A thermostable and organic-solvent tolerant esterase from Pseudomonas putida ECU1011: Catalytic properties and performance in kinetic resolution of α-hydroxy acids

被引:47
|
作者
Ma, Bao-Di [1 ]
Yu, Hui-Lei [1 ]
Pan, Jiang [1 ]
Liu, Jia-Yan [1 ]
Ju, Xin [1 ]
Xu, Jian-He [1 ]
机构
[1] E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biocatalysis & Synthet Biotechnol, Shanghai 200237, Peoples R China
基金
中国国家自然科学基金;
关键词
Esterase; Thermostability; Organic-solvent tolerance; alpha-Hydroxy (carboxylic) acids; Deacetylation; ENANTIOSELECTIVE HYDROLYSIS; SEQUENCE SIMILARITY; CARBOXYLIC-ACIDS; MANDELIC-ACID; LIPASE; BIOCATALYSIS; OVEREXPRESSION; OPTIMIZATION; PURIFICATION; ACYLATION;
D O I
10.1016/j.biortech.2013.01.089
中图分类号
S2 [农业工程];
学科分类号
0828 ;
摘要
A novel esterase, rPPE01, from Pseudomonas putida ECU1011 was heterologously expressed in Escherichia coli and identified for enzymatic resolution of hydroxy acids via O-deacetylation. alpha-Acetoxy carboxylates were converted with approximately 50% yield and excellent enantioselectivity (E > 200) at a substrate concentration of 100 mM. The half-lives of rPPE01 were 14 days at 50 degrees C and 30 days at 30 degrees C, indicating the enzyme has relatively high thermostability. Another remarkable advantage of rPPE01 is that both the activity and thermostability were enhanced significantly in the presence of hydrophobic alkanes and ethers. rPPE01 retained 159% of its initial activity after incubation with 50% (v/v) n-heptane at 30 degrees C for 60 days. The attractive organic-solvent tolerance, good thermostability and high enantioselectivity towards alpha-acetoxy carboxylates endow rPPE01 with the potential of practical application for the production of enantiopure hydroxy acids. (C) 2013 Elsevier Ltd. All rights reserved.
引用
收藏
页码:354 / 360
页数:7
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