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α-Factor directed expression of the human epidermal growth factor in Saccharomyces cerevisiae

✍ Scribed by Steven J. Coppella; Prasad Dhurjati

Publisher
John Wiley and Sons
Year
1989
Tongue
English
Weight
738 KB
Volume
33
Category
Article
ISSN
0006-3592

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✦ Synopsis

Expression kinetics of the human Epidermal Growth Factor (hEGF) from the a-factor prepro region in a 2-pm based plasmid was studied in Saccharomyces cerevisiae. Production o f hEGF was highly medium dependent 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. a-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 700% 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.

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