酿酒酵母表面展示NX1101蛋白酶全细胞催化剂制备火麻多肽  

Preparation of Industrial Hemp Seed Polypeptide by a Whole-cell Biocatalyst Displayed NX1101 Protease on the Surface of Saccharomyces cerevisiae

作　　者：曲亚通 苏宇波 王悦 高钢[1] 赵浩含 陈佳[1] 王晓飞[1] 马智峰 朱爱国[1] 陈继康[1] QU Yatong;SU Yubo;WANG Yue;GAO Gang;ZHAO Haohan;CHEN Jia;WANG Xiaofei;MA Zhifeng;ZHU Aiguo;CHEN Jikang(Institute of Bast Fiber Crops,Chinese Academy of Agricultural Sciences/Key Laboratory of Biological and Processing for Bast Fiber Crops,MARA,Changsha 410221,Hunan,China;Linchuan County Qingliang Taihang Agricultural Products Development Company,Jincheng 048300,Shanxi,China;Economic Crop Station of Zhangjiachuan Hui Autonomous County,Tianshui 741599,Gansu,China)

机构地区：[1]中国农业科学院麻类研究所/农业农村部麻类生物学与加工重点实验室,湖南长沙410221 [2]陵川县清凉太行农产品开发有限公司,山西晋城048300 [3]张家川回族自治县经济作物工作指导站,甘肃天水741599

出　　处：《中国麻业科学》2024年第3期169-174,共6页Plant Fiber Sciences in China

基　　金：财政部和农业农村部:国家麻类产业技术体系建设专项(CARS-16);甘肃省技术创新引导计划“张家川工业大麻提质增效关键技术研发与集成示范”(22CX8NE213)。

摘　　要：酶解法火麻肽制备具有安全性好、能耗低、营养损失小等优点。研究在综合脱脂火麻仁籽粕蛋白提取以及蛋白水解酶筛选的基础上,以穿梭质粒pYD1为表达载体,将源于牛肠道宏基因组文库中的蛋白水解酶NX1101锚定在酿酒酵母菌株(EYB100)表面,并以此重组菌株作为全细胞催化剂制备火麻多肽。酶学性质分析发现:该全细胞催化剂的最适反应pH值为8.0,在pH 8~10酶活稳定性较高,为耐碱性全细胞催化剂;其最适反应温度50℃,在50℃以下时酶活稳定性较高。脱脂火麻蛋白经表面展示表达NX1101的重组酵母全细胞催化剂酶解后,共获得1095个多肽,其中氨基酸数量≤10的短肽数量达140个。全细胞催化剂酶解效率相较常规酶解技术(以碱性蛋白酶粉剂为对照)增加6.5倍以上。The preparation of industrial hemp seed polypeptide by enzymatic hydrolysis has the advantages of good safety,low energy consumption and little nutritional loss.In this study,based on the synthesis of defatted hemp seed meal protein extraction and proteolytic enzyme screening,the shuttle plasmid pYD1 was used as the expression vector to anchor proteolytic enzyme NX1101 from bovine intestinal metgenomic library on the surface of Saccharomyces cerevillus(EYB100),and the recombinant strain was used as the whole cell catalyst to prepare hemp polypeptides.The enzymatic properties analysis of the whole cell catalyst showed that the most suitable reaction pH value was 8.0,and the enzyme activity stability was high between 8 and 10,so it was an alkaline resistant protease.The optimum reaction temperature was 50℃,and the stability of enzyme activity was higher below 50℃.A total of 1095 peptides were obtained by enzymolysis of the defatted hemp protein with the whole cell catalyst of recombinant yeast displaying NX1101,among which 140 short peptides with amino acid number≤10 were obtained.The enzymolysis efficiency of whole cell catalyst was more than 6.5 times higher than that of conventional enzymolysis technology with alkaline protease powder.

关 键 词：火麻肽 蛋白水解酶 酶学性质 全细胞催化剂 表面展示 

分 类 号：S563.3[农业科学—作物学]