期刊
BIOCATALYSIS AND BIOTRANSFORMATION
卷 33, 期 5-6, 页码 260-269出版社
TAYLOR & FRANCIS LTD
DOI: 10.3109/10242422.2016.1168815
关键词
Candida rugosa lipase LIP1; co-expression; Pichia pastoris; resolution; Vitreoscilla hemoglobin
资金
- National Natural Science Foundation of China (NSFC) [31170078, 31070089]
- National High Technology Research Development Program of China (863 Program) [2014AA093510, 2011AA02A204]
- Innovation Foundation of Huazhong University of Science and Technology (HUST) [2014NY007, 2014QN119, 2012SHYJ004]
- Fundamental Research Funds for the Central Universities HUST [2172012SHYJ004]
To improve the productivity of Candida rugosa lipase (CRL) and alleviate respiration limitations during high cell-density fermentation, codon-optimized CRL LIP1, and Vitreoscilla hemoglobin (VHb) were co-expressed in Pichia pastoris. The activity of the recombinant strain that expressed LIP1 and VHb from dual promoters, named GS115/9Klip1FZvgb-lip1(#)1, toward olive oil reached 620U/mL, which was 1.69-fold greater than that of the recombinant strain GS115/9Klip1(#)139 (365U/mL) which only expressed LIP1 from a single promoter, and 1.37-fold greater than that of the recombinant strain GS115/9Klip1FZlip1(#)39 (450U/mL), which only expressed LIP1 from two promoters, in shaking flasks. With FM22 as the basic medium and methanol/D-sorbitol (1:1, v/v) as an inducer, the maximum activity of GS115/9Klip1FZvgb-lip1(#)1 reached 7490 +/- 379.5U/mL, which was 2.65-fold greater than that of GS115/9Klip1(#)139 (2820 +/- 112U/mL) and 1.82-fold greater than that of GS115/9Klip1FZlip1(#)39 (4100 +/- 205U/mL) in 10L fermenters. The conversion ratio (C) and enantiomeric excess (ee(s)) of racemic ibuprofen by immobilized CRL LIP1 reached 35.10 and 31.63%, respectively.
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