Journal
ENZYME AND MICROBIAL TECHNOLOGY
Volume 53, Issue 6-7, Pages 438-443Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2013.09.009
Keywords
Lipase; Gene dosage; Chaperone PDI; Pichia pastoris
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Funding
- National High Technology Research and Development Program of China (863 Program) [2012AA022207]
- National Key Basic Research and Development Program of China (973 Program) [2011CB710800]
- Fundamental Research Funds for the Central Universities [JUSRP11014]
- Program of Introducing Talents of Discipline to Universities (111 Project) [111-2-06]
- NSFC [20802027]
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Pichia pastoris has been successfully used in the production of many secreted and intracellular recombinant proteins, but there is still a large room of improvement for this expression system. Two factors drastically influence the lipase r27RCL production from Rhizopus chinensis CCTCC M201021, which are gene dosage and protein folding in the endoplasmic reticulum (ER). Regarding the effect of gene dosage, the enzyme activity for recombinant strain with three copies lipase gene was 1.95-fold higher than that for recombinant strain with only one copy lipase gene. In addition, the lipase production was further improved by co-expression with chaperone PDI involved in the disulfide bond formation in the ER. Overall, the maximum enzyme activity reached 355 U/mL by the recombinant strain with one copy chaperone gene PDI plus five copies lipase gene proRCL in shaking flasks, which was 2.74-fold higher than that for the control strain with only one copy lipase gene. Overall, co-expression with PDI vastly increased the capacity for processing proteins of ER in P. pastoris. (C) 2013 Elsevier Inc. All rights reserved.
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