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
曲亚通 1苏宇波 2王悦 1高钢 1赵浩含 1陈佳 1王晓飞 1马智峰 3朱爱国 1陈继康1
作者信息
- 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
- 折叠
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pH8～10 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.
Abstract
The preparation of industrial hemp seed polypeptide by enzymatic hydrolysis has the ad-vantages 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 intes-tinal 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 activi-ty 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 pep-tides were obtained by enzymolysis of the defatted hemp protein with the whole cell catalyst of recombi-nant yeast displaying NX1101,among which 140 short peptides with amino acid number≤10 were ob-tained.The enzymolysis efficiency of whole cell catalyst was more than 6.5 times higher than that of con-ventional 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Key words
hemp peptide/proteolytic enzyme/enzymatic property/whole cell catalyst/surface display引用本文复制引用
基金项目
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(22CX8NE213)
出版年
2024
国家科技期刊平台
NSTL
万方数据