Optimization of pro-urokinase secretion from recombinant Saccharomyces cerevisiae

✍ Scribed by Brian G. Turner; George C. Avgerinos; Laurence M. Melnick; Donald T. Moir

• Book ID: 102768695
• Publisher: John Wiley and Sons
• Year: 1991
• Tongue: English
• Weight: 665 KB
• Volume: 37
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.260370911

✦ Synopsis

Secretion of a nonglycosylated form of human prourokinase, also known as single-chain urinary plasminogen activator (scu-PA), from Saccharomyces cerevisiae is de- scribed. A "supersecreting" yeast strain harboring multiple copies of integrated plasmids was grown batchwise and at constant respiratory quotient (RQ) in 20-L fermenters. Because the promoters used to drive expression of the prourokinase genes are not tightly regulated, secretion into the culture supernatant was growth associated. Although the final cell density achieved in the perturbed-batch fermentation (45 g dry wt/L) was less than that observed in the RQ-controlled culture (77g dry wt/L), the scu-PA titer in the perturbed-batch fermentation (1863 IU/mL) was nearly twice that attained at constant RQ (1108 IU/mL). The effects on cell growth and scu-PA titer of other process variables (pH, temperature, phosphate concentration, and medium composition) are also discussed.