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  • 1
    Electronic Resource
    Electronic Resource
    Stimulatory effect of growth in the presence of alcohols on biotransformation of penicillin G into cephalosporin-type antibiotics by resting cells of Streptomyces clavuligerus NP1 (1999)
    Springer
    Applied microbiology and biotechnology 52 (1999), S. 484-488 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Growth of Streptomyces clavuligerus NP1 in the presence of methanol or ethanol resulted in a marked increase in production of cephalosporin(s) from penicillin G by resting cells. The mycelium produced in alcohol-supplemented medium was fragmented and dispersed as compared with growth in control medium. HPLC analysis showed that at least two products were present in the biotransformation supernatant fluid after 1 h incubation. One of them has been identified as deacetoxycephalosporin G (DAOG).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002530051549
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  • 2
    Electronic Resource
    Electronic Resource
    Polystyrene Macroporous Bead Support for Mammalian Cell Culture (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 665 (1992), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1992.tb42581.x
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  • 3
    Electronic Resource
    Electronic Resource
    Interdependence of Gene Expression for Early Steps of Cephalosporin Synthesis in Streptomyces clavuligerus (1994)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 721 (1994), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1994.tb47383.x
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  • 4
    Electronic Resource
    Electronic Resource
    Transition of outgrowing spores of Bacillus brevis into vegetative cells is not dependent on destruction of gramicidin S (1990)
    Springer
    Applied microbiology and biotechnology 32 (1990), S. 708-710 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Certain Bacillus brevis strains produce gramicidin S (GS) during sporulation, and germination of such spores is delayed at the stage of outgrowth by endogenous or exogenous GS. Claims have been made that the transition from germinating spores into vegetative cells is dependent on GS destruction. We observed no such destruction of either exogenous or endogenous GS. Thus, in our hands, the “recovery” of the inhibited germinating spores must be dependent on something other than GS elimination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00164745
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  • 5
    Electronic Resource
    Electronic Resource
    Production of intracellular serine protease during sporulation of Bacillus brevis ATCC9999 (1983)
    Springer
    Applied microbiology and biotechnology 17 (1983), S. 227-230 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary It has been reported (Slapikoff et al. 1971) that Bacillus brevis ATCC9999, the producer of gramicidin S, makes no extracellular or intracellular protease in complex medium which supports sporulation. We have found that intracellular protease is made by this strain; however, the activity requires the presence of the reducing agent dithiothreitol in the extraction buffer. B. brevis intracellular protease appears at the time that refractile spores are visible in the mother cells. Its pH optimum is 8.0. The enzyme is inhibited by ethylenediaminetetraacetic acid and phenylmethylsulfonylfluoride but not by o-phenanthroline, boronic acids, and only slightly by p-chloromercuribenzoate. This inhibitor pattern is similar to that of intracellular proteases of B. megaterium and B. subtilis, classifying the B. brevis protease as an intracellular seryl protease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00510420
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  • 6
    Electronic Resource
    Electronic Resource
    Protein polarization in isotropic membrane hollow-fiber bioreactors (1994)
    Hoboken, NJ : Wiley-Blackwell
    AIChE Journal 40 (1994), S. 321-333 
    Details
    ISSN: 0001-1541
    Keywords: Chemistry ; Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model has been developed to predict the coupled hydrodynamics and high-molecular-weight protein transport in mammalian-cell hollow-fiber bioreactors (HFBRs). The analysis applies to reactors with isotropic ultrafiltration membranes under startup conditions when the extracapillary space (ECS) is essentially unobstructed by cells. The model confirms the experimental finding that secondary ECS flows, engendered by the primary flow in the fiber lumens, can cause significant downstream polarization of ECS proteins at typical mammalian-cell HFBR operating conditions. It also reveals that the osmotic activity of the proteins, by curtailing transmembrane fluid fluxes, can influence strongly the outcome of the polarization process. In fact, at order-of-magnitude higher protein concentrations and/or lower recycle flow rates, the secondary flow velocities can be reduced by as much as six orders-of-magnitude throughout the ECS, thereby virtually eliminating the polarization problem. This result has important implications for improved reactor startup procedures.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aic.690400211
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  • 7
    Electronic Resource
    Electronic Resource
    Controlled release process to recover heterologous glycosylphosphatidylinositol membrane anchored proteins from CHO cells (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 42 (1993), S. 480-486 
    Details
    ISSN: 0006-3592
    Keywords: CHO ; PI-PLC ; heterologous glipiated proteins ; controlled release ; GPI anchor ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A semicontinuous process has been developed to recover heterologous proteins at increased concentrations and purities. Proteins attached to mammalian cell membranes by glycosylphosphatidylinositol (GPI) anchors can be selectively released into the supernatant by the enzyme phosphatidylinositol-phospholipase C (PI-PLC). Chinese hamster ovary (CHO) cells, genetically engineered to express the GPI anchored, human melanoma antigen (p97), were used as a model system. These cells were grown in protein containing growth medium. During a brief harvesting phase the medium was replaced by phosphate buffered saline (PBS) containing 10 mU/mL of PI-PLC and the GPI anchored protein was cleaved from the cell surface and recovered in soluble form at up to 30% purity. After harvesting, the cells were returned to growth medium where the protein was re-expressed within 40 h. The growth rate, viability, and protein production of cells, repeatedly harvested over a 44-day period, were not adversely affected. This continuous cyclic harvesting process allowed recovery of a heterologous protein at high purity and concentrations and could be applied to the recovery of other GPI anchored proteins and genetically engineered GPI anchored fusion proteins. © 1993 John Wiley & Sons, Inc.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260420411
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  • 8
    Electronic Resource
    Electronic Resource
    Effect of cDNA copy number on secretion rate of activated protein C (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 46 (1995), S. 22-27 
    Details
    ISSN: 0006-3592
    Keywords: cDNA copy number ; gene dosage ; recombinant protein production ; posttranslational modification ; BHK ; secretion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The secretion rate of activated protein C (APC) by BHK cells was increased 35-fold by increasing the cDNA copy number per cell from 50 to 240. In this range, the relation between APC secretion and cDNA copy number was not linear and the rate of APC secretion per cDNA copy increased sevenfold. This apparent cooperative effect of multiple cDNA copies could be related to their integration in tandem. For cDNA copy numbers higher than 240, the APC secreation rate per cDNA and per cell decreased dramatically. The γ-carboxylation of glutamic acid residues, a posttranslational modification required for APC biological activity, was also investigated. The proportion of APC that was fully γ-carboxylated decreased as the secretion rate of APC increased. © 1995 John Wiley & Sons, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260460104
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  • 9
    Electronic Resource
    Electronic Resource
    Membrane anchored protein production from spheroid, porous, and solid microcarrier chinese hamster ovary cell cultures (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 47 (1995), S. 550-556 
    Details
    ISSN: 0006-3592
    Keywords: spheroids ; porous and solid microcarriers ; CHO ; controlled release ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The influence of the microcarrier type on the performance of a controlled release process used to produce a recombinant glycosyl-phosphatidylinositol anchored protein was investigated. Chinese hamster ovary (CHO) cells expressing the human melanoma tumor antigen (p97) were cultured in 10% serum on Cultispher-GH porous microcarriers and then, for protein production, maintained in 2% serum. Cells were harvested every 48 h and p97 was recovered at 90 μg/mL and 40% purity. Harvested p97 concentrations were increased by harvestingfrom spheroid (241 μg/mL) and smaller porous microcarrier, Cultispher-G (167 μg/mL) cultures. The low total cell specific p97 production of cells cultured on Cultispher-GH was due to necrosis of cells within the beads, decreased p97 expression of the immobilized cells, dilution by the liquid (up to 40% volume) associated with settled beads, and incomplete recovery of p97 from within the beads. Cells cultured on solid microcarriers, Cytodex-1, had the highest cell viability and cell specific p97 production, It is recommended that a two-stage cyclic harvesting process of cells cultured on small Cultispher-G or on Cytodex-1 beads would minimize protein loss and maximize cell specific protein recovery. © 1995 John Wiley & Sons Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260470507
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  • 10
    Electronic Resource
    Electronic Resource
    Increasing GPI-anchored protein harvest concentrations from suspension and porous microcarrier CHO cell cultures (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 136-147 
    Details
    ISSN: 0006-3592
    Keywords: CHO cells ; glycosyl-phosphatidylinositol p97 concentration ; protein harvest ; controlled release ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Chinese hamster ovary (CHO) cells expressing the human melanoma tumour antigen, p97, were used to develop a controlled release process for the production of recombinant glycosyl-phosphatidylinositol (GPI) anchored proteins. The cells were cultured either in suspension or immobilized on porous microcarriers and p97 was selectively cleaved from the cell surface by the bacterial enzyme, phosphatidylinositol-phospholipase C (PI-PLC). The kinetics of p97 cleavage from the cell surface by PI-PLC was shown to be approximated by Michaelis-Menten kinetics. The recovered p97 concentrations were increased by reusing the PI-PLC enzyme solution to harvest multiple batches of cells. A convenient PI-PLC assay was developed to monitor the harvesting process and to determine the stability of PI-PLC under harvesting conditions. Although the Pl-PLC was stable under harvesting conditions, it rapidly adsorbed to the cell surface and was depleted from the reused enzyme solution. In order to maintain PI-PLC activity, it was necessary to add fresh PI-PLC to the reused enzyme solution before harvesting a fresh batch of cells. The maximum p97 concentration that could be obtained from harvesting CHO cells cultured on porous microcarriers was limited by the dilution effects of sample removal, adding fresh PI-PLC and liquid associated with settled microcarriers. A model was developed that adequately predicted the p97 concentration after each harvest and the maximum p97 concentration that could be achieved by this harvesting method. The dilution effects were minimized by harvesting from centrifuged suspension culture cells and the harvested p97 concentration was increased by over sixfold to 0.64 mg/mL. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 136-147, 1997.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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