Arthrobacter nitroguajacolicus腈水解酶基因的克隆和表达  

Cloning and expression of nitrilase from Arthrobacter nitroguajacoticus

作　　者：唐璐敏[1] 徐玉华[1] 薛建萍[1] 

机构地区：[1]上海市农药研究所生物工程中心,上海200032

出　　处：《工业微生物》2014年第6期13-20,共8页Industrial Microbiology

基　　金：国家高技术研究发展计划(863计划);课题编号SS2014AA022106

摘　　要：腈水解酶是一类能将腈类化合物催化生成酸的氰基水解酶。目前已有多个菌种的腈水解酶基因序列被报道,如敏捷食酸菌Acidovorax facilis,粪产碱菌Alcaligenes faecalis,睾丸丛毛单胞菌Comamonas testoteroni,肺炎克雷伯菌Klebsiella pneumoniae,假单胞菌Pseudomonas属菌株,红球菌Rhodococcus属菌株,但节杆菌属菌株Arthrobacter nitroguajacolicus的腈水解酶基因序列尚未见报道。经由野生型酶的分离纯化,基因文库筛选及侧翼序列扩增等步骤,克隆得到该菌株的腈水解酶基因,从而为进一步研究该酶的特性及构建用于工业生产的重组菌打下基础。Nitrilase can hydrolysis nitrile to produce corresponding organic acid. Many nitrilase genes from different microbial strains, such as Acidovorax facilis , Alcaligenes faecalis , Comarnonas testoteroni , Klebsiella pneumoniae , Pseudo- monas sp. and Rhodococcus sp. were reported. However, the nitrilase gene from Arthrobacter nitroguajacolicus has not been reported yet. In this study, with the aid of enzyme purification, genomic library screening and flanking sequence amplification, the nitrilase gene from Arthrobacter nitroguajacollicus was successfully cloned and expressed. The acquisition of this gene provided a basis for further study of enzyme properties and constructing of recombinant strain for industrial application.

关 键 词：腈水解酶 克隆 重组菌 羟基乙腈 

分 类 号：Q78[生物学—分子生物学] TQ920.1[轻工技术与工程—发酵工程]