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  • 2010-2014
  • 1975-1979  (272)
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  • 1977  (272)
  • Biochemistry and Biotechnology  (167)
  • Life Sciences  (105)
  • 101
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 297-300 
    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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  • 102
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 493-505 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Oxygen transfer from gas to liquid under steady-state cocurrent flow conditions was modeled using the dispersion model, and the oxygen transfer coefficients were estimated from available data for a column with Koch motionless mixers. The dispersion in the column was estimated for several different gas and liquid flow rates using steady-state tracer experiments. The estimated oxygen transfer coefficients were compared with those estimated using complete mixing and plug flow models. The results indicate that the dispersion model is the most appropriate model for estimating the mass transfer coefficient from the available data.
    Additional Material: 8 Ill.
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  • 103
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 555-555 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 104
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 527-538 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Chaetomium cellulolyticum, a newly isolated cellulolytic fungus, showed 50-100% faster growth rates and over 80% more final biomass-protein formation than Trichoderma viride, a well-known high cellulase-producing cellulolytic organism, when cultivated on Solka-floc (a purified, predominantly amorphorous form of cellulose) or partially delignified sawdust (consisting of a mixture of hardwoods) as the sole-carbon source in the fermentation media. However, in both cases, T. viride produced much higher quantities of free cellulases at faster rates and also degraded more substrate than C. cellulolyticum. It is concluded that the synthesis mechanisms and/or the nature of the cellulase complexes of the two types of organisms are quite different such that C. cellulolyticum is more optimal for single-cell protein (SCP) production, while T. viride is more optimal for the production of extracellular cellulases.It was also found that the amino acid composition of C. cellulolyticum is generally better than that of T. viride and compares favorably with those of the FAO reference protein, alfalfa, and soya meal. In addition, preliminary feeding trials on rats have shown no adverse effects of the SCP produced by C. cellulolyticum fermentations.
    Additional Material: 4 Ill.
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  • 105
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 583-589 
    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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  • 106
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 595-598 
    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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  • 107
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 599-603 
    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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  • 108
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 701-714 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The uptake mechanism of liquid hydrocarbons of low solubility in water was investigated, using microorganisms with different affinities for liquid hydrocarbon. Microorganisms which could utilize hydrocarbon were much more adherent to hydrocarbon than those which could not. The adhesive force between Candida intermedia IFO 0761 and hydrocarbon was higher than that of Candida tropicalis ATCC 20336, though both could utilize hydrocarbon. The total hydrocarbon uptake from the drop and accommodation forms of hydrocarbons was much higher than that from dissolved hydrocarbon. The uptake rate of drop-form hydrocarbon was nearly equal to that of accommodation-form hydrocarbon for C. intermedia, but was lower for C. tropicalis which shows lower adhesion to hydrocarbon.
    Additional Material: 13 Ill.
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  • 109
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 983-1008 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The response of a polarographic oxygen electrode to a step change and to an exponential change in bulk oxygen concentration was studied theoretically and experimentally for the case where there is a significant liquid film resistance at the outerside of the membrane-covered electrode. The probe response has been described considering the start-up period of the concentration changes (the period of time that will elapse before the new concentration level is established and/or before the volumetric mass transfer coefficient kLa regains its steady-state value after the gas supply is opened to the fermentor). A linear change of the pertinent characteristics is assumed during this start-up period. It is shown that a substantial error could be introduced by neglecting the start-up period for cases frequently occurring in practice. In addition, the dependences of the probe response on the direct contact of bubbles with an electrode and on the fluid flow field around it were discussed.
    Additional Material: 14 Ill.
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  • 110
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1037-1063 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The overall rate of reaction of a gel-immobilized urease particle necessarily depends upon the hydrogen ion concentrations within the particle. When the particle is unbuffered, the internal hydrogen ion concentrations are a consequence of the local rates of reaction and the rate of egress of the products of hydrolysis. A simple apparatus has been devised which allows a fairly rapid determination of the hydrogen ion concentration in the center of a particle for any given size, enzyme concentration, substrate concentration, and external pH. The products of urea hydrolysis are self-buffering in the region of pH 8.83 and for an external pH less than the self-buffering pH, the pH within the particle is increased because of the reaction. When the external pH is greater than the self-buffering pH, the converse occurs. The pH at the center of the particle approaches the self-buffering pH with an increase in particle size and enzyme concentration. The external pH necessarily differs in effect when above or below the self-buffering pH. An increase in the external substrate concentration has a limited effect, simply rendering the local rates of reaction to be of zero order. The center-line pH and therefore all internal hydrogen ion concentrations depend upon the parameter \documentclass{article}\pagestyle{empty}\begin{document}$ L\sqrt {\rho _e} $\end{document} and the external pH. Differences between the external and center-line pH values of the order of units are unexceptional. The implications of the internal pH profiles on the local and overall rates of reaction are explored.
    Additional Material: 13 Ill.
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  • 111
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 112
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1145-1153 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model is presented that describes the concentration of an amino acid in total cell protein as a function of its concentration in individual cell proteins or in sets of cell proteins. The resulting equation makes it possible to calculate how the makeup of cell proteins must change to obtain a specified alteration in the content of an amino acid in the total cell protein. It is recognized that protein species or sets of proteins that are distinguished by being richer or poorer in a key amino acid than the overall protein must undergo considerable variations in content. The necessary extent of these shifts suggests that the amino acid composition of total cell protein is not likely to be affected significantly by variations in the cultivation conditions.
    Additional Material: 1 Ill.
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  • 113
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1155-1169 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucose-limited chemostat cultures of Candida utilis were cultivated at various pH levels (3.0-7.5), temperatures (15-37.5°C), dilution rates (0.06-0.42 hr-1), and with one of two nitrogen sources (NH4+ or NO3-). Enterobacter aerogenes was also cultivated in the chemostat under nitrogen and phosphorus limitations. The amino acid profile of total cell protein is expressed as the content of each amino acid relative to the sum of all amino acids recovered after acid hydrolysis. Cell residues obtained after hot trichloroacetic acid extraction display small variations in amino acid profile. Some of these variations correlate with the growth rate at satisfactory levels of statistical significance. In C. utilis, the correlations cover increased levels of lysine, arginine, and leucine and decreased levels of serine and glutamic acid with increased “reduced dilution rate” (D/Dc). In E. aerogenes, increased levels of lysine and arginine and a decreased level of glutamic acid correlate with increased dilution rate. The directions of most of these correlations and the extents of those pertaining to lysine and arginine are consistent with the change predicted to occur simultaneously in the relative level of the ribosomal protein group.
    Additional Material: 5 Tab.
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  • 114
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1193-1210 
    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 for the growth of an organism on multiple substrates was developed, assuming that each substrate has a competitive inhibition effect on the uptake of other substrates. The model was extended to examine mixed substrates, showing that the coexistence of several species at steady state in continuous cultures is possible, even when all the organisms all strongly prefer the one substrate. The diversity of nutrient sources in a real system may be a key factor in supporting a heterogeneous microbial population.
    Additional Material: 9 Ill.
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  • 115
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1211-1213 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 1 Ill.
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  • 116
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1239-1244 
    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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  • 117
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977) 
    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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  • 118
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1303-1320 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An extracellular polymer was produced by continuous fermentation of Corynebacterium hydrocarboclastus on kerosene in a 24 liter reactor. This polymer was composed of protein, lipid, and carbohydrates. The polymer possesed surface active properties, and had two critical micelle concentrations. Its effectiveness was quite comparable to the effectiveness of synthetic surface active agents such as Tween 80 and Span 20; however, its efficiency was much lower. The polymer also had emulsifying properties. Maximum emulsification was obtained at pH 6. The emulsifying properties were unaffected by high salt concentration [up to 5% (w/v) in Na+], and tolerated a water hardness up to 5,000 ppm. A 2 hr treatment of the polymer at temperatures higher than 65°C resulted in a loss of its emulsifying properties. Two microorganisms, named SLYS and Y, isolated from soil, were able to grow on the polymer as sole carbon and energy source, thus proving its biodegradability. SLYS was tentatively identified as Flavobacterium breve and Y as Flavobacterium devorans.
    Additional Material: 13 Ill.
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  • 119
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977), S. 1331-1349 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Candida lipolytica (strain ATCC 8662) was grown on a simple defined medium with n-hexadecane as the main carbon Source under batch fermentation conditions. The relative importance of the cells growing in the aqueous phase on the overall kinetics was studied. The effect of interfacial tension, unoccupied interfacial area, and pseudosolubility on the specific growth was also studied. Results are presented and discussed here.
    Additional Material: 10 Ill.
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  • 120
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    Biotechnology and Bioengineering 19 (1977), S. 1351-1361 
    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 approach to preparative organic synthesis in aqueous-organic systems is suggested. It is based on the idea that the enzymatic process is carried out in a biphasic system “water-water-immiscible organic solvent.” Thereby the enzyme is localized in the aqueous phase - this eliminates the traditional problem of stabilizing the enzyme against inactivation by a nonaqueous solvent. Hence, in contrast to the commonly used combinations “water-water-miscible organic solvent,” in the suggested system the content of water may be infinitely low. This allows one to dramatically shift the equilibrium of the reactions forming water as a reaction product (synthesis of esters and amides, polymerization of amino acids, sugars and nucleotides, dehydration reactions, etc.) toward the products. The fact that the system consists of two phases provides another very important source for an equilibrium shift, i.e., free energies of the transfer of a reagent from one phase to the other. Equations are derived describing the dependence of the equilibrium constant in a biphasic system on the ratio of the volumes of the aqueous and nonaqueous phases and the partition coefficients of the reagents between the phases. The approach has been experimentally verified with the synthesis of N-acetyl-L-tryptophan ethyl ester from the respective alcohol and acid. Porous glass was impregnated with aqueous buffer solution of chymotrypsin and suspended in chloroform containing N-acetyl-L-tryptophan and ethanol. In water (no organic phase) the yield of the ester is about 0.01%, whereas in this biphasic system it is practically 100%. The idea is applicable to a great number of preparative enzymatic reactions.
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  • 121
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    Biotechnology and Bioengineering 19 (1977), S. 1387-1403 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simplified procedure for the preparation of 1,4-α-glucan phosphorylase from Klebsiella pneumoniae is described. An 80-fold purification is achieved in two steps with an overall yield of about 50%. The specific activity of the homogeneous enzyme protein is 17.7 units/mg. Compared with glycogen phosphorylase from rabbit muscle the enzyme from K. pneumoniae shows a markedly higher stability against deforming and chaotropic agents. The 1,4-α-glucan phosphorylase was covalently bound to porous glass particles by three different methods. Coupling with glutaraldehyde gave the highest specific activity, i.e., 5.6 units/mg of bound protein or 133 units/g of glass with maltodextrin as substrate. This corresponds to about 30% of the specific activity of the soluble enzyme. With substrates of higher molecular weight, such as glycogen or amylopectin, lower relative activity was observed. The immobilized enzyme preparations showed pH activity profiles which were slightly displaced to higher values and exhibited an increased temperature stability.
    Additional Material: 5 Ill.
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  • 122
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 19 (1977) 
    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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  • 123
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    Biotechnology and Bioengineering 19 (1977), S. 1463-1473 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In a previous article, the method of preparation and the physical properties of porous (75 to 80% porosity) cellulose beads were described (Biotechnol. Bioeng., 18, 1057 (1976)). The present article reports that the chemical procedures employed for immobilizing enzymes on ordinary cellulose can be applied to the porous cellulose beads. The results showed more enzyme loading on the beads than ordinary cellulose. The choice of the procedures might also affect the mechanical strength of the cellulose beads.
    Additional Material: 8 Ill.
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  • 124
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    Biotechnology and Bioengineering 19 (1977), S. 1493-1501 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Lipase (EC 3.1.1.3., from Pseudomonas sp.) was entrapped in collagen membrane containing liquid crystal (4-methoxybenzilidene-4′-n-butylaniline). The activity of the lipase-liquid crystal membrane at an applied voltage of 4 V was 3.4 compared to a membrane tested without imposition of an external electric field. A linear relationship was observed between the activity of the lipase-liquid crystal membrane and the current. The apparent Michaelis constant (K′m) of the lipase-liquid crystal membrane under electric field was identical to that of the membrane under ordinary condition. Activation of the lipase-liquid crystal membrane was observed repeatedly, i.e., activation in the presence of an electric field and reversion to a basal level upon removal of the field occurred cyclically. Activity control of immobilized enzymes is desirable for switching devices of a bioreactor. Possible mechanisms of the lipase activation by electric field are discussed.
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  • 125
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    Biotechnology and Bioengineering 19 (1977), S. 1523-1534 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A continuous high-speed horizontal colloid mill of novel design for use in the microbiological and food industries was tested for the disintegration of cells of Saccharomyces cerevisiae and Candida utilis. The mill consists of a horizontal vessel with round or oval cross sections fitted with a high-speed longitudinal agitator shaft on which are mounted disk agitators, alternating radially and obliquely to the shaft. The mill is partly filled with freely moving grinding elements which, during a continuous operation, are maintained in the vessel by a vibrating annular slot separator. Highly efficient cooling is provided by circulation of cooling fluid through a jacket surrounding the vessel as well as through the agitator shaft and disks. The radial agitator disks impart a radial motion to the grinding elements, while the oblique disks give rise to the axial movement of a substantial part of the elements. The crossing of paths thus achieved gives the mill a very high efficiency. Using a mill of 20 liter nominal capacity, the effects of agitator design, agitator speed, flow rate, and concentration of the cell suspension on the disintegration efficiency and heat production were studied. Ninety per cent of S. cerevisiae cells in a 15% suspension could be broken at a residence time of 2.5 min. The temperature rise did not exceed 8° C. The corresponding figure for C. utilis was 84%. The maximal flow rate was 400 liter/hr. Extrapolation indicates that available industrial mills of 300 liter capacity based on the same design can handle flows of 2000 liter/hr.
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  • 126
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    Biotechnology and Bioengineering 19 (1977), S. 1557-1561 
    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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  • 127
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    Biotechnology and Bioengineering 19 (1977), S. 1563-1621 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 9 Ill.
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  • 128
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    Biotechnology and Bioengineering 19 (1977), S. 1667-1677 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The stereospecific hydrolysis of D,L-phenylalanine methylester with immobilized α-chymotrypsin was carried out as a model reaction for the racemate resolution of aromatic amino acids in a five staged fluidized-bed reactor (FBR). Owing to ester hydrolysis, a pH shift occurred along the reactor. Because of the pH-dependent enzyme activity a particular longitudinal pH profile had to be enforced by a proper entrance pH in order to gain an optimum conversion. In the FBR with optimum pH profile, higher conversions were achieved than in a continuous stirred tank reactor (CSTR) at the pH optimum and at the same contact time. By the application of a proton balance and the results of kinetic measurements a model was developed for the prediction of the optimum longitudinal pH profile with regard to the maximum conversion.
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  • 129
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A continuous stirred tank fermentor (CSTF) used for cultivation of the fungus Morchella crassipes in ammonia base waste sulfite liquor (NH3-WSL) was considered as a multivariable linear system around its operating point. Pulse testing on the inputs (inlet jacket temperature, inlet pH, inlet substrate concentration) and their responses at the outputs (biomass, outlet temperature, outlet jacket temperature, outlet pH, outlet substrate concentration) were used for numerical determination of the transfer function matrix:
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  • 130
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    Biotechnology and Bioengineering 19 (1977), S. 1679-1687 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple, low-priced 30 liter tower-type algal pilot plant for the cultivation of light- and motion-sensitive species is described. Two hundred g wet weight of Microcystis aeruginosa were obtained per harvest. Since the self-shading of denser cultures could be compensated for only to a limited extent by increasing the light intensity without damaging the cells, the efficiency of various culture-vessel widths was determined for the growth of Microcystis : the best results were obtained with a width of 3.5 cm. Light requirements of Microcystis were studied in shadowless suspensions. The compensation point of photosynthesis varied between 200 and 300 lx, depending on the preillumination, whereas the light saturation point was found to be near 4000 Ix. The light optimum for photosynthesis was not identical with that for good growth.
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  • 131
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Papain and lipase were immobilized on derivatized Sepharose 4-B. The activated agarose had a binding capacity of 1.2 μmol amino groups/ml packed agarose or 17 mg proteins/g dry agarose. The immobilized enzyme preparations were tested for the effects of pH of assay, temperature of assay, and substrate concentrations. The effect of 6M urea on the activity of papain was also determined. Soluble forms of the enzymes were used for comparison. Immobilization of the enzymes resulted in slightly different pH and temperature optima for activities. For immobilized papain Km (app) was similar to the one observed with soluble papain. Immobilization of lipase, however, caused a decrease in Km values. The immobilized enzyme preparations were stable when stored at 4°C and pH 7.5 for periods up to eight months. The soluble enzymes lost their activity within 96 hr under similar storage conditions. Immobilized papain did not lose any activity after treatment with 6M urea for 270 min, whereas soluble papain lost 81% of its activity after the urea treatment, indicating that the immobilization of papain imparted structural and conformational stability to this enzyme.
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  • 132
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    Biotechnology and Bioengineering 19 (1977) 
    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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  • 133
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    Biotechnology and Bioengineering 19 (1977), S. 1773-1784 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: It is shown that the mass transfer resistance can significantly distort the linearity of the Lineweaver-Burk plot of the kinetic data for a microbial culture which forms aggregates. For small flocs, the linearity of the Lineweaver-Burk plot is largely retained, but a different slope and intercept will be obtained compared with flocs free from mass transfer resistance. For large flocs, the Lineweaver-Burk plot shows pronounced curvature at high limiting substrate concentrations. Hence, if the true intrinsic kinetic parameters are to be extracted from a highly flocculating microbial culture, sufficient agitation has to be provided to remove the effect of mass transfer resistance. If the behavior of the flocculating microbial culture is to be explored, additional values for some physical parameters, such as the effective diffusion coefficient of the substrate in floc, the floc density, and the mean floc radius, are needed.
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  • 134
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    Biotechnology and Bioengineering 19 (1977), S. 1831-1850 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The estimation of parameters in several dynamic models, which describe growth and substrate consumption, has been carried out using a modified Gauss-Newton-type method. The four models considered are Monod, Contois, linear specific growth rate, and an enzyme kinetic model. The initial values of the differential equations are included in the parameter vector which will be estimated. The efficiency of the method and the confidence limits of the parameters were studied using simulated measurement noise. The experimental results describe Trichoderma viride growing on glucose as the main carbon source.
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  • 135
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    Biotechnology and Bioengineering 19 (1977) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 136
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    Biotechnology and Bioengineering 19 (1977), S. 55-67 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Despite the importance of biomass as a parameter in fermentation processes, there are no commercially available sensors suitable for its measurement. An indirect approach for the assessment of biomass concentration can be based on material balances and on the direct monitoring of fermentation parameters for which there are established sensors (e.g., gaseous oxygen and carbon dioxide). As a consequence, this method requires no assumption of cellular yield coefficients or rate constants. This approach is also readily adaptable to general use since it requires only some knowledge of the compositions of the substrate, cells, and noncellular products.
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  • 137
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    Biotechnology and Bioengineering 19 (1977), S. 69-86 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The economics of yeast production depend heavily upon the cellular yield coefficient on the carbon source and the volumetric productivity of the process. The application of an on-line computer to maximize these two terms during the fermentation requires a continuous method of measuring cell density and growth rate. U fortunately, a direct sensor for biomass concentration suitable for use in industrial fermentations is not available. Material balancing, with the aid of on-line computer monitoring, offers an indirect method of measurement. Laboratory results from baker's yeast production in a 14-liter fermentor (with a PDP-11/10 computer for on-line analyses) show this indirect measurement technique to be a viable alternative. From the oxygen uptake and carbon dioxide production data, gas flow rate, and ammonia addition rate, the cell density during the fermentation has been estimated and found to compare well with actual fermentation data.
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  • 138
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    Biotechnology and Bioengineering 19 (1977), S. 185-198 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The kinetic behavior of a system of multiple enzyme in solution has been studied in a variable volume batch reactor at pH 5, controlled dissolved oxygen concentration, and T = 30°C. The enzymes used were glucoamylase (R. delemar), glucose oxidase (A. niger), and gluconolactonase (A. niger), all of which are important commercial biocatalysts, and a disaccharide was employed as the starting substrate. This study includes the basic kinetic properties of individual enzymes and interactions between components of the reaction mixture. Classical Michaelis-Menten single substrate or two substrate kinetic with parameters based on initial rate data predict correctly the batch time course of the sequential reaction network.
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  • 139
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    Biotechnology and Bioengineering 19 (1977), S. 159-184 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucose isomerase in the form of heat-treated whole-cell enzyme prepared from Streptomyces phaeochromogenus follows the reversible single-substrate reaction kinetics in isomerization of glucose to fructose. Based on the Kinetic constants determined and the mathematical model of the reactor system developed, the preformance of a plug-flow-type continuous-enzyme reactor system was studied experimentally and also simulated with the aid of a computer for the ultimate objective of optimization of the glucose isomerase reactor system.The enzyme decay function for both the enzyme storage and during the use in the continuous reactor, was found to follow the first-order decay kinetics. When the enzyme decay function is taken into consideration, the ideal homogeneous enzyme reactor kinetics provided a satisfactory working model without further complicatin of the mathematical model, and the results of computer simulation were found to be in good agreement with the experimental results. Under a given set of constraints the performance of the continuous glucose isomerase reactor system can be predicted by using the computer simulation method described in this paper.The important parameters studied for the optimization of reactor operation were enzyme loading, mean space time of the reactor, substrate feed concentration, enzyme decay constants, and the fractional conversion, in addition to the kinetic constants. All these parameters have significant effect on the productivity.Some unique properties of the glucose isomerization reaction and its effects on the performance of the continuous glucose isomerase reactor system have been studied and discussed. The reaction kinetics of glucose isomerase and the effects of both the enzyme loading and the changes in reaction rate within a continuous reactor on the productivity are all found to be of particular importance to this enzyme reactor system.
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  • 140
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    Biotechnology and Bioengineering 19 (1977), S. 199-210 
    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 intracellular RNase in disintegrated cell suspensions of Saccharomyces cerevisiae have been studied. The influence of salt addition and/or incubation of the suspension on the activity of RNase and on the degradation of endogenous RNA was determined. No significant change in the RNase activity in the disintegrated suspensions was obtained by addition of 3% NaCl or by incubation at 50°C with 3% NaCl. During the incubation with NaCl the active RNase was able to degrade endogenous RNA. By incubation without salt the RNase was inactivated. Inactivation also occurred after extraction at alkaline pH. The RNase had an optima at pH 5-6 and temperatures between 50-60°C. The main part of the RNase in the unincubated suspension was soluble at pH 5.6 but not at pH 4.0. After incubation with NaCl the RNase was soluble at pH 4.0. No serious protein degradation occurred during the short time incubation needed for RNA reduction. 70% of the protein in the suspensions was recovered in the precipitate at pH 4.0 after 20 min of incubation. The corresponding protein recovery from unincubated suspensions was 77%.
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  • 141
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    Biotechnology and Bioengineering 19 (1977), S. 211-218 
    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 mechanochemical method for enzyme immobilization has been elaborated. The principle of this method consists of the following precepts. Partially hydrolyzed nylon fiber, the surface of which is known to be strewn with micro-cracks, is reversibly stretched (∼25%) and placed into an enzyme solution. Then, in the same solution, the fiber is made to relax and is taken out. The fiber retains considerable enzymatic activity even after numerous thorough washings (in a similar procedure without fiber stretching, equivalent washing removed all the enzymatic activity from the fiber). Immobilization on the fiber proceeds due to trapping of enzyme molecules by the microcavities on the surface of the support. The catalytic activity of mechanochemically immobilized chymotrypsin and trypsin is commensurable with their activity on covalent immobilization on nylon (calculated per unit of the macrosurface). A wide range of commercial polymers may be made of use as supports in the mechanochemical method of immobilization.
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  • 142
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    Biotechnology and Bioengineering 19 (1977), S. 219-233 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Experiments were carried out on dextran-dextranase systems to test the prediction of a mechanistic model recently proposed by us, for the synergistic effect of combined exo/endo enzymic action in the degradation of polymeric substrate. Soluble forms of the substrate were used. Preliminary experiments with an insoluble form of the substrate were also carried out to demonstrate the applicability of the analytical techniques to these cases. Molecular weight distributions of the degradation products were determined (by gel-permeation chromatography) and the rates of production of glucose and of other reducing sugars were also measured. It was found that the exodextranase alone had very little effect on the molecular weight distributions compared to a significant shift towards lower molecular weight obtained with the endodextranase which was synergistically enhanced by the action of the combined enzymes. Glucose was produced more rapidly by the exoenzyme compared to the endoenzyme, but combinations of the two enzymes gave a rate enhancement greater than the linear sum of the effects of the two individual enzymes. In comparing the degradation indices and polydispersities of the various degradation products, similar synergistic effects of the combined enzymes in accordance with the theoretical predictions, were observed. The practical implications of these findings to the design of fermentation processes which depend on the action of endo- and exoenzyme mixtures are noted.
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  • 143
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    Biotechnology and Bioengineering 19 (1977), S. 337-348 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Using ball milled cellulose as the only carbon source Trichoderma viride was grown in a continuous flow culture at pH = 5.0 and T = 30°C. Steady-state values for cell protein, cellulose, and cellulase for different substrate concentrations (4-11 g/liter) and dilution rates (0.033-0.080 hr-1) were obtained. Under steady-state conditions, 50-75% of the cellulose was consumed indicating a critical dilution rate on 0.17 hr-1.Cellulase activity (U/ml) in the fermentation broth increased slightly with increasing substrate concentration and decreased with increasing dilution rate, while the specific cellulase productivity (U/mg cell protein·hr) was fairly independent of the dilution rate, with a maximum around D = 0.05 hr-1.Following step changes in substrate concentration and dilution rate, new steady-state values were reached after three to five residence times (cell protein and cellulose) and four to six residence times (celullase activity).
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  • 144
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    Biotechnology and Bioengineering 19 (1977), S. 445-458 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A theoretical model is developed for continuous multistage enzyme production systems, which consist of a growth fermentor used for growing microorganisms rapidly without enzyme production and a subsequent system of induction reactors in which enzymes induction and production occurs. The model allows the computation of the fraction of induced cells residing in the induction reactor for organisms exhibiting a lag phase in enzyme induction. For this model a general analytical solution was obtained for the cumulative internal residence time distribution of a series of n well-stirred vessels with a recycle. The theoretical results are compared in a preliminary way with experimentally measured cellulase productivities of continuous multistage cellulose fermentations with Trichoderma viride QM 9414.
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  • 145
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    Biotechnology and Bioengineering 19 (1977), S. 467-492 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The kinetics of phosphate limited growth of two green algae Chorella pyrenoidosa and Selenastrum capricornutum have been studied in chemostats. Several kinetic models which express the specific growth rate as a function of the intracellular phosphours content have been examined, and one of the models was found to be significantly better than the other models. The principles of this model were described in a recent paper by Nyholm.The kinetics of phosphate uptake have been investigated by adding pulses of phosphate to the chemostats. The uptake by phosphours deficient cells could be described by Michaelis-Menten kinetics for phosphate concentrations below approximately 500 μg P/liter. Further, with the assumption of a discontinuous adjustment of the uptake rate at the onset of phosphours deficiency, a complete kinetic model for growth and phosphate removal is proposed.The mean cell size and the contents of chlorophyll and RNA per unit dry weight have been measured for C. pyrenoidosa as a function of the dilution rate.
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  • 146
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    Biotechnology and Bioengineering 19 (1977), S. 557-559 
    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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  • 147
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    Biotechnology and Bioengineering 19 (1977), S. 561-564 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 148
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    Biotechnology and Bioengineering 19 (1977), S. 565-573 
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    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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  • 149
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    Biotechnology and Bioengineering 19 (1977), S. 591-593 
    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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  • 150
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    Additional Material: 5 Ill.
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  • 151
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    Biotechnology and Bioengineering 19 (1977) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 152
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    Biotechnology and Bioengineering 19 (1977), S. 611-618 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 153
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    Biotechnology and Bioengineering 19 (1977), S. 605-610 
    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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  • 154
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    Biotechnology and Bioengineering 19 (1977), S. 621-629 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The gas-liquid oxygen transfer rate is a key step in the production of antibiotics in submerged fermentation. If the gas-liquid oxygen mass transfer rate is not equal to the required liquid-solid oxygen mass transfer rate at a particular cell concentration, then productivity of the particular fermentation operation will not be the maximum possible value.One way to increase the productivity of a given fermentation tank installation is to increase the cell concentration and to increase the oxygen transfer by changing the mixer and air supply to match the new requirements. In order to evaluate the cost of making this change to the larger mixing equipment, a typical cost example is given which can easily be modified for other combinations of production cost and mixer cost. As an example, it is seen that a considerable savings can result from a given installation by primarily changing the oxygen transfer ability of the equipment to utilize a given fermentor more efficiently. Production cost savings of 8 to 25% are shown in the example cited.
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  • 155
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    Biotechnology and Bioengineering 19 (1977), S. 649-660 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: On the basis of elastic waves released by imploding cavitation bubbles, a mechanism for biological cell disintegration in high intensity ultrasounds has been proposed. Comparison of this mechanism with the published results on yeast cells shows many points of agreement suggesting that yeast cell disintegration in ultrasonic cavitation occurs by shear stresses developed by viscous dissipative eddies arising from shock waves.
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  • 156
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    Biotechnology and Bioengineering 19 (1977), S. 821-839 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The principles of a method for the continuous manufacture of yogurt, based upon a two stage system, are given. The first stage, the prefermentation of milk at 45°C to a pH of 5.7, is described.The limitations of this continuous prefermentation are experimentally determined. No change in the balance of the yogurt bacteria, Lactobacillus bulgaricus and Streptococcus thermophilus, was observed. A mathematical approach is given for starting and stopping the process.
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  • 157
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    Biotechnology and Bioengineering 19 (1977), S. 801-819 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Rates of CO2 desorption from fermentation broths under actual operating conditions were determined by measuring the CO2 partial pressure in the exit gas. The concentrations of CO2 physically dissolved in the broths were measured by the so-called tubing method. Values of kLa for CO2 desorption calculated from these values agreed well with the kLa values for oxygen absorption corrected for the difference in gas diffusivities. The dissolved CO2 concentration in the broth, which seems to bean important operating parameter, can easily be estimated from the CO2 partial pressure in the exit gas, a more easily measurable quantity, if the kLa value is known. For a given value of kLa, assumption of perfect mixing or plug flow in the gas phase made little difference in the calculated values of the dissolved CO2 concentration, indicating that the gas phase was probably in between perfect mixing and plug flow.In industrial fermentors, the CO2 partial pressure in the exit gas can practically be assumed to be in equilibrium with the dissolved CO2 concentration.
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  • 158
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    Biotechnology and Bioengineering 19 (1977), S. 841-851 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The procedure and apparatus for the continuous coagulation of yogurt are described. The continuous coagulation takes place in a plug-flow fermentor. Prefermented milk is brought into this fermentor with the help of a centrifugal distributor, which avoids any undesirable mixing of the prefermented milk with the acidifying milk. A special stirring plate allows a stirring treatment in the coagulation tank. By this procedure the acidity and the viscosity of the final yogurt can be controlled between certain limits. The organoleptic characteristics of the continuous manufactured yogurt are good.
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  • 159
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    Biotechnology and Bioengineering 19 (1977), S. 853-865 
    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 method to estimate the oxygen transfer coefficient (KLa) from the experimental dynamic response data is presented. Employing a linear model which allows for gas phase, diffusion film, and oxygen electrode dynamics, the first moment of the response curve is simply related to the sum of the model parameters. Two separate experiments are used to characterize the measurement dynamics and to measure the unknown KLa parameter. The simple calculation procedure involves only measuring the area above the response curves.
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  • 160
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    Biotechnology and Bioengineering 19 (1977), S. 923-928 
    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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  • 161
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    Biotechnology and Bioengineering 19 (1977), S. 867-878 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A laboratory-scale research program was undertaken to investigate the kinetics of the mesophilic (37°C) anaerobic digestion of brewery industry by-product. The purpose was to develop data for the design and operation of full-scale units which could be used to generate methane fuel gas from these materials. This is important because the brewery industry has been susceptible to shortages of natural gas in recent years. The minimum SRT is 2.3 days, although for design purposes as much as ten days is recommended. The biomass yield is 0.512 g volatile suspended solids (VSS)/g volatile solids (VS) or 0.421 g VSS/g chemical oxygen demand (COD). The maintenance requirement is 0.052 g VS/g VSS per day or 0.061 g COD/g VSS per day. The specific methane yield is 2.51 liter/g VSS, and the methane productivity is 0.32-0.41 liter/g dry substrate added or 0.69-0.91 liter/g destroyed. The maximum loading rate for which substrate inhibition is not observed is 6 g dry substrate added per liter per day. The results of the entire program indicate that processing brewery by-product in this manner is both technically feasible and economically attractive.
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  • 162
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    Biotechnology and Bioengineering 19 (1977), S. 901-921 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We have demonstrated that a simple electrochemical cell can serve as a detector of NADH concentration in a flow system thereby providing an assay technique for NADH dependent enzymes. When this is applied to NADH produced by enzymatic reaction, then a reproducible measure of enzyme activity is obtained. This method of enzyme activity assay is applicable to a number of oxidoreductase enzymes which employ NAD+ or NADP+ as coenzymes to achieve substrate modification. The presence of electroactive species in samples of human serum has proved a serious problem in the electrochemical analysis of serum activity. These species produce a large background anode current at the anode voltage appropriate for NADH oxidation. The presence of this high current limits the usefulness of amplification of the current output to detect small changes in NADH concentration.
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  • 163
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    Biotechnology and Bioengineering 19 (1977), S. 879-899 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A gram-negative bacterium strongly lytic toward living cells of the food yeast Saccharomyces fragilis was isolated by continuous-flow enrichment from compost. The organism was identified as a species of Arthrobacter. The extracellular lytic enzyme complex produced by this bacterium contained β-1,3-glucanase, mannan mannohydrolase, and proteolytic activities. The polysaccharases were inducible by whole yeast cells. In chemostat cultures on chemically defined media, synthesis of the polysaccharases was very slight and only detectable at dilution rates below 0.02 hr-1. Enzyme production in defined media was not solely dependent on growth rate but also was influenced by the growth limiting substrate and the culture history. The production of individual depolymerases and of the lytic activity was studied in batch and chemostat cultures containing yeast as the limiting substrate. The maximum specific growth rate of the Arthrobacter under these conditions was 0.22 hr-1. β-1,3-Glucanase and proteolytic activities were synthesized by exponentially growing bacteria but maximum lytic titers did not develop until the specific growth rate was declining, at which time mannan mannohydrolase syntheses was induced. In yeast limited chemostats polysaccharase syntheses were greatest at the lowest dilution rates examined, namely 0.02 hr-1. Further optimization of enzyme production was achieved by feeding the Arthrobacter culture to a second-stage chemostat. A comparison of lytic enzyme productivities in batch and chemostat cultures has been made.
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  • 164
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    Biotechnology and Bioengineering 19 (1977), S. 933-935 
    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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  • 165
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    Biotechnology and Bioengineering 19 (1977), S. 929-932 
    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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  • 166
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    Biotechnology and Bioengineering 19 (1977) 
    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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  • 167
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    Biotechnology and Bioengineering 19 (1977), S. 937-937 
    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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  • 168
    ISSN: 0091-7419
    Keywords: insulin ; glucagon ; transport ; amino acids ; diabetes ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The transport of 2-aminoisobutyric acid (AIB) into liver tissue was increased by both insulin and glucagon. We have now shown that these hormones do not stimulate the same transport system. Glucagon, possibly via cAMP, increased the hepatic uptake of AIB by a mechanism which resembled system A. This glucagon-sensitive system could be monitored by the use of the model amino acid MeAIB. In contrast, the insulin-stimulated system exhibited little or no affinity for MeAIB and will be referred to as system B. On the basis of other reports that the hepatic transport of AIB is almost entirely Na+ dependent and the present finding that the uptake of 2-aminobicyclo [2,2,1] heptane-2-carboxylic acid (BCH) was not stimulated by either hormone, we conclude that system B is Na+ dependent. Furthermore, insulin added to the perfusate of livers from glucagon-pretreated donors suppressed the increase in AIB or MeAIB uptake. Depending upon the specificities of systems A and B, both of which are unknown for liver tissue, the insulin/glucagon ratio may alter the composition of the intracellular pool of amino acids.
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  • 169
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    Journal of Supramolecular Structure 6 (1977), S. 215-228 
    ISSN: 0091-7419
    Keywords: reconstitutions of ion pumps ; coupling factors of oxidative phosphorylation ; phospholipids ; role in ion pump activity ; mechanism of ATP-driven Ca2+ pump ; oxidative phosphorylation ; a new hypothesis ; ATPases of membranes ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Reconstitutions of membranous activities can tell us how many components are required and what their functions are. The mitochondrial proton pump is used as an example. Moreover, the biological activity, such as Pi transport, can be used in reconstituted vesicles as an assay during the isolation of the transporter.Reconstitution experiments reveal the importance of membrane asymmetry and allow us to study conditions of vectorial assembly.The mechanism of action of ion pumps has been successfully analyzed in reconstituted liposomes. We can study the movement of ions and the electrogenicity of the system without interference by other unrelated processes.Based on studies with the resolved Ca2+-ATPase of sarcoplasmic reticulum, we propose a novel formulation of the mechanism of ATP-driven ion pumps in which cyclic binding of Mg2+ plays a key role.
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  • 170
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    Journal of Supramolecular Structure 6 (1977), S. 1-12 
    ISSN: 0091-7419
    Keywords: sugar transport ; cell shape ; transformed chick cells ; methyl cellulose ; scanning electron microscopy ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The rate of hexose transport was compared in normal and virus-transformed cells on a monolayer and in suspension. It was shown that: (1) Both trypsin-removed cells and those suspended for an additional day in methyl cellulose had decreased rates of transport and lower available water space when compared with cells on a monolayer. Thus, cell shape affects the overall rate of hexose transport, especially at higher sugar concentrations. (2) Even in suspension, the initial transport rates remained higher in transformed cells with reference to normal cells. Scanning electron micrographs of normal and transformed chick cells revealed morphological differences only in the flat state. This indicates that the increased rate of hexose transport after transformation is not due to a difference in the shape of these cells on a monolayer.
    Additional Material: 6 Ill.
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  • 171
    ISSN: 0091-7419
    Keywords: amoeboid movement ; calcium ions ; cell shape ; Naegleria gruberi ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Amoebae of Naegleria gruberi differentiate to temporary flagellates that have a regular, asymmetric, streamlined body contour. During the hour-long differentiation, amoeboid movement gradually ceases and as a consequence the cells round up. Subsequent elongation to flagellate shape includes the formation of a microtubular cytoskeleton. Both the loss of amoeboid motility and the formation of the flagellate shape require prior transcription and translation, suggesting the possibility that specific syntheses of RNA and protein may be required for each shape change. Flagellates can “revert” to motile amoebae within 20 sec after a suitable stimulus, indicating that the amoeboid motility system remains latent in flagellates. A cell-produced chemical factor extracted from Naegleria, Ψ, triggers a reproducible sequence of rapid shape changes in flagellates when added to their environment. Cells respond to the presence of external Ψ only “transiently,” and the reaction of flagellates to added Ψ requires extracellular Ca+2. Ionophore A23187 produces shape changes in flagellates similar to those produced by Ψ, supporting the conclusion that Ψ is involved in the movement of Ca+2. Normally Ψ is intracellular, and the intracellular distribution of Ψ changes during differentiation.These results lead to and support a working hypothesis to explain the rapid changes in shape and motility in Naegleria. Four elements are postulated: Ca+2; an actin-based amoeboid motility system that depends on free Ca+2 for functioning; a tubulin-based cytoskeleton that assembles and remains assembled only when free Ca+2 is low; and Ψ. The factor Ψ is postulated to regulate the intracellular release of Ca+2. According to the hypothesis, intracellular free Ca+2 is constantly swept up into Ca-reservoirs. Motility of amoebae depends on local release of Ca+2 from these reservoirs, which in turn is caused by the intracellular release of Ψ. During differentiation, Ψ is “compartmentalized” as part of the developmental program, and as a consequence intracellular Ca+2 is swept up into Ca-reservoirs but not released. As free Ca+2 becomes limiting, amoeboid movement stops, and the cells round up. Subsequently, in a process that depends on low free Ca+2, the microtubular cytoskeleton is assembled, and the flagellate shape is formed. During reversion of flagellates to amoebae, release of Ψ from its “compartments” permits local release of Ca+2, which then causes both disassembly of the flagellate cytoskeleton and immediate resumption of amoeboid movement. This testable hypothesis has implications for the study of cell shape, motility, and differentiation.
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  • 172
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    Journal of Supramolecular Structure 6 (1977), S. 291-299 
    ISSN: 0091-7419
    Keywords: rhodopsin ; retinal disk membranes ; galactosyl transferase ; fluorescent probes ; carbohydrate unit ; enzymatic modification ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Galactose was specifically inserted into the carbohydrate moiety of rhodopsin by incubating retinal disk membranes with UDP-galactose: N-acetylglucosamine galactosyltransferase. The stoichiometry of labeling ranged from 1.2 to 1.8 (average = 1.5) residues of galactose per molecule of rhodopsin, indicating that some or all of the oligosaccharide chains of membrane-bound rhodopsin are readily accessible to enzymatic modification. These modified membranes were treated with galactose oxidase to generate an aldehyde at the C-6 position of the inserted galactose units. The enzymatically-oxidized membranes were then reacted with dansyl hydrazide to yield a fluorescent hydrazone which is sufficiently stable to permit spectroscopic analysis. This procedure for the specific attachment of a spectroscopic probe should be applicable to a wide variety of membrane glycoproteins.
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  • 173
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    Journal of Supramolecular Structure 6 (1977), S. 363-374 
    ISSN: 0091-7419
    Keywords: thymidine transport ; nitrobenzylthioinosine ; bromodeoxyuridine resistances ; HeLa cells ; thymidine kinase ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A line of HeLa cells resistant to 5-bromo-2′-deoxyuridine (BUdR) was established by continuous culture in growth medium containing BUdR; during the selection period, BUdR concentrations, initially 15 μM, were gradually increased to 100 μM. Cells of a clone (HeLa/B5) established from this line were also resistant to 5-fluoro-2′-deoxyuridine (FUdR), but not to the free base, 5-fluorouracil. Although extracts of HeLa/B5 cells exhibited levels of thymidine kinase activity comparable to those of parental cells, rates of uptake of BUdR, FUdR, and thymidine into intact cells were much reduced. The kinetics of uptake of uridine and adenosine, nucleosides which appear to be transported independently of thymidine in HeLa cells, were similar for HeLa/B5 and the parental line (HeLa/0). Relative to thymidine uptake by HeLa/0 cells, that by HeLa/B5 cells was distinctly less sensitive to nitrobenzlthionosine (NBMPR), a specific inhibitor of nucleoside transport in various types of animal cells. Despite this difference in NBMPR sensitivity, both cell lines possessed the same number of high affinity NBMPR binding sites per mg cell protein. The altered kinetics of thymidine uptake and the NBMPR insensitivity of that function in HeLa/B5 cells suggest that resistance to BUdR is due to an altered thymidine transport mechanism.
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  • 174
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    Journal of Supramolecular Structure 6 (1977), S. 375-381 
    ISSN: 0091-7419
    Keywords: human erythrocytes ; ATP-dependent Ca uptake ; (Ca+Mg)-ATPase ; spectrin ; inside-out vesicles ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Ghost membranes prepared from human erythrocytes exhibit 2 distinct (Ca+Mg)-ATPase1 activities (Quist and Roufogalis, Arch Biochem Biophys 168:240, 1975). (Ca+Mg)-ATPase activity dependent on a water soluble protein fraction is selectively lost from ghost membranes during preparation of vesicles under low ionic strength, slightly alkaline conditions. In this study, the Ca2+ dependence of the remaining membrane bound (Ca+Mg)-ATPase activity and ATP-dependent Ca uptake in vesicles were compared. The C2+ activation curves for (Ca+Mg)-ATPase activity and Ca uptake into vesicles were parallel over a Ca2+ range of 0.3-330 μM, and both curves have 2 apparent KA values for Ca2+ of 0.45 and 100 μM. Addition of a concentrated soluble protein fraction containing predomintly spectrin to the vesicles increased (Ca+Mg)-ATPase activity over twofold but did not affect the rate of Ca uptake. These findings suggest that the (Ca+Mg)-ATPase activity remaining in vesicles after extraction of the water soluble proteins is associated with the Ca pump whereas (Ca+Mg)-ATPase activity dependent on the soluble protein fraction is associated with some other function.
    Additional Material: 2 Ill.
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  • 175
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    Journal of Supramolecular Structure 6 (1977), S. 179-189 
    ISSN: 0091-7419
    Keywords: valinomycin ; human fibroblast ; amino acid transport ; serum stimulation ; membrane potential ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The Na+-dependent accumulation of α-aminoisobutyric acid (AIB), measured in normal growing and quiescent (serum-deprived) HSWP cells (human diploid fibroblast), was found to be twofold higher (AIBin/AIBout = 20-25) under the normal growing conditions. Serum stimulation of quiescent cells increases their AIB concentrating capacity by approximately 70% within 1 hr. These observations suggest that the driving forces for AIB accumulation may be reversibly influenced by the serum concentration of the growth medium. Addition of valinomycin (Val) to cells preequilibrated with AIB causes an enhanced accumulation of AIB, suggesting that the membrane potential can serve as a driving force for AIB accumulation. After preequilibration with AIB in 6 mM K+, transfer to 94 mM K+ with Val results in a marked and rapid net loss of AIB. The effect of Val on the accumulation of AIB is greatest in quiescent cells, with the intracellular AIB concentrations reaching those seen both in Val-stimulated normal cells and in Val-stimulated serum-stimulated cells. By adjusting [K+]0, in the presence of Val, the membrane potential of growing cells can be matched to that of quiescent cells or vice versa. When this is done, the two accumulate AIB to the same extent. Hence the AIB accumulating capacity is characteristic of the membrane potential rather than of the growth state. In summary, these data suggest that the accumulation of AIB in HSWP cells is influenced by changes in membrane potential and that a serum-associated membrane hyperpolarization could be responsible for the increased capacity for AIB accumulation in serumstimulated cells.
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  • 176
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    Journal of Supramolecular Structure 6 (1977), S. 239-247 
    ISSN: 0091-7419
    Keywords: folate ; thiamine ; transport ; binding proteins ; Triton X-100 ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Two separate binding proteins, one specific for folate and the other for thiamine, have been isolated from membrane fragments of Lactobacillus casei. Purification to homogeneity was achieved by fractionation of the Triton-solubilized proteins with microgranular silica (Quso G-32) and Sephadex G-150. Amino acid analyses revealed that the folate (Mr = 25,000) and thiamine (Mr = 29,000) binders have unusually low polarity constants, 0.32 and 0.26, respectively. Evidence obtained with intact cells has established a direct role for these binding proteins in transport of the corresponding vitamins: (A) In each case, the processes of binding and transport showed similarities in substrate affinities and repression by excess vitamin in the growth medium. (B) Competition studies employing amethopterin, 5-formyl tetrahydrofolate, and 5-methyl tetrahydrofolate (for folate) and thiamine monophosphate and thiamine pyrophosphate (for thiamine) have shown that the ability of these compounds to inhibit the transport of the corresponding vitamins is paralleled by their ability to inhibit binding. (C) Amethopterin-resistant mutants which are defective in folate transport have a comparable defect in ability to bind folate. (D) Amethopterin-resistant cells which (compared with the parent cell line) contain folate transport systems with altered affinities for amethopterin also contain binding proteins whose affinities for amethopterin have changed by equivalent amounts. (E) Both the transport and binding of folate by one of the mutants were stimulated (approximately 3-fold) in parallel by the addition of mercaptoethanol.
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  • 177
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    Journal of Supramolecular Structure 6 (1977) 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 178
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    Journal of Supramolecular Structure 6 (1977), S. 599-616 
    ISSN: 0091-7419
    Keywords: plants ; polysaccharides ; elicitors ; phytoalexins ; Rhizobium ; nitrogen-fixation ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Plants are resistant to almost all of the microorganisms with which they come in contact. In response to invasion by a fungus, bacterium, or a virus, many plants produce low molecular weight compounds, phytoalexins, which inhibit the growth of microorganisms. Phytoalexins are produced whether or not the invading microorganism is a pathogen. The production of phytoalexins appears to be a widespread mechanism by which plants attempt to defend themselves against pests. Molecules of microbial origin which trigger phytoalexin accumulation in plants are called elicitors. Structural polysaccharides from the mycelial walls of several fungi elicit phytoalexin accumlation in plants. Approximately 10 ng of the polysaccharide elicits the accumulation in plants of more than sufficient amounts of phytoalexin to stop the growth of microorganisms in vitro. The best characterized elicitors have been demonstrated to be β-1,3-glucans with branches to the 6 position of some of the glucosyl residues. Oligosaccharides, produced by partial acid hydrolysis of the mycelial wall glucans, are exceptionally active elicitors. The smallest oligosaccharide which is still an effective elicitor is composed of about 8 sugar residues.Bacteria also elicit phytoalexin accumulation in plants, but the Rhizobium symbionts of legumes presumably have a mechanism which allows them to avoid either eliciting phytoalexin accumulation or the effects of the phytoalexins if they are accumulated. The lectins of legumes bind to the lipopolysaccharides of their symbiont, but not of their non-symbiont, Rhizobium. It is not known whether the lectin-lipopolysaccharide interaction is involved with the establishment of symbiosis. However, evidence will be presented that suggests that lectins are, in fact, enzymes capable of modifying the structurs of the lipopolysaccharides of their symbiont, but not of their non-symbiont, Rhizobium. It will also be shown that the lipopolysaccharides isolated from different Rhizobium species and from different strains of individual Rhizobium species have different sugar compositions. Thus, the different strains of a single Rhizobium species are as different from one another as the different species of Salmonella and other gram-negative bacteria. This conclusion is substantiated by experiments demonstrating that antibodies to the lipopolysaccharide from a single Rhizobium strain can differentiate that strain from other strains of the same species as well as from other Rhizobium species. The role in symbiosis of the strain-specific O-antigens is unknown.
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  • 179
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    Journal of Supramolecular Structure 7 (1977), S. 37-48 
    ISSN: 0091-7419
    Keywords: transport ; sulfhydryl oxidants ; p-chloromercuribenzenesulfonate ; glutathione maleimide I ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: At 5 μg/ml, insulin stimulates hexose, A-system amino acid, and nucleoside transport by serum-starved chick embryo fibroblasts (CEF). This stimulation, although variable, is comparable to that induced by 4% serum. The sulfhydryl oxidants diamide (1-20 μM). hydrogen peroxide (500 μM), and methylene blue (50 μM) mimic the effect of insulin in CEF.PCMB-S,1 a sulfhydryl-reacting compound which penetrates the membrane slowly, has a complex effect on nutrient transport in serum- and glucose-starved CEF. Hexose uptake is inhibited by 0.1-1 mM PCMB-S in a time- and concentration-dependent manner, whereas A-system amino acid transport is inhibited maximally within 10 min of incubation and approaches control rates after 60 min. A differential sensitivity of CEF transport systems is also seen in cells exposed to membrane-impermeant glutathione-maleimide I, designated GS-Mal. At 2 mM GS-Mal reduces the rate of hexose uptake 80-100% in serum- and glucose-starved CEF; in contrast A-system amino acid uptake is unaffected. D-glucose, but not L-glucose or cytochalasin B, protects against GS-Mal inhibition. These results are consistent with the hypothesis that sulfhydryl groups are involved in nutrient transport and that those sulfhydryls associated with the hexose transport system and essential for its function are located near the exofacial surface of the membrane in CEF.
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  • 180
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    Journal of Supramolecular Structure 6 (1977), S. 571-577 
    ISSN: 0091-7419
    Keywords: sialic acid uptake ; sialoglycoproteins ; sialoglycolipids ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: BHK cells can be grown in the presence of growth medium to which radiolabeled sialic acid has been added. After 24 h, 85% of the radioactivity in the cells is covalently bound to glycoproteins and glycolipids. No metabolism of the radiolabeled sialic acid could be detected.
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  • 181
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    Journal of Supramolecular Structure 6 (1977), S. 579-589 
    ISSN: 0091-7419
    Keywords: mannosyltransferase ; glycopeptide ; GDP-mannose ; Penicillium ; phosphomannan ; galactofuranosyl ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Membranes from Penicillium charlesii were separated into 6 fractions by sucrose density gradient ultracentrifugation. The least dense fraction (ρ = 1.1 g cm-3) contained GDP-mannose: glycopeptide mannosyltransferases that transferred [14C] mannose onto mannopyranosyl-(seryl/threoyl)-polypeptide and phosphogalactomannan regions of peptidophosphogalactomannan. Approximately 90% of the [14C] mannose incorporated was isolated as mannobiose following treatment of peptidophosphogalactomannan with 0.5 N NaOH. The remainder was located in phosphogalactomannan. About 10% of the membrane-bound mannosyltransferase activity was solubilized with 1% Triton X-100. The soluble mannosyltransferase activity was purified by affinity chromatography on peptidophosphogalactomannan-Sepharose 4B and ammonium sulfate fractionation. Mannose incorporation was shown to be a function of the concentration of added acceptor. No incorporation occurred in the absence of added acceptor or when MgCl2 was substituted for MnCl2. Peptidophosphogalactomannan, phosphogalactomannan, phosphomannan, and mannan, each obtained by appropriate treatment of peptidophosphogalactomannan from P. charlesii, served as mannosyl acceptors. In contrast, α-mannosidase treated peptidophosphogalactomannan did not serve an acceptor of mannosyl residues. Up to 70% of the mannose from GDP-mannose was transferred to added acceptor. Treatment of [14C] mannosyl-labeled peptidophosphogalactomannan with 0.5 N NaOH released 90% of the [14C] mannose as phosphogalactomannan and the remainder was released as mannobiose. [14C] Mannose-labeled phosphogalactomannan was subjected to acetolysis. Mannobiose was the major [14C]-labeled product isolated. Significant quantities of [14C] mannose were isolated also. These results show that soluble mannosyltransferase catalyzes the formation of (1-6)-linked mannosyl residues as well as the transfer of a mannosyl residue to a (1-6)-linked mannosyl residue in the phosphogalactomannan. The specificity of the enzyme is shown by its inability to catalyze mannosyl transfer to α-mannosidase treated peptidophosphogalactomannan, or to incorporate more than 2 mannosyl residues onto the phosphogalactomannan region. Presumably the second mannosyl residue is attached by a (1-2) linkage as the mannan contains only (1-6)- and (1-2)-linked mannosyl residues (Gander et al: J Biol Chem 249:2063, 1974). No evidence was obtained for the participation of a lipid-linked mannosyl-containing intermediate in this system.
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  • 182
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    Journal of Supramolecular Structure 7 (1977), S. 223-234 
    ISSN: 0091-7419
    Keywords: cell surfaces ; carbohydrates ; implantation ; lectin binding ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Preimplantation embryos were obtained from the uteri and oviducts of 2 strains of mice, Swiss CD-1 and B6 CBA. After removal of the zona pellucida by treatment with pronase, FITC-lectins were bound to the embryonic cell surfaces at either 4°C or 37°C. Both morula and blastocyst stage embryos bound the following lectins, FITC-ConA, FITC-WGA, FITC-RCAII and FITC-RCAI. No difference in binding was observed between the morula stage and the blastocyst stage within each mouse strain for each specific lectin. However B6 CBA embryos bound less FITC-ConA and FITC-WGA than the corresponding Swiss CD-1 embryos. The topographical arrangement of the lectin receptors was observed to differ between 4°C and 37°C for FITC-Con A, FITC-RCAII, and FITC-RCAI. While lectins bound at 4°C showed a pattern of continuous labeling, the same lectin at 37°C showed aggregation of lectin receptors into patches indicating lateral mobility of these receptors within the embryonic cell membranes. In contrast FITC-WGA bound at 4°C and 37°C demonstrated continuous labeling of embryos at both temperatures. FITC-fucose binding protein did not bind to Swiss CD-1 embryos.The invasiveness of trophoblastic cells of mouse blastocysts was studied by culturing isolated embryos without prior enzyme treatment on reconstituted collagen gels. After 4 days in BME containing only glutamine and bovine serum albumin as supplements, the embryos shed their zona pellucida and implanted into the collagen gel as indicated by zones of lysis in proximity to the embryonic cells when analyzed by scanning electron microscopy.
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  • 183
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    Journal of Supramolecular Structure 7 (1977), S. 1-97 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 184
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    Journal of Supramolecular Structure 7 (1977), S. 301-306 
    ISSN: 0091-7419
    Keywords: cytoplasmic activator ; red blood cells ; membrane ATPase ; Ca2+ transport ; (Ca2+-Mg2+)ATPase ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Human red blood cells (RBC) contain a cytoplasmic, nonhemoglobin protein which activates the (Ca2+-Mg2+) ATPase of isolated RBC membranes. Results presented in this paper confirm that activation of (Ca2+-Mg2+)ATPase is associated with binding of the cytoplasmic activator to the membrane. Binding of the cytoplasmic activator is reversible and dependent on ionic strength and Ca2+. Cytoplasmic activator is sensitive to trypsin but is not degraded when intact RBC are exposed to trypsin. Cytoplasmic activator does not modify the (Ca2+-Mg2+)-ATPase of membranes from RBC exposed to activator prior to hemolysis. Thus, the activator is located in the cell and appears to act by binding to the inner membrane surface.
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  • 185
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    Journal of Supramolecular Structure 7 (1977), S. 371-379 
    ISSN: 0091-7419
    Keywords: Sindbis ; glycoproteins ; cell surface ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The carbohydrate portions of the Sindbis virus glycoproteins were compared with the carbohydrate portions of cell surface glycoproteins from uninfected host cells. Comparisons of the size of glycopeptides were made using gel filtrations. Comparisons of sugar linkages were made by methylation analysis. The conclusion was that the Sindbis carbohydrate is similar to a portion of the host carbohydrate. Thus, the Sindbis carbohydrate structures appear to be structures normally made in the uninfected host cell, but which are added to the Sindbis glycoproteins in virus-infected cells.
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  • 186
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    Journal of Supramolecular Structure 7 (1977), S. 381-395 
    ISSN: 0091-7419
    Keywords: dolichyl phosphomannose ; glycoproteins ; mannosyltransferases ; polyprenyl phosphosugars ; retina ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Large-scale incubations were carried out with homogenates of the retinas of the 15-16-day-old chick embryo in the presence of GDP[U-14C] mannose, from which there were isolated a mannolipid (Lipid I), oligosaccharide-lipids (Lipid II), and glycoprotein (residue). These incubations were performed in the presence of endogenous acceptors as well as dolichyl phosphate. [14C] Mannolipid I was subjected to chromatography on DEAE cellulose and silicic acid. The response to these, as well as TLC, enzymatic, and chemical treatments, were consistent with the product being dolichyl phosphomannose. [14C] Lipid II was purified by DEAE cellulose chromatography and gel filtration on LH-20. Responses to these treatments, as well as TLC and paper chromatography, were consistent with this product being of the class of the oligosaccharide-pyrophosphate-lipids. The residue remaining after removal of the lipids was shown to contain glycoproteins by conversion of high-molecular-weight radioactive material to low-molecular-weight [14C] mannose-containing glycopeptides by the action of pronase. These reactions and their products are consistent with there being in the retina, the pathway for glycoprotein synthesis involving the participation of the lipid-activated carbohydrates.When the incubations were performed in the presence of ATP or ADP there was a decrease in the labeling of Lipid I, accompanied by an increase in the labeling of Lipid II and glycoprotein. When incubated in the presence of dolichyl phosphate and deergent, however, the stimulatory effect of ATP did not occur. The effect on these activities of a variety of other nucleotide phosphates was also examined.
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  • 187
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    Journal of Supramolecular Structure 7 (1977), S. 435-442 
    ISSN: 0091-7419
    Keywords: lipoprotein structure ; x-ray scattering ; thermal trasnsitions ; interaction arterial proteoglycans ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The structure and thermal behavior of human serum low-density lipoproteins showing either a high or a low reactivity against a proteoglycan isolated from human arteries have been found to be different from each other. It is suggested that modifcations in the lipoprotein surface structure induced by the physical state of the neutral lipids could modulate the affinity of the macromolecule for the arterial component.
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  • 188
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    Journal of Supramolecular Structure 7 (1977), S. 515-530 
    ISSN: 0091-7419
    Keywords: breast ; prostate ; carcinoma ; glycoproteins ; organ culture ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We demonstrate that a technique is available to investigate glycoprotein synthesis in organ cultures of human breast and prostate surgical specimens where the 3-dimensional epithelial cell arrangement remains intact. Malignant breast and prostate epithelium maintained their capacity to synthesize glycoproteins for at least 3 days as followed by the incorporation of [3H] glucosamine into macromolecules. Over 70% of incorporation was by malignant cells as judged by autoradiography. Labeled glycoproteins were released into glandular lumina and consequently into the culture fluid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed predominantly one group of macromolecules released with an apparent molecular weight of 48,000 ± 6,000 daltons. This glycoprotein was found in all of the breast specimens studied, which included 1 medullary, 1 infiltrating lobular, and 8 infiltrating duct carcinomas. The pattern was independent of the availability of estrogen receptors. A similar glycoprotein was also observed in the culture media from a Grade I and a Grade II well-differentiated infiltrating prostate carcinoma. Incorporation was below the level of detection in 4 of 6 cases of benign prostatic hyperplasia. A more complex pattern of labeled glycoproteins was found in the media of a Grade II and a Grade III poorly-differentiated prostate carcinoma. The established human mammary carcinoma cell line MCF-7 synthesized and released a similar 48,000 molecular weight glycoprotein but additional components with larger molecular weights were also released. An intriguing interpretation that 3-dimensional tissue integrity restricts some glycorprotein synthesis is discussed. Cells grown in 2-dimensional monolayers could escape from such a topographic restriction and express additional families of glycoproteins.
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  • 189
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    Journal of Supramolecular Structure 6 (1977), S. 125-133 
    ISSN: 0091-7419
    Keywords: amino acid transport ; gradient hypothesis ; electrogenic cation pump ; electrolyte movements ; ouabain ; furosemide ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The existence of an electrogenic Na+ pump in Ehrlich cells which substantially contributes to the membrane potential, previously derived from the distribution of the lipid soluble cation tetraphenylphosphonium (TPP+), could be confirmed by an independent method based on the quenching of fluorescence of a cyanine dye derivative, after the mitochondrial respiration had been suppressed by appropriate inhibitors. The mitochondrial membrane potential, by adding to the overall potential as measured in this way is likely to cause an overestimation of the membrane potential difference (p.d.). But since this error tends to diminish with increasing pump activity, the true p.d. of the plasma membrane should easily account for the driving force to drive the active accumulation of amino acids in the absence of an adequate Na+ concentration gradient. Accordingly, the F2-aminoisobutyric acid (AIB) uptake rises linearly with the distribution of TPP+ at constant Na+ concentrations, suggesting that each responds directly to membrane potential. There is evidence that the electrogenic (free) movement of Cl- is slow, at least at normal p.d., whereas a major part of the Cl- movement across the cellular membrane appears to occur by an electrically silent Cl--base exchange mechanism. By such a mode Cl-, together with an almost stoichiometric amount of K+, may under certain conditions move into the cell against a high adverse electrical potential difference. This “paradoxical” movement of K+Cl- contributing to the deviation of the Cl- distribution from the electrochemical equilibrium distribution, is not completely understood. It is insensitive towards ouabain but can almost specifically be inhibited by furosemide. As a likely explanation a H+-K+ exchange pump was previously offered, even though unequivocal evidence of such a pump is so far lacking. According to available evidence the electrogenic movement of free Cl- is too small, at least at normal orientation of the p.d., to significantly shunt the electrogenic pump potential so that the establishment of such a potential is plausible. The evidence presented is considered strong in favor of the gradient hypothesis since even in the absence of an adequate Na+ concentration gradient, the electrogenic Na+ pump will contribute sufficient extra driving force to actively transport amino acid into the cells.
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  • 190
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    Journal of Supramolecular Structure 6 (1977), S. 135-153 
    ISSN: 0091-7419
    Keywords: periplasmic proteins ; transport ; precursor ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The cold osmotic shock procedure releases a protein (GLPT) from the cell envelope of Escherichia coli that is related to the transport of sn-glycerol-3-phosphate in this organism. The evidence for this correlation is as follows: (1) GLPT is under the regulatory control of the glpR gene. (2) Some glpT mutants that were isolated as phosphonomycin resistant clones do not synthesize GLPT. Revertants of these mutants (growth on sn-glycerol 3-phosphate) again synthesize GLPT. (3) Some amber mutations in glpT reduce the amount of GLPT while suppressed strains produce normal amounts. (4) Transfer of a plasmid carrying the glpT genes into a strain lacking GLPT and sn-glycerol-3-phosphate transport restores both functions in the recipient. Transport and GLPT synthesis in the plasmid carrying strain are increased 2- to 3-fold over a fully induced wild-type strain, but appear to be constitutive. GLPT is a soluble protein of molecular weight 160,000 composed of 4 identical subunits. The 160,000 molecular weight complex is stable in 1% sodium dodecylsulfate at room temperature. Upon boiling in 1% sodium dodecylsulfate GLPT dissociates into its subunits. Likewise, 8 M urea at room temperature dissociates GLPT into its subunits. Dialysis of dissociated GLPT against phosphate or Tris-HCl buffer, pH 7.0, allows renaturation to the tetrameric form. The protein is acidic in nature (isoelectric point 4.4).In contrast to the typical transport-related periplasmic-binding proteins, no conditions could be found where pure GLPT exhibited binding activity toward its supposed substrate, sn-glycerol-3-phosphate.In vivo new appearance of transport activity for sn-glycerol-3-phosphate transport occurs only shortly before cell division. However, GLPT synthesis does not fluctuate during the cell cycle. The available evidence indicates a cell-division-dependent processing of GLPT in the cell envelope as a reason for the alteration in transport activity.Transport in whole cells is sensitive to the cold osmotic shock procedure, demonstrating the participation of an essential periplasmic component. However, isolated membrane vesicles that are devoid of periplasmic components, including GLPT, are fully active in sn-glycerol-3-phosphate transport. Therefore, we conclude that GLPT is essential in overcoming a diffusion barrier for sn-glycerol-3-phosphate established by the outer membrane. Attempts to isolate mutants that are transport negative in whole cells due to a defect in GLPT but are active in isolated membrane vesicles have failed so far. All GLPT mutants tested, whether or not they synthesize GLPT, are not active in isolated membrane vesicles.Iodination of whole cells with [125I] followed by osmotic shock reveals that several shock-releasable proteins including GLPT become radioactively labeled. This indicates that some portions of GLPT are accessible to the external medium.
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  • 191
    ISSN: 0091-7419
    Keywords: cilia ; dynein ; conformation change ; sulfhydryl groups ; ATPase activity ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Incubation of glycerol-extracted, Triton X-100 demembranated Tetrahymena cilia with 2-10 vol % acetone caused an enhancement of ATPase activity by 2- to 3- fold, depending on concentration and time of incubation. Axonemal ATPase activity was also increased upon incubation with bis (4-fluoro-3-nitrophenyl) sulfone (FNS). Acetone and FNS enhanced the activity of solubilized 30S dynein, but slightly inhibited that of 14S dynein. Heating at 38°C, incubation with FNS, and incubation with acetone activated axonemal ATPase to the same extent. Subsequent studies of (1) the effect of time of preincubation with a spin-labeled maleimide (SLM) at 25°C as a function of pH on the ATPase activity, (2) the concentration dependence of the inhibition of ATPase activity by N-ethylmaleimide or SLM, (3) the ratio of ATPase activity assayed at 25°C to that assayed at 0°C, and (4) the ratio of ATPase activity at pH 8.6 to that at pH 6.9 did not reveal any difference in the properties of the axonemal ATPase after near maximal enhancement by the heat, acetone, or FNS treatments. It was concluded that enhancement of ATPase activity by gentle heat treatment, by incubation with acetone (or other organic solvents), or by FNS results from a conformation change of 30S dynein.The effect of acetone and of FNS on the pellet height response (a measure of the increase in height of the pellet of cilia precipitated by brief centrifugation in the presence of ATP as compared to the absence of ATP) was also determined. Enhancement of ATPase by these reagents did not lead to a decrease in pellet height response. This observation, in conjunction with other data, indicates that there are at least 3 states of the cross-bridge cycle of dynein arms in cilia.
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  • 192
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    Journal of Supramolecular Structure 6 (1977), S. 169-177 
    ISSN: 0091-7419
    Keywords: Halobacterium halobium ; amino acid transport ; sodium-proton exchange ; asymmetry of transport system ; reconstitution of glutamate transport ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Cell envelope vesicles prepared from H. halobium contain bacteriorhodopsin and upon illumination protons are ejected. Coupled to the proton motive force is the efflux of Na+. Measurements of 22Na flux, exterior pH change, and membrane potential, ΔΨ (with the dye 3,3′-dipentyloxadicarbocyanine) indicate that the means of Na+ transport is sodium/proton exchange. The kinetics of the pH changes and other evidence suggests that the antiport is electrogenic (H+/Na+ 〉 1). The resulting large chemical gradient for Na+ (outside 〉 inside), as well as the membrane potential, will drive the transport of 18 amino acids. The 19th, glutamate, is unique in that its accumulation is indifferent to ΔΨ: this amino acid is transported only when a chemical gradient for Na+ is present. Thus, when more and more NaCl is included in the vesicles glutamate transport proceeds with longer and longer lags. After illumination the gradient of H+ collapses within 1 min, while the large Na+ gradient and glutamate transporting activity persists for 10-15 min, indicating that proton motive force is not necessary for transport. A chemical gradient of Na+, arranged by suspending vesicles loaded with KCl in NaCl, drives glutamate transport in the dark without other sources of energy, with Vmax and Km comparable to light-induced transport. These and other lines of evidence suggest that the transport of glutamate is facilitated by symport with Na+, in an electrically neutral fashion, so that only the chemical component of the Na+ gradient is a driving force.The transport of all amino acids but glutamate is bidirectional. Actively driven efflux can be obtained with reversed Na+ gradients (inside 〉 outside), and passive efflux is considerably enhanced by intravesicle Na+. These results suggest that the transport carriers are functionally symmetrical. On the other hand, noncompetitive inhibition of transport by cysteine (a specific inhibitor of several of the carriers) is only obtained from the vesicle exterior and only for influx: these results suggest that in some respects the carriers are asymmetrical.A protein fraction which binds glutamate has been found in cholate-solubilized H. halobium membranes, with an apparent molecular weight of 50,000. When this fraction (but not the others eluted from an Agarose column) is reconstituted with soybean lipids to yield lipoprotein vesicles, facilitated transport activity is regained. Neither binding nor reconstituted transport depend on the presence of Na+. The kinetics of the transport and of the competitive inhibition by glutamate analogs suggest that the protein fraction responsible is derived from the intact transport system.
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  • 193
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    Journal of Supramolecular Structure 6 (1977), S. 259-274 
    ISSN: 0091-7419
    Keywords: conformational analysis ; polysaccharides ; cooperative interactions ; synergistic interactions ; cooperative cation binding ; spectroscopic techniques ; circular dichroism ; nuclear magnetic resonance ; optical rotation ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: For consideration of their conformations and interactions, carbohydrate chains can conveniently be divided into 3 classes on the basis of their covalent structure; namely periodic (a), interrupted periodic (b), and aperiodic (c) types. In aqueous solution carbohydrate chains often exist as highly disordered random coils. Under appropriate conditions, however, polysaccharides of types (a) and (b) can adopt a variety of ordered conformations. Physical methods, and in particular optical rotation, circular dichroism, and nuclear magnetic resonance, provide sensitive probes for the study of the mechanism and specificity of these disorder-order transitions in aqueous solution.Intermolecular interactions between such polysaccharide chains arise from co-operative associations of long structurally regular regions which adopt the ordered conformations. For acidic polysaccharides these cooperative associations may involve alignment of extended ribbons with cations sandwhiched between them. In other systems the interactions involve double belices which may then aggregate further, and geometric “matching” of different polysaccharide chains can also occur. These ordered, associated regions are generally terminated by deviations from structural regularity or by “kinks” which prevent complete aggregation of the molecules.The complex carbohydrate chains which occur at the periphery of animal cells have very different, aperiodic structures and although their conformations are as yet poorly understood, preliminary indications are considered.
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  • 194
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    Journal of Supramolecular Structure 6 (1977), S. 301-311 
    ISSN: 0091-7419
    Keywords: red cell ; erythrocyte ; membrane ; scanning electron microscope ; spectrin ; actin ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A web-like reticulum underlying the human erythrocyte membrane was studied at a resolution of 5-10 nm by means of a scanning electron microscope. The network was visualized in isolated membranes (ghosts) torn open to reveal their interior space and in residues derived from ghosts extracted with Triton X-100. It formed a continuous (rather than patchy) cover over the entire cytoplasmic surface, except where lifted off or torn away. Filaments (5-40 nm in diameter), annular figures (40-60 nm in diameter), and nodes (30-100 nm in diameter) were prominent in different networks. The dimensions of the filaments and the interstices in the reticulum varied with conditions, suggesting that the network has elastic properties. This reticulum is probably related to the erythrocyte membrane proteins spectrin and actin.
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  • 195
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    Journal of Supramolecular Structure 6 (1977), S. 313-323 
    ISSN: 0091-7419
    Keywords: peripheral and integral proteins ; membrane biosynthesis ; hydrophobic and hydrophilic interactions ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Membranes are structures whose lipid and protein components are at, or close to, equilibrium in the plane of the membrane, but are not at equilibrium across the membrane. The thermodynamic tendency of ionic and highly polar molecules to be in contact with water rather than with nonpolar media (hydrophilic interactions) is important in determining these equilibrium and nonequilibrium states. In this paper, we speculate about the structures and orientations of integral proteins in a membrane, and about how the equilibrium and nonequilibrium features of such structures and orientations might be influenced by the special mechanisms of biosynthesis, processing, and membrane insertion of these proteins. The relevance of these speculations to the mechanisms of the translocation event in membrane transport is discussed, and specific protein models of transport that have been proposed are analyzed.
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  • 196
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    Journal of Supramolecular Structure 6 (1977), S. 355-362 
    ISSN: 0091-7419
    Keywords: amino acid transport ; mammary gland ; cell proliferation ; feedback regulation ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The regulation of the uptake of the amino acid analog α-aminoisobutyric acid was studied in diced mammary glands from pregnant mice. Stimulation of uptake by insulin was not prevented by inhibitors of protein synthesis; protein synthesis inhibitors decreased uptake by 20%; this response occurred more promptly in insulintreated tissues. Elimination of extracellular amino acids led to a substantial increase in transport which was not abolished by inhibitors of protein synthesis. These results indicate that insulin does not increase amino acid transport in this system by altering synthesis and degradation of transport protein. They are consistent with a model in which the activity of the existing amino acid transport protein is subject to negative feedback regulation from the intracellular amino acid pool.
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  • 197
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    Journal of Supramolecular Structure 6 (1977), S. 433-440 
    ISSN: 0091-7419
    Keywords: transport ; incorporation ; uptake ; thymidine ; nucleoside ; Novikoff rat hepatoma cells ; rapid sampling technique ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Incorporation of thymidine into Novikoff rat hepatoma cells was analyzed with a rapid sampling technique which allowed collection of 12 time points in 20 sec. Transport was studied in the absence of metabolism by using either ATP-depleted cells or a thymidine kinase negative subline. Transport was a rapid, saturable, nonconcentrative process with a Km of about 85 μM. The intracellular thymidine pool was also rapidly labeled in cells which phosphorylated thymidine, so that a group translocation process involving thymidine kinase can be ruled out. Under all conditions examined, phosphorylation, not the transport, of thymidine was the rate-determining step in its incorporation into the acid-soluble pool. Estimation of transport rates from total incorporation into cells which phosphorylate the substrate is invalid in this cell system and must be questioned in all instances.
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  • 198
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    Journal of Supramolecular Structure 6 (1977) 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 199
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    Journal of Supramolecular Structure 6 (1977), S. 473-484 
    ISSN: 0091-7419
    Keywords: placenta ; brush border ; sialoglycoprotein ; alkaline phosphatase ; two-dimensional electrophoresis ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A brush border membrane enriched fraction was isolated from human, full-term placenta. This membrane fraction exhibited large membrane fragments with microvilli projecting from the basal membrane in electron micrographs and was enriched tenfold in alkaline phosphatase, a brush border enzyme marker. The sialoglycoproteins associated with this membrane fraction were tritiated by mild periodate oxidation of sialic acid and reduction with tritiated NaBH4. The membranes were solubilized in 8 M urea, 2% Triton X-100, and the tritiated glycoprotein subunits were reduced with β-mercaptoethanol and characterized by 2-dimensional polyacrylamide gel electrophoresis using a method similar to that described by O'Farrell and Bhakdi, Knüferman, and Wallach. The tritiated subunits were detected in the gels by autofluorography. The 2-dimensional subunit “maps” resolved at least 17 major sialoglycoprotein subunits whereas only 10 major periodate-Schiff reagent staining components were resolved by 1-dimensional SDS polyacrylamide gel electrophoresis. Placental alkaline phosphatase (PAP) was identified on the subunit maps by inclusion of 32P-labeled PAP in the tritiated membrane sample. The 32P-labeled PAP corresponded to a major tritiated sialoglycoprotein subunit, which was heterogeneous with respect to charge as demonstrated by 3 closely running spots of the same molecular weight.
    Additional Material: 4 Ill.
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  • 200
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    Journal of Supramolecular Structure 6 (1977), S. 503-518 
    ISSN: 0091-7419
    Keywords: L-arabinose-binding protein ; three-dimensional structure ; spectrochemical studies ; active transport ; chemotaxis ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The crystal structure of the L-arabinose-binding protein (ABP), an essential component of the high affinity L-arabinose transport system in E. coli, has been determined at 3.5- and 2.8-Å resolutions. The Fourier maps indicate that the molecule is ellipsoidal with overall dimensions of 70 × 35 × 35 Å (axial ratio ≃ 2:1) and consists of 2 distinct globular domains (designated “P” and “Q”). A tentative trace of the polypeptide backbone is presented. The 2 domains are arranged to create a deep and narrow cleft, the base of which is which is formed by 3 polypeptide chain segments linking the 2 domains. The arrangements of the secondary structure of the 2 domains are remarkably similar and can be related by a pseudo-twofold axis. Each domain has a pleated sheet core with 2 helices on either side of the plane of the β sheet. This secondary structural arrangement is similar to that found in other proteins, specifically the dehydrogenases and kinases. The structural similarity is particularly intriguing in light of the recent finding in this laboratory that the dye 2′,4′,5′,7′-tetraiodofluorescein, an adenine analogue which has been shown to bind to several dehydrogenases and kinases, binds to ABP with a dissociation constant of 30 μM.Experiments performed with protein, modified with the chromophoric probe 2-chloromercuri-4-nitrophenol (MNP), suggest that the binding site is near an essential cysteine residue: modification of the thiol with the mercurial dramatically decreases the ligand-binding affinity of ABP, and conversely, the sugar protects the cysteine from reaction with MNP. The binding of L-arabinose to MNP-labeled protein perturbs the nitrophenol absorbance spectrum. The essential cysteine has been assigned to position 64 in the proposed chain tracing, which is consistent with the amino acid sequence. As an explanation for the failure of the difference Fourier analyses to locate the sugar-binding site, it is postulated that the structure has been solved with the sugar bound. Electron density to which no amino acid residue can be assigned and which could be the sugar molecule is within van der Waals distance of the sulfur atom.
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