变形假单胞菌吡咯喹啉醌合成基因簇的克隆与分析  

Cloning and Bioinformatics Analysis of the Gene Cluster of Pyrroquinoline Quinone from Pseudomonas plecoglossicida

作　　者：王大明 孙雷[1,2] 孙文敬 崔凤杰[3,4] 龚劲松[1,2] 张晓梅[1,2] 史劲松 许正宏[1,2] WANG Daming;SUN Lei;SUN Wenjing;CUI Fengjie;GONG Jinsong;ZHANG Xiaomei;SHI Jinsong;XU Zhenghong(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China;National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China;School of Food and Biological Engineering,Jiangsu University,Zhenjiang 212013,China;Parchn Sodium Isovitamin C Co.,Ltd.,Dexing 334221,China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学粮食发酵工艺与技术国家工程实验室,江苏无锡214122 [3]江苏大学食品与生物工程学院,江苏镇江212013 [4]百勤异VC钠有限公司,江西德兴334221

出　　处：《食品与生物技术学报》2020年第6期47-54,共8页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(31571885);国家高技术研究发展计划项目(2012AA022103);江西省科技计划项目(赣知发[2015]64号);江西省创新团队建设计划项目(20142BCB24024);德兴市科技计划项目(德科发[2015]44号)。

摘　　要：从变形假单胞菌JUIM01中克隆到吡咯喹啉醌(PQQ)合成基因簇,阐明了其基因组成和生物学信息。根据已报道的假单胞菌的基因组进行简并引物设计,采用LA-PCR技术克隆变形假单胞菌的PQQ合成基因簇,对克隆的基因片段进行测序并使用生物信息学方法进行综合分析。结果表明:克隆到的基因片段全长为11659 bp,其中包括pqqF、pqqA、pqqB、pqqC、pqqD、pqqE、pqqM、pqqH和pqqI共9个基因,编码PQQ生物合成的前体短肽PqqA和合成途径的相关酶;这些基因与荧光假单胞菌Pf0-1的PQQ合成基因簇的基因组成类似,相应基因的序列一致性达41%~94%。本研究中首次从变形假单胞菌中克隆到PQQ合成基因簇,并对其进行生物信息学分析,为变形假单胞菌的PQQ生物合成途径和胞内再生机制的研究奠定了基础,进而为提高2KGA的生产强度提供了理论支撑。The gene cluster for pyrroquinoline quinone(PQQ)synthesis was cloned from Pseudomonas plecoglossicida JUIM01.Its genome composition and biological information were subsequently analyzed.According to the reported genome sequence of P.plecoglossicida JUIM01,primers were designed for amplification of the PQQ synthesis gene cluster via LA-PCR.The gene fragments were sequenced and analyzed by bioinformatics approaches.The results showed that the total length of the cloned gene fragment was 11659 bp,which included 9 genes:pqqF,pqqA,pqqB,pqqC,pqqD,pqqE,pqqM,pqqH and pqqI,and encoded the related enzymes in PQQ biosynthesis pathway and precursor peptide PqqA.These encoding genes were similar to the PQQ synthesis gene cluster of P.fluorescens Pf0-1 with sequence identifity of 41~94%.This study,for the first time,reported cloning of the PQQ synthesis gene cluster from P.plecoglossicida JUIM01,and the bioinformatics analysis laid a foundation for the study of PQQ biosynthesis pathway and intracellular regeneration mechanism of Pseudomonas strains.Furthermore,it provided theoretical support for improving the production intensity of 2KGA.

关 键 词：变形假单胞菌 吡咯喹啉醌 基因簇 克隆 生物信息学分析 

分 类 号：Q933[生物学—微生物学]