代谢工程改造枯草芽孢杆菌高效生产MK-7  

Efficient Production of MK-7 by Metabolic Engineering in Bacillus subtilis

作　　者：王永利 付刚[4] 张子梦 林心萍[1,2] 张大伟[4] WANG Yongli;FU Gang;ZHANG Zimeng;LIN Xinping;ZHANG Dawei(School of Food Science and Technology,Dalian Polytechnic University,Dalian 116034,China;National Engineering Research Center of Seafood,Dalian Polytechnic University,Dalian 116034,China;School of Biological Engineering,Dalian Polytechnic University,Dalian 116034,China;Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China)

机构地区：[1]大连工业大学食品学院,辽宁大连116034 [2]大连工业大学国家海洋食品工程技术研究中心,辽宁大连116034 [3]大连工业大学生物工程学院,辽宁大连116034 [4]中国科学院天津工业生物技术研究所,天津300308

出　　处：《食品与生物技术学报》2022年第8期86-94,共9页Journal of Food Science and Biotechnology

基　　金：天津市合成生物技术创新能力提升行动项目(TSBICIP-CXRC-055,TSBICIP-CXRC-004)。

摘　　要：七烯甲萘醌(MK-7)作为一种重要的K族维生素,在保健食品和医药制造领域具有良好的应用前景。为了得到一株高产MK-7菌株,作者以Bacillus subtilis 168为出发菌株,分别进行了代谢分支途径基因敲除、MK-7合成途径关键基因menA的突变体筛选优化、人工操纵子强化部分途径基因的表达来提升MK-7发酵产量。最终构建得到高产菌株MK7-13,发酵产MK-7达到(73.73±1.86)mg/L,与对照菌株相比提高了9.71倍,为后续微生物发酵法工业化生产MK-7奠定了基础。Menaquinone-7(MK-7)has a promising application prospect in the fields of health food and pharmaceutical manufacturing as an important vitamin of the K family.In this work,the yield of MK-7 fermentation was improved by using Bacillus subtilis 168 as the starting strain,knocking out the genes from metabolic branch pathways,screening and optimizing the mutant of speed-limited gene menA of the MK-7 synthesis pathway,and enhancing the expression of several MK-7 biosynthesis pathway genes through an artificial operon.Finally,a high-yield strain MK7-13 was developed with the yield of(73.73±1.86)mg/L,which was 9.71 times higher than that of the original strain.In summary,a high-yield MK-7 strain was successfully constructed by a combinatorial metabolic engineering strategy,which laid a foundation for the subsequent industrial production of MK-7 by microbial fermentation.

关 键 词：七烯甲萘醌 枯草芽孢杆菌 人工操纵子 点突变 

分 类 号：Q81[生物学—生物工程]