来源海洋细菌的产微球茎菌属ALW1降解海藻酸钠生成单糖的外切型褐藻胶裂解酶Alg1815的克隆表达

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• 英文论文题名：Cloning,overexpression and depolymerization of alginate into a monomeric sugar acidusing Alg1815,an exo-alginate lyase from a marine bacterium,microbulbifer sp.ALW1
• 论文作者：王新侠 ; 朱艳冰 ; 倪辉
• 英文论文作者：Wang Xinxia ; Zhu Yanbing ; Ni Hui ; College of Food and Biology Engineering ; Jimei University ; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering Technology ; ResearchCenter of Food Biotechnology of Xiamen City
• 年：2016
• 作者机构：集美大学生物工程学院;福建省高校食品微生物与酶工程技术研究中心;厦门市食品与生物工程技术研究中心;
• 论文关键词：克隆 ; DEH ; 重组褐藻胶裂解酶A1GL1815 ; 外切型褐藻胶裂解酶 ; 产微球茎菌属ALW1
• 英文论文关键词：cloning ; 4-deoxy-L-erythro-5-hexo seulose uronic acidrecombinant alginate lyase A1GL1815 ; exo-type alginate lyase ; microbulbifer sp.ALW1
• 会议召开时间：2016-11-09
• 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
• 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
• 语种：中文
• 分类号：Q943.2
• 学会代码：ZGSP
• 会议名称：中国食品科学技术学会第十三届年会
• 会议地点：中国北京
• 主办单位：中国食品科学技术学会
• 学会名称：中国食品科学技术学会
• 页数：2
• 文件大小：104k
• 原文格式：O
• 会议级别：全国

摘要

藻类作为第三代能源在食品和化工业有广泛用途,褐藻胶作为褐藻中主要物质被褐藻胶裂解酶降解生成糖醛酸单体如古罗糖醛酸和甘露糖醛酸,是利用褐藻糖类的关键步骤。研究褐藻胶裂解酶高酶活性和高表达,利用PCR技术,克隆一个来源Microbulbifer sp的褐藻胶裂解酶的基因(定义为Alg1815),在大肠杆菌中表达。基因大小为2220 bp,编码739氨基酸包括一个假定的29个氨基酸残基的信号肽,属于多糖裂合酶(PL)17家族。该酶的最温度35℃,最适pH是8.0的50 mM磷酸缓冲液,根据TLC和LC-MS结果显示Alg1815能降解海藻酸钠生成海藻酸钠单体-(4-deoxy-L-erythro-5-hexoseulose uronic acid DEH),为外切型褐藻胶裂解酶。重组海藻酸裂解酶A1GL1815高表达和高酶活性可能是一个潜在的研究褐藻胶糖化酶和生物燃料生产。
The algae are considered as third-generation energy are great use in food and chemical industry,and alginate is the main component of brown macroalgae.To utilize brownmacroalgae as biomass,the degradation of alginate,which isthe main carbohydrate of brown macroalgae,into uronate monomers,such as mannuronicacid and guluronic acid,is a critical prerequisite step.Is to report an alginate lyase with high enzymatic activity and high-level expression.Using degenerate PCR,a gene(designated Algl815) encoding a new alginate lyase was cloned from Microbulbifer sp.ALW1 and expressed in Escherichia coli.The gene consisted of 2,220 bp with deduced amino acid size of 739 including a putative signal peptide of 29 amino acid residues belonging to polysaccharide lyase(PL) family 17.The recombinant protein was most active at 35℃ and pH 8.0 in 50 mM phosphate buffer.The recombinant Alg1815 was found to preferentially act on alginate produce the alginate monomer,4-deoxy-L-erythro-5-hexoseulose uronic acid by TLC and LC-MS.With the high-level expression and high enzymatic activity,the recombinant alginate lyase Alg1815 could be a potential enzyme for further research on alginate saccharification and biofuels production.

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