Food and Fermentation Industries

Expression and enzymatic properties of acetaldehyde dehydrogenase

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  • 1(The Key Laboratory of Industrial Biotechnology;Jiangnan University;Wuxi 214122;China) 2(National Engineering Laboratory for Cereal Fermentation Technology;Jiangnan University;Wuxi 214122;China) 3(School of Biotechnology; Jiangnan University;Wuxi 214122;China) 4(Jiangsu Agribusiness Malt Co.Ltd;Sheyang 224300;China) 5(China Resources Snow Breweries Co.Ltd;Qinhuangdao 066001;China)

Online published: 2018-05-25

Abstract

The target geneALDH2 was synthesized according to the sequences of human acetaldehyde dehydrogenase 2 in GeneBank after codon optimization ofSaccharomyces cerevisiae. Then theALDH2 gene expression cassette TRP1L-URA3-TPIp-ALTH2-TPIt-TRP1R was constructed using fusion PCR and electro-transformed intoS.cerevisiaeW303-1A to produce the modified strain W303-ALDH2. The recombinant ALDH2 was obtained after purification by His TrapTMexcel affinity chromatography; and its molecular weight and activity was 56 kDa and 20.70 U/mg; respectively. The results showed that the optimal reaction pH value and temperature for recombinant ALDH2 were 5.0 and 35 ℃; respectively. Except Na+; the K+; Ca2+; Mg2+; Mn2+could increase the activity of ALDH2 in different degrees. When acetaldehyde was used as substrate; theKmvalue and maximum reaction rateVmaxof recombinant ALDH2 was 0.908 mmol/L and 114.94 U/mg; respectively. When NAD+was used as substrate; theKmvalue and maximum reaction rateVmaxof recombinant ALDH2 was 0.282 mmol/L and 58.82 U/mg; respectively.

Cite this article

DOU Xin-xi1;2;3;ZHANG Ming4;LIN Cai5;WU Dian-hui1;2;3; LI Xiao-min1;2;3;SUN Jun-yong1;2;3;CAI Guo-lin1;2;3;LU Jian1;2;3* . Expression and enzymatic properties of acetaldehyde dehydrogenase[J]. Food and Fermentation Industries, 2018 , 44(4) : 22 -28 . DOI: 10.13995/j.cnki.11-1802/ts.016237

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