Construction method and application of squalene synthesis pathway in bacillus subtilis

• Main Authors: SONG HAO, WANG QINGMEI, LI FENG, SUN PANXING, ZHANG JUNQI, XIAO LIN, CHEN YARU
• Format: Patent
• Language: Chinese; English
• Published: 30.09.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a construction method and application of a squalene synthesis pathway in bacillus subtilis, a heterologous expression technology and a genetic engineering means are utilized, a key gene ERG9 for squalene synthesis is connected to a pCE2 plasmid, P43 is used as a promoter to obtain a recombinant plasmid, the recombinant plasmid is amplified and then introduced into bacillus subtilis 168, and the squalene synthesis pathway in the bacillus subtilis is constructed. The bacillus subtilis engineering strain 168-ERG9 capable of synthesizing squalene is formed, a squalene synthesis pathway in the bacillus subtilis is constructed, and the MK-7 synthesis performance of the modified strain is improved. 本发明公开了一种枯草芽孢杆菌中角鲨烯合成通路的构建方法与应用，利用异源表达技术和基因工程手段，将角鲨烯合成的关键基因ERG9连接到pCE2质粒上，以P43为启动子，获得重组质粒，重组质粒扩增后导入枯草芽孢杆菌Bacillus subtilis 168中，形成可以合成角鲨烯的枯草芽孢杆菌工程菌株168-ERG9，构建枯草芽孢杆菌中角鲨烯合成通路，并且该改造菌株在MK-7的合成性能上得到了提升。