食品科学2018，Vol.39Issue(2)：66-72,7.DOI:10.7506/spkx1002-6630-201802011
变形假单胞菌2-酮基葡萄糖酸差向异构酶基因的原核表达
Prokaryotic Expression of a 2-Ketogluconate Epimerase Gene from Pseudomonas plecoglossicida
摘要
Abstract
The complete nucleotide sequence of the gene (kguE) encoding 2-ketogluconate epimerase was cloned by touchdown polymerase chain reaction (TD-PCR) from an industrial 2-ketogluconic acid producer,Pseudomonas plecoglossicida JUIM01.A recombinant vector was constructed by ligating the restriction enzymes digested products of the target gene to pET-28a(+) vector and then transferred into the expression host E.coli BL21(DE3).A specific fusion protein with molecular weight of about 30.5 ku was expressed in the recombinant strain E.coli BL21(DE3)/pET-28a(+)-kguE after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction with a positive Western-blot result.Bioinformatic analysis showed that the protein was predicted to be a hydrophilic protein with molecular weight of 28.5 ku located in the cytoplasm,sharing 78％ amino acid sequence identity with the 2-ketogluconate epimerase from P.putida.The predicted secondary structure consisted of 40.62％ of α-helix,17.19％ of extended strand and 42.19％ of random coil.This study is expected to provide a basis for further elucidating the function of 2-ketogluconate epimerase in P.plecoglossicida.关键词
变形假单胞菌/2-酮基葡萄糖酸差向异构酶/kguE基因/克隆/表达/生物信息学Key words
Pseudomonas plecoglossicide/2-ketegluconate epimerase (KguE)/kguE gene/cloning/expression/bioinformatics分类
生物科学引用本文复制引用
孙文敬,杨荔,栾方,何小用,崔凤杰,王大明,钱静亚,齐向辉..变形假单胞菌2-酮基葡萄糖酸差向异构酶基因的原核表达[J].食品科学,2018,39(2):66-72,7.基金项目
国家自然科学基金面上项目(31571885) （31571885）
国家高技术研究发展计划(863计划)项目(2012AA022103) （863计划）
江西省科技计划项目(赣知发[2015]64号) （赣知发[2015]64号）
江西省创新团队建设计划项目(20142BCB24024) （20142BCB24024）
江西省科技平台建设计划项目(2010DTZ01900) （2010DTZ01900）
德兴市科技计划项目(德科发[2015]44号) （德科发[2015]44号）
