Involvement of a mycothiol-dependent reductase NCgl0018 in oxidative stress response of Corynebacterium glutamicum.

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Keyan Chen, Xiaoyang Yu, Xinyu Zhang, Xiaona Li, Yang Liu, Meiru Si, Tao Su
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引用次数: 2

Abstract

Corynebacterium glutamicum is an important industrial strain for amino acids and a key model organism for human pathogens. The study of C. glutamicum oxidoreductases, such as mycoredoxin 1 (Mrx1), dithiol-disulfide isomerase DsbA, and DsbA-like Mrx1, is helpful for understanding the survival, pathogenic infection, and stress resistance of its homologous species. However, the action mode and enzymatic function of C. glutamicum NCgl0018 preserving the Cys-Pro-Phe-Cys motif, annotated as a putative DsbA, have remained enigmatic. Here, we report that the NCgl0018-deleted strain increased sensitivity to various oxidative stresses. The ncgl0018 expression was induced in the stress-responsive extracytoplasmic function-sigma (ECF-σ) factor SigH- and organic peroxide- and antibiotic-sensing regulator (OasR)-dependent manner by stress. NCgl0018 reduced S-mycothiolated mixed disulfides and intramolecular disulfides via a monothiol-disulfide mechanism preferentially linking the mycothiol/mycothione reductase/NADPH electron pathway. Site-directed mutagenesis confirmed Cys107 was the resolving Cys residue, while Cys104 was the nucleophilic cysteine that was oxidized to a sulfenic acid and then could form an intramolecular disulfide bond with Cys107 or a mixed disulfide with mycothiol under stress. Biochemical analyses indicated that NCgl0018 lacked oxidase properties like the classical DsbA. Further, enzymatic rates and substrate preferences of NCgl0018 were highly similar to those of DsbA-like Mrx1. Collectively, our study presented the first evidence that NCgl0018 protected against stresses by functioning as a novel DsbA-like Mrx1 but not DsbA and Mrx1.

真菌硫醇依赖还原酶NCgl0018参与谷氨酸棒状杆菌氧化应激反应。
谷氨酸棒状杆菌是一种重要的氨基酸工业菌株,也是人类病原体的关键模式生物。研究C. glutamum氧化还原酶,如核氧化还蛋白1 (Mrx1)、二硫醇二硫异构酶DsbA和DsbA样Mrx1,有助于了解其同源物种的生存、致病感染和抗逆性。然而,C. glutamicum NCgl0018保留cys - pro - ph - cys基序(注释为假定的DsbA)的作用模式和酶功能仍然是谜。在这里,我们报道了ncgl0018缺失菌株增加了对各种氧化应激的敏感性。胁迫诱导ncgl0018以应激反应性胞浆外功能-sigma (ECF-σ)因子和有机过氧化物和抗生素敏感调节剂(OasR)依赖的方式表达。NCgl0018通过优先连接菌硫醇/菌硫酮还原酶/NADPH电子途径的单硫-二硫机制还原s -菌硫化混合二硫化物和分子内二硫化物。位点诱变证实Cys107是溶解的Cys残基,而Cys104是亲核半胱氨酸,在胁迫下被氧化成亚磺酸,与Cys107形成分子内二硫键或与真菌硫醇形成混合二硫键。生化分析表明NCgl0018缺乏与经典DsbA类似的氧化酶特性。此外,NCgl0018的酶促率和底物偏好与dbas样Mrx1高度相似。总的来说,我们的研究首次提出了NCgl0018通过作为一种新的DsbA样Mrx1而不是DsbA和Mrx1来保护免受压力的证据。
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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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