高产L-5-甲基四氢叶酸基因工程菌的构建与表达  

Construction of Recombinant Strains Capable of Producing L-5-Methyltetrahydrofolic Acid

作　　者：刘琳[1] 许激扬[1] 卞筱泓[1] 于丽[1] 刘娅梅[1] 

机构地区：[1]中国药科大学生命科学与技术学院生物化学教研室,江苏南京210009

出　　处：《药物生物技术》2011年第3期201-205,共5页Pharmaceutical Biotechnology

摘　　要：L-5-甲基四氢叶酸是叶酸最具生物活性的形式,也是唯一可以穿透血脑屏障的叶酸类药物,有极大的应用价值。为提高菌体内L-5-甲基四氢叶酸的含量,将5,10-亚甲基四氢叶酸还原酶基因metF插入载体质粒,并将重组质粒pET-22b-metF转化至E.coliBL21(DE3),乳糖诱导蛋白表达。成功构建的重组菌经诱导后,L-5-甲基四氢叶酸量比原始菌株中提高28%,达到66μg/g菌体。本研究成功的为生物合成L-5-甲基四氢叶酸寻找到一种新的途径。L-5-methyltetrahydrofolic acid is the most active form of folic acid and is also the only form of folic acid which is able to cross the blood-brain barrier,so it has a great application value.To construct a recombinant strains capable of producing L-5-methyltetrahydrofolic acid,the gene of 5,10-methylenetetrahydrofolate reductase（metF） was cloned into vector pET-22b,and then the recombinant vector pET-22b-metF was transformed into E.coli BL21（DE3）.The recombinant protein was induced by lactose,and finally the amount of L-5-methyltetrahydrofolic acid was improved.After the recombinant strain was induced by lactose,the amount of L-5-methyltetrahydrofolic acid of recombinant strain was increased by 28%,and the yield of L-5-methyltetrahydrofolic acid was up to 66μg/g cell.These results found a new way for the bio-synthesis of L-5-methyltetrahydrofolic acid.

关 键 词：L-5-甲基四氢叶酸 5 10-亚甲基四氢叶酸还原酶 克隆 表达 

分 类 号：Q78[生物学—分子生物学]