钝齿棒杆菌丙酮酸激酶的克隆表达及其对精氨酸合成的扰动影响  

Cloning,Expression of the Pyruvate Kinase Gene from Corynebacterium crenatum and Its Effect on L-Arginine Synthesis

作　　者：卢妍[1] 饶志明[1] 徐美娟[1] 满在伟[1] 张显[1] 耿燕[2] 许正宏[2] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,无锡214122 [2]江南大学药学院制药工程实验室,无锡214122

出　　处：《中国生物工程杂志》2014年第3期48-55,共8页China Biotechnology

基　　金：国家"973"计划(2012CB725202);国家"863"计划(2012AA022102);国家自然科学基金(21276110;31300028);教育部新世纪优秀人才计划(NCET-10-0459);江苏省青年自然科(学基金(BK20130137);中央高校基本科研业务费专项资金(JUSRP51306A;JUSRP1009)资助项目

摘　　要：钝齿棒杆菌(Corynebacterium crenatum)SYPA 5-5是诱变选育的一株高产精氨酸菌株。丙酮酸激酶(pyruvate kinase)是EMP途径的限速酶,对胞内能量产生和代谢流调节方面都有重要作用。为提高葡萄糖消耗速率以缩短发酵周期,以钝齿棒杆菌基因组为模板,克隆得到pyk基因并将其在C.crenatum SYPA 5-5中成功表达,葡萄糖平均消耗速率为1.71 g/L/h较C.crenatum SYPA 5-5提高33.5%,精氨酸产量和最大产率分别为43.32 g/L和0.55 g/L/h较C.crenatum SYPA 5-5分别提高20%和19.5%。结果表明,加强表达丙酮酸激酶能增强EMP途径,提高精氨酸产量。In order to increase glocuse consumption rate and shorten fermentation time, pyruvate kinase（ EC 2.7.1.40, PYK）gene from L-arginine producing mutant Corynebacterium crenatum SYPA 5-5 was expressed in Corynebacterium crenatum SYPA 5-5. The results of fermentive character showed that compared with wild-type strain, the average glocuse consumption rate was improved 33.5% to 1.71 g/L/h, the L-arginine concentration was improved 20% to 43.32 g/L, and the maximum yield of L-Arg was improved 19.5% to 0.55 g/L/h Overexpression of pyruvate kinase could increased the glocuse consumption and improved the yield of L-arginine

关 键 词：L-精氨酸钝齿棒杆菌 丙酮酸激酶 

分 类 号：Q78[生物学—分子生物学]