Production of cytotoxic proteins inEscherichia coli:a fermentation process for producing enzymatically active HIV-1 protease
✍ Scribed by Wayne K. Herber; F. James Bailey; Christine E. Carty; Jill C. Heimbach; Robert Z. Maigetter
- Publisher
- Springer
- Year
- 1991
- Tongue
- English
- Weight
- 562 KB
- Volume
- 36
- Category
- Article
- ISSN
- 1432-0614
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✦ Synopsis
Two fermentation processes for the tryptophan-regulated expression of active HIV protease (HIV-1 prt) in Escherichia coli are described. Since overexpression of HIV-1 prt results in cell death, stringent control of product expression was necessary to attain high enzyme levels. Such control was achieved by separation of growth and production phases in a twostep process or by implementation of nutrient feed in a one-step process. When the two-stage process was used, soluble product was detectable only when induction occurred at low culture density (A55o <3.5). Short induction periods of 1-2 h and rapid harvesting were necessary to recover active product. Similar results were obtained when the single-stage process was operated at 37 ° C; however, cultivation and induction at 28°C resuited in active enzyme formation following induction at increased cell density (A55o = 10).