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Sigma Factor Engineering in Actinoplanes sp. SE50/110: Expression of the Alternative Sigma Factor Gene ACSP50_0507 (σHAs) Enhances Acarbose Yield and Alters Cell Morphology.
Schlüter, Laura; Busche, Tobias; Bondzio, Laila; Hütten, Andreas; Niehaus, Karsten; Schneiker-Bekel, Susanne; Pühler, Alfred; Kalinowski, Jörn.
Afiliación
  • Schlüter L; Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, 33594 Bielefeld, Germany.
  • Busche T; Technology Platform Genomics, Center for Biotechnology, Bielefeld University, 33594 Bielefeld, Germany.
  • Bondzio L; Medical School East Westphalia-Lippe, Bielefeld University, 33594 Bielefeld, Germany.
  • Hütten A; Faculty of Physics, Bielefeld University, 33594 Bielefeld, Germany.
  • Niehaus K; Faculty of Physics, Bielefeld University, 33594 Bielefeld, Germany.
  • Schneiker-Bekel S; Proteome and Metabolome Research, Faculty of Biology, Bielefeld University, 33594 Bielefeld, Germany.
  • Pühler A; Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, 33594 Bielefeld, Germany.
  • Kalinowski J; Genome Research of Industrial Microorganisms, Center for Biotechnology (CeBiTec), Bielefeld University, 33594 Bielefeld, Germany.
Microorganisms ; 12(6)2024 Jun 20.
Article en En | MEDLINE | ID: mdl-38930623
ABSTRACT
Sigma factors are transcriptional regulators that are part of complex regulatory networks for major cellular processes, as well as for growth phase-dependent regulation and stress response. Actinoplanes sp. SE50/110 is the natural producer of acarbose, an α-glucosidase inhibitor that is used in diabetes type 2 treatment. Acarbose biosynthesis is dependent on growth, making sigma factor engineering a promising tool for metabolic engineering. ACSP50_0507 is a homolog of the developmental and osmotic-stress-regulating Streptomyces coelicolor σHSc. Therefore, the protein encoded by ACSP50_0507 was named σHAs. Here, an Actinoplanes sp. SE50/110 expression strain for the alternative sigma factor gene ACSP50_0507 (sigHAs) achieved a two-fold increased acarbose yield with acarbose production extending into the stationary growth phase. Transcriptome sequencing revealed upregulation of acarbose biosynthesis genes during growth and at the late stationary growth phase. Genes that are transcriptionally activated by σHAs frequently code for secreted or membrane-associated proteins. This is also mirrored by the severely affected cell morphology, with hyperbranching, deformed and compartmentalized hyphae. The dehydrated cell morphology and upregulation of further genes point to a putative involvement in osmotic stress response, similar to its S. coelicolor homolog. The DNA-binding motif of σHAs was determined based on transcriptome sequencing data and shows high motif similarity to that of its homolog. The motif was confirmed by in vitro binding of recombinantly expressed σHAs to the upstream sequence of a strongly upregulated gene. Autoregulation of σHAs was observed, and binding to its own gene promoter region was also confirmed.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Microorganisms Año: 2024 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Suiza
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Microorganisms Año: 2024 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Suiza