高等学校化学学报2018，Vol.39Issue(1)：54-63,10.DOI:10.7503/cjcu20170509
复合交联酶聚集体的制备及催化羰基不对称还原合成手性醇
Preparation of Combined Cross-linked Enzyme Aggregates and Its Application in Synthesis of Chiral Alcohols by the Asymmetric Reduction of Carbanyl Group
摘要
Abstract
Using ketoreductase KRED 30 from Corynebacterium sp. and the D-glucose dehydrogenase from Bacillus subtilis as the model enzymes, a combined cross-linked enzyme aggregates( combi-CLEAs) with high activity was prepared by optimization of the precipitant type, glutaraldehyde concentration, cross-linking tem-perature and time. Characterization of the combi-CLEAs showed that the stability and substrate tolerance of free enzymes mixture were significantly improved after combined cross-linking. Investigation of catalytical perform-ance showed that combi-CLEAs could effectively catalyze the reduction of ethyl 4-chloroacetoacetate ( COBE ) and 3′-chloroacetophenone( CPO) for the synthesis of corresponding chiral alcohols in aqueous system. Only a little cofactor was needed to start the reactions. The catalytical activity could retain more than 70％ after con-tinuously catalyzing for 10 batches. The result showed that catalytical performance of combi-CLEAs in the bi-phasic system was much better than that in aqueous system. The total turnover number( TTN) for catalyzing COBE could attend 6595, and TTN for catalyzing CPO could attend 7500. Thus, cofactor regeneration by com-bi-CLEAs is feasible.关键词
酮还原酶/D-葡萄糖脱氢酶/辅酶再生/复合交联酶聚集体/手性醇Key words
Ketoreductase/D-Glucose dehydrogenase/Cofactor regeneration/Combined cross-linked enzyme aggregates( Combi-CLEAs)/Chiral alcohol分类
化学化工引用本文复制引用
杨猛,江惠娟,宁晨曦,魏东芝,苏二正..复合交联酶聚集体的制备及催化羰基不对称还原合成手性醇[J].高等学校化学学报,2018,39(1):54-63,10.基金项目
国家自然科学基金(批准号:31401679)、 江苏省"六大人才高峰"高层次人才项目(批准号:2015-JY-016)、 中国博士后科学基金(批准号:2016M600417,2017T100373)、江苏省第五期"333工程"培养资金资助项目(批准号:BRA2017458)和生物反应器工程国家重点实验室开放基金资助. Supported by the National Natural Science Foundation of China( No. 31401679) ,the Six Talent Peaks Project in Jiangsu Province,China ( No. 2015-JY-016) ,the China Postdoctoral Science Foundation( Nos. 2016M600417,2017T100373) ,the 333 Project of Jiangsu Province,Chi-na( No. BRA2017458) and the Open Funding Project of the State Key Laboratory of Bioreactor Engineering,China. （批准号:31401679）
