组合代谢工程改造提高生物除草剂thaxtomin A的产量  

Combinatorial metabolic engineering modifications to improve the yield of the bioherbicide thaxtomin A

作　　者：王兵 徐冬梅 陈立霞 张林涛 李梦瑶 Wang Bing;Xu Dongmei;Chen Lixia;Zhang Lintao;Li Mengyao(Hebei Vocational University of Industry Technology,Shijiazhuang 050091)

机构地区：[1]河北工业职业技术大学,石家庄050091

出　　处：《中国抗生素杂志》2024年第12期1392-1398,共7页Chinese Journal of Antibiotics

基　　金：河北省教育厅高等学校自然科学研究重点项目(No.ZD2022084)。

摘　　要：以thaxtomin A产生菌酸性疮痂链霉菌S8为出发菌株,利用组合代谢工程的方法,通过增加S-腺苷甲硫氨酸还原酶MetK和途径特异性正调控因子TxtR的基因拷贝数,使用组成型kasOp*启动子和RBS核糖体结合位点,分别构建了txtR和metK单基因以及两基因共表达的重组质粒。通过接合转移的方法将质粒整合至菌株基因组,筛选获得阳性基因工程菌。对菌株发酵产物进行HPLC分析,结果显示菌株thaxtomin A的发酵单位比出发菌株分别提高了105%、38%和154%。组合表达调控元件和txtR和(或)metK基因的遗传改造可显著促进菌株thaxtomin A的生物合成。Streptomyces acidscabies S8,a thaxtomin A-producing strain,was used as the starting strain.By increasing the gene copy number of S-adenosine methionine reductase MetK and positive regulatory factor TxtR,using the constitutive promoter element kasOp*and ribosome binding site(RBS),respectively,the combinational metabolic engineering method was applied to construct the recombinant plasmides of txtR,metK and their co-expression.The plasmids were integrated into the genome of the strain by conjugation transfer,and the positive genetically engineered bacteria were screened.The fermentation products were analyzed by HPLC,and the results showed that the fermentation units of thaxtomin A were increased by 105%,38%and 154%,respectively.The combination of gene expression regulatory elements and the genetic modification of txtR and(or)metK could significantly improve the biosynthesis of thaxtomin A.

关 键 词：生物除草剂 Thaxtomin A 酸性疮痂链霉菌 txtR metK 

分 类 号：R9[医药卫生—药学]