解淀粉芽胞杆菌WS-8中LanM基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of LanM Gene in Bacillus Amyloliquefaciens WS-8

作　　者：崔丽婷 刘洪伟[1] 程辉彩[1] 王雅娜 王琦[3] 李清扬 张丽萍[1] Cui Liting;Liu Hongwei;Cheng Huicai;Wang Yana;Wang Qi;Li Qingyang;Zhang Liping(Institute of Biology,Hebei Academy of Science,Shijiazhuang,050081;Hebei Normal University,Shijiazhuang,050024;China Agricultural Uni- versity,Beijing,100083;College of Food Science and Engineering,South China University of Technology,Guangzhou,510640)

机构地区：[1]河北省科学院生物研究所,石家庄050081 [2]河北师范大学,石家庄050024 [3]中国农业大学,北京100083 [4]华南理工大学食品科学与工程学院,广州510640

出　　处：《基因组学与应用生物学》2019年第2期644-649,共6页Genomics and Applied Biology

基　　金：河北省科技计划项目(No.16292904D);河北省科学院科技计划项目(No.16304);河北省科学院科技计划项(No.2017G02)共同资助

摘　　要：克隆解淀粉芽胞杆菌WS-8 (Bacillus amyloliquefaciens WS-8)中的第二类羊毛硫肽合成酶LanM基因,并对LanM编码蛋白的理化性质及结构特征进行分析。设计LanM基因(登录号:APQ49580.1)扩增引物,提取解淀粉芽胞杆菌WS-8基因组DNA,以其作为模板进行PCR扩增。综合多种软件预测和分析LanM编码蛋白的理化性质、结构域和二级结构等。采用邻位连接法(Neighbor-Joining, NJ)构建系统发育树来分析LanM所编码蛋白与同源蛋白的亲缘关系,以及各蛋白结构域的亲缘关系。PCR扩增出的目的条带约为2 840 bp,通过测序鉴定,序列信息与基因组数据一致。LanM基因编码961个氨基酸,等电点为6.07,相对分子质量为111.485 1 kD。LanM编码的蛋白属于亲水性蛋白,无信号肽。该蛋白含有LANC_like结构域,其二级结构主要由螺旋结构和环状结构组成。系统发育树的分析结果表明LANC_like结构域和LanM同源蛋白的亲缘关系一致。解淀粉芽胞杆菌WS-8中存在第二类羊毛硫肽合成酶LanM基因。揭示了羊毛硫肽合成酶LanM发挥脱水和环化作用的理化性质和结构基础,以期为进一步阐明羊毛硫肽合成酶的生物学功能提供参考。The study was aimed to clone the LanM gene of type Ⅱ lanthipeptide synthetases from Bacillus amyloliquefaciens WS-8 and to analyze the physicochemical property and structure characteristics of LanM encoded protein. Primer for LanM gene(Accession: APQ49580.1) amplification was designed to extract genome DNA from B. amyloliquefaciens WS-8, which was later taken as template for PCR amplification. The physicochemical properties, function domains and secondary structures of LanM encoded protein were analyzed and predicted by various software. Neighbor-Joining method(NJ) was applied to construct the phylogenetic trees so as to analyze the genetic relationship between LanM encoded proteins and homologous proteins, and the genetic relationship of their function domains. The size of the target band amplified by PCR was about 2 840 bp, of which the sequencing results were consistent with the genome data. LanM gene encoded 961 amino acids and the relativemolecular mass was 111.485 1 kD, with the isoelectric point of 6.07. LanM encoded protein belonged to the hydrophilic protein. The encoded protein had a LANC_like domain but no signal peptide, and the helical structure and the cyclic structure dominated secondary structure of the encoded protein. The phylogenetic trees analysis indicated that the genetic relationship of LANC_like domains were consistent to LanM homologous proteins. There is a LanM gene of type Ⅱ lanthipeptide synthetases in B. amyloliquefaciens WS-8. The study revealed the physicochemical properties and structural basis of lanthipeptide synthetases for dehydration and cyclization, which would provide reference for further elucidating the biological function of lanthipeptide synthetases.

关 键 词：解淀粉芽胞杆菌WS-8 第二类羊毛硫肽 生物信息学 蛋白质结构 

分 类 号：R378[医药卫生—病原生物学]