Regulation of heterologous subtilin production in Bacillus subtilis W168

Publication type:
Journal article
Metadata:
Autoren
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000779367100001&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1186/s12934-022-01782-9
eISSN
1475-2859
Externe Identifier
  • Clarivate Analytics Document Solution ID: 0I4BR
  • PubMed Identifier: 35392905
Ausgabe der Veröffentlichung
1
Zeitschrift
MICROBIAL CELL FACTORIES
Schlüsselwörter
  • Lantibiotic subtilin
  • Heterologous expression
  • Bacillus subtilis W168
  • spa-locus
  • Biosynthesis regulation
  • Two-component system (TCS)
  • Transition state regulator AbrB
Artikelnummer
ARTN 57
Datum der Veröffentlichung
2022
Status
Published
Titel
Regulation of heterologous subtilin production in <i>Bacillus subtilis</i> W168
Sub types
  • Article
Ausgabe der Zeitschrift
21

Data source: Web of Science (Lite)

Other metadata sources:
Abstract
<jats:title>Abstract</jats:title><jats:sec> <jats:title>Background</jats:title> <jats:p>Subtilin is a peptide antibiotic (lantibiotic) natively produced by <jats:italic>Bacillus subtilis</jats:italic> ATCC6633. It is encoded in a gene cluster <jats:italic>spaBTCSIFEGRK</jats:italic> (<jats:italic>spa</jats:italic>-locus) consisting of four transcriptional units: <jats:italic>spaS</jats:italic> (subtilin pre-peptide), <jats:italic>spaBTC</jats:italic> (modification and export), <jats:italic>spaIFEG</jats:italic> (immunity) and <jats:italic>spaRK</jats:italic> (regulation). Despite the pioneer understanding on subtilin biosynthesis, a robust platform to facilitate subtilin research and improve subtilin production is still a poorly explored spot.</jats:p> </jats:sec><jats:sec> <jats:title>Results</jats:title> <jats:p>In this work, the intact <jats:italic>spa</jats:italic>-locus was successfully integrated into the chromosome of <jats:italic>Bacillus subtilis</jats:italic> W168, which is the by far best-characterized Gram-positive model organism with powerful genetics and many advantages in industrial use. Through systematic analysis of <jats:italic>spa</jats:italic>-promoter activities in <jats:italic>B. subtilis</jats:italic> W168 wild type and mutant strains, our work demonstrates that subtilin is basally expressed in <jats:italic>B. subtilis</jats:italic> W168, and the transition state regulator AbrB strongly represses subtilin biosynthesis in a growth phase-dependent manner. The deletion of AbrB remarkably enhanced subtilin gene expression, resulting in comparable yield of bioactive subtilin production as for <jats:italic>B. subtilis</jats:italic> ATCC6633. However, while in <jats:italic>B. subtilis</jats:italic> ATCC6633 AbrB regulates subtilin gene expression via SigH, which in turn activates <jats:italic>spaRK,</jats:italic> AbrB of <jats:italic>B. subtilis</jats:italic> W168 controls subtilin gene expression in SigH-independent manner, except for the regulation of <jats:italic>spaBTC</jats:italic>. Furthermore, the work shows that subtilin biosynthesis in <jats:italic>B. subtilis</jats:italic> W168 is regulated by the two-component regulatory system SpaRK and strictly relies on subtilin itself as inducer to fulfill the autoregulatory circuit. In addition, by incorporating the subtilin-producing system (<jats:italic>spa</jats:italic>-locus) and subtilin-reporting system (P<jats:sub><jats:italic>psdA</jats:italic></jats:sub>-<jats:italic>lux</jats:italic>) together, we developed “online” reporter strains to efficiently monitor the dynamics of subtilin biosynthesis.</jats:p> </jats:sec><jats:sec> <jats:title>Conclusions</jats:title> <jats:p>Within this study, the model organism <jats:italic>B. subtilis</jats:italic> W168 was successfully established as a novel platform for subtilin biosynthesis and the underlying regulatory mechanism was comprehensively characterized. This work will not only facilitate genetic (engineering) studies on subtilin, but also pave the way for its industrial production. More broadly, this work will shed new light on the heterologous production of other lantibiotics.</jats:p> </jats:sec>
Autoren
DOI
10.1186/s12934-022-01782-9
eISSN
1475-2859
Ausgabe der Veröffentlichung
1
Zeitschrift
Microbial Cell Factories
Sprache
en
Artikelnummer
57
Online publication date
2022
Status
Published online
Herausgeber
Springer Science and Business Media LLC
Herausgeber URL
http://dx.doi.org/10.1186/s12934-022-01782-9
Datum der Datenerfassung
2022
Titel
Regulation of heterologous subtilin production in Bacillus subtilis W168
Ausgabe der Zeitschrift
21

Data source: Crossref

Abstract
<h4>Background</h4>Subtilin is a peptide antibiotic (lantibiotic) natively produced by Bacillus subtilis ATCC6633. It is encoded in a gene cluster spaBTCSIFEGRK (spa-locus) consisting of four transcriptional units: spaS (subtilin pre-peptide), spaBTC (modification and export), spaIFEG (immunity) and spaRK (regulation). Despite the pioneer understanding on subtilin biosynthesis, a robust platform to facilitate subtilin research and improve subtilin production is still a poorly explored spot.<h4>Results</h4>In this work, the intact spa-locus was successfully integrated into the chromosome of Bacillus subtilis W168, which is the by far best-characterized Gram-positive model organism with powerful genetics and many advantages in industrial use. Through systematic analysis of spa-promoter activities in B. subtilis W168 wild type and mutant strains, our work demonstrates that subtilin is basally expressed in B. subtilis W168, and the transition state regulator AbrB strongly represses subtilin biosynthesis in a growth phase-dependent manner. The deletion of AbrB remarkably enhanced subtilin gene expression, resulting in comparable yield of bioactive subtilin production as for B. subtilis ATCC6633. However, while in B. subtilis ATCC6633 AbrB regulates subtilin gene expression via SigH, which in turn activates spaRK, AbrB of B. subtilis W168 controls subtilin gene expression in SigH-independent manner, except for the regulation of spaBTC. Furthermore, the work shows that subtilin biosynthesis in B. subtilis W168 is regulated by the two-component regulatory system SpaRK and strictly relies on subtilin itself as inducer to fulfill the autoregulatory circuit. In addition, by incorporating the subtilin-producing system (spa-locus) and subtilin-reporting system (P<sub>psdA</sub>-lux) together, we developed "online" reporter strains to efficiently monitor the dynamics of subtilin biosynthesis.<h4>Conclusions</h4>Within this study, the model organism B. subtilis W168 was successfully established as a novel platform for subtilin biosynthesis and the underlying regulatory mechanism was comprehensively characterized. This work will not only facilitate genetic (engineering) studies on subtilin, but also pave the way for its industrial production. More broadly, this work will shed new light on the heterologous production of other lantibiotics.
Addresses
  • Institute of Microbiology, Technische Universität Dresden, 01217, Dresden, Germany.
Autoren
DOI
10.1186/s12934-022-01782-9
eISSN
1475-2859
Externe Identifier
  • PubMed Identifier: 35392905
  • PubMed Central ID: PMC8991943
Funding acknowledgements
  • Technische Universität Dresden:
  • China Scholarship Council:
Open access
true
ISSN
1475-2859
Ausgabe der Veröffentlichung
1
Zeitschrift
Microbial cell factories
Schlüsselwörter
  • Bacillus subtilis
  • Peptides
  • Bacterial Proteins
  • Bacteriocins
  • Gene Expression Regulation, Bacterial
Sprache
eng
Medium
Electronic
Online publication date
2022
Open access status
Open Access
Paginierung
57
Datum der Veröffentlichung
2022
Status
Published
Publisher licence
CC BY
Datum der Datenerfassung
2022
Titel
Regulation of heterologous subtilin production in Bacillus subtilis W168.
Sub types
  • research-article
  • Journal Article
Ausgabe der Zeitschrift
21

Files

https://microbialcellfactories.biomedcentral.com/track/pdf/10.1186/s12934-022-01782-9 https://europepmc.org/articles/PMC8991943?pdf=render

Data source: Europe PubMed Central

Abstract
BACKGROUND: Subtilin is a peptide antibiotic (lantibiotic) natively produced by Bacillus subtilis ATCC6633. It is encoded in a gene cluster spaBTCSIFEGRK (spa-locus) consisting of four transcriptional units: spaS (subtilin pre-peptide), spaBTC (modification and export), spaIFEG (immunity) and spaRK (regulation). Despite the pioneer understanding on subtilin biosynthesis, a robust platform to facilitate subtilin research and improve subtilin production is still a poorly explored spot. RESULTS: In this work, the intact spa-locus was successfully integrated into the chromosome of Bacillus subtilis W168, which is the by far best-characterized Gram-positive model organism with powerful genetics and many advantages in industrial use. Through systematic analysis of spa-promoter activities in B. subtilis W168 wild type and mutant strains, our work demonstrates that subtilin is basally expressed in B. subtilis W168, and the transition state regulator AbrB strongly represses subtilin biosynthesis in a growth phase-dependent manner. The deletion of AbrB remarkably enhanced subtilin gene expression, resulting in comparable yield of bioactive subtilin production as for B. subtilis ATCC6633. However, while in B. subtilis ATCC6633 AbrB regulates subtilin gene expression via SigH, which in turn activates spaRK, AbrB of B. subtilis W168 controls subtilin gene expression in SigH-independent manner, except for the regulation of spaBTC. Furthermore, the work shows that subtilin biosynthesis in B. subtilis W168 is regulated by the two-component regulatory system SpaRK and strictly relies on subtilin itself as inducer to fulfill the autoregulatory circuit. In addition, by incorporating the subtilin-producing system (spa-locus) and subtilin-reporting system (PpsdA-lux) together, we developed "online" reporter strains to efficiently monitor the dynamics of subtilin biosynthesis. CONCLUSIONS: Within this study, the model organism B. subtilis W168 was successfully established as a novel platform for subtilin biosynthesis and the underlying regulatory mechanism was comprehensively characterized. This work will not only facilitate genetic (engineering) studies on subtilin, but also pave the way for its industrial production. More broadly, this work will shed new light on the heterologous production of other lantibiotics.
Date of acceptance
2022
Autoren
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/35392905
DOI
10.1186/s12934-022-01782-9
eISSN
1475-2859
Externe Identifier
  • PubMed Central ID: PMC8991943
Ausgabe der Veröffentlichung
1
Zeitschrift
Microb Cell Fact
Schlüsselwörter
  • Bacillus subtilis W168
  • Biosynthesis regulation
  • Heterologous expression
  • Lantibiotic subtilin
  • Transition state regulator AbrB
  • Two-component system (TCS)
  • spa-locus
  • Bacillus subtilis
  • Bacterial Proteins
  • Bacteriocins
  • Gene Expression Regulation, Bacterial
  • Peptides
Sprache
eng
Country
England
Paginierung
57
PII
10.1186/s12934-022-01782-9
Datum der Veröffentlichung
2022
Status
Published online
Datum, an dem der Datensatz öffentlich gemacht wurde
2022
Titel
Regulation of heterologous subtilin production in Bacillus subtilis W168.
Sub types
  • Journal Article
Ausgabe der Zeitschrift
21

Data source: PubMed

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