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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 8 (1966), S. 473-488 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An improved system is presented for measurement of interactions between a number of individual bacterial species. In the procedure, steady-state populations are fed into a common-mixed-culture vessel. Generation times of each species are determined under these conditions and contrasted with generation time in an identical situation in pure culture. Populations for generation time are determined with the aid of differential media. The mechanical system includes three types of peristaltic pumps for media feed and a unit for measurement, recording, and/or control of pH. A new type of anaerobic continuous-culture vessel which can be inoculated, sampled, and fed continuously is also described. A functional test of a three-part system including Streptococcus salivarius, Veilloncella alcalescens, and Staphylococcus aureus is presented. An unusual feature was the finding that, under certain conditions, the generation time of S. Salivarius was less than 10 min.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 12
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 36 (1990), S. 467-475 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: β-Galactosidase served as a model system to explore the feasibility of enhancing the selectivity of a low-cost, easily scaled separation method - precipitation. Enhanced selectivity was sought by fusing the enzyme with polypeptide tails including 5 and 11 aspartaies. The unfused protein could not be selectively removed from the Escherichia coli cell extract by precipitation with polyethylenimine (PEI), but the longest fusion could be selectively removed. The presence of nucleic acids limited the purification attainable. Pretreatment with nuclease followed by diafiltration resulted in an extract from which the same fusion could be precipitated with greater than fivefold enrichment, while the untailed enzyme remained unenriched by the same precipitation step. Selectivitiy is attributed to the binding strength of the polyanionic tails to the polycationic PEI.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 13
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 30 (1987), S. 724-735 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Determination of glucose concentrations in fluids frequently requires the application of immobilized glucose oxidase. An accurate description of the immobilized enzyme kinetics is critical for such applications. In this study, the overall rate of reaction of immobilized glucose oxidase is investigated theoretically. A novel steady-state model based on a ping-pong kinetic mechanism for glucose oxidase is developed. Numerical studies are used to examine the parameter sensitivity of this model. The enzyme loading, matrix thickness and geometric con figuration are found to have a significant influence on substrate uptake by insolubilized glucose oxidase.Aditionally, this new model is compared with a previously developed model based on an alternative ping-pong kinetic mechanism. Under steady-state conditions, no significant difference between the two models is apparent when appropriate kinetic parameters are applied to each of the models. The model developed herein is also compared with models utilizing the simplifying assumption of Michaelis-Mented kinetics for substrate reaction. Numerical studies indicate that under most realizable biological conditions, a model based on ping-pong kinetics should be applied to accurately describe substrate uptake by immobilized glucose oxidase.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 14
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 29 (1987), S. 215-221 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An auxotrophic mutant of Saccharomyces cerevisiae, containing a recombinant 2-μ-based plasmid, was grown in selective media in continuous culture. The plasmid retained the ability to synthesize acid phosphatase as product, which was deleted from the host. Plasmid loss was followed at various dilution rates, and the level of plasmid expression was controlled by changing the β-glycero/inorganic phosphate ratio.Some interesting trends were observed. As the level of plasmid expression was raised, the stability dropped markedly. Since acid phosphatase expression is regulated at the level of transcription, it is possible that increased transcription interfered with plasmid replication, hindered segregation, or overburdened the cell's DNA repair capability. It was also observed that plasmid stability was substantially increased at high growth rates. At dilution rates of 0.3 and 0.37 h-1, feeding only inorganic phosphate, the plasmid was completely stable.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 15
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 46 (1995), S. 452-458 
    ISSN: 0006-3592
    Keywords: protein hydration ; enzymes in organic solvents ; adsorption isotherms ; essential water ; water activity ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A very sensitive NMR method has been developed for measuring deuterated water bound to proteins suspended in nonpolar solvents. This has been used to determine the amount of bound water as a function of water activity for subtilisin Carlsberg suspended in hexane, benzene, and toluene and for α-chymotrypsin in hexane. The adsorption isotherms for subtilisin in the three solvents are very similar showing that water activity can be usefully employed to predict the amount of water bound to proteins in nonpolar organic media. Comparison of the degree of enzyme hydration reached in nonpolar solvents with that obtained in air shows that adsorption of strongly bound water is hardly affected by the low dielectric medium, but adsorption of loosely bound water is significantly reduced. This suggests that the hydrophobic regions of the protein surface are preferentially solvated by solvent molecules, and that in a nonpolar environment formation of a complete monolayer of water over the protein surface is thermodynamically unfavorable. © 1995 John Wiley & Sons, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 16
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 60 (1998), S. 541-550 
    ISSN: 0006-3592
    Keywords: biofilm ; dual substrate limitation ; cometabolism ; secondary substrate ; biofilm modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A dynamic model was developed to describe the behaviour of primary and secondary substrates in a biofilm reactor. The model incorporates structured kinetics to describe the generation and consumption of reducing power in the catabolic and respiratory subsystems, respectively. Secondary substrate transformation through oxygenolytic or reductive mechanisms can be modelled under either single or dual limitation of the electron donor and electron acceptor substrates. An example simulation of a theoretical biofilm system was performed. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 541-550, 1998.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 17
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 13 (1998), S. 117-123 
    ISSN: 0884-3996
    Keywords: electrochemiluminescence ; metals ; melanins ; luminol ; binding ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Electrochemiluminescence (ECL) studies of the chemiluminescent (CL) polymer diazoluminomelanin (DALM) biosynthesized in nitrate reductase transfected Escherichia coli JM109 bacteria revealed noteworthy anodic ECL and even more intense cathodic ECL. Bacterial DALM (BD) ECL was also assessed in the presence of 100 ppm of 33 different metal and non-metal ions which revealed specific anodic, but not cathodic, enhancements of BD ECL with Ag+, Hg2+ and Ru3+. The precursors and intermediate polymers which comprise DALM, such as luminol, 3-amino-L-tyrosine (3-AT), aminomelanin (AM) and diazomelanin (DM) were screened for ECL enhancement against the same set of elemental ions. Significant anodic ECL enhancements were observed for luminol with Hg2+ in the presence of tripropylamine (TPA), but not for any other DALM component in combination with other elemental ions, either anodically or cathodically. Comparison of BD with luminol in the presence and absence of TPA and Hg2+ revealed very different ECL activity patterns and suggested different mechanisms for BD and luminol ECL. © 1998 John Wiley & Sons, Ltd.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 18
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 12 (1991), S. 687-688 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: This report describes the incorporation of an alkyl maltoside detergent in two-dimensional polyacrylamide gel Electrophoresis (2-D PAGE) sample lysis buffer in order to improve resolution of protein patterns separated by nonequilibrium pH gradient Electrophoresis. Membrane-associated proteins with alkaline isoelectric points form horizontal streaks on two-dimensional electrophoretograms when solubilized with conventional nonionic detergent. Dodecyl maltoside enhances protein delipidation during solubilization and improves pattern resolution and protein mobility.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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  • 19
    ISSN: 0173-0835
    Keywords: Peptide mass fingerprinting ; Yeast ; Two-dimensional polyacrylamide gel electrophoresis ; Keratin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: With the complete sequence of the yeast genome now available, efforts by many laboratories are underway to identify each of the spots on two-dimensional (2-D) gels corresponding to the most abundant yeast proteins. The high mass accuracy now attainable using matrix assisted laser desorption/ionization (MALDI)-mass spectrometry equipped with delayed extraction simplifies the process of identification, such that many spots can be unambiguously identified in a short period of time merely by using peptide mass fingerprinting and generally available database matching programs. Although it is not always possible to match spots between gels run by different laboratories, proteins generally yield the same abundant proteolytic fragments when tryptic digestions are performed. Databases containing these signature peptides not only simplify the task of reidentifying proteins from different gels, but also make it possible to identify small amounts of cross-contaminating proteins from different spots, as well as common extraneous contaminants such as human keratins. In this paper, we present data on the identification of 〉 20 previously unreported yeast proteins from 2-D gels. Some novel proteins were identified from randomly analyzed spots. Focusing on 14 spots in a narrow-pH-range gel, we demonstrate how organizing peak-table data and peptide match-list data into databases enables the identification of a larger percentage of the peaks.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 20
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Capillary electrophoresis has been used in conjunction with electrospray mass spectrometry using both full-scan and selected ion monitoring modes to supply as much information as possible about the venom of Dendroaspis polylepis polylepis (Black Mamba). As an example of the application of capillary electrophoresis/electrospray mass spectrometry (CE/ESI/MS) to the analysis of a complex mixture of small proteins, we have analyzed the venom of Dendroaspis polylepis polylepis using the combined techniques. Both full-scan and selected ion monitoring modes were employed. CE/ESI/MS provides a rapid and extremely sensitive method for molecular weight determination, particularly when selected ion monitoring is employed. It has been utilized to provide sequence confirmation for those toxins which have already been described in the literature. Our methodology indicates the presence of at least 70 peptides in the molecular weight range 6000-9000.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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