以重组菌E.coli Rosetta/pETDuet-rcr-fdh全细胞为催化剂,2-羟基苯乙酮和甲酸钠为双底物,研究了生物催化反应体系中温度、pH值、底物2-羟基苯乙酮的初始浓度、反应时间、细胞浓度和状态等对产物(R)-苯基乙二醇生成效率的影响规律.结果表明,在温度35℃和pH7.0、底物初始浓度6g/L、时间36h、湿细胞浓度10%(ω)的条件下,产物的光学纯度高达98.37%e.e.,得率达79.14%.在上述反应体系中添加5mmol/LZnSO4后,重组菌催化的不对称还原反应效率显著提高,产物(R)-苯基乙二醇光学纯度达到100%e.e.,得率高达86.3%.
The effects of reaction temperature,pH value,initial substrate concentration,reaction duration,cell concentration and forms on biotransformation reaction for preparation of(R)-1-phenyl-1,2-ethanediol were studied with 2-hydroxyacetophenone and sodium formate as substrates and the whole cells of E.coli Rosetta/pETDuet-rcr-fdh as catalyst.The results showed that when the concentration of 2-hydroxyacetophenone was 6 g/L,(R)-1-phenyl-1,2-ethanediol was produced with high optical purity of 98.37% enantiomeric excess and yield of 79.14% at the conditions of temperature 35 ℃,pH 7.0,reaction duration 36 h and wet cell concentration 10%(ω).Further research showed that the biotransformation efficiency could be well improved by addition of Zn2+.In the presence of Zn2+ with a final concentration of 5 mmol/L,the enantiomeric excess and yield reached 100% and 86.3%,respectively.