摘要
将来源于7种微生物的13个氧化还原酶基因分别与表达载体pET21c连接后,转化入Escherichia coli BL21(DE3)中,得到13株重组菌。重组菌在IPTG诱导下进行表达,并对2-羟基苯乙酮进行不对称催化还原。研究发现,在17℃下诱导表达的重组酶比活明显高于30℃和37℃下诱导表达的比活。另外,来源于Candida parapsilosis CCTCCM203011羰基还原酶SCR1能够较好地不对称催化还原2-羟基苯乙酮,在5 g/L底物浓度下,2-羟基苯乙酮被还原得到(S)-PED,光学纯度大于99%,产率达到90.3%。通过对有机溶剂的筛选,构建了水/十二酸乙酯两相反应体系,可以还原底物浓度达8 g/L的2-羟基苯乙酮,光学纯度大于99%,产率为86.9%。
13 recombinant Escherichia coli BL21 （DE3） overexpressing 13 different oxidoreductases were constructed respectively. 2-hydroxyacetophenone was applied as substrate to investigate the biocatalytic asymmetric reduction by the recombinants. The results indicated that relative enzyme activity of the expressed enzyme was much higher at the induc- tion temperature of 17℃ than that at 30℃ and 37℃. Besides, SCR1, which was from Candida parapsilosis CCTC- CM203011 showed best performance. The effects of substrate concentration on asymmetric conversion of 2-hydroxyacetophenone were investigated. It could asymmetrically reduce a-hydroxyacetophenone to produce （S）-PED with 99.9 % ee and 90.3 % yield with substrate concentration of 5 g/L. By introducing ethyl laurate as organic phase to form biphasic system, 8 g/L 2-hydroxyacetophenone was reduce to （S）-PED with 99.9% ee and 86.9% yield.
出处
《工业微生物》
CAS
CSCD
2011年第5期1-5,共5页
Industrial Microbiology
基金
国家自然科学基金(20776060
30800017)
863国家高技术研究发展计划(2006AA020104
2007AA02Z226)
973国家重点基础研究发展规划(2009CB724706)
高等学校学科创新引智计划(111计划
111-2-06)
江苏省自然科学基金(BK2008528)
