Scale-up fermentation of recombinant Candida rugosa lipase expressed in Pichia pastoris using the GAP promoter
- 18 December 2007
- journal article
- Published by Oxford University Press (OUP) in Journal of Industrial Microbiology & Biotechnology
- Vol. 35 (3), 189-195
- https://doi.org/10.1007/s10295-007-0283-8
Abstract
The high-cell-density fermentation of Candida rugosa lipase in the constitutive Pichia pastoris expression system was scaled up from 5 to 800 l in series by optimizing the fermentation conditions at both lab scale and pilot scale. The exponential feeding combined with pH-stat strategy succeeded in small scale studies, while a two-stage fermentation strategy, which shifted at 48 h by fine tuning the culture temperature and pH, was assessed effective in pilot-scale fermentation. The two-stage strategy made an excellent balance between the expression of heterogeneous protein and the growth of host cells, controlling the fermentation at a relatively low cell growth rate for the constitutive yeast expression system to accumulate high-level product. A stable lipase activity of approximately 14,000 IU ml−1 and a cell wet weight of ca. 500 g l−1 at the 800-l scale were obtained. The efficient and convenient techniques suggested in this study might facilitate further scale-up for industrial lipase production.Keywords
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