生物加工过程2016，Vol.14Issue(5)：37-44,8.DOI:10.3969/j.issn.1672-3678.2016.05.008
嗜热踝节菌脂肪酶（TTL）的重组表达及催化合成普瑞巴林手性中间体
Overexpression of Talaromyces thermophilus lipase (TTL) and its application in biosynthesis of pregabalin intermediate
摘要
Abstract
(3S)-2-carboxyethyl-3-cyano-5-methylhexanoic acid is the key intermediate of pregabalin. In this study,a novel lipase( TTL) was exploited from genome of Talaromyces thermophiles by gene mining for efficient synthesis of ( 3S )-2-carboxyethyl-3-cyano-5-methylhexanoic acid. TTL was overexpressed in Escherichia coli BL21 at low temperature. TTL was purified to electrophoretic homogeneity and characterized. The obtained TTL exhibited the maximum activity at 50 ℃ and showed excellent thermostability with a half-life of 14. 5 d.TTL was stable in the pH range from 7. 0 to 8. 5 and the optimal pH was 8. 0. Kinetic resolution of 2-carboxyethyl-3-cyano-5-methylhexanoic acid ethyl ester by TTL was carried out at pH 8. 0,50 ℃.A conversion of 42% with e. e.>94% was obtained after 20 h at a substrate loading of 100 mmol/L. The obtained results demonstrate that TLL is a promising biocatalyst in chemoenzymatic synthesis of pregabalin.关键词
嗜热踝节菌/脂肪酶/低温表达/酶学性质/普瑞巴林Key words
Talaromyces thermophilus/lipase/cold-inducible expression/enzymatic properties/pregabalin分类
生物科学引用本文复制引用
吴欣玮,郑仁朝,郑裕国..嗜热踝节菌脂肪酶（TTL）的重组表达及催化合成普瑞巴林手性中间体[J].生物加工过程,2016,14(5):37-44,8.基金项目
浙江省重大科技专项社会发展项目 （）
