饲料研究2024，Vol.47Issue(14)：100-106,7.DOI:10.13557/j.cnki.issn1002-2813.2024.14.017
蛋白酶枯草芽孢杆菌工程菌株WB800N/pHT43-npr-PrsA的发酵工艺优化
Optimization of fermentation process for Bacillus subtilis WB800N/pHT43-npr-PrsA recombinant strain with protease activity
摘要
Abstract
The experiment aimed to optimize the fermentation protease activity of the engineered Bacillus subtilis strain WB800N/pHT43-npr-PrsA,providing technical support for domestic feed protease enterprises.The fermentation process was monitored for glucose content and specific consumption rate to determine the dynamic impact of carbon and nitrogen sources on the glucose consumption of this strain.The components of the fermentation medium were optimized through single-factor and response surface experiments.The results showed that optimizing the fermentation medium with the OD600nm value of the cell body from 0 to 10 hours as a parameter can promote cell growth and achieve a higher cell quantity.After 10 hours,supplementing with 5 g/L yeast extract and 70 g/L peptone could extend the stationary phase to achieve the maximum specific enzyme activity.The optimized fermentation medium conditions were:60 g/L yeast extract,30 g/L peptone,10 g/L racemic glucose,and 3 g/L MgSO4.At 8 h of fermentation,adding 0.5‰ IPTG and 2‰ protease inhibitor,the final fermentation protease activity reached 1 156 U/mL.The study indicates that delayed stable period can increase the peptone content in the medium,which is beneficial for improving the specific glucose consumption rate of the cells.Peptone promotes the adaptation of the cells to the surrounding environment.Maintaining a low glucose concentration during the fermentation process can effectively avoid the formation of acetic acid,promote cell growth,and enhance enzyme production efficiency,keeping the cell consumption of the substrate at its most vigorous state.关键词
蛋白酶/工程菌株/酶活性/发酵工艺Key words
protease/engineering strains/enzyme activity/fermentation process分类
农业科技引用本文复制引用
曹倩荣,刘春卯,郑翔,贾振华,吴芳彤..蛋白酶枯草芽孢杆菌工程菌株WB800N/pHT43-npr-PrsA的发酵工艺优化[J].饲料研究,2024,47(14):100-106,7.基金项目
河北省科学院科技计划项目(项目编号:23315) （项目编号:23315）
