生物加工过程2016，Vol.14Issue(2)：33-40,8.DOI:10.3969/j.issn.1672-3678.2016.02.007
一种新型醛酮还原酶的克隆表达、性质研究以及在不对称合成（R）CHBE中的应用
Overexpression,characterization of a novel aldo-keto reductase for asymmetric synthesis of (R)-CHBE
摘要
Abstract
An NADPH⁃dependent aldo⁃keto reductase ( CAK) gene from Candida albicans SC5314 was discovered for reducing 4⁃chloroacetoacetate ethyl ester ( COBE ) to ethyl ( R )⁃4⁃chloro⁃3⁃hydroxybutanoate ( ( R )⁃CHBE ) . The CAK gene was cloned and overexpressed in Escherichia coli Rosetta. The catalytic properties of purified CAK were studied. Furthermore,an enzyme⁃coupled coenzyme regeneration system with co⁃substrate glucose was constructed, and its capability of reducing COBE to ( R)⁃CHBE was studied. The results showed that CAK had catalytic activity for a variety of aldehydes and ketones. CAK had a maximum activity at 40℃ and pH 5. The enzyme was stable below 40℃ and under acidic conditions. Mg2+, Na+ and K+ could enhance the activity of CAK. When Cu2+ was added, no enzyme activity was detected. The inhibit effect of ethyl acetate and dibutyl⁃O⁃phthalate was relatively light. When the coenzyme regeneration system for conversion reaction was applied,the molar conversion yield reached 80�6% with 600 mmol/L COBE,and the e�e� value was over 99%.关键词
醛酮还原酶/NADPH再生/蛋白纯化/酶学性质/(R)4氯3羟基丁酸乙酯Key words
aldo-keto reductase/NADPH regeneration/protein purifying/enzyme property/( R)-4-chloro-3-hydroxybutanoic acid ethyl ester分类
生物科学引用本文复制引用
张瑾成,魏淼,许琳,严明..一种新型醛酮还原酶的克隆表达、性质研究以及在不对称合成（R）CHBE中的应用[J].生物加工过程,2016,14(2):33-40,8.基金项目
国家重点基础研究发展计划（973计划）（2011CBA00804）；国家高技术研究发展计划（863计划）（2012AA022101）；江苏高校优势学科建设工程 （）
