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

作　　者：付声亮 邹诗瑶 高娃 赵筱 王金华 王永泽[1,2,3] Fu Shengliang;Zou Shiyao;Gao Wa;Zhao Xiao;Wang Jinhua;Wang Yongze(Cooperative Innovation Center of Industrial Fermentation(Ministry of Education&Hubei Province),Wuhan 430068;Hubei Key Laboratory of Industrial Microbiology,Wuhan 430068;Hubei University of Technology,School of Bioengineering and Food Science,Wuhan 430068)

机构地区：[1]工业发酵省部共建协同创新中心,武汉430068 [2]湖北省工业微生物重点实验室,武汉430068 [3]湖北工业大学生物工程与食品学院,武汉430068

出　　处：《中国食品学报》2024年第7期219-228,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金青年科学基金项目(31501677)。

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。It is proposed to establish a synthetic route of pyruvate obtained from malic acid by whole cell biotransformation 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 concentration,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 conditions were as follows,the cell concentration of bacterial OD600nm value was 30,the mass concentration of substrate malic acid was 30 g/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 

分 类 号：TS201.2[轻工技术与工程—食品科学]