Bioinformatics analysis of extracellular subtilisin E from <i>Bacillus subtilis</i>

<i>Bacillus</i> spp. are the main sources of subtilisin E, which has several applications in biotechnology. The 3D structure of subtilisin E has a significant impact on its efficacy. In this study, we evaluated subtilisin E from <i>Bacillus subtilis</i> subsp. <i>subtilis</i> str. 168 by bioinformatic methods. The results revealed that the subtilisin E sequence from <i>B. subtilis</i> contains highly conserved amino acids, including histidine (H), aspartic acid (D) and serine (S). Subtilisin E cleaves the bonds between hydrophobic and polar amino acids in keratin-associated proteins. The effects of point mutations on the crystal structure of subtilisin E (PDB ID: 1SCJ) showed that changes of asparagine 123 (N123) to valine (V) and serine 331 (S331) to leucine (L) respectively, were the most stabilizing. Genomic analysis of the subtilisin E-coding gene (<i>aprE</i>) indicated that this gene and the <i>yhfN</i> gene are expressed through a σA promoter. The analysis of TBFs revealed AbrB, ScoC, DegU, Hpr, σA, SinR, TenA, and DegU as relevant regulators of <i>aprE</i> expression. Phylogenetic analysis showed that subtilisin Es have highly conserved structures among <i>Bacillus</i> spp., sharing a common ancestor, where their coding genes were duplicated and evolved within the <i>Bacillus</i> spp. As the conclusion, our <i>in silico</i> study demonstrated that the overexpression of the <i>aprE</i> gene and stability of the produced subtilisin E can be improved though system biology methods such as point mutations and identifying the involved transcription factors (TFs) or/and TBFs. Communicated by Ramaswamy H. Sarma芽孢杆菌属（*Bacillus*）菌株是枯草杆菌蛋白酶E（subtilisin E）的主要来源，该酶在生物技术领域拥有诸多应用场景。枯草杆菌蛋白酶E的三维结构对其催化效能具有显著影响。本研究通过生物信息学方法，对枯草芽孢杆菌枯草亚种（*Bacillus subtilis* subsp. *subtilis*）菌株168所产的枯草杆菌蛋白酶E开展分析评估。结果显示，枯草芽孢杆菌（*B. subtilis*）所编码的枯草杆菌蛋白酶E序列中存在高度保守的氨基酸残基，包括组氨酸（H）、天冬氨酸（D）与丝氨酸（S）。枯草杆菌蛋白酶E可水解角蛋白关联蛋白中疏水氨基酸与极性氨基酸之间的肽键。针对枯草杆菌蛋白酶E晶体结构（PDB编号：1SCJ）的点突变效应分析表明，将天冬酰胺123（N123）突变为缬氨酸（V）、丝氨酸331（S331）突变为亮氨酸（L）的突变组合，可实现最显著的稳定性提升。对编码枯草杆菌蛋白酶E的基因*aprE*进行基因组分析后发现，该基因与*yhfN*基因均通过σA启动子完成转录表达。对转录因子结合因子（Transcription Factor Binding Factors, TBFs）的分析结果显示，AbrB、ScoC、DegU、Hpr、σA、SinR、TenA及DegU是*aprE*基因表达的关键调控因子。系统发育分析结果表明，芽孢杆菌属（*Bacillus*）菌株所产的枯草杆菌蛋白酶E结构高度保守，拥有共同的祖先，其编码基因在芽孢杆菌属内通过基因复制事件完成演化。综上，本计算机模拟（*in silico*）研究证实，通过点突变、鉴定相关转录因子（Transcription Factors, TFs）及/或转录因子结合因子（TBFs）等系统生物学手段，可实现*aprE*基因的过表达，并提升所产枯草杆菌蛋白酶E的稳定性。本文由Ramaswamy H. Sarma转交刊发。

搜集汇总

背景与挑战

背景概述

该数据集基于生物信息学方法，对枯草芽孢杆菌产生的细胞外枯草杆菌蛋白酶E进行深入研究，分析了其结构保守性、点突变对稳定性的影响以及基因表达调控机制，旨在通过系统生物学方法优化酶的过表达和稳定性。数据集包含相关研究文档，覆盖生物化学、微生物学和分子生物学等多个科学领域，适用于生物技术应用和基础研究参考。