杨捷1, , ,钱晋1,叶江1,吴海珍1,张惠展1, , 1.
华东理工大学生物反应器工程国家重点实验室
通讯作者: 杨捷, yangyang22222@hotmail.com ; 张惠展, huizhzh@ecust.edu.cn
Genetic Manipulation of the Genes Related with Shikimate metabolism of Escherichia coli
Corresponding author: , yangyang22222@hotmail.com ;, huizhzh@ecust.edu.cn
摘要
摘要: 为了调查大肠杆菌中相关基因的遗传操作对莽草酸途径和莽草酸积累的影响，利用温度敏感型质粒敲除了大肠杆菌JM83染色体DNA上编码莽草酸激酶的aroL基因，获得了该基因缺陷株JDL02；同时从大肠杆菌JM83染色体中分别克隆了aroG、ppsA和tktA 3个基因，构建了多个表达质粒导入JDL02，得到了一系列重组菌。摇瓶发酵显示aroG基因的作用最为明显，各重组菌莽草酸产量均有不同程度的提高；在等生物量情况下，JDL02/pTrc-aroG-tktA的莽草酸产量是JM83的18.35倍，其摇瓶产量为94.33 mg/L。
关键词:
莽草酸； 基因敲除； 强化表达； 发酵
Abstract: In order to investigate how the genetic manipulation with related genes in Escherichia coli impacts the shikimate metabolism and the accumulation of shikimic acid, shikimate kinase II gene (aroL) of Escherichia coli JM83 was disrupted using temperature sensitive plasmid and the mutant was named as JDL02. Meanwhile, ppsA, tktA and aroG gene were cloned from Escherichia coli JM83. A series of expression plasmids were constructed and transferred into JDL02. Flask fermentation verified that the yields of all the recombinant strains are improved. In the same bacterial density condition, the shikimic acid yield of JDL02/pTrc-aroG-tktA strain was 18.35 times of the control JM83, and the flask fermentation yield is 94.33 mg/L.
Key words:
shikimic acid
