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  • 1985-1989  (457)
  • 1985  (457)
  • Biochemistry and Biotechnology  (457)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 171-176 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hemicellulosic sugars, predominantly D-xylose, comprise about one-half the total carbohydrate that can be obtained from hardwoods and agricultural residues through dilute acid hydrolysis. Because rates and yields in the xylose fermentation are low, economic utilization of these materials as fermentation feedstocks is difficult. Pachysolen tannophilus formed 5.5% ethanol from 12% glucose but only 2% ethanol from 12% xylcose. Aeration doubled the specific rate of D-glucose fermentation by P. tannophilus, as compared to anaerobic fermentation, but the specific rate of the xylose fermentation remained unchanged. Periodic additions of 0.5% D-glucose to aerobic fermentations of 3% xylose increased the yield of ethanol from 0.28 g/g xylose to greater than 0.41 g/g xylose utilized. The rate of xylose utilization remained unchanged, and radiotracer studies showed that addition of 0.5% glucose did not inhibit xylose utilization under aerobic or anaerobic conditions. No enhancement was observed anaerobically, nor was enhancement observed with acid hydrolysates, apparently because of the presence of acetic acid which inhibited growth and fermentation.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 182-186 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mathematical analyses of a diffusion-limited hollow fiber reactor for the measurement of effective substrate diffusivities are presented. An analytical solution to the mathematical model with a first order substrate consumption rate is used to show that the procedure of Webster and Shuler1 is incorrect. A rigorous analysis that requires numerical solution is also outlined for any form of the substrate consumption rate. These analyses allow for more accurate estimations of effective substrate diffusivities since they should be used in conjunction with integral reactor behavior.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 177-181 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Cellulose samples from cotton and wood pulps with varying low degrees of crystallinity (mechanically decrystallized) were studied. The influence of initial cellulose crystallinity on sugar yield after enzymatic hydrolysis was determined by two different methods. As expected, samples with low crystallinity were much more accessible to enzymatic attack and glucose yields were higher than were samples of high initial crystallinity. Hydrolysis of cellulose seems more dependent on cellulose crystallinity than on the source of cellulose. It is known that decrystallized or amorphous cellulose can recrystallize under proper conditions, e.g., during acid hydrolysis. The data reported here also reveal some recrystallization during enzymatic hydrolysis which probably occurs simulataneously with a selective enzymatic attack on the amorphous regions of cellulose. In all cases, the amorphous celluloses recrystallized in the original lattice form, that of native cellulose.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 187-191 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Stable steady-state growth of Escherichia coli B limited by succinate, phosphate, or sulfate ion over the range of specific growth rates of 0.025-0.51 h-1 was achieved using pH-controlled auxostasis in the phauxostat. The concentration of the growth-limiting substrate in the growth vessel could be varied at will in the region of the Monod half-maximal saturation constants by adjusting the concentration of that substrate in the reservior (at constant buffering capacity) or by varying the population density of the culture through changing the buffering capacity of the medium in the reservoir.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 208-213 
    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.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 192-201 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Automatic constant-value control of mineral ions was attempted in semibatch culture of high cell mass concentration (more than 150 g dry cell/L) with ethanol and ammonia feeds. Equations were derived from the mass balance principle to calculate the required concentration of each mineral ion in the mineral feed solution, taking into account both the decrease in the volume of the culture supernatant as a proportion of the whole culture broth and the increase in the volume of the whole culture broth during the cultivation. The mineral solution was supplied automatically, linked either with ethanol feed or ammonia water feed. The actual concentrations of mineral ions could be kept within small variations. To adjust the supplementation in accordance with the culture change from oxygen sufficiency (early growth phase) to oxygen deficiency (later growth phase), the concentration of each mineral ion was altered stepwise when the dissolved oxygen concentration fell to zero. The mineral supplementation gave better results coupled with ethanol feed than with ammonia feed. The mineral ions studied were K+, Mg2+, Na+, Fe2+, Zn2+, Ca2+, Co2+, Cu2+, Mn2+, NH+4, PO43- and SO42-.
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  • 7
    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 process for the production of extracellular carboxymethylcellulase (CMCase) and xylanase by fermentation under nonaseptic or nonsterile conditions is described. The fermentation process is carried out under very acidic conditions of pH 2.0 by using a acidophilic cellulolytic fungus. Microbial contamination is avoided or minimized to an insignificant level under this acid pH condition. The culture medium for this production consists of a carbon source from cellulosics or lignocellulosics, such as Na-CMC, xylan, Avicel cellulose, cellulose powder, α-cellulose, sawdust, etc., or a mixture of the forementioned together with simple ingredients such as (NH4)2SO4, K2HPO4, MgSO4 and NaNO3. The fermentation is carried out at room temperature (28-30°C), under aerobic conditions, and without controlling the pH. The CMCase and xylanase produced are stable under very simple storage conditions, such as in the fresh culture medium not containing the substrate for a period of 3 days, at any temperature from 0 to 30°C. These extracellular enzymes have an optimum pH around 3, with the best range of pH from 2.0 to 3.6, for any temperature between 15 and 60°C. The optimum temperatures are 55°C for CMCase activity and 25-50°C for xylanase activity, at any pH between 2.0 and 5.2. The apparent Michaelis constants Km are 2.6 and 1.5 mg/mL for CMCase and xylanase of the culture filtrate, respectively.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 463-470 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The enzymatic hydrolysis of cellulose to glucose involves the formation of cellobiose as an intermediate. It has been found necessary1 to add cellobiase from Aspergillus niger (NOVO) to the cellobiase component of Trichoderma reesei mutant Rut C-30 (Natick) cellulase enzymes in order to obtain after 48 h complete conversion of the cellobiose formed in the enzymatic hydrolysis of biomass. This study of the cellobiase activity of these two enzyme sources was undertaken as a first step in the formation of a kinetic model for cellulose hydrolysis that can be used in process design. In order to cover the full range of cellobiose concentrations, it was necessary to develop separate kinetic parameters for high- and low-concentration ranges of cellobiose for the enzymes from each organism. Competitive glucose inhibition was observed with the enzymes from both organisms. Substrate inhibition was observed only with the A. niger enzymes.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 471-481 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Three strains of Erwinia rhapontici especially suitable for use in the form of nongrowing immobilized cells were selected by screening strains of cells for high activity and operational stability in an immobilized form. Immobilization in calcium alginate gel pellets was easily the best method of immobilizing E. rhapontici. Much greater operational stabilities were obtained than when other immobilization methods were used. Conditions of operation which optimize the activity, stability, and yield and the ease of operation of the immobilized cell columns working in a steady state are described. These include the effects of substrate concentration, diffusional restrictions and water activity, the concentration of cells immobilized, and the type of reactor used. Thus, the immobilized cells produce about 1500 times their own weight of isomaltulose during one half-life of use (ca. 1 year). Loss of activity was most closely correlated with the volume of substrate processed and so presumably is due to the presence of low concentrations of a cummulative inhibitor in the substrate. Methods for regenerating the activity of the immobilized cells by the periodic administration of nutrients, of forming isomaltulose by continuously supplying nutrients to growing immobilized cells, and of crystallizing isomaltulose from the column eluate are also described.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 490-497 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The multispecies biological treatment model including the consequent substrates consumption is suggested. The model describes the adaptation in the distribution of species that are included in the composition of activated sludge. It was shown that the effects of adaptation are similar for the consequent as well as simultaneous mechanisms of substrate oxidation.
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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 296-301 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Continuous cultivation of Pelobacter acidigallici on gallic acid is accompanied by biofilm formation. Steady states can be obtained at dilution rates nearly twice the maximum growth rate. A two-compartment model is presented which allows us to determine rate constants of attachment and detachment. The model is based on the assumption that the biofilm is fully penetrated by the substrate. At high dilution rates the outcome of our experiments is mainly determined by the attachment rate. The influence of the thickness of the biofilm is discussed in order to demonstrate the limitations of the model.
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  • 12
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 308-315 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: High-level yeast inocula was investigated as a means of overcoming the toxicity problem in ethanol fermentation of acid hydrolyzate of wood cellulose. When the inoculum level exceeded 108 initial cells/mL, 50% of the yeast cells survived the initial cell death period during which furfural and HMF were depleted. The fermentation thus proceeded to completion by virtue of cell regrowth. The specific ethanol productivity in batch fermentation on the basis of viable cells was comparable to that of pure glucose fermentation. Continuous fermentation with cell recycle was superior to batch fermentation in that there was no overall cell decline and the ethanol yield was substantially higher. The maximum ethanol productivity in continuous fermentation was 4.9 g/L h and it occurred at a dilution rate of 0.24 hr-1.
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  • 13
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    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 519-524 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article deals with the effect of P-control and Plcontrol on the dynamic behavior of a continuous stirred tank fermentor (CSTF). Several conditions concerning the stability of steady states and the existence of limit cycles of the closed-loop system are derived. The application is made to the substrate inhibition model with variable yield coefficient. It is shown how the unfavorable phase-plane trajectories of the uncontrolled CSTF can be altered to favorable ones by the appropriate choice of control parameters based on the conditions derived.
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  • 14
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two endo-xylanases (1,4-β-D-xylan xylanohydrolase, EC 3.2.1.8) were purified to homogeneity from a crude Aspergillus niger pentosanase preparation by Ultrogel AcA 54 gel permeation chromatography, SP-Sephadex C-25 cation exchange chromatography at pH 4.5, Sephadex G-50 gel permeation chromatography, and a second SP-Sephadex C-25 step, this one at pH 5.8. The two xylanases hydrolyzed soluble xylan more rapidly than insoluble branched xylan, but attacked each substance to an equal extent. Their low activity on a linear xylooligosaccharide mixture and absence of activity on insoluble xylan freed of branches suggest that the xylanases require a branch point nearby for significant attack. No xylose or L-arabinose was produced, the major products of low molecular weight being tri- and pentasaccharides and smaller amounts of di-, tetra-, and hexasaccharides. There was low activity on untreated and crystalline cellulose and on carboxymethylcellulose and no activity on other polysaccharides tested. These two xylanases had molecular weights of ca. 1.3 × 104 and similar amino acid profiles, high in acidic and low in sulfur-containing residues. Isoelectric points were 8.6 for I and 9.0 for II. Optimum pH values for activity were 6.0 and 5.5, respectively. In a 20-min assay at pH 5.5, each was most active at 45°C, with activation energies up to 40°C of 30.4 and 38.8 kJ/ mol, respectively. Optimum pH levels for stability were 5.0 and 6.0, with half-lives at 60°C and those pHs of 20 and 75 min, respectively.
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  • 15
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 334-344 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Wheat straw was treated with NaOH and anaerobically digested for methane production. Alkaline treatment resulted in a greater than 100% increase in biodegradability of wheat straw. The potential of a process flow scheme employing high alkali concentration at ambient temperature with solids separation and recycle of filtrate containing residual alkali was explored. The effect of NaOH on the solubilization of cell wall constituents and potential problems of toxicity are discussed. A solubilization model was developed which is used to predict biodegradability of whole samples based on solids and filtrate biodegradabilities. Energy requirements and chemical costs are also addressed.
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  • 16
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 327-333 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Cell growth and lincomycin production were measured in batch cultures of Streptomyces lincolnensis in chemically defined media. In these fermentations the specific rate of antibiotic production was maximal during growth and always declined at the end of the growth phase. It was found that both phosphate and ammonium salts, while required for cell growth, had negative effects on antibiotic production. By increasing the concentration of magnesium sulfate, it was possible to increase both the production rates and final titers of lincomycin. The mechanism for this effect was found to be the reduction of soluble phosphate in the medium through the precipitation of ammonium magnesium phosphate. Lincomycin production rates were not inhibited by glucose at concentrations of up to 30 g/L.
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  • 17
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 345-354 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The alkali consumption during alkaline treatment of wheat straw at ambient temperature was measured as a function of time, solids concentration, and alkali concentration. The maximum measured alkali consumption was 5.5 g NaOH/100 g TS over a period of 30 days of treatment. Chemical functional groups (e.g., acyl and carboxyl groups) were measured and compared with the observed alkali consumption. The kinetics of alkali consumption were studied and a model was developed which predicts alkali consumption reasonably well. Use of this model was made to predict biodegradability of alkali-treated wheat straw, since a strong correlation was found to exist between alkali consumption and observed biodegradability. The method of bioconversion used was anaerobic digestion for methane production.
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  • 18
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 792-799 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The balances of reductivity and high-energy bonds (HEB) during microbial growth on glucose (a standard substrate), methanol, and ethanol are reported. Also, numerical values for the quantities of HEB formation in the respiratory metabolism, HEB consumption in the constructive metabolism, as well as in a number of the other intracellular processes are evaluated. Estimations of maximum cell yields by mass and energy are made during growth on methanol and ethanol with regard to peculiar features of different microbe metabolism.
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  • 19
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    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 800-806 
    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 hybrid reactor, the upflow blanket filter (UBF), which combined on open volume in the bottom two-thirds of the reactor for a sludge blanket and submerged plastic rings (Flexiring, Koch Inc., 235 m2/m3) in the upper one-third of the reactor volume, was studied. This UBF reactor was operated at 27°C at loading rates varying from 5 to 51 g chemical oxygen demand (COD)/L d with soluble sugar wastewater (2500 mg COD/L). Maximum removal rates of 34 g COD/L d and CH4 production rates of 7 vol/vol d [standard temperature and pressure (STP)] were obtained. The biomass activity was about 1.2 g COD/g volatile suspended solids per day. Conversion (based on effluent soluble COD) was over 93% with loading rates up to 26 g COD/L d. At higher loading rates conversion decreased rapidly. The packing was very efficient in retaining biomass.
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  • 20
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 786-791 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The properties of two isozymes of β-glucosidase of Penicillium funiculosum (part I of this series) are described. The molecular weights of isozyme 1 was 2.3 × 105 by gel filtration and 1.2 × 105 by SDS gel electrophoresis, indicating two subunits. The molecular weight of isozyme 2 was unusually low for a fungal β-glucosidase: 1.6 × 104 by gel filtration and 3.7 × 104 in the presence of isopropanol. The two enzymes differed from other fungal β-glucosidases in their substrate specificities. They showed high activity with pNPG, cellobiose, cellotriose, cellotetraose, cellopentaose, gentiobiose, and laminarin, but were inactive with filter paper, CM cellulose, or derivatives or stabilized by bovine serum albumin and several alcohols such as butanol and propanol. It was inhibited by glucono-δ-lactone (Ki = 0.67μM) and glucose (Ki = 0.92mM).The enzyme was quantitatively adsorbed by P. funiculosum mycelium at pH 4 and the immobilized enzyme was as enzymically active as the free enzyme, but more heat stable. The binding efficiency was very high (5000 IU enzyme/g mycelium). It could be quantitatively eluted with buffers at pH 7 or by 0.02M Ca, Mg, or Al chlorides. The binding was selective, since mycelium grown on lactose could produce and also bind only β-glucosidase isozyme 1, whereas mycelium grown on cellulose could produce as well as bind both β-glucosidase isozymes as well as cellulases. Mycelial binding was unaffected by washing with EDTA or trypsinization, but was totally lost by washing with dilute KOH, HCl, or ethylenediamine.
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  • 21
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1005-1011 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A newly designed batch calorimeter was used to investigate the degradability of some celluloses having varying degrees of crystallinity. The PTC of an aerobic culture of Cellulomonas uda ATCC 21399 obtained revealed a diauxic growth which is attributed to the presence of hemicellulose contaminating Avicel and MN300 cellulose. The microcrystalline celluloses used were not completely utilized, whereas amorphous cellulose was easily metabolized, indicating that under the growth conditions used here, the physical structure of cellulose strongly influenced its microbial degradability. An equivalent growth yield of ca. 0.44 g/g was found with all the substrates used. The heat evolved by metabolism of 1 g cellulose was -5.86 kJ/g, a value similar to that obtained with glucose culture. The growth rate was the only variable parameter. The data obtained showed as expected that the hydrolysis product of cellulose was consumed in the same way as that of glucose and that the only limiting factor to the biodegradability of cellulose was the breakdown of the polymeric substrate. It is concluded that data obtained with glucose metabolism can be used to evaluate the extent of cellulose degradation.
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  • 22
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1012-1020 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Streptomyces aureofaciens (ATCC 12416c) was grown in the interstitial region formed by a parallel arrangement of three hollow silicone tubules contained within a microporous polypropylene hollow fiber. Liquid-soluble nutrients were supplied by diffusion across the polypropylene fiber to the interstitial cell-containing region whereas air or oxygen was provided by diffusion from the silicone tubule lumina to the cell mass. In this bioreactor, S. aureofaciens grew to high cell densities (greater than 1011 cells/cm3) and the culture so-obtained continously synthesized the secondary metabolite tetracycline. The volumetric productivity of tetracycline based on the interstitial volume was 90 μg/ml/h and based on the total reactor volume was 5.5 μg/mL/h. The high surface area-to-volume ratio afforded by the cylindrical configuration together with spatially distinct conduits to continuously transport liquids and gases, each of which may be nutrients or products of biosynthesis, to or from a tissuelike cell mass provides an alternative to the conventional air- or oxygen-sparged fermentation vessel. High volumetric reactor productivities may be achieved by virute of the concentrated stationary cell mass and by the appropriate selection of fiber sizes and materials so as to ensure adequate supplies of liquid and gaseous substrates to, as well as removal of metabolites from, most cells in the culture. This reactor topology is quite general and may be adapted to most microbial as well as mammalian and plant cell systems.
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  • 23
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    Biotechnology and Bioengineering 27 (1985), S. 1021-1026 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Human embryonic kidney cells grown as an attached, confluent monolayer on a flat substrate were subjected to steady, uniform laminar flow of medium in a specially designed chamber in which flow patterns and shear stress are accurately defined and controlled. Experiments were performed for shear stress levels ranging from 0.2 to 6.0 N/m2 with times of exposure to the shear stress ranging from 2 to 24 h. The influence of the shear field was slight at low shear stress (0.26 N/m2). Higher stress levels (0.65 N/m2 and higher) had significant effects on cell morphology, and on the post-shear release of urokinase enzyme. Still higher stress levels (2.6 N/m2 and higher) caused marked reduction in cell viability. These results may be of interest in addressing practical problems in developing commercial biosynthesis reactors.
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  • 24
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    Biotechnology and Bioengineering 27 (1985), S. 41-49 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: More than 90% of initial pyritic sulfur was removed from bituminous coal samples (containing 2.1% pyritic sulfur) using the thermophilic organism Sulfolobus acidocaldarius. Microbial desulfurization rate was improved nearly ten fold by adjusting the N/P and N/Mg ratios in the nutrient medium. Environmental conditions were optimized. The optimal values of temperature and pH were 70°C and 1.5, respectively. The influence of certain process variables (such as coal pulp density, particle size, and initial cell number density) on the rate of pyritic sulfur removal were determined. A pulp density of 20%, particle size of D¯p 〈 48 μm, and an initial cell number density of 1012 cells/g pyrite in coal were found to be optimal. The carbon dioxide enriched air did not improve the rate of pyritic sulfur removal compared to pure air at 10% pulp density of coal samples containing 2.1% pyritic sulfur. The kinetics of microbial leaching of pyritic sulfur from coal was investigated. The rate of leaching was found to be first order with respect to pyritic sulfur concentration in the reaction medium.
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  • 25
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    Biotechnology and Bioengineering 27 (1985), S. 1036-1043 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The partitioning of endo-β-glucanase, exo-β-glucanase, and β-glucosidase from Trichoderma reesei QM 9414 in aqueous two-phase systems has been studied with the object of designing a phase system for continuous bioconversion of cellulose. The partitioning of the enzymes in two-phase systems composed of various water soluble polymeric compounds were studied. Systems based on dextran and polyethylene glycol (PEG) were optimal for one-sidedly partitioning the enzymes to the bottom phase. The influence of polymer molecular weights, polymer concentration, ionic composition of the medium, pH, temperature, and adsorption of the enzymes to cellulose on the enzyme partition coefficients (K) were studied. By combining the effects of polymer molecular weight and adsorption to cellulose, K values could be reduced for endo-β-glucanase to 0.02 and for β-glucosidase to 0.005 at 20°C in a phase system of Dextran 40-PEG 40000 in the presence of excess cellulose, At 50°C, K values were increased by a factor of two. In a phase system based on inexpensive crude dextran and PEG, the partition coefficient for endo-β-glucanase was 0.16 and for β-glucosidase was 0.14 at 20°C with excess cellulose present.
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  • 26
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A model substrate, Solka Floc BW 200, was semicontinuously hydrolyzed in an aqueous two-phase system based on crude dextran and polyethylene glycol over a period of more than 450 h. With an initial concentration of 75 g/L and intermittent addition of cellulose an average concentration of 50 g/L sugar was semicontinuously produced at dilution rates of 0.006-0.012 h-1. The conversion of substrate varied between 49 and 66%. The enzyme consumption measured as FPU/g reducing sugar (RS) produced could be reduced by a factor two when compared to a batch process since, in the aqueous two phase system investigated, the enzyme could be recycled two times.
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  • 27
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    Biotechnology and Bioengineering 27 (1985), S. 861-869 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two rate equations have been developed to model the hydrolysis of ground lean meat protein by Alcalase. The first equation was based on classical Michaelis-Menten kinetics and the second on the adsorption of enzyme to the protein prior to reaction. It was assumed that this adsorption could be modelled by a Langmuir-type adsorption isotherm. Each equation considered the enzyme to be competitively inhibited by reaction product, and considered enzyme inactivation to be first order. Both rate equations have been fitted to experimental data obtained from the hydrolysis of meat protein by Alcalase. Initial rate data indicated that the adsorption model was more appropriate. However, both equations gave satisfactory fits to 11 reaction progress curves determined over a wide range of enzyme and substrate concentrations.
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  • 28
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    Biotechnology and Bioengineering 27 (1985), S. 883-886 
    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.
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  • 29
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    Biotechnology and Bioengineering 27 (1985), S. 128-131 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The aqueous effluent generated by the Fischer--Tropsch process, containing a total of 13 g/L C2-C5 monocarboxylic acids, was investigated as a potential substrate for the production of single-cell protein (SCP). A bacterial isolate, Acinetobacter calcoaceticus, could utilize all the acids in the effluent simultaneously in chemostat cultures, and no residual acids were detected in the culture below a dilution rate of 0.78 h-1. The critical dilution rate was 1.04 h-1. The maintenance energy requirement of the cells growing on the monocarboxylic acid mixture was considerably lower than that of cells growing on acetate as the sole carbon source. Enrichment of the effluent with ethanol to increase the biomass concentration was successful and still allowed the simultaneous and efficient utilization of all the carbon sources, but resulted in a decrease of the critical dilution rate by ca. 20%.
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  • 30
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    Biotechnology and Bioengineering 27 (1985), S. 137-145 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Dried spheres made from an alginate solution containing magnetite particles have excellent potential as a support for enzyme immobilization and chromatographic applications. The beads were found to be much stronger than gels such as polyacrylamide and dextran, indicating that high flow rates and pressures could be used in column separations. The support withstood not only temperatures of up to 120°C, but also most pH values and common solvents. While some solutions, such as phosphate buffers, dissolved the spheres, stabilization with Tyzor TE® eliminated this problem. The physical properties of the beads include a glasslike density of 2.2 g/mL, excellent sphericity, low porosity, and a narrow size distribution. The magnetite present in the support allows the beads to be used for magnetic separations such as high gradient magnetic filtration. Their high degree of microroughness provides a large exposed surface area for enzyme and ligand binding. Mixed Actinomyces fradiae proteases and Aspergillus niger α-amylase, two enzymes representative of classes which attack large substrates, were immobilized on the bead's surface with high activity and stability. A cyanuric dye which can be used in chromatographic applications (Cibacron Blue F3GA®) was also readily coupled to the surface of this support with good yield. The support should have a wide range of applications in bioseparation and immobilized biochemical technology.
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  • 31
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    Biotechnology and Bioengineering 27 (1985), S. 146-150 
    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 membrane has been developed for the microencapsulation of islets of Langerhans which protects these cells from the immune system. These requirements were accomplished through the optimization of important microencapsulation parameters and through the improved biocompatibility of a new alginate-poly-l-lysine (PLL)-alginate capsule membrane. Spherical and smooth microcapsules could be formed by utilizing a purer sodium alginate and by keeping the viscosity of the sodium alginate solution above 30 cps. The strength of the capsule membrane was enhanced by increasing the alginate-PLL reaction time as well as the PLL concentration. The permeability of the membrane [4 μm thick, 93% (w/w) water] was a function of the viscosity average molecular weight (Mv) of the PLL (Mv = 4000-4 × 105) used in the encapsulation procedure. Microcapsules prepared with PLL with Mv = 1.7 × 104 were the least permeable, being impermeable to normal serum immunoglobulin, albumin, and haemoglobin. The microencapsulation procedure, by protecting transplanted tissue from the components of the immune system, has great clinical potential as a new form of treatment for diseases such as diabetes and liver disease.
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  • 32
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    Biotechnology and Bioengineering 27 (1985), S. 555-561 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two photosynthetic algal cultures, one Chlorella vulgaris, and the other a Chlorogonium sp., were cultured under light limitations in chemostats. The effects of growth temperature on their energy yield and maintenance energy requirement were studied. It was observed that a lowering in temperature resulted in a lower maximum growth yield from the light energy, YG. This was attributed to two reasons. First, at low temperatures there was a change in the algal cell composition with more energy being expended to synthesize a higher biomass protein content. Secondly, at low temperatures, a cyanide-resistant respiratory pathway became operative which led to a decrease in the number of ATP being generated. The maintenance energy coefficient was a function of temperature increasing with decreasing temperature. This might reflect energy wastage by the cell at low temperatures. The maximum specific growth rate dropped with decreasing temperature, and can be described by an Arrhenius type rate-temperature model up to the optimal temperature for growth; i.e., activation energy remained constant.
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  • 33
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    Biotechnology and Bioengineering 27 (1985), S. 1102-1107 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucose oxidase, invertase, and amyloglucosidase were entrapped in calcium alginate gels as concanavalin A complexes in order to prevent the leaching out of the enzymes from the porous matrix. The free as well as the gel-entrapped concanavalin A-glycoenzyme complexes exhibited a relatively high effectiveness factor, η, indicating good accessibility to the substrates. Concanavalin A-invertase complex exhibited marked broadening of pH-activity and temperature-activity profiles and was highly resistant to temperature inactivation even after entrapment in the alginate beads. It was possible to entrap considerable quantities of invertase as concanavalin A complex in the beads without a marked decrease in η. A column containing crosslinked concanavalin A-invertase complex entrapped in alginate beads retained the ability to completely hydrolyze 1M sucrose even after continuous operation for over four months.
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  • 34
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    Biotechnology and Bioengineering 27 (1985), S. 953-960 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In enzymatic synthesis of cephalexin from D-α-phenylglycine methyl ester (PGM) and 7-amino-3-deacetoxy-cephalosporanic acid (7-ADCA) using α-acylamino-β-lactam acylhydrolase from Xanthomonas citri, it was found that this enzyme catalyzes all three reactions including PGM hydrolysis, cephalexin synthesis, and cephalexin hydrolysis. Based on our experimental results, a mechanistic kinetic model for cephalexin synthesizing enzyme system having acyl-enzyme intermediate was proposed. From this kinetic model, the reaction rate equations for three reactions were derived, and the kinetic parameters were evaluated. A good agreement between the simulation results and the experimental results was found.
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  • 35
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    Biotechnology and Bioengineering 27 (1985), S. 1146-1151 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A classical quantitative (analytical and/or statistical) analysis is not appropriate for some ill-defined and/or very complex bioengineering problems. Therefore, a new form of analysis using fuzzy mathematics has been developed. The fuzzy model can utilize semiqualitative and linguistic data which are to a certain level inconsistent. Bioengineering interpretation of the basic concept of fuzzy mathematics is given together with a numerical algorithm for a fuzzy evaluation of a model. A simple fuzzy model of a fermentor is studied in detail.
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  • 36
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    Biotechnology and Bioengineering 27 (1985), S. 1136-1145 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An analytical model for dissolved oxygen concentration in an algal minipond was used to develop a new method for estimating, on-line, the net O2 production rate (OPR) of the biological process. The method was tested experimentally and was found to provide crucial information on the vitality of the biological process and to provide an early warning of a possible forthcoming collapse of the ecosystern. It is suggested that the newly developed model and measurement method could provide investigators with useful tools for optimization of algal cultivation in the laboratory and plant.
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  • 37
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    Biotechnology and Bioengineering 27 (1985), S. 943-952 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The simultaneous separation of volatile fermentation products from product-inhibited fermentations can greatly increase the productivity of a bioreactor by reducing the product concentration in the bioreactor, as well as concentrating the product in an output stream free of cells, substrate, or other feed impurities. The Immobilized Cell Reactor-Separator (ICRS) consists of two column reactors: a cocurrent gas-liquid “enricher” followed by a countercurrent “stripper” The columns are four-phase tubular reactors consisting of (1) an inert gas phase, (2) the liquid fermentation broth, (3) the solid column internal packing, and (4) the immobilized biological catalyst or cells. The application of the ICRS to the ethanol-from-whey-lactose fermentation system has been investigated. Operation in the liquid continuous or bubble flow regime allows a high liquid holdup in the reactor and consequent long and controllable liquid residence time but results in a high gas phase pressure drop over the length of the reactor and low gas flow rates. Operation in the gas continuous regime gives high gas flow rates and low pressure drop but also results in short liquid residence time and incomplete column wetting at low liquid loading rates using conventional gas-liquid column packings. Using cells absorbed to conventional ceramic column packing (0.25-in. Intalox saddles), it was found that a good reaction could be obtained in the liquid continuous mode, but little separation, while in the gas continuous mode there was little reaction but good separation. Using cells sorbed to an absorbant matrix allowed operation in the gas continuous regime with a liquid holdup of up to 30% of the total reactor volume. Good reaction rates and product separation were obtained using this matrix. High reaction rates were obtained due to high density cell loading in the reactor. A dry cell density of up to 92 g/L reactor was obtained in the enricher. The enricher ethanol productivity ranged from 50 to 160 g/L h while the stripper productivity varied from 0 to 32 g/L h at different feed rates and concentrations. A separation efficiency of as high as 98% was obtained from the system.
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  • 38
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    Biotechnology and Bioengineering 27 (1985), S. 1173-1184 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The leaching of a zinc sulfide concentrate by Thiobacillus ferrooxidans was investigated in continuous stirred tank reactors. A mathematical model for the growth of T. ferrooxidans on this solid substrate is presented and tested. Experimental leaching studies were done using two reactors in series with and without recycle of solids from the outlet of the first reactor back to its inlet. The proposed model fits the authors' experimental data well. However, comparison of the parameters calculated with those calculated from the data of others showed that a wide variation can exist; thus, the parameters seem to depend on the nature of the substrate. The area occupied on the sulfide surface by a bacterium was found to be 5.4 μm2. The calculated maximum specific growth rates ranged from 0.20 to 0.31 h-1, and these values were dependent on whether they were observed in the first or second of the two reactors in series.
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  • 39
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    Biotechnology and Bioengineering 27 (1985), S. 1185-1191 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The hollow fiber enzyme reactor with pulsation developed by Kim and Chang (1983) was operated in a differential mode by recycling a substrate solution, in order to assess the efficiency of ultrafiltration swing. The rates of lactose conversion by β-galactosidase contained in the shell side of the reactor were measured to determine the effects of recirculation rate, pulsation period, and amplitude. The conversion increased with the increase of recirculation flow rate and the amplitude while variation in period affected the conversion relatively little. The maximum increase of 113% in the activity was observed in the reactor with pulsation as compared to that without pulsation. The two-compartment model well described the experimental data obtained in this study. Square-wave pulsation was theoretically more effective in increasing conversion than sine wave pulsation. However, in experimental operation the damping effect of the hollow fiber wall narrowed the difference between these two wave forms.
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  • 40
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    Biotechnology and Bioengineering 27 (1985), S. 638-649 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Anaerobic digestion of biological organic particulates to methane has been described by a structured mathematical model based on multiple-reaction stoichiometry, conventional material balances, and liquid phase equilibrium chemistry. A general stoichiometric treatment for any set of multiple biological reactions is derived based on a unit mass of oxggen equivalents of the reactions limiting substrate. The model agrees well with two existing experimental studies of anaerobic digestion of biomass particulates. Hypothetical computer simulations are presented to illustrate possible instabilities of the anaerobic process under various operating scenarios.
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  • 41
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    Biotechnology and Bioengineering 27 (1985), S. 660-674 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 42
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    Biotechnology and Bioengineering 27 (1985), S. 1591-1595 
    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.
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  • 43
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    Biotechnology and Bioengineering 27 (1985), S. 1603-1607 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 3 Ill.
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  • 44
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    Biotechnology and Bioengineering 27 (1985), S. 1634-1639 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A formate-selective biocatalytic potentiometric electrode system based on whole cells Pseudomonas oxalaticus has been developed. Permeation of the Gram-negative Pseudomonas oxalaticus microbial cells by EDTA is shown to improve the response slope from 38 mV/decade to 49 mV/decade, resulting in increased sensitivity. Out of 13 possibly interfering compounds tested, only pyruvate and lactate showed moderate response.
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  • 45
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    Biotechnology and Bioengineering 27 (1985), S. 1623-1633 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A technique using C-14 isotope tracers to probe the branching of carbon flow in methylotrophic bacteria has been devised and applied to continuous steady-state cultures. Methylomonas L3, a strain which utilizes the KDPG/TA variant of the ribulose monophosphate cycle for carbon fixation, was employed in the experimental studies. The actual in vivo rates of substrate-carbon incorporation into biomass, both direct and via CO2, and of the two carbon oxidation schemes were determined in three different steady-state cultures. The results show that the carbon substrate is oxidized predominantly via formate (the linear oxidation scheme), and that the cyclic scheme of oxidation is minimally, if at all, utilized. The carbon incorporation and oxidation patterns appear to vary considerably with the dilution rate and the inoculum history. The experimental accuracy of the new technique is discussed in detail.
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  • 46
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    Biotechnology and Bioengineering 27 (1985), S. 1229-1235 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: During the steady-state continuous culture of Saccharomyces cerevisiae on sugar cane blackstrap molasses under different experimental conditions, oscillatory variations of the invertase activity of the intact yeast cells were observed. The continuous morphological changes of the cells wall and of the periplasmic space affecting the interaction between invertase and sucrose molecules could be responsible by the observed oscillatory phenomena. The average invertase activity at the steady state is linearly correlated to the cell's growth rate.
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  • 47
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    Biotechnology and Bioengineering 27 (1985), S. 1246-1257 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A fermentation system has been designed to demonstrate the use of gas chromatography (GC) for on-line monitoring of the butanol-acetone and other complex saccharolytic fermentations. Tangential flow ultrafiltration was used to sterilely and continuously obtain a cell-free filtrate from the fermentation broth for on-line GC analysis of butanol, butyrate, acetate, acetone, ethanol, and acetoin. The liquid injection system consists of a phosphoric acid contactor, a slider-type injection valve, and a heater to address the difficulties (ghosting) encountered in the analysis of carboxylic acids. The fermentation headspace gas was also analyzed by on-line GC for nitrogen and carbon dioxide, while hydrogen was measured by difference. Raw chromatographic data were analyzed by a chromatography data system. Both raw and processed data were transmitted to a VAX 11/750 computer for further processing (using the fermentation equation) and archiving. The fermentation equation, which has recently been derived and tested on completed fermentation data, was also found to be valid during transient fermentations and thus useful as a gateway sensor for calculating various fermentation parameters on-line. Such parameters include glucose concentration and gas composition, as well as a number of unobservable parameters (such as YATP, excess ATP, and NAD reduced by FdH2), which characterize the state of the fermentation.
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  • 48
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    Biotechnology and Bioengineering 27 (1985), S. 1258-1269 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The microbial epoxidation of propene and 1-butene was used to study some fundamental aspects of two-liquid-phase biocatalytic conversions. Introduction of a water-immiscible organic solvent phase in a free-cell suspension gave rise to a series of undesired phenomena, e.g., inactivation by the solvent, clotting of biomass, and aggregation of cells at the liquid-liquid interface. Immobilization of the cells in hydrophilic gels, e.g., calcium alginate, prevented direct cell-organic solvent contact and the related clotting and aggregation of biomass. However, the gel entrapment did not seem to provide additional protection against the organic solvent. The influence of various organic solvents on the retention of immobilized-cell activity was related to solvent properties like the polarity (as expressed by the Hildebrand solubility parameter) and the molecular size (as expressed by the molecular weight or molar volume). High activity retention was favored by a low polarity in combination with a high molecular weight. The solubility parameter also proved useful to describe the capacity of various organic solvents for oxygen and alkene oxides. This facilitated the optimization of the solvent polarity.
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  • 49
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    Biotechnology and Bioengineering 27 (1985), S. 1721-1725 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The advant of a new range of high-protein capacity cellulosic ion exchangers suitable for use on an industrial scale made it worthwhile investigating the conditions necessary to remove the contaminating enzyme glucosyltransferase from a commercial preparation of crude glucoamylase. The potential of the SP derivative, SP Indion®, for achieving this separation is shown. At pH 2.5 the glucosyltransferase was selectively adsorbed by the ion exchanger, and 99% of the glucoamylase was recovered in the eluate from the column. Purification of an Aspergillus culture filtrate by this method will require careful control of the ionic strength of the culture medium if it is to be used without the additional step of cation exchange to lower than pH to 2.5.
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  • 50
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    Biotechnology and Bioengineering 27 (1985), S. 1276-1276 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 51
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    Biotechnology and Bioengineering 27 (1985), S. 1274-1275 
    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 of the effect of glucose addition to the kinetics of maltodextrin hydrolysis catalyzed with free and immobilized Aspergillus niger glucoamylase does not show any significant glucose inhibitory effect. This result is in contradiction with data previously reported in the literature. On the contrary, a slight glucose-activating effect was observed. This effect was greater in the case of the immobilized enzyme. The glucose inhibitory effect may thus not be involved in the case of practical saccharification conditions catalyzed with glucoamylase when maltodextrins are used as substrate.
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  • 52
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    Biotechnology and Bioengineering 27 (1985) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 53
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1710-1716 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The feasibility of applying the UASB concept for the anaerobic treatment of stillage of distilleries in the sugar producing area of Argentina was subject to study. Results obtained in a 100-L UASB reactor treating stillages with COD values between 35 and 100 g COD/L are presented. Loading rates of up to 24 g COD/L/day were applied with an average COD removal of 75% and a biogas production of more than 9 L/L/day, with an average methane content of 58%. The settling velocity distribution of sludge particles would indicate a good formation of biomass pellets. System interruptions of months without feed and at ambient temperature (20-24°C) were well tolerated.
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  • 54
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1726-1729 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 7 Ill.
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  • 55
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1403-1410 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 8 Ill.
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  • 56
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1411-1417 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The feasibility of the upflow anaerobic sludge bed (UASB) process for the treatment of potato starch wastewater at low ambient temperatures was demonstrated by operating two 5.65-L reactors at 14°C and 20°C, respectively. The organic space loading rates achieved in these laboratory-scale reactors were 3 kg COD/m3/day at 14°C and 4-5 kg COD/m3/day at 20°C. The corresponding sludge loading rates were 0.12 kg COD/kg VSS/day at 14°C and 0.16-0.18 kg COD/kg VSS/day at 20°C.These findings are of considerable practical importance because application of anaerobic treatment at low ambient temperatures will lead to considerable savings in energy needed for operating the process. As compared with various other anaerobic wastewater treatment processes, a granular sludge upflow process represents one of the best options developed so far. Although the overall sludge yield under psychrophilic conditions is slightly higher than under optimal mesophilic conditions, this doesn't seriously hamper the operation of the process. The extra sludge yield, due to accumulation of slowly hydrolyzing substrate ingredients, was 4.75% of the COD input at 14°C and 1.22% of the COD input at 20°C.
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  • 57
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985), S. 1418-1426 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Assessment was made to evaluate the effect of hydrogen peroxide pretreatment on the change of the structural features and the enzymatic hydrolysis of rice straw. Changes in the lignin content, weight loss, accessibility for Cadoxen, water holding capacity, and crystallinity of straw were measured during pretreatment to express the modification of the lignocellulosic structure of straw. The rates and the extents of enzymatic hydrolysis, cellulase adsorption, and cellobiose accumulation in the initial stage of hydrolysis were determined to study the pretreatment effect on hydrolysis. Pretreatment at 60°C for 5 h in a solution with 1% (w/w) H2O2 and NaOH resulted in 60% delignification, 40% weight loss, a fivefold increase in the accessibility for Cadoxen, an one times increase in the water-holding capacity, and only a slight decrease in crystallinity as compared with that of the untreated straw. Improvement on the pretreatment effect could be made by increasing the initial alkalinity and the pretreatment temperature of hydrogen peroxide solution. A saturated improvement on the structural features was found when the weight ratio of hydrogen peroxide to straw was above 0.25 g H2O2/g straw in an alkaline H2O2 solution with 1% (w/w) NaOH at 32°C. The initial rates and extents of hydrolysis, cellulase adsorption, and cellobiose accumulation in hydrolysis were enhanced in accordance with the improved structural features of straw pretreated. A four times increase in the extent of the enzymatic hydrolysis of straw for 24 h was attributed to the alkaline hydrogen peroxide pretreatment.
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  • 58
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    Biotechnology and Bioengineering 27 (1985), S. 1477-1481 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The semicontinuous operation of affinity chromatography columns charged and eluted alternately is studied mathematically. A strongly nonlinear adsorption equilibrium relation is utilized to examine the periodic mode of adsorption and desorption. The governing differential equations are solved by the method of characteristics. The time interval between desorption and adsorption periods is estimated, and implications for separations are discussed to show the advantages of semicontinuous cyclic operation.
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  • 59
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    Biotechnology and Bioengineering 27 (1985), S. 1500-1506 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 8 Ill.
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  • 60
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    Biotechnology and Bioengineering 27 (1985), S. 1512-1515 
    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.
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  • 61
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    Biotechnology and Bioengineering 27 (1985), S. 1507-1511 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 6 Ill.
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  • 62
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    Biotechnology and Bioengineering 27 (1985), S. 1516-1519 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 3 Ill.
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  • 63
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    Biotechnology and Bioengineering 27 (1985), S. 1520-1523 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 3 Ill.
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  • 64
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 27 (1985) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 65
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    Biotechnology and Bioengineering 27 (1985), S. 1525-1532 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Under certain simplifying hypotheses, a chemostat in which one microbial population feeds on another can be described by a system of three ordinary differential equations. A study is conducted to find which features of the equations are the most important to the dynamics of the system; it is found that the main influence comes from the from the particular form of the specific growth rate of the feeding population. Some conditions are derived that relate the form of the specific growth rate of the feeding population to the dynamic behaviour of the system.
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  • 66
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    Electrophoresis 6 (1985), S. 53-56 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Examples of increased resolution obtained with non-linear pH gradients are presented. Discussed in detail is the “ideal” pH course in the range 4-10, based on the statistical distribution of isoelectric points of protein from data in the literature.
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  • 67
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    Electrophoresis 6 (1985), S. 96-97 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A heating-cooling plate for open casting of agarose gels is described. Use of this plate provides a flawless and convenient means of producing 0.8 mm or thicker gels of any size up to 25 × 30 cm on glass plates (0.5 mm thick gels tend to pull slightly away from the edge of the glass plate during pouring). Gels of of any thickness (0.5 mm or greater) can be prepared if GelBond film is used.
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  • 68
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    Electrophoresis 6 (1985), S. 97-99 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: An ultrathin-layer polyacrylamide gel isoelectric focusing method is described for subtyping transferrin. After a one-hour pretreatment at room temperature of transferrin in serum with ferrous ammonium sulfate, the samples were subjected to isoelectric focusing for 140 min with a final voltage of 3500 V. Thus, transferrin subtyping was achieved in one-half of a working day. Population data on transferrin in Black and White samples from Baltimore, Maryland, were obtained using this method. The data were similar to that previously reported on these races from other geographical locations.
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  • 69
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    Electrophoresis 6 (1985), S. 100-101 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 70
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 71
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    Electrophoresis 6 (1985) 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 72
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    Electrophoresis 6 (1985), S. 145-145 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 73
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The effect of protein complexing on quantitation by rocket immunoelectrophoresis was studied using group-specific component (Gc), a serum protein which undergoes 1:1 interactions with both sterols (Vitamin D metabolites) and protein (G-actin). Gc purified from serum was used as a reference. Complex formation with 25-〈OH〉 D3 did not significantly affect the measurement of Gc. However, addition of excess G-actin to purified Gc led to substantially higher rockets at every concentration of Gc tested. In dose-response experiments performed with a constant quantity of Gc and increasing quantities of added G-actin, the increment in rocket height was progressive up to equimolarity, at which point a plateau was reached. Further demonstration that the increased rocket heights were directly related to 1:1 complex formation between Gc and G-actin (Gc:G-actin) was obtained by [125I]G-acting and isoelectric focusing (IEF) with autoradiography. Examination of the relative charge of Gc and of Gc:G-actin by crossed immunoelectrophoresis showed that complexes exhibited faster mobility, whereas very little evidence of alteration in antigenicity of Gc:G-actin complexes was apparent. These results suggest that Gc:G-actin complexes exhibit increased electrophoretic mobility and that their presence in samples containing Gc causes an increase in the height of Gc rockets. The findings also indicate that interactions with specific ligands could cause artefacts in immunoelectrophorestic quantitation of other proteins present in biological fluids.
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  • 74
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    Electrophoresis 6 (1985), S. 175-178 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Esterase-19 (ES-19) of the house mouse, following specific visualization in electrophoretic and isoelectric focusing gels, is described. A list of diagnostic features was compiled including substrate specificity, inhibition and activation characteristics, temporal and spatial patterns of expression, distribution among laboratory and wild mouse strains. ES-19 was classified as arylesterase (E.C. 3.1.1.2) and compared with ES-16 and ES-25, two other mouse arylesterases. Homology with brain esterases of rat, guinea pig, rabbit and man was found, and a uniform disegnation “arylesterase I” is proposed.
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  • 75
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    Electrophoresis 6 (1985), S. 186-188 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Amino acid analysis shows that commercial grades of agarose contain peptides. The quantity varies with the purity of the agarose. These findings agree with our observations that the extent of the background after silver-staining for agarose gels or where agarose was used in the system (as in two-dimensional polyacrylamide gel electrophoresis) was determined mainly by the quality of agarose used.
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  • 76
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Non-histone proteins (NHP) extracted from chromatin of two plasmocytoma cell lines, X63-Ag8-653 and Sp2/O-Ag 14, with 0.35 M NaCl are analyzed by two-dimensional polyacrylamide gel electrophoresis. These NHP have been fractionated into a high mobility group (HMG) and a low mobility group (LMG) using 2 % trichloroacetic acid (TCA). The HMG which is well-characterized is soluble in 2 % TCA whereas the LMG precipitates. LMG resolubilization is difficult. Only a buffer containing glycerol, sodium dodecyl sulfate (SDS) and β-mercaptoethanol enables them to be studied by polyacrylamide SDS electrophoresis. The LMG is clearly differentiated from the HMG by the predominance of high molecular weight proteins and by the high number of molecular species in a wide pH range. In the LMG there are also some diffrences between the two plasmocytoma cell lines. For example, a high molecular weight protein of 100 000 and pI 7.2-8.2 is present in Sp2/O-Ag 14 but absent in X63-Ag8-653. Also noticeable in the LMG proteins is the presence of low molecular weight proteins with high electrophoretic mobility.
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  • 77
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: An overlay grid was constructed to facilitate the measurement of densiometric scans of proteins in pH gradients formed by electrofocusing. The overlay procedure provides a rapid way for data collection. The relationship between pH and gel distance closely fit a straight line, (R 〉0.99). The reliability of the system was shown by the close agreement of the linear regressions from six different sets of data. Also, the feasibility of using a similar grid for a nonlinear system was established.
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  • 78
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Glucocorticoids are known to enhance fetal lung maturation. Previous studies with whole lung have shown that specific proteins are enhanced by these hormones. At least two effects of glucocorticoids on fetal lung epithelia require the presence of mesenchymal elements. Since the lung consists of several different cell types, further understanding of glucocorticoid effects on fetal lung protein synthesis requires that such studies be carried out with specific cell types. Using two-dimensional gel electrophoresis, we have examined the synthesis and secretion of proteins by fetal rat lung fibroblasts in the presence and absence of glucocorticoid. Two proteins, fsa and fsb, are enhanced by dexamethasone, appear to be organ specific, and are enhanced only during prenatal and early postnatal life. They are not enhanced by a variety of other hormones.
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  • 79
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    Electrophoresis 6 (1985), S. 242-246 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Analysis of two-dimensional electrophoresis patterns demands computer assistance. Specialized machines to read and store the pattern of an autoradiograph or of a “wetgel” into a digital memory are rare. A fast and flexible opto-mechanical picture reading and storage machine, specially designed for the computer-aided analysis of two-dimensional electrophoresis patterns, has been constructed. It is now possible to digitize, for example, a picture of 250 × 400 mm into pixels of ca. 250 × 250 μm with a grey level resolution of 8-bit within 70 s. Digitizing at significant pixel sizes down to 64 μm is possible at correspondingly longer acquisition times for the areas to be digitized. The design is a flat-bed scanner. One axis is scanned optomechanically (Galvanometer scanned laser beam), the other axis is scanned mechanically. The area covered by the scanning laser beam has a size of 250 × 400 mm or smaller. The control of the scanner, including pixel density and sampling synchronization is executed by a microprocessor system.
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  • 80
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    Electrophoresis 6 (1985), S. 247-247 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 81
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    Electrophoresis 6 (1985) 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 82
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    Electrophoresis 6 (1985), S. 249-258 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Additional Material: 9 Ill.
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  • 83
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    Electrophoresis 6 (1985), S. 259-267 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Photography under ultraviolet illumination is used routinely to record the results of gel electrophoresis of DNA when a fluorescent dye, which binds to DNA, is present in the gel. The distribution of DNA in the gel can be determined accurately from the photographic negative when the absorbance is interpreted according to the nonlinear film darking function, when the contrast of the film is calculated, and when the images are corrected for uneven ultraviolet transillumination. The contrast can be calculated from the images of a fluorescent plastic intensity standard. The local intensity of the unbound-dye fluorescence can be used to correct images for uneven illumination. Using this approach, relative abundances of DNA fragments, compared within each electrophoretic track, were measured within an average error of 6.6% of the expected value with no evidence of systematic error. Quantitative comparisons were valid anywhere on the film for a 50:1 range of intensities, with errors distributed normally with a standard deviation of ± 10 % of the expected value.
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  • 84
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A commercially available electrooptical device, coupled to a microcomputer, is capable of analyzing densitometric values with a resolution of 280 × 128 points (35 840 individual points). A complex autoradiogram of a two-dimensional electrophoretic gel is completely sorted into X and Y coordinates and exposure values differeing by as little as 0.01 O.D. (optical density) units or over an O.D. range of 0 to 4.00 in less than 2 min. For other purposes such as comparison of two gels, immunoelectrophoretic patterns, enzyme linked immunosorbent assay or microtitration plates, dot-blot assays, or tissue samples, a comparison may be made and all differences stored in the computer in less than one second and further decisions or computer-driven actions decided on that basis.
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  • 85
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A new fast, precise and reproducible method for lanes detection and profiles evaluation of one-dimensional gel electrophoretic autoradiograms is presented. It is fully automated, and its principles are based on image and signal processing techniques. The operator usually has only to specify the number of lanes that are present in the preparation, but still may retain overall control. In the first phase, the program automatically locates the lateral position of the lanes, and approximates their left and right boundaries by polynomials (which allow curved as well as straight lanes). The operator can then intervene, if necessary, to adjust these positions. In the second phase, the background variations are corrected; two types of density profiles are computed, representing the average density as well as the total amount of material in the lanes. This calculation is carried out with correction for tilt or curvature for each lane. Finally, prinouts are produced that contain graphs of the profiles together with quantitative measurements. Use of this method is illustrated by analysis of electrophoretic gels containing proteins and lipopolysaccharides of varying mobilities. Experiments made on these gels as well as others yield accurate quantitation of the relative amount of each component.
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  • 86
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    Electrophoresis 6 (1985), S. 277-282 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The abundance of protein Mitcon: 5, a major component of monocytes and granulocytes as observed by high-resolution two-dimensional electrophoresis, is increased in human fibrolasts treated with interferon or poly I:C. Production of this polypeptide is eliminated in cells treated with dinitrophenol, but not in cells treated with chloramphenicol, indicating that it is a cytoplasmically synthesized mitochondrial protein. Neither chloroquine nor tunicamycin, both of which interfere with proper targeting of lysozomal enzymes, affects Mitcon: 5 production. Actinomycin D prevents induction, indicating that new transcription is required. Cell fractionation studies show that Mitcon: 5 is found in the mitochondria/lysosome fraction, and that it is released from this fraction by treatments designed to liberate matrix enzymes. Mitcon: 5 thus appears to be the first example of a direct effect of interferon treatment on the composition of mitochondria. Furher characterization of this effect may help to explain side-effects of interferon treatment, including fever.
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  • 87
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    Electrophoresis 6 (1985), S. 282-286 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Separator isoelectric focusing is a simple technique for creating narrow pH gradients (less than 1 pH unit wide). It has proved useful elsewhere for the enhanced separation of phosphoglucomutase, esterase D, group specific component (Gc) haemoglobin, transferrin and for distinguishing between seminal and vaginal acid phosphatases. The properties of 10 different zwitterionic buffers have been studied in order to determine pH gradient profiles, their stability, experimental reproducibility and the effect of temperature. The effect of adding non-ionic constituents (glycerol or sucrose) to the gel has also been investigated. The buffers examined produced narrow pH gradients within the pH 4-6 range. It was shown that specific pH gradients could be produced by mixing different buffers and carrier ampholytes together.
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  • 88
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    Electrophoresis 6 (1985), S. 296-298 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A simple method of stain fixation of polyacrylamide isoelectric focusing tube gels is described. Gel handling and storage are simplified by fixation and second-dimension quality is markedly improved over that from the original procedure involving quick freezing and frozen storage of the gel.
    Additional Material: 3 Ill.
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  • 89
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 301-306 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Isolated human monocytes showed an increased electrophoretic mobility (EM) during incubation in vitro. However, when the monocytes were isolated after activation in a mixed leukocyte culture, they tended to maintain their original EM during culturing. Activated and non-activated monocytes were compared with respect to their activities during lymphocyte transformation. The lymphocytes which were used to test the activities of both kinds of monocytes had been depleted of all accessory cells by a two-step physical isolation procedure including coutercurrent centrifugal elutriation and cell electrophoresis. Non-activated monocytes supported concanavalin A (Con A) - induced transformation of the purified lymphocytes optimally when the culture medium was supplemented by fetal calf serum (FCS). If, however, activated monocytes were used in lymphocyte transformation tests, Con A induced optimal [14C]thymine-deoxyribose incorporation, even if FCS was replaced by autologous human plasma. The results suggest that there is a relationship between the stability of the EM of human monocytes and their ability to support lymphocyte proliferation during an in vivo immune response, when FCS is obviously not present.
    Additional Material: 6 Ill.
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  • 90
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Partially hepatectomized mice were injected intraperitoneally with isoproterenol (IPR), sodium phenobarbital (PB), or a combination of the two, and the regenerating lobe of the liver was removed. Nuclei were isolated, electrostatically sorted from various phases of the cell cycle, and the acid-soluble proteins were extracted. Synthesis of several high-molecular weight non-histone proteins were observed in two-dimensional polyacrylamide gel electrophoretic autoradiographs after independent administration of IPR and PB. The histones revealed alterations in synthesis patterns and the histone variants demonstrated concentration changes in response to these drugs. A protein designated as “a” appeared in G0 + G1 phase nuclei after IPR treatment and showed an increase in concentration after PB was included in the dosing regime. This protein displayed homology with a non-histone protein from calf thymus when its mobility and amino acid content was examined.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 91
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Dextran Blue incorporated in concentrations up to 1% w/v into a gel layer buffered with Immobiline chemicals to pH 9.1 does not interfere with isoelectric focusing within an adjacent immobilized pH gradient (IPG) and retains its ability to strongly bind human serum albumin. With minor modifications of the procedure, it is possible to use Dextran Blue shelves in connection with either non-denaturing or denaturing (i. e. containing 8 M urea) analysis media. Examples include easier typing of group-specific component in an ultranarrow IPG against a completely clear background and improved resolution of transferrin and hemopexin in two-dimensional separations.
    Additional Material: 5 Ill.
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  • 92
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 349-350 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The preparation of 0.5 mm fabric reinforced polyacrylamide (FRP) gels is described. After isoelectric focusing, egg white and isoelectric point marker proteins were quantitatively transferred in only 20 min from the FRP gels to nitrocellulose membranes by electroblotting. FRP gels combine the advantages of previously described supports for polyacrylamide gels with penetrability for electric current during blotting. The gels may be conveniently stored and afford operational versatility.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 93
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have examined methods for broadening and stabilising pH gradients used in the first dimension of two-dimensional gel electrophoresis. The replacement of typical strong electrolytes with weak electrolytes as reservoir anolyte and catholyte allows the generation of broad (3.5 pH unit) gradients that are stable for at least 28 000 volt × hours (Vh). Protein patterns form within 10 000 Vh are stable throughout the subsequent focusing. Large quantities of protein, up to 6 mg, can beloaded onto such gels by mixing samples into the gel prior to polymerisation. We found no evidence for extensive modification of proteins because of exposure to the polymerisation reaction. The pH gradient formed in strong electrolytes in the typical O'Farrell gel system (J. Biol. Chem. 1975, 250, 4007-4021) could also be broadened if the samples were again mixed into the gel before polymerisation. This last effect appears to be due to an effect of the sample buffer used upon end loading of samples.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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  • 94
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 351-352 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Non-heme iron proteins were stainned in the gel with bathophenanthroline disulfonate after polyacrylamide gel electrophoresis. This staining method proved more resistant to color fading than iron detection by α,α′-dipyridyl, with 〉50% of the color being retained after 24 h by three ferredoxins of different origin. A linear relationship was found for the amount of iron in non-heme iron proteins, applied to the gel, and the areas of densitometric tracings.
    Additional Material: 3 Ill.
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  • 95
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 357-358 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A new method is described for staining proteins after high resolution two-dimensional gel electrophoresis. The polyacrylamide gel matrix is stained blue, by photoreduction of Nitro Blue Tetrazolium, while the spots of polypeptides remain unstained. The method is much more sensitive than Coomassie Blue staining and its sensitivity is close to that obtained by silver staining.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 96
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A simple procedure is described to double-stain sodium dodecyl sulfate-polyacrylamide gels both with Coomassie Blue and by the concanavalin A - horseradish peroxidase method to detect glycoproteins among the protein bands. Double staining proves useful in cases where many bands are present on the gels and where a comparison of banding patterns from separate gels or even separate lanes of the same gel cannot be carried out with sufficient accuracy.
    Additional Material: 4 Ill.
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  • 97
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985) 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 98
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 468-469 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A routine method for species' determination of raw muscle tissues is described, Defrosted raw muscle tissues were evenly spread on filter paper applicators which were then placed on a thin-layer agarose plate. A pH 4-8 gradient was obtained by mixing equal parts of Ampholine 4-6 and 6-8. Tissue homogenates of bovine, equine, porcine, ovine and other species were compared with spread tissue applications. The proposed method is useful in the routine control for quick identification of cut meat in cases of fraud where an exchange of species may have taken place.
    Additional Material: 4 Ill.
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  • 99
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 471-474 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A linear equation representing the ionic mobility as a function of the ionic strength square root was worked out on the basis of our hypothesis for the existence of two ionic radii. The equation contains a proposed parameter, pi. Comparisons between this equation and the equations of Onsager, and Robinson-Stokes were made, and the absolute mobilities of 20 ions at 0 and 25 °C were calculated according to it. It was demonstrated that equations derived from the proposed equation can be used for calculating electrolyte dissociation degrees and electrolyte concentration dissociation constants.
    Additional Material: 1 Ill.
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  • 100
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 483-488 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In the design of continuous electrophoresis devices, the main variables are the gap width, the flow rate and inlet temperature of the buffer, the electric field strength, the point of injection of the proteins, and the orientation of the flow to the gravitational field. Best values for many of these can be derived from first principles by requiring that the flow be stable, and that streak spreading be minimized. The result is a downflow device, with larger gap width than is usual, warm inlet buffer solution, and protein injection slightly downstream of the start of the electrodes. If cooling is improved by increasing the thermal conductivity of the cooling walls, the gap width must be reduced and the buffer flow increased. The performance possible with perfect cooling walls is similar to that obtained in space because, in both cases, the limiting factor is not thermal convection but the high temperature generated at the mid-point between the cooling walls.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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