耦合透性化辅酶循环制备手性(R)-苯基乙醇  

Coupling Permeabilized Cofactor Recycling for Chiral(R)-Phenethanol Preparation

作　　者：彭益强[1] 陈李玲 

机构地区：[1]华侨大学工业生物技术福建省高等学校重点实验室,福建厦门361021

出　　处：《精细化工》2016年第11期1219-1226,共8页Fine Chemicals

基　　金：福建省高校(华侨大学)科研基金资助项目(14BS317)~~

摘　　要：通过耦合透性化博伊丁假丝酵母(Candida boidinii,C.boidinii)加强了近平滑假丝酵母(Candida parapsilosis,C.parapsilosis)内羰基还原酶(Carbonyl reductase,CR)催化反应中的辅酶循环与反复利用,构建了有效的手性(R)-苯基乙醇透性化细胞耦合制备体系。实验结果如下:透性化的C.parapsilosis细胞底产物渗透效果增强,底物转化率由33.5%提升到43.8%;进一步外添加辅酶后,底物转化率由43.8%提高到了52.1%;耦合透性化C.boidinii细胞并同时外添加辅酶可大大增强辅酶循环效率,底物转化率可提高至71.8%。优化透性化细胞耦合体系反应条件,当以0.2 g近平滑假丝酵母(C.parapsilosis)为基准时,在C.parapsilosis与C.boidinii细胞质量比为1.5∶1,底物浓度为10 mmol/L,辅酶浓度为0.1 mmol/L时,底物转化率可达78.2%。离心法收集细胞进行多批次反应,当反应12批次时,耦合体系仍然保持有55%底物转化率,此时总的底物转化率达70.3%,产物经高效液相色谱测定为(R)-苯基乙醇,对映体过量值(e.e.值)达98.2%。Cofactor recycling and reutilization of carbonyl reductase catalysis in Candida parapsilosis（ C. parapsilosis） was reinforced by coupling with permeabilized cell of Candida boidinii（ C. boidinii）.Thus,an effective permeabilized cell coupling system was constructed for preparation of chiral（ R）-phenethanol. The experimental results showed that the substrate and product infiltration ability of permeabilized C. parapsilosis enhanced. The substrate conversion rate improved from 33. 5% to 43. 8%and it further improved from 43. 8% to 52. 1% after addition of the additional cofactor. The cofactor recycling efficiency could greatly enhance due to the coupling of permeabilized C. boidinii with the additional cofactor,and the substrate conversion rate could reach 71. 8%. Under the optimized reaction conditions of 0. 2 g C. parapsilosis,C. parapsilosis to C. boidinii cell mass ratio of 1. 5 ∶ 1,10 mmol / L substrate,0. 1 mmol / L cofactor,the substrate conversion rate could be up to 78. 2%. The cells were recollected by centrifugation for the multi-batch reaction. The substrate conversion rate could still maintain 55% when the coupling system was reacted for 12 batches,the total conversion rate of the substrate was 70. 3%.（ R）-phenethanol was obtained with an enantiomeric excess（ e. e.） value of98. 2%.

关 键 词：透性化细胞 辅酶再生 羰基还原酶 甲酸脱氢酶 (R)-苯基乙醇 生物工程 

分 类 号：Q552[生物学—生物化学]