期刊
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
卷 35, 期 3, 页码 189-195出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s10295-007-0283-8
关键词
scale-up; candida rugosa lipase; Pichia pastoris; GAP promoter; high-cell-density
The high-cell-density fermentation of Candida rugosa lipase in the constitutive Pichia pastoris expression system was scaled up from 5 to 800 1 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 800-1 scale were obtained. The efficient and convenient techniques suggested in this study might facilitate further scale-up for industrial lipase production.
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