黄杆菌中浒苔多糖降解有关基因的异源表达及酶活测定  

Heterologous expression and enzyme activity assays of gene related to the degradation of enteromorpha polysaccharides in Flavobacteria

作　　者：徐展 刘芳芳 徐玉雯 闫达中[1] 晁红军 陈静[1] XU Zhan;LIU Fangfang;XU Yuwen;YAN Dazhong;CHAO Hongjun;CHEN Jing(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China)

机构地区：[1]武汉轻工大学,生命科学与技术学院,武汉430023

出　　处：《武汉轻工大学学报》2024年第3期59-65,共7页Journal of Wuhan Polytechnic University

基　　金：国家自然科学基金青年基金项目(编号:31901081),中国博士后科学基金项目(编号:2019M662746).

摘　　要：浒苔绿潮爆发带来了一系列环境问题,也对浒苔资源的开发利用提出了新挑战。从生物酶法降解浒苔的主要成分浒苔多糖入手,筛选出能降解浒苔多糖的菌株,对其进行16S rRNA基因鉴定,同时对其中表达量上调的木聚糖酶基因(GH43)进行异源表达,利用紫外-可见分光光度计比色法进行酶活测定及酶学性质研究。结果表明筛选到的浒苔多糖降解菌为黄杆菌(Algibacter pacificus),克隆表达的木聚糖酶的最适温度为50℃,最适pH约为5,粗酶液酶活为65.71 U/mL。实验可为由浒苔引起的海洋污染治理问题提供理论依据,同时为该菌株及木聚糖酶的工业化利用提供了参考,有利于实现对浒苔的资源化利用。The outbreak of the green tide of Enteromorpha prolifera has brought a series of environmental problems and also posed new challenges to the development and utilization of Enteromorpha prolifera resources.Polysaccharides from Enteromorpha prolifera are the main components of Enteromorpha prolifera.Starting with the enzymatic degradation of Enteromorpha prolifera polysaccharides,strains capable of degrading Enteromorpha prolifera polysaccharides were screened,and their 16S rRNA genes were identified.At the same time,the upregulated xylanase gene(GH43)was heterologously expressed.Enzyme activity and enzymatic properties were determined using UV visible spectrophotometer colorimetry.The results showed that the screened Enteromorpha prolifera polysaccharide degrading bacteria were Algibacter pacificus.The optimal temperature for cloning and expressing xylanase was 50℃,the optimal pH was about 5,and the crude enzyme activity was 65.71 U/mL.The experiment can provide a theoretical basis for the treatment of marine pollution caused by Enteromorpha prolifera,and provide a reference for the industrial utilization of this strain and xylanase,which is conducive to realizing the resource utilization of Enteromorpha prolifera.

关 键 词：浒苔多糖 黄杆菌 多糖降解 木聚糖酶 

分 类 号：Q14[生物学—生态学]