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Escherichia coli Uses Separate Enzymes to Produce H₂S and Reactive Sulfane Sulfur From L-cysteine.

ABSTRACT: Hydrogen sulfide (H₂S) has been proposed to have various physiological functions, and it may function through reactive sulfane sulfur. Since the two sulfur forms often coexist, they are normally considered interchangeable. Here, we characterized the production of H₂S and reactive sulfane sulfur in Escherichia coli MG1655 and found that they are not readily interchangeable. They are primarily produced from L-cysteine via different enzymes. L-Cysteine desulfhydrases consumed L-cysteine and directly generated H₂S. The produced H₂S was mainly lost through evaporation into the gas phase, as E. coli does not have enzymes that easily oxidize H₂S to reactive sulfane sulfur. L-Cysteine desulfhydrases were also responsible for the degradation of exogenous L-cysteine, which is toxic at high levels. Conversely, L-cysteine aminotransferase and 3-mercaptopyruvate sulfurtransferase sequentially metabolized endogenous L-cysteine to produce cellular reactive sulfane sulfur; however, it was not a major route of H₂S production during normal growth or during the metabolism of exogenous L-cysteine by the resting cells. Noticeably, the 3-mercaptopyruvate sulfurtransferase mutant contained less reactive sulfane sulfur and displayed a greater sensitivity to H₂O₂ than did the wild type. Thence, reactive sulfane sulfur is likely a common cellular component, involved in protein sulfhydration and protecting cells from oxidative stress.

SUBMITTER: Li K

PROVIDER: S-EPMC6401616 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Sulfane sulfur, a collective term for hydrogen polysulfide and organic persulfide, often damages cells at high concentrations. Cells can regulate intracellular sulfane sulfur levels through specific mechanisms, but these mechanisms are unclear in Corynebacterium glutamicum. OxyR is a transcription factor capable of sensing oxidative stress and is also responsive to sulfane sulfur. In this study, we found that OxyR functioned directly in regulating sulfane sulfur in C. glutamicum. OxyR binds to the promoter of katA and nrdH and regulates its expression, as revealed via in vitro electrophoretic mobility shift assay analysis, real-time quantitative PCR, and reporting systems. Overexpression of katA and nrdH reduced intracellular sulfane sulfur levels by over 30% and 20% in C. glutamicum, respectively. RNA-sequencing analysis showed that the lack of OxyR downregulated the expression of sulfur assimilation pathway genes and/or sulfur transcription factors, which may reduce the rate of sulfur assimilation. In addition, OxyR also affected the biosynthesis of L-cysteine in C. glutamicum. OxyR overexpression strain Cg-2 accumulated 183 mg/L of L-cysteine, increased by approximately 30% compared with the control (142 mg/L). In summary, OxyR not only regulated sulfane sulfur levels by controlling the expression of katA and nrdH in C. glutamicum but also facilitated the sulfur assimilation and L-cysteine synthesis pathways, providing a potential target for constructing robust cell factories of sulfur-containing amino acids and their derivatives. IMPORTANCE C. glutamicum is an important industrial microorganism used to produce various amino acids. In the production of sulfur-containing amino acids, cells inevitably accumulate a large amount of sulfane sulfur. However, few studies have focused on sulfane sulfur removal in C. glutamicum. In this study, we not only revealed the regulatory mechanism of OxyR on intracellular sulfane sulfur removal but also explored the effects of OxyR on the sulfur assimilation and L-cysteine synthesis pathways in C. glutamicum. This is the first study on the removal of sulfane sulfur in C. glutamicum. These results contribute to the understanding of sulfur regulatory mechanisms and may aid in the future optimization of C. glutamicum for biosynthesis of sulfur-containing amino acids.

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Escherichia coli Uses Separate Enzymes to Produce H₂S and Reactive Sulfane Sulfur From L-cysteine.

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Escherichia coli Uses Separate Enzymes to Produce H₂S and Reactive Sulfane Sulfur From L-cysteine.