重组大肠杆菌全细胞转化苹果酸合成丙酮酸
Biosynthesis of Pyruvate from Malic Acid by Whole-cell Biotransformation Using Recombinant Escherichia coli
付声亮 1邹诗瑶 1高娃 1赵筱 1王金华 1王永泽1
作者信息
- 1. 工业发酵省部共建协同创新中心 武汉 430068;湖北省工业微生物重点实验室 武汉 430068;湖北工业大学生物工程与食品学院 武汉 430068
- 折叠
摘要
拟建立一条以苹果酸为原料,通过大肠杆菌全细胞转化获得丙酮酸的合成途径.以大肠杆菌BL21(DE3)为宿主,在pET28a上共表达内源的苹果酸酶(ME)和来自博伊丁假丝酵母(Candida boidinii)的醛糖还原酶(AR),并对细胞浓度、底物浓度、温度和pH值等催化关键因素进行研究.共表达后大肠杆菌苹果酸酶和博伊丁假丝酵母醛糖还原酶酶活分别为(2.8±0.21),(3.1±0.34)U/mL.适宜的催化条件为:细胞浓度OD600nm值为30,底物苹果酸质量浓度为30 g/L,木糖和苹果酸物质的量比为1.0,温度为40℃,反应体系pH值为7.8,丙酮酸产量最高可达23.16 g/L.本研究为生物法合成丙酮酸提供了一种新的方法.
Abstract
It is proposed to establish a synthetic route of pyruvate obtained from malic acid by whole cell biotransforma-tion using Escherichia coli.With Escherichia coli BL21(DE3)as the host,endogenous malice enzyme(ME)and aldose reductase(AR)from Candida boidinii were co-overexpressed on pET28a,and catalytic key factors such as cell concen-tration,substrate concentration,temperature and pH were investigated.Activities of malic enzyme from Escherichia coli and aldose reductase from Candida boidinii were(2.8±0.21),(3.1±0.34)U/mL respectively.The suitable catalytic condi-tions were as follows,the cell concentration of bacterial OD600nm value was 30,the mass concentration of substrate malic acid was 30g/L,the molar ratio of xylose to malic acid was 1.0,temperature was 40 ℃,the pH value of the reaction system was 7.8,pyruvate yield was up to 23.16 g/L.This study provides a new method for the biological production of pyruvate.
关键词
丙酮酸/全细胞催化/苹果酸/苹果酸酶/醛糖还原酶Key words
pyruvate/whole cell catalysis/malic acid/malic enzyme/xylose reductase引用本文复制引用
基金项目
国家自然科学基金青年科学基金项目(31501677)
出版年
2024
NETL
NSTL
万方数据