Optimization of conditions for expression of Rhizopus oryzae lipase in Pichia pastoris and its high-density fermentation

HE Xiaoyun,LI Xun,SHI Hao,LI Zhilin, WANG Fei

Journal of Nanjing Forestry University (Natural Sciences Edition) ›› 2013, Vol. 37 ›› Issue (03) : 117-122.

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Journal of Nanjing Forestry University (Natural Sciences Edition) ›› 2013, Vol. 37 ›› Issue (03) : 117-122. DOI: 10.3969/j.issn.1000-2006.2013.03.021

Optimization of conditions for expression of Rhizopus oryzae lipase in Pichia pastoris and its high-density fermentation

  • HE Xiaoyun,LI Xun,SHI Hao,LI Zhilin, WANG Fei*
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Abstract

The expression conditions of lipase from genetically engineering Pichia pastoris were optimized and its high-density fermentation in a 10 L fermenter was investigated. The results showed that the optimal pH was in range of 5.5-6.0, suitable methanol dosage was 1%, the optimal fermentation time was 96 h after induction with methanol and temperature was 28-30 ℃, and it was also indicated that 2% of corn steep powder could substitute yeast extract and peptone as nitrogen source. By using glycerol feeding strategy with DO control in high-density fermentation, the dry cell weight was up to 120.1 g/L, and the total protein concentration reached at 2.5 g/L in the fed batch fermentation phase. 54 hours after the induction with methanol, the lipase activity got to be 564 μmol/(min·mL), which is as much as two times of the lipase activity in the shake flask fermentation.

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HE Xiaoyun,LI Xun,SHI Hao,LI Zhilin, WANG Fei. Optimization of conditions for expression of Rhizopus oryzae lipase in Pichia pastoris and its high-density fermentation[J]. Journal of Nanjing Forestry University (Natural Sciences Edition). 2013, 37(03): 117-122 https://doi.org/10.3969/j.issn.1000-2006.2013.03.021

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