The Old Yellow Enzyme OfrA Fosters Staphylococcus aureus Survival via Affecting Thiol-Dependent Redox Homeostasis

Ibrahim, Eslam S.; Ohlsen, Knut

doi:10.3389/fmicb.2022.888140

Cited by 3 publications

(2 citation statements)

References 62 publications

(72 reference statements)

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“…In addition, the paralogous protein YqiG, of which the function is not yet understood, was also more abundant in all three strains treated with diamide (Table S3). While its function in B. subtilis is still unclear, its ortholog in Staphylococcus aureus has been linked to remediating oxidative damage as caused by diamide ( 39 ). Notably, YqiG was also more abundant in the ribT deletion strain 4 h after diamide addition, when cell growth had recovered from diamide inhibition.…”

Section: Resultsmentioning

confidence: 99%

Diamide-based screening method for the isolation of improved oxidative stress tolerance phenotypes in Bacillus mutant libraries

Walgraeve,

Ferrero-Bordera,

Maaß

et al. 2023

Microbiol Spectr

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The bacterium Bacillus subtilis is of high importance both as a model organism for Gram-positive bacteria and as an industrial workhorse in the production of biomolecules. In recent years, advancements have been made to engineer the bacterium even further toward industrial applications. In this study, we present a novel screening method for mutant libraries using diamide, an oxidizing agent that binds free thiols and creates disulfide bonds between them, thereby causing a so-called “disulfide stress” in bacteria. The method shows promise to selectively identify phenotypes in B. subtilis with improved tolerance toward oxidative and disulfide-associated stress. Phenotypes initially identified by transposon mutagenesis were recreated through targeted gene deletions. Among the resulting deletion mutants, the largest difference in diamide tolerance compared to the parental strain was observed for pfkA and ribT deletion strains. A proteomics analysis showed that diamide tolerance can be achieved through different routes involving increased expression of stress management proteins and reduced availability or activity of the RNA degradosome. We conclude that our screening method allows the facile identification of Bacillus strains with improved oxidative stress tolerance phenotypes. IMPORTANCE During their life cycle, bacteria are exposed to a range of different stresses that need to be managed appropriately in order to ensure their growth and viability. This applies not only to bacteria in their natural habitats but also to bacteria employed in biotechnological production processes. Oxidative stress is one of these stresses that may originate either from bacterial metabolism or external factors. In biotechnological settings, it is of critical importance that production strains are resistant to oxidative stresses. Accordingly, this also applies to the major industrial cell factory Bacillus subtilis . In the present study, we, therefore, developed a screen for B. subtilis strains with enhanced oxidative stress tolerance. The results show that our approach is feasible and time-, space-, and resource-efficient. We, therefore, anticipate that it will enhance the development of more robust industrial production strains with improved robustness under conditions of oxidative stress.

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Section: Resultsmentioning

confidence: 99%

Diamide-based screening method for the isolation of improved oxidative stress tolerance phenotypes in Bacillus mutant libraries

Walgraeve,

Ferrero-Bordera,

Maaß

et al. 2023

Microbiol Spectr

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“…FMN-dependent OYEs reduce a wide range of activated C=C bonds in α,β-unsaturated carbonyl compounds to their saturated counterparts [ 41 ] and were reported to participate in stress response. Thus, the expression of ofrA gene encoding OYE of Staphylococcus aureus was induced by hypochlorite, oxidative, and electrophilic stresses [ 42 ]. YqiG appears to be important in maintaining an intracellular redox balance reducing the electrophilic carbonyl compounds imposed by intensive aeration in strain CM MSU 529.…”

Section: Discussionmentioning

confidence: 99%

Adaptation of Lacticaseibacillus rhamnosus CM MSU 529 to Aerobic Growth: A Proteomic Approach

et al. 2023

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The study describes the effect of aerobic conditions on the proteome of facultative homofermentative lactic acid bacterium Lacticaseibacillus rhamnosus CM MSU 529 grown in a batch culture. Aeration caused the induction of the biosynthesis of 43 proteins, while 14 proteins were downregulated as detected by label-free LC-MS/MS. Upregulated proteins are involved in oxygen consumption (Pox, LctO, pyridoxine 5’-phosphate oxidase), xylulose 5-phosphate conversion (Xfp), pyruvate metabolism (PdhD, AlsS, AlsD), reactive oxygen species (ROS) elimination (Tpx, TrxA, Npr), general stress response (GroES, PfpI, universal stress protein, YqiG), antioxidant production (CysK, DkgA), pyrimidine metabolism (CarA, CarB, PyrE, PyrC, PyrB, PyrR), oligopeptide transport and metabolism (OppA, PepO), and maturation and stability of ribosomal subunits (RbfA, VicX). Downregulated proteins participate in ROS defense (AhpC), citrate and pyruvate consumption (CitE, PflB), oxaloacetate production (AvtA), arginine synthesis (ArgG), amino acid transport (GlnQ), and deoxynucleoside biosynthesis (RtpR). The data obtained shed light on mechanisms providing O2-tolerance and adaptation to aerobic conditions in strain CM MSU 529. The biosynthesis of 39 from 57 differentially abundant proteins was shown to be O2-sensitive in lactic acid bacteria for the first time. To our knowledge this is the first study on the impact of aerobic cultivation on the proteome of L. rhamnosus.

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Evolutionary and molecular basis of ADP-ribosylation reversal by zinc-dependent macrodomains

Ariza,

Liu,

Cowieson

et al. 2024

Journal of Biological Chemistry

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The Old Yellow Enzyme OfrA Fosters Staphylococcus aureus Survival via Affecting Thiol-Dependent Redox Homeostasis

Cited by 3 publications

References 62 publications

Diamide-based screening method for the isolation of improved oxidative stress tolerance phenotypes in Bacillus mutant libraries

Diamide-based screening method for the isolation of improved oxidative stress tolerance phenotypes in Bacillus mutant libraries

Adaptation of Lacticaseibacillus rhamnosus CM MSU 529 to Aerobic Growth: A Proteomic Approach

Evolutionary and molecular basis of ADP-ribosylation reversal by zinc-dependent macrodomains

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