%0 Journal Article
%T Glycine-aspartic _ acid-serine-leucine esterase Xcc_ est from Xanthomonas campestris pv. campestris 8004 and its esterase domain: gene expression in Escherichia coli, refolding and characterization<br>野油菜黄单胞菌8004甘天丝亮特征序列酯酶及其酯酶结构域在大肠杆菌中的表达，包涵体复性及性质
%A Jianjun Wang
%A Liu Yang
%A Yanping Cao
%A Guojun Zheng
%A <br>王建军
%A 杨柳
%A 曹燕萍
%A 郑国钧
%J 微生物学报
%D 2009
%I 
%X Objective] To characterize the GDSL (glycine, aspartic acid, serine and leucine motif in protein sequence) esterase Xcc_ est from Xanthomonas campestris pv. campestris (Xcc) 8004. Methods] Xcc _ est gene and different domains of Xcc _ est gene were PCR amplified and expressed in Escherichia coli, the HIS-Tagged fusion proteins were pttrified by Ni-NTA chromatography. Results] The optimum pH and temperature of partly purified Xcc_ est were 8.0 and 52℃ when pNPB (4-nitrophonylbutyrate) was used as substrate. The Km and Vmax value of Xcc _ est and the passenger domain (Xcc _ estN1-334) for pNPB were 47.6±4.6 mol/L, 67.6±7.8 U/mg and 469.4 ±9.8 mol/L, 2.5±0.9 U/mg respectively. Inclusion bodies of mature domain Xcc _ est (Xcc _ estN26-606) could be refolded but inclusion bodies of the passenger domain (Xcc _ estN26-334) could not be refolded. Refolded mature domain had broad substrate spectrum and showed higher stability than Xcc_ est when stored at 25 ℃. Conclusions] Refolded Xcc_ estN26-606 can be a candidate for biotransformation application.
%K campestris)
%K GDSL esterase
%K Xanthomonas campestris
%K refolding<br>GDSL酯酶
%K 野油菜黄单胞菌(Xanthomonas
%K 复性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=508F706625A5EC0FDE5250A2E2FB6186&yid=DE12191FBD62783C&vid=2A3781E88AB1776F&iid=0B39A22176CE99FB&sid=AC1578C6BB9EBDEF&eid=2BA123C6EB9D54C2&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=20