Expression and characterization of carbonyl reductase CpCR from Candida parapsilosis ATCC 7330
SONG Ting1,2, WANG Shuaijing1,2, WANG Chen1,2, LYU Yucai1,2, LUO Huajun1,2, GUO Jingling1,2, GONG Dachun1,2*
1(China Key Laboratory of Light Industry Functional Yeast, China Three Gorges University, Yichang 443002, China) 2(College of Biological and Pharmaceutical, China Three Gorges University, Yichang 443002, China)
Abstract: Carbonyl reductases from Candida parapsilosis are valuable to the synthesis of hydroxyl compounds. By constructing recombinant E. coli BL21 (DE3)/pACYCDuet-1-cpcr, the carbonyl reductase CpCR gene (cpcr) from C. parapsilosis. ATCC 7330 was expressed. The recombinant CpCR was separated and purified by Ni-Agarose affinity chromatography from His-tag on the pACYCDuet-1 and its enzymatic properties were investigated. The results showed that the full length of the carbonyl reductase gene cpcr was 1 107 bp,containing 368 amino acids with molecular weight around 41 kDa and specific enzyme activity of 20 U/mg. The recombinant CpCR was stable in the temperature range of 4-33 ℃ with the relative enzyme activity above 80%, and the T50 value was 37 ℃. The suitable pH range was 6.2-7.5, and optimal stability was obtained under neutral conditions. Cu2+ had a strong inhibition on the recombinant CpCR, and lowered the relative enzyme activity by 30% at 1 mmol/L, and by 50% at a 5 mmol/L. The affinity of the enzyme to benzaldehyde and n-butyraldehyde was stronger than that to 4-chlor-3-keto- butyrate-ethyl ester (COBE). The catalytic efficiency for benzaldehyde or n-butyraldehyde was about 20 times of that for COBE. The recombinant CpCR was an NADPH-dependent carbonyl reductase. This study provided basic data for the molecular modification and catalytic application of carbonyl reductase CpCR.
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2021, Vol. 47 
