Cloning of a Novel Pyrethroid-Hydrolyzing Carboxylesterase Gene from Sphingobium sp Strain JZ-1 and Characterization of the Gene Product
Cloning of a Novel Pyrethroid-Hydrolyzing Carboxylesterase Gene from Sphingobium sp Strain JZ-1 and Characterization of the Gene Product
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DOI：
10.1128/aem.01298-09
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发表时间：
2009-09-01
影响因子：
4.4
通讯作者：
Li, Shun-peng
中科院分区：
文献类型：
--
作者：
Wang, Bao-zhan;Guo, Peng;Li, Shun-peng
从鞘氨醇杆菌JZ-1菌株中克隆了一个新的酯酶基因pytH，编码拟除虫菊酯水解羧酸酯酶。该基因含有一个840 bp的开放阅读框。序列同一性搜索显示，推导的酶与许多α/β-水解酶折叠蛋白（20%至24%的同一性）具有最高的相似性。在大肠杆菌BL 21中表达PytH，并使用Ni-次氮基三乙酸亲和层析纯化。它是一种单体结构，分子量约为31 kDa，pI为4.85。PytH能够转化短链脂肪酸和广泛的拟除虫菊酯农药的对硝基苯基酯，异构体的选择性没有观察到。酶活性不需要辅因子。
A novel esterase gene, pytH, encoding a pyrethroid-hydrolyzing carboxylesterase was cloned from Sphingobium sp. strain JZ-1. The gene contained an open reading frame of 840 bp. Sequence identity searches revealed that the deduced enzyme shared the highest similarity with many alpha/beta-hydrolase fold proteins ( 20 to 24% identities). PytH was expressed in Escherichia coli BL21 and purified using Ni-nitrilotriacetic acid affinity chromatography. It was a monomeric structure with a molecular mass of approximately 31 kDa and a pI of 4.85. PytH was able to transform p-nitrophenyl esters of short-chain fatty acids and a wide range of pyrethroid pesticides, and isomer selectivity was not observed. No cofactors were required for enzyme activity.