%0 Journal Article
%T Cloning, expression and characterization of a gamma-butyrobetaine hydroxylase gene bbh from Pseudomonas sp. L-1<br>一株假单胞菌(Pseudomonas sp.)L-1菌株γ-丁基甜菜碱羟化酶基因bbh 的克隆、表达及酶学性质
%A Xiangfeng Lu
%A Pengcheng Zhang
%A Qianyan Li
%A Hui Liu
%A Xiaoqing Lin
%A Xiaohang Ma
%A <br>卢向锋
%A 张鹏程
%A 李倩延
%A 刘辉
%A 林小清
%A 马晓航
%J 微生物学报
%D 2012
%I 
%X Objective] Gamma-butyrobetaine hydroxylase is an enzyme that catalyzes the last step in the biosynthesis of L-carnitine.We cloned,expressed and characterized a gamma-butyrobetaine hydroxylase gene bbh from Pseudomonas sp.L-1,to facilitate the production of L-carnitine using microorganisms.Methods] We cloned bbh gene by PCR,and then cloned the open reading frame of bbh into pET-15b vector and expressed by Isopropyl β-D-1-thiogalactopyranoside(IPTG) induction.After His-Bind Resin purification,the characteristics of BBH were studied.The three-dimensional structure of BBH monomer was modeled by SWISS-MODEL Workspace and resting cells were used for L-carnitine transformation.Results] We cloned a gamma-butyrobetaine hydroxylase gene bbh(GenBank: JQ250036) from Pseudomonas sp.L-1 and expressed the gene in Escherichia coli BL21(DE3).BBH fusion protein was a homodimer,and the molecular weight of subunit was about 46.5kDa.The optimal temperature and pH was 30 ℃ and pH 7.5.The enzyme was stable below 45 ℃.The enzyme was most stable at pH 6.0.We used resting cells of recombinant E.coli for L-carnitine biotransformation,after incubated at 30 ℃ and pH 7.0 for 31 h,the concentration of L-carnitine reached 12.7 mmol/L.Conclusion] The bbh gene from Pseudomonas sp.L-1 strain is remarkably different from that of reported one.The gamma-butyrobetaine hydroxylase expressed by this gene could effectively transform γ-butyrobetaine for L-carnitine production.Beside by reporting of a bbh gene from bacteria,this research also provided a new process for biotransformation of L-carnitine.
%K Pseudomonas sp
%K gamma-butyrobetaine hydroxylase
%K bbh
%K cloning
%K expression<br>假单胞菌
%K γ-丁基甜菜碱羟化酶
%K bbh
%K 克隆
%K 表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=4D1F54CAF2A20CB3F0A666A73094F842&yid=99E9153A83D4CB11&vid=286FB2D22CF8D013&iid=94C357A881DFC066&sid=3B2BF7AC5674E8E2&eid=44E78A5D1B37D836&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=0