Journal article
Activation and identification of a griseusin cluster in streptomyces sp. Ca-256286 by employing transcriptional regulators and multi-omics methods
Natural Products Genome Mining, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Strain Design Teams, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
Fundación MEDINA4
Analytics, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
Reconstruction, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark6
University of Tübingen7
Center for Microbial Secondary Metabolites, Centers, Technical University of Denmark8
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.
Language: | English |
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Publisher: | MDPI AG |
Year: | 2021 |
Pages: | 6580 |
ISSN: | 14203049 and 14315157 |
Types: | Journal article |
DOI: | 10.3390/molecules26216580 |
ORCIDs: | Beck, Charlotte , Gren, Tetiana , 0000-0002-9410-3718 , 0000-0003-4481-8157 , 0000-0001-6984-2941 , 0000-0002-0373-3731 , 0000-0002-1017-5459 , 0000-0002-4202-1219 , Weber, Tilmann , Jørgensen, Tue Sparholt , Kontou, Eftychia E. and Mohite, Omkar S. |
Biosynthetic gene cluster Forosamine Genome mining Griseusin Heterologous expression Mycothiol Pyranonaphtoquinone Transcriptional regulators
Computational Biology Molecular Structure Naphthoquinones Organic chemistry QD241-441 Streptomyces Transcription Factors Transcription, Genetic biosynthetic gene cluster forosamine genome mining griseusin heterologous expression mycothiol pyranonaphtoquinone transcriptional regulators