report the implementation of a finite-difference algorithm which solves the linearized Poisson-Boltzmann equation for molecules of arbitrary shape and charge distribution and which includes the screening effects of electrolytes. The microcoding of the algorithm on an ST-100 array processor allows us to obtain electrostatic potential maps in and around a protein, including the effects of ionic strength, in about 30 minutes. We have applied the algorithm to a dimer of the protein Cu-Zn superoxide dismutase (SOD) and compared our results to those obtained from uniform dielectric models based on coulombic potentials. We find that both the shape of the protein-solvent boundary and the ionic strength of the solvent have a profound effect on the potentials in the solvent. For the case of SOD, the cluster of positive charge at the bottom of the active site channel produces a strongly enhanced positive potential due to the focusing of field lines in the channel - a result that cannot be obtained with any uniform dielectric model. The remainder of the protein is surrounded by a weak negative potential. The electrostatic potential of the enzyme seems designed to provide a large cross-sectional area for productive collisions. Based on the ionic strength dependence of the size of the positive potential region emanating from the active site and the repulsive negative potential barrier surrounding the protein, we are able to suggest an explanation for the ionic strength dependence of the activity of the native and chemically modified forms of the enzyme.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010109
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  • 10
    Electronic Resource
    Electronic Resource
    A domain of the klenow fragment of Escherichia coli DNA polymerase I has polymerase but no exonuclease activity (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 66-73 
    Details
    ISSN: 0887-3585
    Keywords: protein domain ; polymerase ; 3′-5′ exonuclease ; artificial gene ; expression vector ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The Klenow fragment of DNA polymerase I from Escherichia coli has two enzymatic activities: DNA polymerase and 3′-5′ exonuclease. The crystal structure showed that the fragment is folded into two distinct domains. The smaller domain has a binding site for deoxynucleoside monophosphate and a divalent metal ion that is thought to identify the 3′-5′ exonuclease active site. The larger C-terminal domain contains a deep cleft that is believed to bind duplex DNA. Several lines of evidence suggested that the large domain also contains the polymerase active site. To test this hypothesis, we have cloned the DNA coding for the large domain into an expression system and purified the protein product. We find that the C-terminal domain has polymerase activity (albeit at a lower specific activity than the native Klenow fragment) but no measurable 3′-5′ exonuclease activity. These data are consistent with the hypothesis that each of the three enzymatic activities of DNA polymerase I from E. coli resides on a separate protein structural domain.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010111
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  • 11
    Electronic Resource
    Electronic Resource
    PseqIP: A nonredundant and exhaustive protein sequence data bank generated from 4 major existing collections (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 60-65 
    Details
    ISSN: 0887-3585
    Keywords: computerized data bank ; sequence comparison heuristics ; databank access ; data bank merging ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Four major protein sequence data collections (NBRF-PIR, PSD-Kyoto, PGtrans, and NEWAT) have been merged into a single nonredundant data bank called PseqIP. The data bank entries were automatically matched by a heuristic computer program relying on the fast computation of the number of tetrapeptides shared by two sequences. PseqIP 1.0 includes 6,068 different protein sequences for a total of 1,357,067 residues, representing most of the available sequence information to date. During the course of this work, we found about 600 occurrences course of a protein sequence recorded with a one-amino-acid variation in at least two different data banks. A flat file (ASCII computer-readable format) version of PseqIP 1.0, well-suited for exhaustive homology searches and statistical sequence analysis, is available from our laboratory.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010110
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  • 12
    Electronic Resource
    Electronic Resource
    The galactan-binding immunoglobulin Fab J539: An x-ray diffraction study at 2.6-Å resolution (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 74-80 
    Details
    ISSN: 0887-3585
    Keywords: antibody ; crystal structure ; anti-galactan ; J539 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The crystal structure of the Fab of the galactan-binding immunoglobulin J539 (a mouse IgA,κ) has been determined at a resolution of approximately 2.6 Å by X-ray diffraction. The starting model was that obtained from the real space search described previously (Navia, M.A., Segal, D.M., Padlan, E.A., Davies, D.R., Rao, D.N., Rudikoff, S. and Potter, M. “Crystal structure of galactan-binding mouse immunoglobulin J539 Fab at 4.5 Å resolution.” Proc. Nat. Acad. Sci. USA, 76:4071-4074, 1979). This Fab structure has now been refined by restrained least-squares procedures to an R-value of 19% for the 11,690 unique reflections between 8.0 Å and 2.6 Å. The rms deviation from ideal bond lengths is 0.025 Å. The overall structure differs from McPC603 Fab, another mouse IgA,κ antibody, in that the elbow bend, relating the variable and constant parts of the molecule, is 145° vs. 133° for McPC603. The region of the molecule expected to be the antigen binding site contains a large cavity with two clefts leading away from it. This has been fitted with a model of an oligo-galactan.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010112
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  • 13
    Electronic Resource
    Electronic Resource
    Reciprocal control of retinal rod cyclic GMP phosphodiesterase by its γ subunit and transducin (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 90-99 
    Details
    ISSN: 0887-3585
    Keywords: visual excitation ; rhodopsin ; enzyme regulation ; cyclic nucleotide cascade ; G-proteins ; inhibitory subunit ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The switching on of the cGMP phosphodiesterase (PDE) in retinal rod outer segments by activated transducin (Tα-GTP) is a key step invisual excitation. The finding that trypsin activates PDE (αβγ) by degrading its γ subunit and the reversal of this activation by γ led to the proposal that Tα-GTP activates PDE by relieving an inhibitory constraint imposed by γ (Hurley and Stryer: J. Biol. Chem. 257:11094-11099, 1982). We report here studies showing that the addition of γ subunit also reverses the activation of PDE by Tα-GTP-γS. A procedure for preparing γ in high yield (50-80%) is presented. Analyses of SDS polyacrylamide gel slices confirmed that inhibitorya activity resides in the γ subunit. Nanomolar γ blocks the activation of PDE by micromolar Tα-GTPγS. The degree of activation of PDE depends reciprocally on the concentrations of γ and Tα-GTPγS. γ remains bound to the disk membrane during the activation of PDE by transducin. The binding of γ to the αβ subunits of native PDE is very tight; the dissociation constant is less than 10 pM, indicating that fewer than 1 in 1,700 PDE molecules in rod outer segments are activated in the absence of Tα-GTP.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010114
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  • 14
    Electronic Resource
    Electronic Resource
    Mutant forms of staphylococcal nuclease with altered patterns of guanidine hydrochloride and urea denaturation (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 81-89 
    Details
    ISSN: 0887-3585
    Keywords: protein stability ; protein denaturation ; denatured state ; structural intermediates ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Eleven mutant forms of staphylococcal nuclease with one or more defined amino acid substitutions have been analyzed by solvent denaturation by using intrinsic fluorescence to follow the denaturation reaction. On the basis of patterns observed in the value of m-the rate of change of log Kapp (the apparent equilibrium constant between the native and denatured states) with denaturant concentration - these proteins can be grouped into two classes. For class I mutants, the value of m with guanidine hydrochloride is less than the wild-type value and is either constant or increases slightly with increasing denaturant; the value of m with urea is also less than wild type but shows a marked increase with increasing denaturant concentration, often approaching but never exceeding the wild-type value. For class II mutants, m is constant and is greater than wild type in both denaturants, with the increase being consistently larger in guanidine hydrochloride than in urea. When double or triple mutants are constructed from members of the same mutant class, the change in m is usually the sum of the changes produced by each mutation in isolation. One plausible explanation for these altered patterns of denaturation is that chain-chain or chain-solvent interactions in the denatured state have been modified - interactions which appear to involve hydrophobic groups.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010113
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  • 15
    Electronic Resource
    Electronic Resource
    Masthead (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010201
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  • 16
    Electronic Resource
    Electronic Resource
    A novel method for selective isolation of C-terminal peptides from tryptic digests of proteins by immobilized anhydrotrypsin: Application to structural analyses of the tail sheath and tube proteins from bacteriophage T4 (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 100-107 
    Details
    ISSN: 0887-3585
    Keywords: affinity chromatography ; high-performance liquid chromatography ; bacteriophage T4 tail sheath protein ; bacteriophage T4 tail tube protein ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A novel method useful for selective isolation of the C-terminal peptide from a tryptic digestion mixture of a protein has been developed by taking advantage of a unique property of anhydrotrypsin, which has a strong specific affinity for the peptides containing arginine or lysine at their C-termini. Briefly, peptides produced by tryptic digestion of a protein are fractionated by affinity chromatography on a column of immobilized anhydrotrypsin. The C-terminal peptide is recovered in a breakthrough fraction, which the remainders are adsorbed on the column (unless the protein ends in arginine or lysine). The breakthrough fraction is then subjected to reversed-phase high-perfomance liquid chromatography in order to purify the C-terminal peptide. Using this method, we have successfully isolated the C-terminal peptides from tryptic digests of the sheath protein (gp 18) and the tube protein (gp 19) of bacteriophage T4. The analytical results on these peptides, together with the information on the N-terminal structures of the original proteins and on the nucleotide sequences of genes 18 and 19, allowed us to establish the complete primary structures of the two proteins.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010115
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  • 17
    Electronic Resource
    Electronic Resource
    Protein fluorescence quenching by small molecules: Protein penetration versus solvent exposure (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 109-115 
    Details
    ISSN: 0887-3585
    Keywords: proteins ; protein dynamics ; tryptophan exposure ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Experiments were done to test the thesis that acrylamide and similar small molecules can penetrate into proteins on a nanosecond time scale. The approach taken was to measure the pattern of fluorescence quenching exhibited by quenching molecules differing in molecular character (size, polarity, charge) when these are directed against protein tryptophans that cover the whole range of tryptophan accesibility. If quenching involves protein penetration and internal quencher migration, one expects that larger quenchers and more polar quenchers should display lesser quenching. In fact, no significant dependence on quencher character was found. For proteins that display measurable quenching, the disparate quenchers studied display very similar quenching rate constants when directed against any particular protein tryptophan. For several proteins having tryptophans known to be buried, no quenching occurs. These results are not consistent with the view that the kinds of small molecules studied can quite generally penetrate into and diffuse about within proteins at near-diffusion-limited rates. Rather the results suggest that when quenching is observed, the pathway involves encounters with tryptophans that are partially exposed at the protein surface. Available crystallographic results support this conclusion.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010202
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  • 18
    Electronic Resource
    Electronic Resource
    Translational repression in vitro by the bacteriophage T4 regA protein (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 116-124 
    Details
    ISSN: 0887-3585
    Keywords: translational repressor ; in vitro transcription-translation ; gene expression ; protein purification ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The bacteriophage T4 translational represor regA protein has been purified from an overproducing strain, and its activity has been studied in simple in vitro protein synthesis reactions. RegA protein was found to inhibit the translation of T4 genes 44, 45, and ORF45-1 in a concentration-dependent fashion. Expression of two other T4 genes which are insensitive to regA protein in vivo, genes 32 and 43, was unaffected by the presence of regA protein. Specific inhibition of synthesis of genes 44, 45, and ORF 45-1 proteins was achieved with 5-20 μM concentrations of regA protein, without the addition of any other T4 encoded proteins or cofactors. When in vitro protein synthesis was performed in two steps, uncoupling translation from transcription, regA protein had an inhibitory effect regardless of whether it was added at the initiation of transcription or only at the translation step. This indicates that regA protein functions during the translation step of protein synthesis in vitro in agreement with previous in vivo studies of regA protein.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010203
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  • 19
    Electronic Resource
    Electronic Resource
    Cloning, expression, and nucleotide sequence of livR, the repressor for high-affinity branched-chain amino acid transport in Escherichia coli (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 125-133 
    Details
    ISSN: 0887-3585
    Keywords: regulation ; prokaryotic bacteria ; leucine uptake ; leucyl tRNA corepressor ; cell physiology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The livR gene encoding the repressor for high-affinity branched-chain amino acid transport in Escherichia coli has been cloned from a library prepared from the episome F106. The inserted DNA fragment from the initial cloned plasmid, pANT1, complemented two independent, spontaneously derived, regulatory mutations. Subcloning as well as the creation of deletions with Bal31 exonuclease revealed that the entire regulatory region is contained within a 1.1-kb RsaI-SalI fragment. Expression of the pANT plasmids in E. coli minicells showed that the regulatory region encodes one detectable protein with an apparent molecular weight of 21,000. DNA sequencing revealed one open reading frame of 501 bp encoding a protein with a calculated MW of 19,155. The potential secondary structure of the regulatory protein has been predicted and it suggests that the carboxy terminus may fold into three consecutive alpha helices. These results suggests that the livR gene encodes a repressor which plays a role in the regulation of expression of the livJ and the livK transport genes.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010204
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  • 20
    Electronic Resource
    Electronic Resource
    Peptide and ester synthesis in organic solvents catalyzed by seryl proteases linked to alumina (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 134-138 
    Details
    ISSN: 0887-3585
    Keywords: enzymatic transesterification ; peptide synthesis ; trypsin ; chymotrypsin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Trypsin and α-chymotrypsin were immobilized to alumina-phosphocolamine complex, activated by glutaraldehyde. The immobilized enzymes show a great stability toward organic solvents miscible or immiscible with water. In the presence of a low concentration of water, the immobilized enzymes catalyzed transesterification reactions as well as peptide synthesis. The synthesized peptides were stable toward the immobilized enzymes.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010205
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  • 21
    Electronic Resource
    Electronic Resource
    Isolation and characterization of native human renin derived from Chinese hamster ovary cells (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 139-145 
    Details
    ISSN: 0887-3585
    Keywords: hypertension ; renin production ; mammalian expression ; affinity chromatography ; genetic engineering ; prorenin secretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Transfection of Chinese hamster ovary (CHO) cells with a plasmid containing the cDNA for human preprorenin has provided cell lines that secrete 15 mg of native prorenin per liter of culture medium. Tryptic activation of the prorenin occurs by selective cleavage of the Arg66-Leu67 bond (numbering as in preprorenin). The renin product, purified in a single step and in high yield by affinity chromatography, is fully stable for as long as 8 months when stored in solution at 4°C and pH 6.5. Purity of the renin was judged to be greater than 95% by gel electrophoresis, compositional and N-terminal sequence analyses, and specific enzyme activity. An important aspect of the present work is the development of a direct assay for renin which permits accurate and reproducible evaluation of enzyme units and kinetic parameters. Application of methods described herein, combined with appropriate scale-up fermentation capabilities, provides the means for generating gram quantities of human renin and its zymogen.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010206
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    Electronic Resource
    An algorithm for determining the conformation of polypeptide segments in proteins by systematic search (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 146-163 
    Details
    ISSN: 0887-3585
    Keywords: protein conformation ; energy calculations ; protein modeling ; loop conformation ; surface area ; homologous proteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The feasibility of determining the conformation of segments of polypeptide chain up to six residues in length in globular proteins by means of systematic search through the possibles conformations has been investigated. Trial conformations are generated by using representative sets of φ, ψ, and χ angels that have been derived from an examination of the distributions of these angles in refined protein structures. A set of filters based on simple rules that protein structures obey is used to reduce the number of conformations to a manageable total. The most important filters are the maintenance of chain integrity and the avoidance of tooshort van der Waals contacts with the rest of the protein and with other portions of the segment under construction. The procedure is intended to be used with approximate models so that allowance is made throughout for errors in the rest of the structure. All possible main chains are first constructed and then all possible side-chain conformations are built onto each of these. The electrostatic energy, including a solvent screening term, and the exposed hyrophobic area are evaluated for each accepted conformation. The method has been tested on two segments of chain in the trypsin like enzyme from Streptomyces griseus. It is found that there is a wide spread of energies among the accepted conformations, and the lowest energy ones have satisfactorily small root mean square deviations from the X-ray structure.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010207
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    Electronic Resource
    Electronic Resource
    pH Dependence of 2,3-diphosphoglycerate binding to human hemoglobin Ao at 21.5°C (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 164-175 
    Details
    ISSN: 0887-3585
    Keywords: DPG ; organophosphates ; ligand interactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Rate equilibrium dialysis was used to measure the binding of 2,3-diphosphoglycerate (DPG) to human oxy- and deoxyhemoglobin AO over the range pH 5-9, at 21.5°C. This approach yielded an accurate, precise, and self-consistent set of model-independent association constants. These data were successfully fitted to a thermodynamic model which is fuctionally similar to a Hill equation. The isotherms generated by this fitting procedure appear to intersect at low pH and converge at high pH. This apparent convergence at high pH is consistent with results obtained by oxygen equilibria studies performed under conditions of saturating DPG. These calculated isotherms were used to determine the enhancement of the Bohr effect as a function of pH. These results are consistent with data obtained by pH stat measurements by other investigators.This paper presents the first in a series of studies that will provide a systematic characterization of the interaction between hemoglobin and DPG.
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    URL: http://dx.doi.org/10.1002/prot.340010208
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  • 24
    Electronic Resource
    Electronic Resource
    The restoration of electron micrographs blurred by drift and rotation (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 176-187 
    Details
    ISSN: 0887-3585
    Keywords: drifted micrographs ; rotational blur ; thin sections ; Wiener filtering ; image analysis ; sickle hemoglobin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have investigated the restoration of electron micrographs exhibiting blurring due to drift and rotation. Blurring due to drift arises in micrographs taken of a specimen which is moving relative to the image plane. A related problem is that of rotational blurring which arises in micrographs of thin sections of helical particles viewed in cross section. The twist of the particle within the finite thickness of the section causes the image to appear rotationally blurred about the helical axis.Restoration algorithms were evaluated by applying them to the restoration of blurred model images degraded by additive Gaussian noise. Model images were also used to investigate how an incorrect estimate of the point spread function function describing the blur would effect the restoration. Images were, if necessary, geometrically transformed to a space in which the point spread function of the blur can be considered as linear and space invariant as, under these conditions, the restoration algorithms are greatly simplified. In the case of the rotationally blurred images this procedur was accomplished by transforming the image to polar coordinates.The restoration techniques were successfully applied to blurred micrographs of bacteriophage T4 and crystals of catalase. The quality of the restoration was judged by comparisons of the restored images to undegraded images. Application to micrographs of rotationally blurred cross sections of helical macrofibers of sickle hemoglobin resulted in a reduction in the amount of rotational blurring.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010209
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  • 25
    Electronic Resource
    Electronic Resource
    Activation of retinal rod cyclic GMP-phosphodiesterase by transducin: Characterization of the complex formed by phosphodiesterase inhibitor and transducin α-subunit (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 188-193 
    Details
    ISSN: 0887-3585
    Keywords: G-protein ; phototransduction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The GTP-binding subunit of transducin (Tα) activates the cGMP phosphodiesterase (PDE) of bovine retinal rods by relieving the constraint imposed by the inhibitory subunit PDEγ. We have isolated and characterized the complex Tα.GTPγS-PDEγ formed when Tα is activated by the nonhydrolyzable analog GTPγS. Sedimentation and light-scattering techniques demonstrate that, in contrast to free Tγ.GTPγS, which is soluble, the Tα.GTPγS-PDEγ complex, as well as Tα.GTP-PDEγ, is membrane bound at cytosolic ionic strength. It is eluted from the membrane at low ionic strength as a monomeric and 1:1 stoichiometric complex. The relative affinities of PDEγ for PDEαβ and for Tα.GTP are discussed.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010210
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    Electronic Resource
    Electronic Resource
    Masthead (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010301
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  • 27
    Electronic Resource
    Electronic Resource
    Set of novel, conserved proteins fold pre-messenger RNA into ribonucleosomes (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 195-210 
    Details
    ISSN: 0887-3585
    Keywords: protein domains ; oligomeric assembly ; RNA splicing ; multiple binding sites ; RNP core proteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010302
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  • 28
    Electronic Resource
    Electronic Resource
    Stabilization of the ribonuclease S-peptide α-helix by trifluoroethanol (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 211-217 
    Details
    ISSN: 0887-3585
    Keywords: protein folding ; α-helix stabilization ; peptide structural stability ; circular dichroism ; protein electrostatics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The effects of trifluoroethanol (TFE) on the stability of the α-helix formed by ribonuclease S-peptide, residues 1-19 of ribonuclease A, were studied by measuring circular dichroism as a function of TFE concentration, pH, and temperature. The S-peptide forms an unusually stable α-helix, which is known to be stabilized by TFE. The magnitude of the effect of charged groups on the peptide, manifested by the change in α-helix stability as a function of pH, was not altered significantly by either TFE concentration or temperature, indicating that the lower dielectric constant of TFE is not important in the stabilization of this α-helix. This suggests that the α-helix might be stabilized by many interactions in addition to the effects of charges. The titration curve of circular dichroism vs. TFE concentration appears to be cooperative at 0°C, but becomes progressively less cooperative at temperatures between 25 and 75°C. The properties of the TFE stabilization indicate that TFE might be a useful probe with which to measure the stability of marginally stable peptides and small proteins.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010303
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  • 29
    Electronic Resource
    Electronic Resource
    The DNA replication inhibitor microcin B17 is a forty-three-amino-acid protein containing sixty percent glycine (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 230-238 
    Details
    ISSN: 0887-3585
    Keywords: microcins ; peptide antibiotic ; protein processing ; HPLC ; protein secretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Microcin B17 is a low-molecular-weight protein that inhibits DNA replication in a number of enteric bacteria. It is produced by bacterial strains which harbor a 70-kilobase plasmid called pMccB17. Four plasmid genes (named mcbABCD) are required for its production. The product of the mcbA gene was identified by labelling minicells. The mcbA gene product was slightly larger when a mutation in any of the other three production genes was present. This indicates that these genes are involved in processing the primary mcbA product to yield the active molecule. The mcbA gene product predicted from the nucleotide sequence has 69 amino acids including 28 glycine residues. Microcin B17 was extracted from the cells by boiling in 100 mM acetic acid, 1 mM EDTA, and purified to homogeneity in a single step by high-performance liquid chromatography through a C18 column. The N-terminal amino acid sequence and amino acid composition demonstrated that mcbA is the structural gene for microcin B17. The active molecule is a processed product lacking the first 26 N-terminal residues. The 43 remaining residues include 26 glycines. While microcin B17 is an exported protein, the cleaved N-terminal peptide does not have the characteristic properties of a “signal sequence,” which suggests that it is secreted by a mechanism different from that used by most secreted proteins of E. coli.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010305
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  • 30
    Electronic Resource
    Electronic Resource
    Properties of a second sensory receptor protein in Halobacterium halobium phototaxis (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 239-246 
    Details
    ISSN: 0887-3585
    Keywords: halobacteria ; photosensory receptor ; retinal ; slow-cycling rhodopsins ; sensory rhodopsins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A second slow-cycling retinylidene protein, in addition to slow-cycling (sensory) rhodopsin (SR), can be bleached with hydroxylamine and regenerated with all-trans retinal in photosensory signaling Halobacterium halobium membranes. Flash photolysis shows this protein undergoes a photochemical reaction cycle characterized by photoconversion of its ground state (λmax 480 nm) to a species with λmax ≤ 360 nm, which thermally regenerates the 480-nm species with a t½ of 260 msec at 25°C, under conditions in which SR photocycles at 650 msec in the same membranes. Mutants characterized with respect to their phototaxis behavior are identified which contain SR and the 480-nm pigment, the latter ranging from undetectable to a concentration equal to that of SR. Receptor mutants lacking all phototaxis sensitivity lack both of the photochemically reactive proteins. The mutant properties contribute to an accumulation of behavioral and spectroscopic evidence that the 480-nm pigment is a second sensory photoreceptor in H. halobium. NaDodSO4-polyacrylamide gel electrophoresis of [3H]retinal-labeled membrane proteins from the mutants indicates SR and the 480-nm pigment contain distinct chromophoric polypeptides differing in their migration rates. The data implicate polypeptides of 25,000 Mr and 23,000 Mr as retinal-binding polypeptides of SR and the 480-nm protein, respectively.
    Additional Material: 8 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010306
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  • 31
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    A very short peptide makes a voltage-dependent ion channel: The critical length of the channel domain of colicin E1 (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 218-229 
    Details
    ISSN: 0887-3585
    Keywords: colicin E1 ; site-directed mutagenesis ; ion channel ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cleavage of colicin E1 molecules with a variety of proteases or with cyanogen bromide (CNBr) generates COOH-terminal fragments which have channel-forming activity similar to that of intact colicin in planar lipid bilayer membranes. The smallest channel-forming fragment obtained by CNBr cleavage of the wild-type molecule consists of the C-terminal 152 amino acids. By the use of oligonucleotide-directed mutagenesis, we have made nine mutants along this 152 amino acid peptide, in which an amino acid was replaced by methionine in order to create a new CNBr cleavage site. The smallest of the CNBr-cleaved C-terminal fragments with channel-forming activity, in planar bilayer membranes, was generated by cleavage at new Met position 428 and has 94 amino acids, whereas a 75 amino acid peptide produced by cleavage of a new Met at position 447 did not have channel activity. The NH2-terminus of the channel-forming domain of colicin E1 appears therfore to lie between residues 428 and 447. Since, however, the last six C-terminal residues of the colicin can be removed without changing activity, the number of amino acids necessary to form the channel is 88 or less. In addition, the unique Cys residue in colicin E1 was replaced by Gly, and nine mutants were then made with Cys placed at sequential locations along the peptide for eventual use as sulfhydryl attachment sites to determine the local environment of the replaced amino acid. In the course of making 21 mutants, eight charged residues have been replaced by uncharged Met or Cys without changing the biological activity of the intact molecule.It has been proposed previously that the conformation of the colicin E1 channel is a barrel formed from five or six α-helices, each having 20 amino acids spanning the membrane and two to four residues making the turn at the boundary of the membrane. Our finding that 88 amino acids can make an active channel, combined with recently reported stoichiometric evidence that the channel is a monomer excludes this model and adds significant constraints which can be used in building a molecular model of the channel.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010304
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  • 32
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    Kinetic characterization of early intermediates in the folding of E. coli tryptophan-synthase β2 subunit (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 247-255 
    Details
    ISSN: 0887-3585
    Keywords: protein folding ; domain interactions ; fluorescence transfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This report describes the use of fluorescence energy transfer between an intrinsic energy donor (tryptophan 177) and two chemically added acceptors to study intermediates in the folding of the β2 subunit of E. coli tryptophan-synthase. Two early folding steps are thus identified and characterized. One is very rapid (its rate constant at 12°C is 0.02 sec-1) and corresponds to the folding of the N-terminal domain into a structure whose overall features approximate well those of the native domain. The second step is somewhat slower (its rate constant at 12°C is 0.008 sec-1) and involves a conformational rearrangement of the N-terminal domain brought about by the interactions between the N-and C-terminal domains within a monomeric β chain. This brings to five the number of intermediates which have been identified and ordered on the folding pathway of the dimeric β2 subunit.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010307
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  • 33
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    Preliminary x-ray diffraction analysis of HhaII endonuclease-DNA cocrystals (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 263-266 
    Details
    ISSN: 0887-3585
    Keywords: protein-DNA interaction ; overproducer clone ; sequence-specific recognition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: HhaII restriction endonuclease purified from an overproducing recombinant E. coli clone has been cocrystallized with a heptanucleotide duplex, d-GGAGTCC:GGACTCC. The cocrystals are monoclonic and belong to the space group C2. The unit cell dimensions are a = 199.0±1.0 Å, b = 100.0±0.5 Å, c = 80.3±0.4 Å, and β = 101.0±1.0°. There appear to be two dimers per asymmetric unit and the crystals diffract to 4-Å resolution.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010309
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  • 34
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    Calculating three-dimensional changes in protein structure due to amino-acid substitutions: The variable region of immunoglobulins (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 267-279 
    Details
    ISSN: 0887-3585
    Keywords: molecular model building ; energy minimization ; homology modeling ; site-specific mutagenesis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A procedure (coupled perturbation procedure, CPP) is introduced as a specific method for calculating the detailed three-dimensional structure of a protein molecule which has a nummber of amino-acid substitutions relative to some previously determined “parent” protein structure. The accuracy of the procedure is tested by calculating the conformation of a region of the human immunoglobulin fragment Fab Kol based on the analogous region of the human immunoglobulin fragment Fab New. Both structures have previously been determined crystallographically. The calculated model is accurate to the extent that both of the sequence differences in the region are modeled correctly and that conformational changes in a number of nearby residues are correctly identified. CPP is shown to give better results than other commonly used modeling procedures when applied to the same problem.
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    URL: http://dx.doi.org/10.1002/prot.340010310
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  • 35
    Electronic Resource
    Electronic Resource
    Proteolytic dissection of a hapten binding site (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 256-262 
    Details
    ISSN: 0887-3585
    Keywords: FV fragment ; space filling hapten ; reconstitution ; scratched analysis ; equilibrium dialysis ; contact residues ; hypervariable loops ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: IgG Gar, a human myeloma protein that binds riboflavin with a high affinity, was used to derive variable region fragments from the heavy chain and the light chain. Riboflavin binding ability of the active site generated by V(H) and light chain and the active site generated by V(H) and V(L) was compared to riboflavin binding by the F(ab) fragment. The riboflavin binding ability of the F(ab) fragment is the same as the intact molecule, while the binding ability of the active site formed by V(H) and light chain is lowered by two to three orders of magnitude, indicating that the removal of C(H1) domain decreases the interaction between riboflavin and the amino acids that is important in tight binding of riboflavin. Removal of the third hypervariable region and the constant region domain from the light chain further lowers the binding constant by one order of magnitude. The results indicate that the V(H) and V(L) segments of IgG Gar can reconstitute a riboflavin binding site. The decrease in affinity probably reflects a decrease in the rigidity with which the hypervariable loops are held together to place the contact amino acid residues in optimal contact with the hapten.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010308
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  • 36
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    Affinity of nonhomologous amphiphilic peptides toward a monoclonal antibody raised against apolipoprotein A-I (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 280-286 
    Details
    ISSN: 0887-3585
    Keywords: antibody affinity ; ELISA ; β-endorphin models ; melittin model ; amphiphilic α-helix ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Monoclonal antibodies against human apolipoprotein A-I (apoA-I) were generated by the hybridoma technique. Clone G-10 was selected on the basis of its highest titer. The affinity of this antibody toward a series of synthetic peptides differing in length, amino acid composition, and amphiphilicity was tested by using both the indirect and the competitive enzyme-linked immunosorbent techniques (ELISA). From these measurements we calculated dissociation constants of the complexes of the antibody with apoA-I bound to the surface of the microtiter plate, apoA-I in solution, and any of the several peptides in solution. The dissociation constant (Kd) of the immobilized apoA-I/anti-apoA-I-complex, Kd = 2 × 10-9 M, was significantly lower than that of the complex resulting from the interaction between anti-apoA-I and either apoA-I in solution or any of the several amphiphilic helical peptides in solution. Peptides devoid of amphiphilic secondary structure were inert. These data are consistent with the proposal that monoclonal G-10 recognizes in antigenic peptides an α-helical secondary structure of defined hydrophilic-lipophilic balance and comparatively less the specific amino acid side chains. We propose that the highest contribution to the free energy of binding (8 Kcal/mole) is derived from the docking of the helix to the antibody. It follows that in probing the specificity of a monoclonal antibody the conformation and the physical environment of the interacting antigen must be taken into account.
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    URL: http://dx.doi.org/10.1002/prot.340010311
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  • 37
    Electronic Resource
    Electronic Resource
    Masthead (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010401
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  • 38
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    Editorial (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. i 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010402
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  • 39
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    Isolation and analysis of arc repressor mutants: Evidence for an unusual mechanism of DNA binding (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 302-311 
    Details
    ISSN: 0887-3585
    Keywords: mutant proteins ; protein stability ; operator binding ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have isolated 64 different missense mutations at 36 out of 53 residue positions in the Arc repressor of bacteriophage P22. Many of the mutant proteins with substitutions in the C-terminal 40 residues of Arc have reduced intracellular levels and probably have altered structures or stabilities. Mutations in the N-terminal ten residues of Arc cause large decreases in operator DNA binding affinity without affecting the ability of Arc to fold into a stable three-dimensional structure. We argue that these N-terminal residues are important for operator recognition but that they are not part of a conventional helix-turn-helix DNA binding structure. These results suggest that Arc may use a new mechanism for sequence specific DNA binding.
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    URL: http://dx.doi.org/10.1002/prot.340010404
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  • 40
    Electronic Resource
    Electronic Resource
    Molecular structure and evolution of adrenergic and cholinergic receptors (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 287-301 
    Details
    ISSN: 0887-3585
    Keywords: primary sequence homology ; primary structure ; secondary structure ; quaternary structure ; gene cloning ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010403
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    Primary structure of the reaction center from Rhodopseudomonas sphaeroides (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 312-325 
    Details
    ISSN: 0887-3585
    Keywords: membrane proteins ; nucleotide sequence ; evolution ; photosynthesis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The reaction center is a pigmentprotein complex that mediates the initial photochemical steps of photosynthesis. The amino-terminai sequences of the L, M, and H subunits and the nucleotide and derived amino acid sequences of the L and M structural genes from Rhodopseudomonas sphaeroides have previously been determined. We report here the sequence of the H subunit, completing the primary structure determination of the reaction center from R. sphaeroides. The nucleotide sequence of the gene encoding the H subunit was determined by the dideoxy method after subcloning fragments into single-stranded M13 phage vectors. This information was used to derive the amino acid sequence of the corresponding polypeptide. The termini of the primary structure of the H subunit were established by means of the amino and carboxy terminal sequences of the polypeptide. The data showed that hte H subunit is composed of 260 residues, corresponding to a molecular weight of 28,003. A molecular weight of 100,858 for the reaction center was calculated from the primary structures of the subunits and the cofactors. Examination of the genes encoding the reaction center shows that the codon usage is strongly bviased towards codons ending in G and C. Hydropathy analysis of the H subunit sequence reveals one stretch opf hydrophobic residues near the amino terminus; the L and M subunits contain five such stretches. From a comparison of the sequences of homologous proteins found in bacterial reaction centers and photosystem II of plants, an evolutionary tree was contructed. The analysis of evolutionary relationships showed that the L and M subunits of reaction centers and D1 and D2 proteins of photosystem II are descended from a common ancestor, and that the rate of change in these proteins was much higher in the first billion years after the divergence of the reaction center and photosystem II than in the subsequent billion years represented by the divergence of the species containing these proteins.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010405
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    Sequence conservation and region shuffling in an endoglucanase and an exoglucanase from Cellulomonas fimi (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 335-341 
    Details
    ISSN: 0887-3585
    Keywords: cellulases ; active sites ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cellulomonas fimi produces an endoglucanase and an exoglucanase which bind strongly to cellulose. Each enzyme contains three distinct regions: a short sequence of about 20 amino acids containing only proline and threonine (the ProThr box); an irregular region, rich in hydroxyamino acids, of low charge density, and which is predicted to have little secondary structure; and an ordered region of higher charge density which contains a potential active site, and which is predicted to have secondary structure; and an ordered region of higher charge density which contains a potential active site, and which is predicted to have secondary structure. The Pro-Thr box is conserved almost perfectly in the two enzymes. The irregular regions are 50% conserved, and the conserved sequences include four Asn-Xaa-Ser/Thr sites. The ordered regions appear not to be conserved, but the potential active sites both have the sequence Glu-Xaa7-Asn-Xaa6-Thr; they occur at widely separated sites in the two regions. The order of the regions is reversed in the two enzymes: irregular-Pro-Thr box-ordered in the endoglucanase; ordered-Pro-Thr box irregular in the exoglucanase. The genes for the two enzymes appear to have arisen by shuffling of two conserved sequences and either one or two other sequences.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010407
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    Proteases of enhanced stability: Characteization of a thermostable variant of subtilisin (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 326-334 
    Details
    ISSN: 0887-3585
    Keywords: thermal stability ; protein engineering ; mutagenesis ; plate assay ; thermophilic enzymes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A Procedure has been developed for the isolation and identification of mutants in the bacterial serine protease subtilisin that exhibit enhanced thermal stability. The cloned subtilisin BPN'gene from Bacillus amyloliquefaciens was treated with bisulfite, a chemical mutagen that deaminates cytosine to uracil in single-stranded DNA. Strains containing the cloned, mutagenized subtilisin gene which produced subtilisin with enhanced thermal stability were selected by a simple plate assay procedure which screens for esterase activity on nitrocellulose filters after preincubation at elevated temperatures. One thermostable subtilisin variant, designated 7150, has been fully characterized and found to differ from wild-type subtilisin by a single substitution of Ser for Asn at position 218. The 7150 enzyme was found to undergo thermal inactivation at onefourth the rate of the wild-type enzyme when incubated at elevated temperatures. Moreover, the midpoint in the thermally induced transition from the folded to unfolded state was found to be 2.4-3.9°C higher for 7150 as determined by differential scanning calorimetry under a variety of conditions. The refined, 1.8-Å crystal structures of the wild-type and 7150 subtilisin have been compared in detail, leading to the conclusion that slight improvements in hydrogen bond parameters in the vicinity of position 218 result in the enhanced thermal stability of 7150.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010406
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  • 44
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    Predicting antibody hypervariable loop conformations II: Minimization and molecular dynamics studies of MCPC603 from many randomly generated loop conformations (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 342-362 
    Details
    ISSN: 0887-3585
    Keywords: antibodies ; immunoglobulins ; conformation prediction ; energy minimazation ; random stranting conformations ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We describe a method for predicting the conformations of loops in proteins and its application to four of the complementarity determining regions [CDRs] in the crystallographically determined structure of MCPC603. The method is based on the generation of a large number of randomly generated conformations for the backbone of the loop being studied, followed by either minimization or molecular dynamics followed by minimization starting from these random structures. The details of the algorithm for the generation of the loops are presented in the first paper in this series (Shenkin etal.[submitted]). The results of minimization and molecular dynamics applied to these loops is presented here. For the two shortest CDRs studied (H1 and L2, which are five and seven amino acids long), minimizations and dynamics simulations which ignore interactions of the loop amino acids beyond the carbon ture closely. This suggests that these loops fold independently of sequence variation. For the third CDR (L3, which is nine amino acids), those portions of the CDR near its base which are hydrogen bonded to framework are well replicated by our procedures, but the top of the loop shows singificant conformational variability. This variability persists when side chain interactions for the MCPC603 sequence are included. For a fourth CDR (H3, which is 11 amino acids long), new low-energy backbone conformations are found; however, only those which are close to the crystal are compatible with the sequence when side chain interactions are taken into account. Results from minimuzation and dynamics on single CDRs with all other CDRs removed are presented. These allow us to explore the extent to which individual CDR conformations are determined by interactions with framework only.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010408
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  • 45
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    Multiple conformations of amino acid residues in ribonuclease A (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 370-375 
    Details
    ISSN: 0887-3585
    Keywords: enzyme structure ; disorder ; refinement ; high resolution ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The highly refined 1.26 Å structure (R = 0.15) of phosphate-free bovine pancreatic ribonuclease A was modeled with 13 residues having discrete multiple conformations of side chains. These residues are widely distributed over the protein surface, but only one of them, Lys 61, is involved in crystal packing interactions. The discrete conformers have no unusual torsion angles, and their interactions with the solvent and with other atoms of the protein are similar to those residues modeled with a single conformation. For three of the residues-Val 43, Asp 83, and Arg 85-two correlated conformations are found. The observed multiple conformations on the protein surfaces will be of significance in analyzing structure-function relationships and in performing protein engineering.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340010410
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  • 46
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    Electronic Resource
    Structural features of ribonucleotide reductase (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 376-384 
    Details
    ISSN: 0887-3585
    Keywords: ribonucleotide reductase ; unique N-terminal domain ; nucleotide binding site ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Herpes simplex virus type 1 (HSV-1) encodes a ribonucleotide reductase which comprises two polypeptides with sizes of 136,000 (RR1) and 38,000 mol. wt. (RR2). We have determined the entire DNA sequence specifying HSV-1 RR1 and have identified two adjacent open reading frames in varicella-zoster virus (VZV) which have homology to HSV RR1 and RR2; the predicted sizes for the VZV RR1 and RR2 polypeptides are 87,000 and 35,000 mol. wt. respectively. Amino acid comparisons with RR1 and RR2 polypeptides from other organisms indicate that HSV-1 RR1 contains a unique N-terminal domain which is absent from other RR1 polypeptides apart from HSV-2 RR1. These N-terminal amino acid sequences are poorly conserved between HSV-1 and HSV-2 in contrast to the remainder of the protein which shows greater than 90% homology. Polypeptide structural predictions suggest that the HSV-1 N-terminal domain may be separated into two regions, namely, a β-sheet structure followed by a nonstructured area. Across the remainder of RR1 and RR2, comparisons also reveal blocks of amino acids conserved between the different ribonucleotide reductases, and these may be important for enzyme activity. From predictions on the structure of these conserved blocks, we have proposed that the location of a substrate binding site within RR1 is centered on three conserved glycine residues in a region which is predicted to adopt a β-sheet/turn/α-helical structure;this approximates to the structure for ADP nucleotide binding folds. Finally, we propose that the promoters for the HSV and Eptein-Barr virus (EBV) RR2 transcripts have evolved by separate evolutionary routes.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010411
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  • 47
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    Primary structure of a precursor to the asprartic proteinase from Rhizomucor miehei shows that the enzyme is synthesized as a zymogen (1986)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 363-369 
    Details
    ISSN: 0887-3585
    Keywords: cDNA library ; disulphide bridges ; prosegment homology ; mRNA structure ; N-glycosylation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In order to characterize the zymogen of the milk-clotting enzyme from Rhizomucor miehei, we constructed a cDNA library on pBR327 in Escherichia coli. Aspartic proteinase-specific recombinants were isolated by colony hybridization to a specific oligonucleotide mixture, and the cDNA sequence corresponding to a precursor form of the enzyme was determined.The decuced amino sequence shows that this secreted fungal proteinase is synthesized as a precursor. The first 22 amino acid residues in this precursor constitute a typical signal peptide. The amino acid sequence of the following 47-amino-acid-long prosegment shows homology to the prosegments from both the extracellular and intracellular vertebrate aspartic proteinases, and to the prosegments from the yeast and Mucor pusillus aspartic proteinases as well. These observations suggest that all aspartic proteinases are synthesized with a prosegment and that this prosegment is essential for the correct folding of all the mature enzymes. The active Rhizomucor miehei enzyme consists of 361 amino acide residues with a total molecular weight of 38,701. Clusters of idendtities around the active site cleft support the assumption that these proteinasess have a common folding of their peptide chains. The disulphide bridges were localized in the fungal enzyme, and 2 N-glycosylation sites were identified.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340010409
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  • 48
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    The application of a novel heat flux calorimeter for studying growth of Escherichia coli W in aerobic batch culture (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1780-1793 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The modification and principle of a novel heat flux calorimeter for the in situ, on-line measurement of the heat generated during microbial growth is described. Data concerning the physical characterization of the calorimeter as a fermentor, including stability and sensitivity of the heat signal, are presented. The calorimeter has been successfully applied to the study of the aerobic batch culture of Escherichia coli W on glucose under carbon and nitrogen limitation. A direct correlation between growth and heat evolution was obtained. Quantitative analysis of the data suggests that the new calorimetric technique could be used for monitoring growth and specific metabolic events, for convenient medium optimization, and as a basis for a novel fermentation process control system.
    Additional Material: 12 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260281205
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  • 49
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    A method for estimating the rejection coefficient-molecular weight relationship of ultrafiltration membrane for a chain polymer by using gel permeation chromatography (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1809-1813 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An improved method is presented for estimating rejection coefficient-molecular weight relationship of an ultrafiltration membrane for a polydisperse chain polymer. It is based on the basic idea using gel permeation chromatography originally developed by Cooper and Van Derveer. The method, in which peak spreading of an elution curve of the polymer was taken into consideration, is available for evaluating the relationship over a wide range of the molecular weight through only one experiment in analyses of the retentate and filtrate.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281208
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  • 50
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    Production of penicillin in a fluidized-bed bioreactor using a carrier-supported mycelial growth (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1838-1844 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A carrier-supported mycelial growth of Penicillium chrysogenum was applied to penicillin fermentation system using celite as a support material. Hyphal growth through the pore matrices of the material showed strong anchorages and provided highly stable biofilm growth. With bioparticles developed in such a manner, both cell growth and penicillin production were observed to increase significantly compared to the conventional dispersed filamentous cultures. Maximum values of specific penicillin production rate were found to be constant regardless of the growth form. A three-phase fluidized-bed fermentor was designed and tested for penicillin production using the bioparticles. Two modes of operation, semicontinuous and repeated fed batch, of the fermentor were tried. It was noted that the overgrowth of free mycelia and the development of fluffy loose bioparticles caused poor mixing and made the fermentor operation quite difficult. Control of the bioparticle size and the extension of production phase were therefore considered important to maintain the reactor productivity at a desired level. From the results of repeated fed-batch operation it was found that the control of bioparticle size could be successfully achieved by phosphate-limiting culture condition. Penicillin production under this condition was also observed to be maintained at a high level (about 80% of the maximum) for at least 1 month.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281211
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  • 51
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    Pretreatment of wheat straw for fermentation to methane (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1857-1866 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of pretreating wheat straw with gamma-ray irradiation, ammonium hydroxide, and sodium hydroxide on methane yield, fermentation rate constant, and loss of feedstock constituents were evaluated using laboratory-scale batch fermentors. Results showed that methane yield increased as pretreatment alkali concentration increased, with the highest yield being 37% over untreated straw for the pretreatment consisting of sodium hydroxide dosage of 34 g OH-/kg volatile solids, at 90°C for 1 h. Gamma-ray irradiation had no significant effect on methane yield. Alkaline pretreatment temperatures above 100°C caused a decrease in methane yield. After more than 100 days of fermentation, all of the hemi-cellulose and more than 80% of the cellulose were degraded. The loss in cellulose and hemicellulose accounted for 100% of the volatile solids lost. No consistent effect of pretreatments on batch fermentation rates was noted. Semicontinuous fermentations of straw-manure mixtures confirmed the relative effectiveness of sodium and ammonium-hydroxide pretreatments.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281213
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  • 52
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    Enhancement of growth and ferrous iron oxidation rates of T. Ferrooxidans by electrochemical reduction of ferric iron (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1867-1875 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Thiobacillus ferrooxidans, the bacterium most widely used; in bioleaching or microbial desulfurization studies, was grown in an electrolytic bioreactor containing a synthetic, ferrous sulfate medium. Passage of current through the medium reduced the bacterially generated ferric iron to the ferrous iron substrate. When used in conjunction with an inoculum that had been adapted to the electrolytic growth conditions, this technique increased the protein (cell) concentration by 3.7 times, increased the protein (cell) production rate by 6.5 times, increased the yield coefficient (cellular efficiency) by 8.0 times, and increased the ferrous iron oxidation rate by 1.5 times at 29°C, compared with conventional cultivation techniques. A Monod-type equation with accepted values for the maximum specific growth rate could not account for the increased growth rate under electrolytic conditions.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281214
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  • 53
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    Electrochemical ion control in batch cultures of Saccharomyces cerevisiae NCYC 1018 (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1884-1888 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281217
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  • 54
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    Monitoring of protein product formation during fermentation with fast protein liquid chromatography (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 16-20 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A method is described using fast protein liquid chromatography (FPLC) for the monitoring of protein formation during fermentation. The procedure consists of centrifugation to recover the cells, sonication of the cells, centrifugation to remove cell debris, and analysis of supernatant on a column of Mono Q (a strong anion exchanger). Analysis of peak areas provides quantitative determination of product concentration. Maintenance and life of the Mono Q column is discussed. We find that FPLC is a convenient method for measuring products in cell homogenates because it gives rapid, highly resolved separations.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280104
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  • 55
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    Performance evaluation of an anoxic/oxic activated sludge system: Effects of mean cell residence time and anoxic hydraulic retention time (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 7-15 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A laboratory investigation has been undertaken to asses the effects of two operating parameters, mean cell residence time (MCRT) and anoxic hydraulic retention time (HRT), on the performance of an anoxic/oxic activated sludge system. The performance of the system was evaluated in terms of its COD, nitrogen, and biomass characteristics. An activated sludge system is capable of producing a better effluent, in terms of COD and nitrogen characteristics, when it is operated in an anoxic/oxic fashion. A longer MCRT and an adequate anoxic HRT are desirable in the operation of an anoxic/oxic activated sludge system. For the wastewater used in this investigation, the anoxic/oxic unit was capable of producing an effluent with the following characteristics when it was operated at MCRT = 20 days, total system HRT = 10 h, and anoxic HRT = 3-5 h: COD = 15 mg/L; VSS = 10 mg/L; TKN = 1.30 mg/L; NH3 - N = 0.60 mg/L; and NO2 + NO3 - N = 5.0 mg/L. A uniform distribution of biomass is achievable in an anoxic/oxic activated sludge system because of the intensive recirculation/convection maintained. The provision of an anoxic zone in the aeration tank promotes a rapid adsorption of feed COD into the biomass without an immediate utilization for cell synthesis. This, in turn, results in a high microbial activity and a lower observed biomass yield in the system. A tertiary treatment efficiency is achievable in an anoxic/oxic activated sludge system with only secondary treatment operations and costs. A conventional activated sludge system can be easily upgraded by converting to the anoxic/oxic operation with minor process modifications.
    Additional Material: 10 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280103
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  • 56
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    The production of cellulase in a spouted bed fermentor using cells immobilized in biomass support particles (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 41-50 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Continuous cellulase production by Trichoderma viride QM 9123, immobilized in 6 mm diameter, spherical, stainless steel biomass support particles, has been achieved using a medium containing glucose as the main carbon source. Experiments were carried out in a 10-L spouted bed fermentor. In this type of reactor-recycled broth is used to create a jet at the base of a bed of particles, causing the particles to spout and circulate. During the circulation, particles pass through a region of high shear near the jet inlet. This effectively prevents a buildup of excess biomass and thus enables steady-state conditions to be achieved during continuous operation. Continuous production of cellulase was achieved at significantly higher yield and productivity than in conventional systems. At a dilution rate of 0.15 h-1 (nominal washout rate for freely suspended cells is 0.012 h-1), the yield of cellulase on glucose was 31% higher than that measured during batch operation, while the volumetric productivity (31.5 FPA U/L· h) was 53% greater than in the batch system. The specific cellulase productivity of the immobilized cells was more than 3 times that of freely suspended cells, showing that diffusional limitations can be beneficial. This offers significant opportunity for the further development of biomass support particles and associated bioreactors.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280107
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  • 57
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    Urokinase bound to fibrocollagenous tubes: An in vitro kinetic study (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 58-63 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Urokinase (UK) has been immobilized to the inner surfaces of fibrocollagenous tubes (FCT) in an attempt to develop a fibrinolytic biomaterial which may be suitable for use as a small diameter vascular prosthesis. The enzyme was bound by adsorption followed by glutaraldehyde crosslinking. An in virto kinetic study of immobilized urokinase was conducted by employing the tubular material as a flow through reactor operated in a batch recycle mode in which the esterolysis of the model substrate, N-α-acetyl-L-lysine methyl ester (ALME), was monitored as a function of substrate concentration, recycle flow rate, and temperature. Results were compared with data from the soluble enzyme reaction, which was conducted in the presence and absence of 10% swine skin gelatin, in order to identify the specific effects of a collagenous microenvironment. Observed rates for the UK-FCT catalyzed reaction were observed to be dependent on recycle flow rates below 12 mL/min (Re = 107). Apparent Michaelis-Menten rate parameters were determined by a nonlinear search technique for two flow rates: one above the critical point for external diffusion effects (Re = 282) and one within the mass-transfer-limited region (Re = 71). When the latter data were corrected for external diffusion by applying the Graetz correlation for laminar flow in tubes to estimate themass transfer coefficient, the corrected Km of 6.45 ± 0.38 mM agreed very closely with the diffusion free parameter (i.e. 6.13 ± 0.63). Furthermore, this value was observed to be an order of magnitude higher than that of the soluble enzyme but approximately equal to the Km of the soluble enzyme in a 10% gelatin environment (8.13 ± 1.53 mM). It is postulated that the difference in kinetic parameters between soluble and collagen immobilized UK is due to an inherent interaction between collagen and enzyme rather than to mass transfer effects. Such aninteraction is supported by the effects of collagen on thermal stability and energy of activation.
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    URL: http://dx.doi.org/10.1002/bit.260280109
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    Membrane separation of protein precipitates: Unstirred batch studies (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 88-96 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The study examines the use of ultrafiltration and microfiltration membranes for concentrating isoelectric soya protein. Experiments with an unstirred batch cell indicate that the flux is limited by the protein which remains in solution after precipitation of the major proportion. The porosityof the precipitate cake formed is shown to be a second important factor. A significant improvement in flux can be obtained by using membranes which permit passage of the soluble protein and by increasing the precipitate particle size. The results are shown to be within the range predicted theoretically by the two limiting cases of a particulate model and a soluble protein model.
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    URL: http://dx.doi.org/10.1002/bit.260280112
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  • 59
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    An automatic and sterilizable sampler for laboratory fermentors: Application to the on-line control of glucose concentration (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 119-121 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Absract.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280119
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  • 60
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    Release of cellulases from penicillium janthinellumNCL Communication No. 3517. (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 129-132 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280122
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  • 61
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    Acid hydrolysis of Jerusalem artichoke for ethanol fermentation (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 138-141 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280124
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  • 62
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    Masthead (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986) 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280201
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  • 63
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    Maltodextrin hydrolysis in a fluidized-bed immobilized enzyme reactor (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 151-159 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The present work deals with maltodextrin hydrolysis by glucoamylase immobilized onto corn stover in a fluidized bed reactor. An industrial enzyme preparation was covalently grafted onto corn stover, yielding an activity of up to 372 U/g and 1700 U/g for support particle sizes of 0.8 and 0.2 mm, respectively. A detailed kinetic study, using a differential reactor, allowed the characterization of the influence of mass transfer resistance on the reaction catalyzed by immobilized glucoamylase. A simple and general mathematical model was then developed to describe the experimental conversion data and found to be valid.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280202
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  • 64
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    The relationship between hydrogen gas and butanol production by Clostridium saccharoperbutylacetonicum (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 305-310 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two simultaneous fermentations were performed at 26°C with simultaneous inocula using Clostridium saccharoperbutylacetonicum. Fermentation 1 prevented the gas formed by the biomass from escaping the fermentor while 2 allowed the gas formed to escape. Fermentor 1 provided for the production of butanol, acetone, and ethanol, while when the H2 formed was allowed to escape with fermentor 2, neither butanol nor acetone were produced. Ethanol was also formed in both fermentors and began along with the initial growth of biomass and continued until the fermentations were complete. Butanol and acetone production began after biomass growth had reached a maximum and began to subside. The butanol-acetone-ethanol millimolar yields and ratios were 38:1:14 respectively. The fermentor 2 results show that a yield of 2.1 L H2, 93 or 370 mmol H2/mol glucose, was formed only during the growing stage of growth; neither butanol nor acetone were produced; ethanol was formed throughout the fermentation, reaching a yield of 15.2 mmolar. It appears that hydrogen gas is required for butanol production during the resting stage of growth.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280302
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  • 65
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    A computer model for the growth and differentiation of a fungal colony on solid substrate (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 405-416 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mold growth and differentiation are closely related to the formation of secondary products. In solid-substrate fermentations this interrelationship is often more completely realized than in submerged cultures. Solid substrate reactions are used commercially in a limited manner in the western world, but are relatively common in Asia. Basic studies in solid-substrate fermentation should yield results applicable to all types of commercial mold fermentations for the production of a secondary product. This paper presents a relatively simple model for the growth of a mold colony on a solid surface with a defined medium utilizing glucose. Unlike submerged cultures the model must account for both cellular differentiation and the spatial heterogeneity in the system. Model parameters were estimated independently using literature values. The results of the simulation studies suggest that mass transfer limitations are at least partially responsible for the proliferation of differentiated structures on solid substrates as compared to liquid cultures. Since the concentration profile depends on the depth of the substratum, conditions that enhance conidia production can be achieved by controlling the depth of the solid medium.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280314
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  • 66
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    Heat and microbial treatments for nutritional upgrading of wheat straw (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 381-386 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The ligninolytic activities of four cellulolytic organisms were compared using straw. Only Aspergillus japonicus and Polyporous versicolor appreciably degraded lignin with A. japonicus yielding the most protein. In solid culture, most protein was produced by P. versicolor, closely followed by A. japonicus. Pretreatment of the straw by hot water facilitated biodegradation and protein production. The nutritional value of the residual straw was also increased by some fungal cultures. The greatest amount of degradable polysaccharide in the straw was made available by A. japonicus in liquid media and Pleurotus ostreatus in solid media.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280311
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  • 67
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    Evaluation of startup and operation of four anaerobic processes treating a synthetic meat waste (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 372-380 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two continuous stirred tanks reactors (CSTR) and four anaerobic fluidized bed reactors (AFBR) were used to study the treatment of a synthetic meat waste during single-and two-stage anaerobic treatment. Four configurations were investigated; a single-stage CSTR and AFBR and the two-stage systems CSTR-AFBR and AFBR-AFBR. Startup of the anaerobic reactors was achieved within 50 days by use of a regime that included stepped increases in influent COD, methanol substitution of the substrate, and addition of essential trace metals such as cobalt and nickel. Two-stage reactors removed up to 85% of influent COD concentrations of 5000 mg/L, whereas the single-stage AFBR and CSTR removed 76 and 9%, respectively. The proportion of methane in the effluent gases increased as the influent COD concentration was increased. Volumetric production of methane was greatest for the first stage of the AFBR-AFBR system. Solids retention times calculated for the AFBRs ranged from 7 to 12 days, sufficient to support methanogenesis. The AFBRs and two-stage systems were more resistant to an influent pH shock from the operating value of pH 6.8 down to pH 3 than the CSTRs and single-stage reactors. It was concluded that high-rate anaerobic treatment systems were applicable to meat industry wastewaters and that two-stage digestion produced a better quality effluent.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280310
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  • 68
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    Enzymatic analyses in organic solvents (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 417-421 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Horseradish peroxidase has been found to vigorously act as a catalyst in a number of water-immiscible organic solvents. The rates of peroxidase-catalyzed oxidation of p-anisidine with H2O2 in toluene, benzene, ethyl and butyl acetates, and ether are in the range of 10-25% of that in water (pH 7.0) at the same reactant concentrations. Per oxidase was coupled with cholesterol oxidase (which was also found to be catalytically active in organic media), and the bienzymic system was successfully used for accurate, reliable, and reproducible determination of cholesterol in toluene.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280315
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  • 69
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    Membrane separation of protein precipitates: Studies with cross flow in hollow fibers (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 422-431 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hollow fiber ultrafiltration and microfiltration membranes are examined for the processing of isoelectric soya protein precipitate suspensions. A model based on the various resistances to permeate flux is used to describe membrane performance. The main resistance to permeate flux is due to the interaction between the active membrane and the soluble and precipitated protein; that is, as compared with resistances due to the active membrane itself or the membrane support structure, or arising from concentrated soluble or precipitated protein layers over the membrane surface. Soluble protein rejection and precipitate mean particle diameter are correlated with observed values of this main resistance.In contract to the ultrafiltration of soluble proteins, the flux rates observed when processing protein precipitate suspensions under a similar range of operating conditions do not approach a limiting value with increased transmembrane pressure. At high protein concentrations, greater flux rates may be achieved for precipitated as compared with soluble proteins. The use of a microfiltration membrane does not give further improvement in flux rate; this may be attributed to problems of pore fouling with precipitate particles.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280316
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  • 70
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    Pressure drop in a packed bed with a liquid of variable viscosity: The case of dextrin hydrolysis by immobilized glucoamylase (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 452-455 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The activity of an immobilized enzyme in a packed bed is monitored for the change in a substrate conversion with time. But pressure drop in the packed bed with immobilized glucoamylase can serve as an indirect indicator for the changes in the conversion and activity of the immobilized enzyme. The method is simple and the change can be monitored continuously. This method can be generally applicable to systems where the viscosity of a substrate changes with its conversion.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280318
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  • 71
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    Masthead (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986) 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280401
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  • 72
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    Control of nitrate concentration in fermentations of Corynebacterium glutamicum (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 659-662 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A nitrate control system has been devised for the maintenance of stable nitrate concentrations throughout fed-batch fermentations of Corynebacterium glutamicum. The feedback control system was based on the use of a nitrate-ion-selective electrode to directly monitor the nitrate levels in the fermentor and an automatic controller to activate a nitrate feed pump. The electrode which was used for controlling the nitrate level was stable through-out the fermentation period. The apparent maximum specific growth rate, biomass production, protein production, biomass yields on glucose and nitrate, and amino acid production were all optimal at approximately 50mM nitrate.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280504
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  • 73
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    Sugar cane bagasse as a possible source of fermentable carbohydrates. II. Optimization of the xylose isomerase reaction for isomerization of xylose as well as sugar cane bagasse hydrolyzate to xylulose in laboratory-scale units (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 684-699 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Both the forward and backward reactions of xylose isomerase (Sweetzyme Q) with xylose and glucose as substrates have been studied in terms of kinetics and thermodynamics. The relationship between the two reactions can thus be determined. Much attention has been given to the reaction with xylose as substrate. The optimal conditions of the xylose reaction in terms of pH, buffer, metal ions, substrate concentration, temperature, and ionic strength have been determined. These findings did not differ much from those reported for the glucose reaction. Equilibrium constants for the aldose to ketose conversion were more favorable in the case of glucose. The results obtained with continuous isomerization of xylose in columns packed with either Sweetzyme Q or Taka-Sweet were very similar to those obtained from batch isomerization processes. Particle size had a definite effect on reaction rate, which indicates that diffusion limitations do occur with the immobilized enzyme particles. Heat stability of Sweetzyme Q was good with t1/2 of 118, 248, and 1200 h at 70, 55, and 40°C, respectively. A novel method for the separation of xylose-xylulose mixtures with water as eluant on a specially prepared Dowex 1 × 8 column was developed. This technique has the capability of producing pure xylulose for industrial or research applications. A writ for a patent regarding this technique is at present prepared.
    Additional Material: 16 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280508
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  • 74
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    Granulation of biomass in thermophilic upflow anaerobic sludge blanket reactors treating acidified wastewaters (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 718-727 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The development of granular sludge in thermophilic (55°C) upflow anaerobic sludge blanket reactors was investigated. Acetate and a mixture of acetate and butyrate were used as substrates, serving as models for acidified waste-waters. Granular sludge with either Methanothrix or Methanosarcina as the predominant acetate utilizing methanogen was cultivated by allowing the loading rate to increase whenever the acetate concentration in the effluent dropped below 200 and 700 mg COD/L, respectively. The highest methane generation rates, up to 162 kg CH4-COD/m3 day, or 2.53 mole CH4/L day, were achieved at hydraulic retention times down to 21 min, with granules consisting of Methanothrix. The formation of Methanothrix granules did not depend on the type of seed material, nor on the addition of inert support particles. The growth of granules proceeded rapidly with adapted seed material, even when the reactors were inoculated with low concentrations. With mesophilic seed materials growth of granules took much longer. Thermophilic Methanothrix granules strongly resemble mesophilic granules of the “filamentous” type. Some factors governing the thermophilic granulation process are discussed.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280511
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  • 75
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    Particle size effects in the microbiological leaching of sulfide concentrates by thiobacillus ferrooxidans (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 751-755 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280516
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  • 76
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    A mathematical model for cell separation technique of centrifugal elutriation (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 570-577 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model for the cell separation technique of centrifugal elutriation is developed. The model simulates both steady and non-steady-state operation of the elutriator. The model can be used to predict the required set of flow rates of elutriating liquid necessary to fractionate a cell culture, the required time of sampling before steady state is achieved, and the range of cell size/cell density combinations contained in any fraction. The model predictions were verified experimentally. Variations in cell density and cell size due to the suspending environment have a significant effect on the accuracy (although not the trends) of the model predictions. Quantification of these variations will lead to significantly more accurate model predictions. An enhanced separation method was developed using the model, to yield finer separation of a cell culture than previously possible. The use of the centrifugal elutriator may now be given a firm theoretical basis, with the quality of separation understood in terms of the basic theory of operation.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280414
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  • 77
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    Lipid-enhanced ethanol production from xylose by Pachysolen tannophilus (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 605-608 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280418
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  • 78
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    Hexokinase activity as marker to assess time of harvest of foot-and-mouth disease virus in BHK21 Cl13 cell cultures (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 613-615 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hexokinase (B.C. 2.7.1.1) activity as a marker enzyme during FMD viral infection has been observed spectrophotometrically in a system coupled with glucose-6-phosphate dehydrogenase, in supernatants of BHK21Cl13 suspension as well as anchored cell culture at a minimum of 104 infective virus particles/ml. Specific activity increased with virus concentration in culture supernatants and abruptly decreased with a fall in virus titer, as has been noted by TCID/50,146 S concentration, and enzyme-linked immunosorbent assay (ELISA) readings.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280420
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  • 79
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    Immobilization of trypsin on alginic acid-poly (glycidyl methacrylate) graft copolymer (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 609-612 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Trypsin was immobilized onto alginic acid-poly(glycidyl methacrylate) graft copolymer (AAGMA). The resulting immobilized enzyme showed 65% of the soluble enzymatic activity. The temperature optimum was shifted by 5°C to a higher value. The pH optimum of immobilized enzyme has also been shifted by 0.5 units toward the alkaline side when compared to that of soluble enzyme. The pH stability and thermal stability are better than that of soluble enzyme.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280419
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  • 80
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    Synergism of endoenzyme and exoenzyme on hydrolysis of soluble cellulose derivatives (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 878-882 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A kinetic model that represents the reaction of hydrolysis of water-soluble cellulose derivatives by a mixed endo- and exoenzyme system is proposed with the following assumptions: at an early stage of the reaction, endoenzymes split the substrate molecule in order to supply the newly formed nonreducing ends to exoenzymes until the molecular weight of the substrates reaches a low value; after that point, the reaction kinetics obeys only the rate equation of the reaction of the exoenzymes in which the reaction parameters change linearly with decrease of the molecular weight of the substrates. Hydrolysis experiments of soluble cellulose derivatives, carboxymethyl cellulose and hydroxyethyl cellulose, were carried out with endo-and exoenzymes separated from Trichoderma Koningii cellulase. The critical molecular weight of the substrate, from that point the action of endoenzyme can be neglected, was determined from the experimental data. That was ca. 4000 D. With that value, the model fits well the experimental data. Synergism of both enzymes appears as enhancement of the rate of the reaction at the early stage of the reaction.
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    URL: http://dx.doi.org/10.1002/bit.260280615
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  • 81
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    Induction and elimination of oscillations in continuous cultures of Saccharomyces cerevisiae (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 700-710 
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    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Continuous cultures of Saccharomyces cerevisiae are known to exhibit oscillatory behavior in the oxidative region. Important findings of a series of experiments conducted to identify the causes for initiation of and the means for elimination of oscillations in these cultures are reported in this paper. These oscillations are seen to be connected to the growth kinetics of the microorganism and are induced at very low glucose concentrations and at dissolved oxygen (DO) levels that are neither high nor low (DO values between 20 and 78% air saturation at a dilution rate of 0.2 h-1 and pH of 5.5 at 30°C). The oscillatory behavior is encountered over a range of dilution rates (0.09-0.25 h-1 at 30°C for pH = 5.5 and DO = 50% air saturation). The oscillations can be eliminated by raising the DO level above a critical value or by lowering the DO level below a critical value.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/bit.260280509
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    Masthead (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986) 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280701
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    Growth of Saccharomyces cerevisiae is controlled by its limited respiratory capacity: Formulation and verification of a hypothesisDedicated to Prof. A. Fiechter on the occasion of his 60th birthday. (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 927-937 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A novel mechanistic model for the growth of baker's yeast on glucoseis presented. It is based on the fact that glucose degradation proceeds via two pathways under conditions of aerobic ethanol formation. Part is metabolized oxidatively and part reductively, with ethanol being the end product of reductive energy metabolism. The corresponding metabolic state is designated oxidoreductive. Ethanol can be used oxidatively only. Maximum rates of oxidative glucose and ethanol degradation are governed by the respiratory capacity of the cells. The model is formulated by using the stoichiometric growth equations for pure oxidative and reductive (fermentative) glucose and ethanol metabolism. Together with the experimentally determinable yield coefficients (YX/S) for the respective metabolic pathways, the resulting equation system is sufficiently determined. The superiority of the presented model over hitherto published ones is based on two essential novelities. (1) The model was developed on experimentally easily accessible parameters only. (2) For the modeling of aerobic ethanol formation, the substrate flow was split into two simultaneously operating (i.e., in parallel) metabolic pathways that exhibit different but constant energy-generating efficiencies (respiration and fermentation) and consequently different and constant biomass yields (YX/S). The model allows the prediction of experimental data without parameter adaption in a biologically dubious manner.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280620
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  • 84
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    Optimal control of substrate concentrations in bioreactors with separated sensors (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 952-959 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Many of the sophisticated sensors desirable for monitoring bioreactors cannot be placed in the bioreactor either because they are not steam sterilizable or because they require nonphysiological operating conditions. Such sensors can be used if they are separated from the bioreactor. Separation of the sensor from the bioreactor causes a time lag in data acquisition. This results in several complexities in the development of an appropriate and stable feedback control system based on a separated sensor. This paper analyzes the optimal control of a bioreactor with a separated sensor without a time lag and analyzes the feedback control (but not necessarily the optimal control) with a time lag. Simulation results indicate that this type of analysis could be extended to more general bioreactor operating conditions.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280704
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  • 85
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    Vapor liquid equilibrium behavior of aqueous ethanol solution during vacuum coupled simultaneous saccharification and fermentation (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 972-976 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The data on ethanol-water vapor-liquid equilibrium in the presence of cellulase enzyme, nutrients, yeast, and rice straw indicated a substantial increase in ethanol concentration in vapor phase at reduced pressures. Maximum relative volatility of ethanol in the presence of added components is approximately twice that of a pure ethanol-water system. The equation correlating the activity coefficient and ethanol concentration in the liquid phase adequately represents the equilibrium behavior.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280707
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  • 86
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    Breakthrough behavior of 17.5 mol% water in methanol, ethanol, isopropanol, and t-butanol vapors passed over corn grits (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 960-964 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Ground corn is now used in industry as an adsorbent to remove water from ethanol vapors. It is stable and inexpensive at 10 cents/lb (22 cents/kg). For regeneration it requires less than 2000 Btu/gal of 190 proof ethanol processed. If necessary, it could be readily saccharified and fermented into ethanol after use. This renewable resource has further exciting potential as an inexpensive adsorbent for water removal from other alcohols, including methanol, isopropanol, and t-butanol. Water sorption capacity in a fixed bed, nonisothermal adsorption column appears to be a function of the heat capacity of the non-adsorbed alcohol vapor, relative to the heat capacity of the corn adsorbent. Methanol, ethanol, isopropanol, and t-butanol containing 17.5 mol% water gave 105,151, 284, and 358 g anhydrous product/kg adsorbent, respectively, per adsorption cycle. This adsorbent, having operational temperature ranges between 80 and 100°C, is indicated to be of potential utility in solvent recycle processes using these industrially important alcohols. Observed adsorption characteristics are discussed in terms of the alcohol properties of molecular size, heat capacity, and diffusivity. The adsorption mechanism is hypothesized to include transport of water molecules into the structure of adjacent starch molecules present in small spherical bodies (diameter of several microns) immobilized on the surface of the corn grit particles.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280705
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  • 87
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    Photosynthesis in modulated light: Quantitative dependence of photosynthetic enhancement on flashing rate (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 988-995 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In order to predict the potential benefit associated with mixing devices designed to introduce periodic light modulations in dense cultures of microalgae, it is necessary to develop a quantitative understanding of the relationship between the frequency of the modulations and the resulting photosynthetic efficiency enhancement. To explore this relationship, the photosynthetic rate of cells of Phaeodactylum tricornutum from a dense steady state culture was determined as a function of modulation frequency, intensity of light received, and the proportion of the total cycle period during which the cells were illuminated. At high flash frequencies, the photosynthetic rate was determined by the average intensity received by the cells (full light intensity integration), while at low frequencies the cells responded to the instantaneous intensity (no light intensity integration). Full integration was approached asymptotically with increasing flash frequency. The frequency response could be described by a rectangular hyperbola, and the parameters of this hyperbola were nearly independent of the illumination intensity and the flash proportion. The saturation constant of the hyperbola, at which the response is one-half of the maximum, was 0.67 Hz.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280709
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  • 88
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    Enzymatic saccharification of pretreated rice straw and biomass production (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1503-1509 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A comparative study on the saccharification of pretreated rice straw was brought about by using cellulase enzyme produced by Aspergillus terreus ATCC 52430 and its mutant strain UNGI-40. The effect of enzyme and substrate concentrations on the saccharification rate at 24 and 48 were studied. A syrup with 7% sugar concentration was obtained with a 10% substrate concentration for the mutant case, whereas a syrup with 6.8% sugar concentration was obtained with 3.5 times concentrated enzyme from the wild strain. A high saccharification value was obtained with low substrate concentration; the higher the substrate concentration used, the lower the percent saccharification. The glucose content in the hydrolysate comprised 80-82% of total reducing sugars; the remainder was cellobiose and xylose together. The hydrolysate supported the growth of yeasts Candida utilis and Saccharomyces cerevisiae ATCC 52431. A biomass with a 48% protein content was obtained. The essential amino acid composition of yeast biomass was determined.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281008
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  • 89
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    Optimal substrate feeding policy for a fed batch fermentation with substrate and product inhibition kinetics (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1421-1431 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The optimal substrate feeding policy for the fed batch fermentation which is governed by product and substrate inhibited kinetics is presented. The conjunction point between nonsingular and singular arcs and the feeding policy along the singular arc are derived analytically in terms of the concentrations of substrate and product and the liquid volume. Thus, it is possible to determine the feeding rate by monitoring the state variables (i.e., closed loop control). As a specific example, an optimization study of the fed batch fermentation for ethanol production by Saccharomyces cerevisiae is presented. It is shown that the optimal feeding patterns are heavily dependent upon the initial conditions. The point selectivity provides the guideline for predicting the optimal feeding patterns and explaining the results of rigorous mathematical analysis.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260280916
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  • 90
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    Deactivation of free and stabilized acid phosphatase by urea (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1647-1652 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Tests on acid phosphatase (E.G. 3.1.3.2) deactivation by urea have been performed at two pH values. Two conditions have been used: native enzyme operating batch-wise in dilute solution and stabilized enzyme in continuous flow ultrafiltration membrane reactor. Stabilization is achieved by confining the enzyme within a concentrated solution of a linear chain polymer that forms a polarization layer over the membrane. The results provide significant information on the kinetics and thermodynamics of the complex phenomena taking place during deactivation. Deactivation by urea is also compared with thermal deactivation.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281108
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  • 91
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    An integral dynamic model for the UASB reactor (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1621-1636 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this article a dynamic model of a continuous working UASB reactor is described. It results from the integration of the fluid flow pattern in the reactor, the kinetic behavior of the bacteria (where inhibition and limitation were taken into account), and the mass transport phenomena between different compartments and different phases. The mathematical equations underlying the model and describing the important mechanisms were programmed and prepared for computations and simulations by computer. The settler efficiency has to be over 99% to prevent the reactor from wash-out. When the settler efficiency is over 99%, the total sludge content of the reactor increases steadily, so the reactor is hardly ever in a steady state. This implies dynamic modeling. The model is able to predict the various observable and nonobservable or difficult to observe state variables, e.g., the sludge bed height, the sludge blanket concentration, the short-circuiting flows over bed and blanket, and the effluent COD concentration as a function of the hydrodynamic load, COD load, pH, and settler efficiency. The optimal pH value is between 6.0 and 8.0; fatty acid shock loadings are difficult to handle outside this optimal pH range.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281106
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  • 92
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    Substrate conditions that influence the assays used for determining the β-glucosidase activity of cellulolytic microorganisms (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1653-1656 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Culture filtrates from Trichoderma harzianum E58, T. reesei CL 847 and Penicillium sp. C 462 were assayed for β-glucosidase activity using a range of substrates and sugar analysis methods. Although sugar analyses by the dinitrosalicylic acid (DNS) and Nelson-Somogyi methods gave a similar profile, when increasing concentrations of salicin were assayed, considerably higher values were obtained with the DNS assay. The salicin concentration used for the assay greatly influenced the final β-glucosidase values with higher values obtained for T. harzianum E58 and T. reesei CL 847 at substrate concentrations of 1 mg/mL while optimum values for Penicillium sp. C 462 were obtained at substrate concentrations greater than 3 mg/mL. Low concentrations of salicin and p-nitro-phenyl-β-D-glucopyranoside (PNPG) gave the same response as cellobiose. Cellobiose should be used at concentrations greater than 3.74 mg/mL to avoid substrate limitation of the β-glucosidase assay.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281109
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  • 93
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    Kinetics and mechanism of dissimilative Fe(III) reduction by Pseudomonas sp. 200 (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1657-1671 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The kinetics and mechanism of Fe(III) reduction to Fe(II) were studied in pure batch cultures of Pseudomonas sp. 200. The rate of iron reduction has been mechanistically related to aqueous phase iron speciation. In the absence of microbial activity the iron reduction rate was negligible. Initial rates of microbial iron reduction were accelerated more than 20-fold by the addition of equimolar quantities of nitrilotriacetic acid (NTA) to media initially containing 1.86 × 10-3M total Fe(III). Numerical techniques were utilized to quantify relationships between the observed rate of Fe(II) production and the calculated (equilibrium) aqueous phase speciation. These results indicate that soluble ferric iron species are not equivalent in terms of their susceptibility to bacterial (dissimilative) iron reduction. The concentration of Fe(NTA)(OH)22- correlated strongly with observed iron reduction rates. Ferrous iron species appeared to inhibit the reduction process.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281110
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  • 94
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    Phosphorylation of yeast protein: Reduction of ribonucleic acid and isolation of yeast protein concentrate (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1690-1698 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Yeast nucleoproteins were chemically phosphorylated with phosphorus oxychloride (POCL3). Studies using 31P nuclear magnetic resonance (NMR) spectroscopy, stability to pH and lysine estimation all indicated that the ∊-amino group of lysine was the principal functional group phosphorylated. Phosphorylation of ca. 30% of the lysine residues resulted in removal of more than 85% of contaminant ribonucleic acid from protein precipitated at pH 4.2. Phosphorylation did not alter the amino acid composition of yeast proteins and was reversible under acidic conditions. Based on the data, a method for the preparation of phosphorylated yeast protein with low levels of nucleic acid is proposed.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281112
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  • 95
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    Mechanistically detailed model of cellular metabolism for glucose-limited growth of Escherichia coli B/r-A (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1672-1689 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A structured mathematical model for cellular metabolism in Escherichia coli has been extended to encompass the mechanistic structure surrounding the kinetics and control of transcription and translation. The dependence of transcription on RNA polymerase and the mechanism of translation initiation have been explicitly included. This model correctly simulates cell growth, cell composition, and the timing of chromosome synthesis as a function of extracellular substrate concentration for glucose-limited balanced growth. Simulation results for the subpopulation of RNA polymerase engaged in transcription and for the distribution of this subpopulation among different promoter sites agree closely with experimental findings, as do calculated estimates of the active ribosomal fraction. In addition, the existence of an antitermination system for transcription of stable RNA operons is supported by model results. This model should provide a useful framework for investigating metabolic perturbations to E. coli, such as those resulting from insertion of extra-chromosomal vectors into the cells.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281111
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  • 96
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    Electronic Resource
    Modeling active mass in aerobic sludge digestion (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1699-1706 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Aerobic digestion of waste-activated sludge was carried out in lab-scale reactors for both batch and semicontinuous flow patterns. The reactors were monitored at three different temperatures: 10, 20, and 30°C. During the course of digestion, significant solubilization of volatile suspended solids was observed, and its effect on the magnitude of kinetic coefficients was examined. Differences in metabolic activity and sludge stabilization were found between batch and semicontinuous flow patterns. An Arrhenius-type relationship was not found to apply to rate constants for the semicontinuous reactors.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281113
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  • 97
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    A model for the filterability of activated sludge supported by mixed-liquor biochemical data (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1801-1808 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A predictive model was developed to estimate the dewatering characteristics of waste-activated sludges. This model utilizes the COD-nitrogen ratio of the wastewater and the organic loading rate of the process to predict sludge filterability in terms of specific resistance. A completely mixed, continuous flow secondary treatment process with solids recycle was used for the cultivation of activated sludges. The sludge wasted from this process was used in Buchner funnel specific resistance determinations. The basic concepts involved in the development of the model were supported by sludge carbohydrate, protein, and surface charge data.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281207
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  • 98
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    Coimmobilization of malate dehydrogenase and formate dehydrogenase in polyethyleneglycol(#4000)diacrylate gel by droplet gel-entrapping method (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1794-1800 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A new immobilization technique suitable for coupled enzymes requiring cofactors was established. This is a droplet gel-entrapping method in which many small droplets including the enzymes are fixed in the gel. The first emulsion was prepared by mixing of a solution containing thermostable malate dehydrogenase (MDH) and formate dehydrogenase (FDH) with benzene containing a surfactant. The first emulsion was added to a solution containing polyethyleneglycol(#4000)diacrylate and N,N′-methylenebisacrylamide to prepare the second emulsion (w/o/w). After the second emulsion was gelled by addition of potassium persulfate and 3-dimethylaminopropionitrile, the benzene was removed. The expressed MDH and FDH activities of the MDH-FDH immobilized gel were 7.1 and 13.9% of the initial activities, respectively. The Km values of the gel were 0.60mM for formate and 1.5μM for NAD, respectively. The Km for formate and NAD were found to be extremely low. By using the column packed with 30 g gel having the MDH activity of 41.7 units and the FDH activity of 11.1 units, 13.8mM oxalacetate was completely converted to malate at 30°C. The malate production rate was not affected by the concentration of more than 50mM formate, more than 2mM oxalacetate, and more than 0.1 mM NAD, respectively. Long-term malate production was demonstrated at 30°C by passing the substrate solution containing the two substrates and NAD through the column. The maximum conversion ratio (7.8%) was obtained at the fifth day, and 83% of maximum productivity was maintained even after 3 weeks. The expressed FDH activity at the fifth day was calculated to be 20.5% of the initial activity.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281206
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  • 99
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    Mode of action and properties of xylanase and β-Xylosidase from Neurospora crassaNCL Communications No. 3772. (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1832-1837 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Extracellular β-xylosidase (1,4-β-D-xylan xylohydrolase, EC 3.2.1.37) from culture filtrates of Neurospora crassa was purified to homogeneity by preparative isoelectric focusing followed by gel electrophoresis. The molecular weight of the purified xylosidase was 83,000 D and the Km on p-nitrophenyl-β-D-xyloside was 0.047mM. The homogeneous xylanase (1,4-β-D-xylan xylanohydrolase, EC 3.2.1.8) and β-xylosidase showed differences in their mode of action towards xylooligosaccharides. The degree of hydrolysis of D-xylan by xylanase of N. crassa was 18%. Supplementation of β-xylosidase from the same organism resulted in 48% hydrolysis. The synergistic effect was more pronounced, with the hydrolysis of 68%, when a homogeneous preparation of β-xylosidase from Sclerotium rolfsii was added to the saccharification system.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281210
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  • 100
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    Flow and oxygen transfer in a plunging water jet system using inclined short nozzles and performance characteristics of its system in aerobic treatment of wastewater (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 28 (1986), S. 1845-1856 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: For the plunging water jet system using inclined short nozzles, the flow characteristics such as the bubble penetration depth and the gas entrainment rate, which changed depending on the jet velocity, the nozzle diameter, the jet length, and the jet angle were first evaluated in an air-water system. A comparable investigation between our results and those of existing studies used the long nozzles on those characteristics revealed that both the bubble penetration depth and the gas entrainment rate differed depending on the nozzle length; that is, the nozzle-length-to-diameter ratio LN/DN and that of these characteristics the gas entrainment rate affected considerably by its magnitude and tended to be high when the nozzle of a large LN/DN ratio was used. It was also confirmed from the oxygen transfer experiments that the transfer efficiency at low jet velocities in the present water jet system was not inferior to the ones of other types of existing aeration systems; that is, the utilization of this jet aeration system to a high rate reactor for wastewater treatment or fermentation was sufficiently possible. The applicability of the plunging jet aeration method to microbial processes was then examined. As a typical example of microbial processes to be tested, the continuous treatment of an organic wastewater using activated sludge microorganisms was carried out, and the performance and related problem when this type of aeration system was applied to such a microbial process were investigated. Experimental results showed that, when viewed from the removal ability of dissolved organic matters, the plunging jet aeration system was capable of treating a wastewater of considerable high loading without the rate of oxygen transfer becoming the biooxydation-rate-limiting factor. Special attention was necessary for the choice of the liquid pump to be employed, however, due to the increased amount of fine suspended solids in the treated water caused by the shearing action between sludge flocks and pump blades.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260281212
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