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  • 1
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    Isolation of high-molecular-weight nucleic acids for copy number analysis using high-performance liquid chromatography (1987)
    Elsevier
    Details
    Publication Date: 1987-01-01
    Print ISSN: 0021-9673
    Electronic ISSN: 1873-3778
    Topics: Chemistry and Pharmacology
    Published by Elsevier
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    PAPER CURRENT
  • 2
    Electronic Resource
    Electronic Resource
    Improved production of heterologous protein from Streptomyces lividans (1990)
    Springer
    Applied microbiology and biotechnology 33 (1990), S. 395-400 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Protein-secreting procaryotic host organisms are currently being sought as alternatives to Escherichia coli for recombinant processing. In this study we examined how manipulation of the cultivation conditions can enhance heterologous protein production by Streptomyces lividans. The recombinant S. lividans used in this study expressed and excreted a Flavobacterium enzyme capable of hydrolyzing organophosphates. Initial shake-flask studies demonstrated that supplementing Luria-Bertani medium with moderate amounts of glucose (30 g/l), led to improved enzyme production. In fermentor studies with controlled pH, a further twofold increase in production was observed when glucose was fed continuously as compared to batch cultivation. This improved production in the glucose-fed culture may be related to a reduced accumulation of acids. Continuous feeding of both glucose and tryptone led to a further sixfold increase in production. In addition to enhancing production 25-fold, the efficiency of enzyme production and the specific activity of the excreted enzyme were also improved by glucose and tryptone feeding. These results demonstrate that in addition to genetic manipulations, optimization of cultivation conditions can lead to significant improvements in the production of heterologous proteins from Streptomyces.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00176653
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Low-cost computer-coupled fermentor off-gas analysis via quadrupole mass spectrometer (1987)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 29 (1987), S. 679-689 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260290604
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    α-Factor directed expression of the human epidermal growth factor in Saccharomyces cerevisiae (1989)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 33 (1989), S. 976-983 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Expression kinetics of the human Epidermal Growth Factor (hEGF) from the α-factor prepro region in a 2-μm based plasmid was studied in Saccharomyces cerevisiae. Production of hEGF was highly medium de pendent as a chemically defined, nonenriched media had a significantly lower yield than did enriched media. Also cells grown on yeast nitrogen base without amino acids with casamino acids degraded the hEGF after cell growth as opposed to a yeast extract, peptone, and dextrose (YEPD) medium, which elicited no measurable extracellular proteolysis of the hEGF. α-factor directed production kinetics of hEGF on the YEPD medium were growth associated, secretion limitations and extracellular degradation were negligible, and the hEGF was nearly 100% selectively secreted. With sufficient agitation, shake flask experiments were representative of aerated controlled batch fermentations. No effect of high cell density was observed on cell growth or hEGF production kinetics. The hollow fiber bioreactor had no direct effect on the substrate or protein yields of S. cerevisiae, however the low oxygen transfer capacity of the membrane was not sufficient to support respiration.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260330806
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  • 5
    Electronic Resource
    Electronic Resource
    Measurement of copy number using HPLC (1987)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 29 (1987), S. 646-647 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260290514
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  • 6
    Electronic Resource
    Electronic Resource
    A mathematical description of recombinant yeast (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 35 (1990), S. 356-374 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A model was formulated to examine specific experimental data of growth and heterologous product formation with recombinant Saccharomyces cerevisiae while incorporating available literature. The model simulated dry cell weight, glucose, ethanol, dissolved oxygen, human Epidermal Growth Factor (hEGF) production, fraction of recombinant cells, oxygen uptake rate, and carbon dioxide production rate for batch, fed batch, and hollow fiber bioreactor configurations. Nineteen differential equations, 24 analytical equations, and 48 parameters were required. Due to the lack of detailed studies needed for the ADH-II and the TCA enzyme pool, 8 of the 48 parameters were adjustable. Simulation results are presented for verification of the model which successfully described the observed phenomena for the fermentations of S. cerevisiae strain AB103. 1 pYαEF-25. Also presented is a statistical analysis of the model's fit and model parameter sensitivity.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260350405
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