基于PTS缺陷型大肠杆菌构建莽草酸生产菌  

Construction of shikimic acid-producing engineered Escherichia coli strains based on ptsHIcrr mutants

作　　者：邹永康[1,2] 周军智[1,2,3] 孙旭[2,4] 蔡亚非[1] 戴红梅[2] 李树龙[2] 周长林[4] 方宏清[2] 

机构地区：[1]安徽师范大学生命科学学院,安徽芜湖241000 [2]军事医学科学院生物工程研究所,北京100071 [3]北京师范大学生命科学学院,北京100875 [4]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《微生物学通报》2011年第8期1186-1192,共7页Microbiology China

基　　金：国家自然科学基金项目(No.30780049);国家973计划项目(No.2011CBA00801)

摘　　要：对大肠杆菌芳香族氨基酸合成途径进行代谢流改造,以实现高效的生物制备莽草酸。以磷酸烯醇式丙酮酸-糖磷酸转移酶系统(PTS系统)敲除菌DH5α△ptsHIcrr(DHP)为基础,特异性敲除aroL、ydiB基因并转入受阿拉伯糖诱导表达的T7-RNA聚合酶基因,最终构建一系列产莽草酸宿主菌。再将aroE、aroB、tktA、glk、aroFfbr组成的系列基因串联起来置于质粒上,在T7启动子控制下表达,经摇瓶培养检测得知,不同重组菌产莽草酸能力与对照相比均有明显提高,其中DHPYA-T7/pAOC-TGEFB菌株产量最高,可达到392 mg/L。为进一步构建高表达莽草酸工程菌奠定基础。Metabolic engineering for aromatic amino acid biosynthesis pathway in Escherichia coli to acquire high-level biosynthesis of shikimic acid was reported. Knockout of aroL, ydiB genes and knock-in of T7-RNA-Polymerase gene which expression was controlled by L-arabinose based on the initial strain DHSα△ptsHIcrr （DHP）, resulted in a series of shikimic acid-producing host strains. A series of tandem genes consisting of aroE, aroB, tktA, glk or aroF^fbr were controlled by T7 promoter on plasmids were transformed into these host strains. According to the concentration of shikimic acid in shake-flask culture, all the engineered strains displayed high-potentiality compared to the control strain DHP, and the strain DHPYA-T7/pAOC-TGEFB synthesized the highest yield of 392 mg/L of shikimic acid. This study layed a strong foundation for constructing a high-level shikimic acid-producing engineered strain.

关 键 词：ptsHIcrr操纵子 莽草酸 aroL ydiB 

分 类 号：TQ921.7[轻工技术与工程—发酵工程]