改造枯草芽孢杆菌底盘细胞高效生产乙偶姻
Modified Bacillus subtilis chassis cells for efficient production of acetoin
任可心 1陈慧骢 1张依凡 1王强 1张显1
作者信息
- 1. 江南大学生物工程学院,江南大学工业生物技术教育部重点实验室,江苏无锡 214122
- 折叠
摘要
以实验室前期保藏的经代谢工程改造的枯草芽孢杆菌(Bacillus subtilis)QRC-1 为出发菌株,其在 5 L发酵罐平均发酵生产乙偶姻的产量达到 50.7 g/L.然而,枯草芽孢杆菌存在的自溶与生长速率较慢的问题限制了其在发酵过程中的生物量增长,同时发酵产泡多的特点也影响了该菌在放大培养过程中的实时调控.因此本实验基于生理特性对枯草芽孢杆菌QRC-1 进行优化改造,利用分子遗传技术开发一种新的策略,达到提高生物量和乙偶姻产量的目标.最终获得的菌株 RKCR-8,比生长速率比原菌提高了 30%,生物量增加了1.36 倍,乙偶姻产量达到 72.3 g/L.使用本研究构建的重组枯草芽孢杆菌生产乙偶姻具有更强的生产效率和工业稳定性.这一结果不仅为乙偶姻的生产提供更多参考价值,也为未来工业微生物底盘的构建提供了一种新的思路.
Abstract
Starting from the metabolically engineered Bacillus subtilis strain QRC-1 preserved in the early stages of laboratory cultivation,its average yield of acetoin was verified to reach 50.7 g/L in a 5 L fermentation vessel.However,issues such as self-dissolution and slow growth rate limit the biomass of Bacillus subtilis,while the foaming characteristics during fermentation affect real-time control in high-density liquid cultivation.Therefore,this experiment is based on the physiological characteristics to optimize and modify the B.subtilis parental cells,using molecular genetic techniques to develop a new strategy aimed at preventing cell lysis,enhancing biomass,and increasing acetoin production.Ultimately,the obtained strain,RKCR-8,exhibited a 30%increase in growth rate compared to the original strain,with biomass increased by 1.36 times,and acetoin yield reaching 72.3 g/L.The recombinant B.subtilis constructed in this study has stronger production efficiency and industrial stability in producing acetoin.This result not only provides more reference value for the production of acetoin but also provides a new approach for the construction of future industrial microbial chassis.
关键词
微生物学/乙偶姻高效生产/分子遗传操作/枯草芽孢杆菌/高密度发酵/底盘细胞Key words
microbiology/efficient production of acetoin/molecular genetic manipulation/Bacillus subtilis/high-density fermentation/chassis cells引用本文复制引用
基金项目
江苏省研究生科研与实践创新计划(SJCX1241)
出版年
2024
NETL
NSTL
万方数据