Scale-up fermentation of recombinant Candida rugosa lipase expressed in Pichia pastoris using the GAP promoter.
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.
Zhao
Wei
W
State Key Laboratory of Biocontrol, School of Life Science, Sun Yat-Sen (Zhong Shan) University, Guangzhou, PR China.
Wang
Jinwen
J
Deng
Riqiang
R
Wang
Xunzhang
X
D002175:Candida / Q000201:enzymology* / Q000235:genetics; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D005656:Fungal Proteins / Q000096:biosynthesis; D008049:Lipase / Q000096:biosynthesis*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014170:Transformation, Genetic
D003470:Culture Media; D005656:Fungal Proteins; D011994:Recombinant Proteins; D008049:Lipase