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Regulation of heterologous subtilin production in Bacillus subtilis W168.
Zhang, Qian; Kobras, Carolin M; Gebhard, Susanne; Mascher, Thorsten; Wolf, Diana.
Afiliação
  • Zhang Q; Institute of Microbiology, Technische Universität Dresden, 01217, Dresden, Germany.
  • Kobras CM; Department Biology I, Ludwig-Maximilians-Universität München, 82152, Planegg-Martinsried, Germany.
  • Gebhard S; School of Biosciences, The University of Sheffield, Sheffield, S10 2TN, UK.
  • Mascher T; Department Biology I, Ludwig-Maximilians-Universität München, 82152, Planegg-Martinsried, Germany.
  • Wolf D; Department of Biology & Biochemistry, Milner Centre for Evolution, University of Bath, Bath, BA2 7AY, UK.
Microb Cell Fact ; 21(1): 57, 2022 Apr 07.
Article em En | MEDLINE | ID: mdl-35392905
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bacillus subtilis / Bacteriocinas Idioma: En Revista: Microb Cell Fact Assunto da revista: BIOTECNOLOGIA / MICROBIOLOGIA Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Alemanha

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bacillus subtilis / Bacteriocinas Idioma: En Revista: Microb Cell Fact Assunto da revista: BIOTECNOLOGIA / MICROBIOLOGIA Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Alemanha