Wenyingzhuangia属海洋细菌产β-紫菜多糖酶的克隆表达及性质研究  

Studies on Cloning Expression and Characterization ofβ-Porphyranase from Marine Bacterium Wenyingzhuangia aestuarii

作　　者：冯瑞方 张玉莹 申晶晶[1] 常耀光[1,2] Feng Ruifang;Zhang Yuying;Shen Jingjing;Chang Yaoguang(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,Shandong;Laboratory for Marine Drugs and Bioproducts,Pilot National Laboratory for Marine Science and Technology,Qingdao 266237,Shandong)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]青岛海洋科学与技术试点国家实验室,海洋药物与生物制品功能实验室,山东青岛266237

出　　处：《中国食品学报》2022年第2期98-106,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31671883)。

摘　　要：从海洋细菌Wenyingzhuangia aestuarii OF219中挖掘到一种新颖的β-紫菜多糖酶编码基因por16Z,对其进行生物信息学分析。利用分子生物学技术进行异源表达,研究其生化性质,采用高效液相色谱-质谱联用技术对降解产物进行解析。生化性质研究表明:β-紫菜多糖酶Por16Z_Wa的最适反应温度为45℃,最适反应pH 6.5,在pH 4.0~9.0范围具有较好的稳定性,且该酶具有适冷性。作用方式结果表明:Por16Z_Wa为内切酶,其降解终产物主要由紫菜二糖构成,含有少量的四糖及六糖,且它们均为被修饰或取代后的紫菜寡糖。Por16Z_Wa作为紫菜多糖的新型工具酶,能够高效、定向制备紫菜二糖,对紫菜多糖的应用研究具有推动作用。A novel GH16β-porphyranase gene por16Z was cloned from marine bacterium Wenyingzhuangia aestuarii OF219 and expressed in E.coli,and the biochemical properties and hydrolytic patterns of the recombinant protein Por16Z_Wa were characterized.Por16Z_Wa showed the maximum activity at 45℃and pH 6.5 and keep stable at a wide pH range from 4.0 to 9.0.Por16Z_Wa exhibited a cold-adapted property.Por16Z_Wa was an endo-acting enzyme.The final products of Por16Z_Wa were majorly composed of porphyran disaccharides L6S-G,with a minor portion of tetrasaccharides and hexasaccharides which were substituted by agarose moieties and(or)modified by methyl groups.Por16Z_Wa could be utilized as a promising tool for degradation of porphyran and directional preparation of porphyran disaccharides,which would facilitate the investigation of porphyran polysaccharides.

关 键 词：紫菜多糖 GH16家族 β-紫菜多糖酶 紫菜寡糖 克隆表达 

分 类 号：S917.3[农业科学—水产科学]