通过失活Sec途径阻遏蛋白和胞外蛋白酶提高地衣芽孢杆菌产碱性蛋白酶的能力  

Improving the Ability of Bacillus licheniformis to Produce Alkaline Protease by Inactivating Sec Pathway Repressor Protein and Extracellular Proteases

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作  者:郝漫 惠威 邵岚莹 史超硕 路福平[1] 张会图[1] HAO Man;HUI Wei;SHAO Lanying;SHI Chaoshuo;LU Fuping;ZHANG Huitu(Laboratory of Applied Microbiology and Enzyme Engineering,College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区:[1]天津科技大学生物工程学院应用微生物与酶工程实验室,天津300457

出  处:《中国生物工程杂志》2024年第2期39-47,共9页China Biotechnology

基  金:绿色生物制造国家重点研发专项(2021YFC2100400)资助项目。

摘  要:为了探究Sec分泌途径对地衣芽孢杆菌(Bacillus licheniformis)碱性蛋白酶产量的影响,对地衣芽孢杆菌TCCC11470(BLΔuppΔepsΔpgs)中的分子伴侣阻遏蛋白基因hrcA和基因组中3个Sec途径分泌的胞外蛋白酶基因epr、bpr和vpr进行叠加敲除。通过对比分析基因缺失前后的碱性蛋白酶酶活力发现,敲除菌株TCCC11470ΔhrcA和TCCC11470ΔhrcAΔeprΔbprΔvpr在42 h的碱性蛋白酶酶活力分别达到18521.2 U/mL和20048.5 U/mL,分别高出对照菌株BLΔuppΔepsΔpgs(14478.6 U/mL)27.9%和38.5%。这一结果指出,通过改进Sec分泌通路可以显著提升碱性蛋白酶的催化效能,为构建优化的工业酶生产宿主提供了新思路和研究方向。In order to investigate the influence of the Sec secretion pathway on alkaline protease production in Bacillus licheniformis,the molecular chaperone-blocking protein gene hrcA and the three extracellular protease genes epr,bpr,and vpr in the genome of B.licheniformis TCCC11470(BLΔuppΔepsΔpgs)were sequentially deleted.By comparing the alkaline protease activities before and after gene deletion,it was found that the knockout strains TCCC11470ΔhrcA and TCCC11470ΔhrcAΔeprΔbprΔvpr reached alkaline protease activities of 18521.2 U/mL and 20048.5 U/mL after 42 hours,respectively,which were 27.9%and 38.5%higher than the control strain BLΔuppΔepsΔpgs(14478.6 U/mL),respectively.These results indicate that optimizing the Sec secretion pathway can effectively enhance the enzymatic activity of alkaline protease,providing new insights and research strategies for the construction of industrial enzyme production hosts.

关 键 词:地衣芽孢杆菌 Sec分泌途径 碱性蛋白酶 HRCA 胞外蛋白酶 

分 类 号:Q814[生物学—生物工程]

 

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