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在Bacillus subtilis中β-葡萄糖酶的重组细胞外表达及其潜在的分泌机制

He Li¹, Wenjing Wang², Longwei Que²

¹School of Life Sciences, Anhui University, Hefei 230601, China; Anhui Key Laboratory of Biocatalysis and Modern Biomanufacturing, Hefei 230601, China; Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis, Hefei 230601, China; School of Environmental Engineering and Chemistry, Luoyang Institute of Science and Technology, Henan Province, Luoyang 471023, China.

International journal of biological macromolecules

|March 26, 2025

概括

研究人员在Bacillus subtilis中增强了β-葡萄糖酶分泌. 不经介导的Bgl2A分泌显示出更高的活性和稳定性,揭示了对酶表达至关重要的非经典分泌途径.

科学领域:

生物化学生物化学
分子生物学分子生物学
微生物生物技术微生物生物技术

背景情况:

β-葡萄糖酶 (Bgl2A) 是一种用于化β-1,4糖键的关键酶.
在Bacillus subtilis中有效的细胞外分泌酶对于工业应用至关重要.

研究的目的:

研究和优化Bacillus subtilis中β-葡萄糖酶Bgl2A的细胞外分泌.
阐明Bgl2A非经典细胞外分泌的机制.

主要方法:

对Bgl2A分泌的信号优化 (SP_AmyQ) 进行了优化.
具有和没有信号介导的Bgl2A活性的比较分析.
确定影响分泌的关键氨基酸残留物 (I266-Q269).
使用淘汰和细胞完整性试验分析非经典分泌途径.

主要成果:

信号的优化增加了Bgl2A细胞外活动的3.35倍.
没有信号的Bgl2A表现出5.05倍高的细胞外活性和增强的稳定性.
在Bacillus subtilis中发现了Bgl2A非经典的细胞外分泌物.
疏水区域I266-Q269中的突变显著影响了Bgl2A的分泌和活性.

结论:

在Bacillus subtilis中,Bgl2A可以通过一种非经典的途径,独立于传统信号,在细胞外分泌.
疏水区I266-Q269对于这种非经典的分泌机制至关重要.
这项研究为改善重组β-葡萄糖酶表达和理解Bacillus subtilis中非经典分泌的基础.

关键词:

这种细菌是 Bacillus subtilis.非经典的分泌表达方式这是一种β-Glucosidase.

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