海洋细菌Bacillus mojavensis 1A00437 β-1,3-1,4-葡聚糖酶基因的克隆、表达及功能研究  

Study on Gene Cloning,Expression and Function of β-1,3-1,4-Glucanase fromBacillus Mojavensis 1A00437

作　　者：左怀雨 王茂淋[1] 邵宗泽[2] 李光玉[2] 徐柳[1] 喻子牛[1] 张吉斌[1] 

机构地区：[1]华中农业大学生命科学技术学院农业微生物学国家重点实验室微生物农药国家工程研究中心,湖北武汉430070 [2]国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,福建厦门361005

出　　处：《化学与生物工程》2014年第9期29-35,共7页Chemistry & Bioengineering

基　　金：农业微生物学国家重点实验室资助项目

摘　　要：Bacillus mojavensis 1A00437是分离自太平洋的一株芽胞杆菌,成功克隆了该菌的β-1,3-1,4-葡聚糖酶基因并在原核系统获得高效表达。测序结果表明,该基因的ORF全长681bp,编码一条226个氨基酸的蛋白,蛋白大小25.3kDa。该酶的最适pH值及最适反应温度分别为6.5、60℃。研究表明,1mmol·L-1 Ca2+、Mn2+可显著抑制该重组酶活性,该重组酶对稻瘟病菌无抗性,但对小麦纹枯病菌及水稻纹枯病菌具有明显抑制作用。饲料体外消化实验表明,该重组酶对稻糠、玉米粉具有显著降解效果。为该酶用作抗真菌剂及饲料添加剂提供了理论基础。Bacillus mojavensis 1A00437 was isolated from Pacific Ocean.Theβ-1,3-1,4-glucanases gene of strain 1A00437 was successfully cloned.The open reading frame（ORF）of the gene contained 681 bp coding for a 226amino-acid matured protein,and the molecular weight of the expressed Bgl protein was 25.3kDa.The optimal pH value and temperature of the recombinant enzyme were 6.5and 60 ℃,respectively.1mmol·L-1Ca2＋,Mn2＋can significantly inhibit the enzyme activity.The recombinant enzyme had no activity on Magnaporthe oryzae,but had strong inhibitory effects on Rhizoctonia solani and Rhizoctonia solani kühn.The feed digestion experiment in vitro showed that the recombinant enzyme had significant digestion effect of rice bran and corn flour.This study provided theoretical basis for the enzyme to be used as antifungal agent and feed additives.

关 键 词：BACILLUS mojavensis 葡聚糖酶 基因克隆 表达 功能 饲料添加剂 

分 类 号：Q785[生物学—分子生物学]