PXD041512

PXD041512 is an original dataset announced via ProteomeXchange.

Title	Diamide-based screening method for isolation of improved oxidative stress tolerance phenotypes in Bacillus mutant libraries
Description	The bacterium Bacillus subtilis is of high importance both as a model organism for Gram-positive bacteria and as an industrial work horse in the production of biomolecules. In recent years, advancements have been made to engineer the bacterium even further towards industrial applications. In this study, we present a novel screening method for mutant libraries using diamide, an oxidising 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 towards 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 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
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:08:33.117.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sandra Maass
SpeciesList	scientific name: Bacillus subtilis subsp. subtilis; NCBI TaxID: 135461;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2023-04-13 04:05:17	ID requested
1	2023-09-25 06:06:30	announced
⏵ 2	2023-11-14 08:08:33	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: mutagenesis, diamide,Bacillus subtilis, screening, library, bacteria

Dörte Becher
contact affiliation	Department of Microbial Proteomics, University of Greifswald, 17489 Greifswald, Germany
contact email	dbecher@uni-greifswald.de
lab head
Sandra Maass
contact affiliation	University of Greifswald, Department for Microbial Proteomics
contact email	sandra.maass@uni-greifswald.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD041512
     1. Label: PRIDE project
     2. Name: Diamide-based screening method for isolation of improved oxidative stress tolerance phenotypes in Bacillus mutant libraries