Scale-up fermentation of recombinant Candida rugosa lipase expressed in Pichia pastoris using the GAP promoter
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- 作者：Wei Zhao ; Jinwen Wang ; Riqiang Deng and Xunzhang Wang
- 关键词：Scale ; up ; Candida rugosa lipase ; Pichia pastoris ; GAP promoter ; High ; cell ; density
- 刊名：Journal of Industrial Microbiology and Biotechnology
- 出版年：2008
- 出版时间：March, 2008
- 年：2008
- 卷：35
- 期：3
- 页码：189-195
- 全文大小：379.6 KB
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Chemistry
Biotechnology
Genetic Engineering
Biochemistry
Bioinformatics
Microbiology
Microbial Genetics and Genomics - 出版者：Springer Berlin / Heidelberg
- ISSN：1476-5535
文摘
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.