Regulation mechanism of nitrite degradation in Lactobacillus plantarum WU14 mediated by Fnr

IF 2.3 3区生物学 Q3 MICROBIOLOGY

Archives of Microbiology Pub Date : 2024-11-04 DOI:10.1007/s00203-024-04183-1

Shaoxian Chen, Hao Zeng, Hulin Qiu, Aiguo Yin, Fengfei Shen, Ying Li, Yunyi Xiao, Jinping Hai, Bo Xu

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Abstract

Fumarate and nitrate reduction regulatory protein (Fnr)—a global transcriptional regulator—can directly or indirectly regulate many genes in different metabolic pathways at the top of the bacterial transcription regulation network. The present study explored the regulatory mechanism of Fnr-mediated nitrite degradation in Lactobacillus plantarum WU14 through gene transcription and expression analysis of oxygen sensing and nir operon expression regulation by Fnr. The interaction and the mechanism of transcriptional regulation between Fnr and GlnR were also examined under nitrite stress. After Fnr and GlnR purification by glutathione S-transferase tags, they were successfully expressed in Escherichia coli by constructing an expression vector. The results of electrophoresis mobility shift assay and qRT-PCR indicated that Fnr specifically bound to the PglnR and Pnir promoters and regulated the expression of nitrite reductase (Nir) and GlnR. After 6–12 h of culture, the expressions of fnr and nir under anaerobic conditions were higher than under aerobic conditions; the expression of these two genes increased with sodium nitrite (NaNO₂) addition during aerobic culture. Overall, the present study indicated that Fnr not only directly participated in the expression of Nir and GlnR but also indirectly regulated the expression of Nir through GlnR regulation.

Graphical Abstract

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植物乳杆菌 WU14 在 Fnr 介导下降解亚硝酸盐的调节机制

富马酸盐和硝酸盐还原调控蛋白（Fnr）是一种全球性转录调控因子，可直接或间接调控细菌转录调控网络顶端不同代谢途径中的许多基因。本研究通过对Fnr调控氧传感和nir操作子表达的基因转录和表达分析，探讨了Fnr介导的植物乳杆菌WU14亚硝酸盐降解的调控机制。本研究还考察了亚硝酸盐胁迫下Fnr和GlnR之间的相互作用及其转录调控机制。Fnr和GlnR经谷胱甘肽S-转移酶标记纯化后，通过构建表达载体在大肠杆菌中成功表达。电泳迁移率测定和 qRT-PCR 结果表明，Fnr 与 PglnR 和 Pnir 启动子特异性结合，并调控亚硝酸还原酶（Nir）和 GlnR 的表达。培养 6-12 h 后，厌氧条件下 fnr 和 nir 的表达量高于有氧条件下；在有氧培养过程中，随着亚硝酸钠（NaNO2）的加入，这两个基因的表达量增加。总之，本研究表明，Fnr不仅直接参与了Nir和GlnR的表达，还通过GlnR的调控间接调节了Nir的表达。

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来源期刊

Archives of Microbiology 生物-微生物学

CiteScore

4.90

自引率

3.60%

发文量

601

审稿时长

3 months

期刊介绍： Research papers must make a significant and original contribution to microbiology and be of interest to a broad readership. The results of any experimental approach that meets these objectives are welcome, particularly biochemical, molecular genetic, physiological, and/or physical investigations into microbial cells and their interactions with their environments, including their eukaryotic hosts. Mini-reviews in areas of special topical interest and papers on medical microbiology, ecology and systematics, including description of novel taxa, are also published. Theoretical papers and those that report on the analysis or ''mining'' of data are acceptable in principle if new information, interpretations, or hypotheses emerge.