Please use this identifier to cite or link to this item: https://doi.org/10.15480/882.4538
DC FieldValueLanguage
dc.contributor.authorKrüger, Anna-
dc.contributor.authorWelsch, Norma-
dc.contributor.authorDürwald, Alexandra-
dc.contributor.authorBrundiek, Henrike-
dc.contributor.authorWardenga, Rainer-
dc.contributor.authorPiascheck, Henning-
dc.contributor.authorMengers, Hendrik G.-
dc.contributor.authorKrabbe, Jana-
dc.contributor.authorBeyer, Sandra-
dc.contributor.authorKabisch, Johannes-
dc.contributor.authorPopper, Lutz-
dc.contributor.authorHübel, Tanno-
dc.contributor.authorAntranikian, Garabed-
dc.contributor.authorSchweder, Thomas-
dc.date.accessioned2022-08-11T08:48:08Z-
dc.date.available2022-08-11T08:48:08Z-
dc.date.issued2022-07-08-
dc.identifier.citationApplied Microbiology and Biotechnology 106 (13/16): 5137-5151 (2022-08-01)de_DE
dc.identifier.issn1432-0614de_DE
dc.identifier.urihttp://hdl.handle.net/11420/13429-
dc.description.abstractAbstract: Target proteins in biotechnological applications are highly diverse. Therefore, versatile flexible expression systems for their functional overproduction are required. In order to find the right heterologous gene expression strategy, suitable host-vector systems, which combine different genetic circuits, are useful. In this study, we designed a novel Bacillus subtilis expression toolbox, which allows the overproduction and secretion of potentially toxic enzymes. This toolbox comprises a set of 60 expression vectors, which combine two promoter variants, four strong secretion signals, a translation-enhancing downstream box, and three plasmid backbones. This B. subtilis toolbox is based on a tailor-made, clean deletion mutant strain, which is protease and sporulation deficient and exhibits reduced autolysis and secondary metabolism. The appropriateness of this alternative expression platform was tested for the overproduction of two difficult-to-produce eukaryotic model proteins. These included the sulfhydryl oxidase Sox from Saccharomyces cerevisiae, which forms reactive hydrogen peroxide and undesired cross-linking of functional proteins, and the human interleukin-1β, a pro-inflammatory cytokine. For the best performing Sox and interleukin, overproducing and secreting variants of these new B. subtilis toolbox fermentation strategies were developed and tested. This study demonstrates the suitability of the prokaryotic B. subtilis host-vector system for the extracellular production of two eukaryotic proteins with biotechnological relevance. Key points: • Construction of a versatile Bacillus subtilis gene expression toolbox. • Verification of the toolbox by the secretory overproduction of two difficult-to-express proteins. • Fermentation strategy for an acetoin-controlled overproduction of heterologous proteins.en
dc.language.isoende_DE
dc.publisherSpringerde_DE
dc.relation.ispartofApplied microbiology and biotechnologyde_DE
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/de_DE
dc.subjectBacillus subtilisde_DE
dc.subjectExpression systemde_DE
dc.subjectFed batchde_DE
dc.subjectHuman interleukin-1βde_DE
dc.subjectOverproductionde_DE
dc.subjectProtease deficiencyde_DE
dc.subjectProtein secretionde_DE
dc.subjectYeast sulfhydryl oxidasede_DE
dc.subject.ddc610: Medizinde_DE
dc.titleA host-vector toolbox for improved secretory protein overproduction in bacillus subtilisde_DE
dc.typeArticlede_DE
dc.identifier.doi10.15480/882.4538-
dc.type.diniarticle-
dcterms.DCMITypeText-
tuhh.identifier.urnurn:nbn:de:gbv:830-882.0194456-
tuhh.oai.showtruede_DE
tuhh.abstract.englishAbstract: Target proteins in biotechnological applications are highly diverse. Therefore, versatile flexible expression systems for their functional overproduction are required. In order to find the right heterologous gene expression strategy, suitable host-vector systems, which combine different genetic circuits, are useful. In this study, we designed a novel Bacillus subtilis expression toolbox, which allows the overproduction and secretion of potentially toxic enzymes. This toolbox comprises a set of 60 expression vectors, which combine two promoter variants, four strong secretion signals, a translation-enhancing downstream box, and three plasmid backbones. This B. subtilis toolbox is based on a tailor-made, clean deletion mutant strain, which is protease and sporulation deficient and exhibits reduced autolysis and secondary metabolism. The appropriateness of this alternative expression platform was tested for the overproduction of two difficult-to-produce eukaryotic model proteins. These included the sulfhydryl oxidase Sox from Saccharomyces cerevisiae, which forms reactive hydrogen peroxide and undesired cross-linking of functional proteins, and the human interleukin-1β, a pro-inflammatory cytokine. For the best performing Sox and interleukin, overproducing and secreting variants of these new B. subtilis toolbox fermentation strategies were developed and tested. This study demonstrates the suitability of the prokaryotic B. subtilis host-vector system for the extracellular production of two eukaryotic proteins with biotechnological relevance. Key points: • Construction of a versatile Bacillus subtilis gene expression toolbox. • Verification of the toolbox by the secretory overproduction of two difficult-to-express proteins. • Fermentation strategy for an acetoin-controlled overproduction of heterologous proteins.de_DE
tuhh.publisher.doi10.1007/s00253-022-12062-2-
tuhh.publication.instituteTechnische Mikrobiologie V-7de_DE
tuhh.identifier.doi10.15480/882.4538-
tuhh.type.opus(wissenschaftlicher) Artikel-
dc.type.driverarticle-
dc.type.casraiJournal Article-
tuhh.container.issue13/16de_DE
tuhh.container.volume106de_DE
tuhh.container.startpage5137de_DE
tuhh.container.endpage5151de_DE
dc.identifier.pmid35802157de_DE
dc.rights.nationallicensefalsede_DE
dc.identifier.scopus2-s2.0-85133588660de_DE
local.status.inpressfalsede_DE
local.type.versionpublishedVersionde_DE
local.funding.infoThis study was financially supported by the German Federal Ministry of Education and Research (BMBF; reference 0315400A/B) in the frame of the BIOCATALYSIS2021 network and by the Ministry of Economy, Labour and Tourism of Mecklenburg Vorpommern and the European Fund for Regional Development (V-630-S-177–2013/213; V-630-F-177–2013/212; V630-VB305-2013/211).de_DE
datacite.resourceTypeArticle-
datacite.resourceTypeGeneralJournalArticle-
item.grantfulltextopen-
item.cerifentitytypePublications-
item.openairetypeArticle-
item.creatorOrcidKrüger, Anna-
item.creatorOrcidWelsch, Norma-
item.creatorOrcidDürwald, Alexandra-
item.creatorOrcidBrundiek, Henrike-
item.creatorOrcidWardenga, Rainer-
item.creatorOrcidPiascheck, Henning-
item.creatorOrcidMengers, Hendrik G.-
item.creatorOrcidKrabbe, Jana-
item.creatorOrcidBeyer, Sandra-
item.creatorOrcidKabisch, Johannes-
item.creatorOrcidPopper, Lutz-
item.creatorOrcidHübel, Tanno-
item.creatorOrcidAntranikian, Garabed-
item.creatorOrcidSchweder, Thomas-
item.languageiso639-1en-
item.creatorGNDKrüger, Anna-
item.creatorGNDWelsch, Norma-
item.creatorGNDDürwald, Alexandra-
item.creatorGNDBrundiek, Henrike-
item.creatorGNDWardenga, Rainer-
item.creatorGNDPiascheck, Henning-
item.creatorGNDMengers, Hendrik G.-
item.creatorGNDKrabbe, Jana-
item.creatorGNDBeyer, Sandra-
item.creatorGNDKabisch, Johannes-
item.creatorGNDPopper, Lutz-
item.creatorGNDHübel, Tanno-
item.creatorGNDAntranikian, Garabed-
item.creatorGNDSchweder, Thomas-
item.fulltextWith Fulltext-
item.openairecristypehttp://purl.org/coar/resource_type/c_6501-
item.mappedtypeArticle-
crisitem.author.deptTechnische Mikrobiologie V-7-
crisitem.author.deptTechnische Mikrobiologie V-7-
crisitem.author.deptTechnische Biokatalyse V-6-
crisitem.author.orcid0000-0001-8007-4955-
crisitem.author.orcid0000-0001-6934-5603-
crisitem.author.parentorgStudiendekanat Verfahrenstechnik (V)-
crisitem.author.parentorgStudiendekanat Verfahrenstechnik (V)-
crisitem.author.parentorgStudiendekanat Verfahrenstechnik (V)-
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