β-胡萝卜合成途径的不同启动子组合的比较研究  

Comparison of promoters' efficiency in β-carotene biosynthetic pathway

作　　者：童阳阳 高书良[2] 蒋宇[3] 杨晟[3] 戈梅[4] 陈代杰[5] 

机构地区：[1]上海师范大学生命与环境科学学院,上海200234 [2]华东理工大学生物工程学院,上海200237 [3]中国科学院上海生命科学研究院,上海200031 [4]上海来益生物药物研究开发中心,上海201203 [5]上海医药工业研究院,上海200040

出　　处：《工业微生物》2017年第3期9-13,共5页Industrial Microbiology

摘　　要：以耶氏解脂酵母菌(Yarrowialipolytica)MYA2613为底盘菌,表达来源于卷枝毛霉菌(Mucorcircinelloides)的基因car B,carRP后能有效生产β-胡萝卜素。启动子作为调控基因的重要顺式元件影响着基因的表达。为了优化基因GGS1,car B,carRP的表达强度组合,将上述3个基因、终止子与两组不同的启动子通过DNA assemble方法拼接后,得到了组合TEF1p-GGS1-xpr2t-GPD1pcar B-mig1t-EXP1p-carRP-lip2t和YAT1p-GGS1-xpr2t-GPD1p-car B-mig1t-FBAin1p-carRP-lip2t。经发酵表型验证,两株菌都产β-胡萝卜素,其中前者的产量是后者的4.18倍,具有明显优势。Yarrowia lipolytica MYA2613 was used as a host to efficiently produce B-carotene with expression of exogenetic genescarB and carRP from Mucorcircinelloides. Pathway could be optimized through dynamic regulation of genes expression. Here, two sets of promoters which control three genes of B-carotene biosynthetic pathway, TGE, TEF1p- GGSI-xpr2t-GPD1 p-carB-migl t-EXP1 p-carRP-lip2t and YGF, YAT1 p-GGS1-xpr2t-GPD1 p-carB-migl t-FBAinl p-carRP- lip2t. Strains carrying each of these two pathways were investigated through fermentation. Results indicated that TGE was a better combination of promoters to express β-carotene pathway. The production of β-carotene by TGE strains was 4.1- fold of that by YGF strains.

关 键 词：Β-胡萝卜素 DNA ASSEMBLE 耶氏解脂酵母 启动子组合 

分 类 号：Q78[生物学—分子生物学]