文献类型：期刊文献

中文题名：解淀粉芽孢杆菌β-N-乙酰氨基葡萄糖苷酶基因的克隆表达及其酶学表征

英文题名：Gene Cloning,Expression and Characterization ofβ-N-Acetylglucosaminidase from Bacillus amyloliquefaciens

作者：陈宝莉[1];秦臻[1];赵黎明[1]

机构：[1]华东理工大学生物工程学院,发酵工业分离提取技术研发中心,生物反应器工程国家重点实验室,上海200237

年份：2020

卷号：41

期号：8

起止页码：123

中文期刊名：食品科学

外文期刊名：Food Science

收录：CSTPCD;;EI(收录号：20202408811685);Scopus;北大核心:【北大核心2017】；CSCD:【CSCD_E2019_2020】；

基金：上海市教育委员会和上海市教育发展基金会“曙光计划”项目(15SG28);高等学校学科创新引智计划项目(B18022)。

语种：中文

中文关键词：β-N-乙酰氨基葡萄糖苷酶;解淀粉芽孢杆菌;克隆;酶学性质;糖苷水解酶3家族

外文关键词：β-N-acetylglucosaminidase;Bacillus amyloliquefaciens;cloning;characterization;glycoside hydrolase family 3

摘要：从解淀粉芽孢杆菌(Bacillus amyloliquefaciens YX-01)中克隆得到一个新型β-N-乙酰氨基葡萄糖苷酶基因(BaNagase),并成功在大肠杆菌中异源表达。BaNagase编码616个氨基酸,与B.subtilis来源的β-N-乙酰氨基葡萄糖苷酶(AIY91451.1)氨基酸序列相似性最高,为76.97%。通过对比分析,BaNagase属于糖苷水解酶3家族。纯化获得的BaNagase比活力为16.42 U/mg,最适温度65℃,最适pH 6.0,且在低于55℃条件下能保持高于70%的酶活力。BaNagase展现出高的热稳定性和严格的底物特异性,为其高效制备N-乙酰氨基葡萄糖提供了理论支持。
In this study,a novelβ-N-acetylglucosaminidase gene was cloned from Bacillus amyloliquefaciens YX-01 and successfully expressed in Escherichia coli BL21.The recombinant protein(defined as BaNagase)was purified by Ni-IDA affinity chromatography and analyzed.The results showed that it encoded 616 amino acids,with the highest similarity(76.97%)to the sequence ofβ-N-acetylglucosidase(AIY91451.1)from B.subtilis.According to the phylogenetic analysis and multiple sequence alignments,BaNagase belonged to glycoside hydrolase(GH)family 3.The recombinant BaNagase showed specific activity of 16.42 U/mg toward the substrate pNP-GlcNAc.Its optimal pH value was 6.0 and the optimal reaction temperature was 65℃.BaNagase showed good stability below 55℃at which more than 70%of initial activity was still retained.BaNagase exhibited strong thermostability and strict substrate specificity.This study provides a theoretical basis for efficient preparation of N-acetylglucosamine.