2. Academic Validation
  3. Activation and Identification of a Griseusin Cluster in Streptomyces sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods

Activation and Identification of a Griseusin Cluster in Streptomyces sp. CA-256286 by Employing Transcriptional Regulators and Multi-Omics Methods

  • Molecules. 2021 Oct 30;26(21):6580. doi: 10.3390/molecules26216580.
Charlotte Beck 1,  Tetiana Gren 1,  Francisco Javier Ortiz-López 2,  Tue Sparholt Jørgensen 1,  Daniel Carretero-Molina 2,  Jesús Martín Serrano 2,  José R Tormo 2,  Daniel Oves-Costales 2,  Eftychia E Kontou 1,  Omkar S Mohite 1,  Erik Mingyar 3 4,  Evi Stegmann 3 4,  Olga Genilloud 2,  Tilmann Weber 1
Affiliations

Affiliations

  • 1 The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800 Kongens Lyngby, Denmark.
  • 2 Fundación MEDINA, Centro de Excelencia en Investigación de Medicamentos Innovadores en Andalucía, Parque Tecnológico de Ciencias de la Salud, Av. Conocimiento, 34, 18016 Granada, Spain.
  • 3 Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.
  • 4 German Center for Infection Research (DZIF), Partner Site Tübingen, 72076 Tübingen, Germany.
PMID: 34770989 DOI: 10.3390/molecules26216580
Abstract

Streptomyces are well-known producers of a range of different secondary metabolites, including Antibiotics and Other bioactive compounds. Recently, it has been demonstrated that "silent" biosynthetic gene clusters (BGCs) can be activated by heterologously expressing transcriptional regulators from Other BGCs. Here, we have activated a silent BGC in Streptomyces sp. CA-256286 by overexpression of a set of SARP family transcriptional regulators. The structure of the produced compound was elucidated by NMR and found to be an N-acetyl cysteine adduct of the pyranonaphtoquinone polyketide 3'-O-α-d-forosaminyl-(+)-griseusin A. Employing a combination of multi-omics and metabolic engineering techniques, we identified the responsible BGC. These methods include genome mining, proteomics and transcriptomics analyses, in combination with CRISPR induced gene inactivations and expression of the BGC in a heterologous host strain. This work demonstrates an easy-to-implement workflow of how silent BGCs can be activated, followed by the identification and characterization of the produced compound, the responsible BGC, and hints of its biosynthetic pathway.

Keywords

biosynthetic gene cluster; forosamine; genome mining; griseusin; heterologous expression; mycothiol; pyranonaphtoquinone; transcriptional regulators.

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