Impact of membrane gene overexpression on endoglucanase secretion in ruminal cellulolytic Escherichia coli

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Process Biochemistry Pub Date : 2024-11-06 DOI:10.1016/j.procbio.2024.11.006

Long Zhang , Jinna Cui , Wei Liu , Zhanying Liu

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Abstract

Various protein secretion systems of gram-negative Escherichia coli (E. coli) are well known, but the secretion pathway of endoglucanase (BcsZ) is unknown. To further elucidate the secretion mechanism, we analyzed the transcriptome data of five ruminal cellulolytic E. coli strains. Gene Ontology (GO) enrichment analysis revealed that the differentially expressed genes (DEGs) upregulated in all five strains were significantly enriched in the membrane. Nine DEGs related to membrane transporters were overexpressed in E. coli JBZ-DH5α (overexpressing BcsZ in the genome of E. coli DH5α). The results revealed that the overexpression of the b4332 (yjiJ), b2142 (yohK), and b2377 (yfdY) genes significantly reduced the growth and BcsZ concentration of the strain. The overexpression of the genes b3533 (bcsA), b3009 (yghB), and b3522 (yhjD) did not significantly affect strain growth or the BcsZ concentration. The overexpression of the b1859 (znuB), b0458 (ylaC), and b1195 (ymgE) genes increased the secretion of BcsZ but had no significant effect on strain growth. In conclusion, the yjiJ, yohK, yfdY, znuB, ylaC, and ymgE genes are involved in the secretion of E. coli BcsZ. These findings provide insight into the secretion mechanism of E. coli BcsZ.

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膜基因过表达对瘤胃纤维素分解大肠杆菌分泌内切葡聚糖酶的影响

革兰氏阴性大肠杆菌（大肠杆菌）的各种蛋白质分泌系统已广为人知，但内切葡聚糖酶（BcsZ）的分泌途径尚不清楚。为了进一步阐明其分泌机制，我们分析了五株瘤胃纤维素溶解大肠杆菌的转录组数据。基因本体（Gene Ontology，GO）富集分析表明，五株菌株中上调的差异表达基因（Differentially expressed genes，DEGs）都显著富集在膜上。在大肠杆菌 JBZ-DH5α（在大肠杆菌 DH5α 基因组中过表达 BcsZ）中，有 9 个与膜转运体相关的 DEGs 过表达。结果发现，过表达 b4332（yjiJ）、b2142（yohK）和 b2377（yfdY）基因会显著降低菌株的生长和 BcsZ 浓度。基因 b3533（bcsA）、b3009（yghB）和 b3522（yhjD）的过表达对菌株的生长和 BcsZ 浓度没有明显影响。过表达 b1859（znuB）、b0458（ylaC）和 b1195（ymgE）基因会增加 BcsZ 的分泌，但对菌株的生长没有明显影响。总之，yjiJ、yohK、yfdY、znuB、ylaC 和 ymgE 基因参与了大肠杆菌 BcsZ 的分泌。这些发现有助于深入了解大肠杆菌 BcsZ 的分泌机制。

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阿拉丁

isopropyl-β-D-thiogalactopyranoside (IPTG)

来源期刊

Process Biochemistry 生物-工程：化工

CiteScore

8.30

自引率

4.50%

发文量

374

审稿时长

53 days

期刊介绍： Process Biochemistry is an application-orientated research journal devoted to reporting advances with originality and novelty, in the science and technology of the processes involving bioactive molecules and living organisms. These processes concern the production of useful metabolites or materials, or the removal of toxic compounds using tools and methods of current biology and engineering. Its main areas of interest include novel bioprocesses and enabling technologies (such as nanobiotechnology, tissue engineering, directed evolution, metabolic engineering, systems biology, and synthetic biology) applicable in food (nutraceutical), healthcare (medical, pharmaceutical, cosmetic), energy (biofuels), environmental, and biorefinery industries and their underlying biological and engineering principles.