代谢工程改造酿酒酵母合成葡萄糖二酸  

Metabolic engineering of Saccharomyces cerevisiae for production of glucaric acid

作　　者：巩旭[1,2] 刘叶[1,2] 王毳[1,2] 李江华[1,2] 康振[1,2] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《生物工程学报》2017年第2期228-236,共9页Chinese Journal of Biotechnology

基　　金：江苏省科技支撑计划项目(No.BE2014607);长江学者和创新团队发展计划(No.IRT_15R26);江苏省自然科学基金(No.BK20141107)~~

摘　　要：葡萄糖二酸是一种高附加值的有机酸,广泛用于食品、医药和化工领域。为获得生产葡萄糖二酸的微生物细胞工厂,通过共表达小鼠来源的肌醇加氧酶(MIOX)及恶臭假单胞菌来源的醛酸脱氢酶(Udh),在酿酒酵母Saccharomyces cerevisiae CEN.PK2-1C中构建了葡萄糖二酸合成途径,产量为(28.28±3.15)mg/L。在此基础上,通过调控前体肌醇的合成途径,发现肌醇-1-磷酸合成酶(INO1)是葡萄糖二酸合成途径的限速酶,过量表达INO1,葡萄糖二酸产量达到(107.51±10.87)mg/L,提高了2.8倍。进一步弱化竞争支路中磷酸果糖激酶(PFK1)的表达,最终葡萄糖二酸的产量达到(230.22±10.75)mg/L,为进一步获得高产葡萄糖二酸细胞工厂提供基础。Glucaric acid, a high value-added organic acid, is widely used in food, pharmaceutical and chemical industries. For microbial production of glucaric acid in Saccharomyces cerevisiae, we constructed a synthetic glucaric acid biosynthetic pathway by coexpressing the genes encoding myo-inositol oxygenase from mice and uronate dehydrogenase from Pseudomonas putida. Moreover, myo-inositol-1-phosphate synthase was identified as a rate-limiting enzyme in glucaric acid pathway and was upregulated, resulting in the production of glucaric acid of（107.51± 10.87） mg/L, a 2.8-fold increase compared to the parent strain. Then, by repressing the activity of phosphofructokinase, the concentration of glucaric acid further increased to（230.22± 10.75） mg/L. The strategy could be further used to construct cell factories for glucaric acid production.

关 键 词：葡萄糖二酸 肌醇-1-磷酸合成酶 磷酸果糖激酶 酿酒酵母 代谢工程 

分 类 号：Q936[生物学—微生物学]