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Efficient secretion of lipase r27RCL in Pichia pastoris by enhancing the disulfide bond formation pathway in the endoplasmic reticulum

  • Biotechnology Methods
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The lipase r27RCL from Rhizopus chinensis CCTCC M201021 was heterologously expressed in Pichia pastoris GS115 by simultaneous co-expression with two secretion factors ERO1p and PDI involved in the endoplasmic reticulum (ER). Compared to the expression of the lipase alone (12,500 U/ml), co-expression with these two proteins resulted in the production of larger total quantities of enzymes. The largest increase was seen when the combined ERO1p/PDI system was co-expressed, resulting in approximately 30 % higher enzyme yields (16,200 U/ml) than in the absence of co-expressed secretion factors. The extracellular protein concentration of the recombinant strain Co XY RCL-5 reached 9.39 g/l in the 7-l fermentor. Simultaneously, the fermentation time was also shortened by about 8 h compared to that of the control. The substrate-specific consumption rate (Qs) and the product-specific production rate (Qp) were both investigated in this research. In conclusion, the space–time yield was improved by co-expression with ERO1p and PDI. This is a potential strategy for high level expression of other heterologous proteins in P. pastoris.

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Acknowledgments

Financial support from the National High Technology Research and Development Program of China (863 program; no. 2012AA022207), the National Key Basic Research and Development Program of China (973 program; no. 2011CB710800), the Program of Introducing Talents of Discipline to Universities (111 project; 111-2-06), and the Ministry of Education, China, are greatly appreciated.

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 lihu Avenue, Wuxi, 214122, Jiangsu, China

    Chong Sha, Xiao-Wei Yu, Meng Zhang & Yan Xu

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  1. Chong Sha
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  2. Xiao-Wei Yu
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  3. Meng Zhang
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  4. Yan Xu
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Correspondence to Xiao-Wei Yu or Yan Xu.

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Sha, C., Yu, XW., Zhang, M. et al. Efficient secretion of lipase r27RCL in Pichia pastoris by enhancing the disulfide bond formation pathway in the endoplasmic reticulum. J Ind Microbiol Biotechnol 40, 1241–1249 (2013). https://doi.org/10.1007/s10295-013-1328-9

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