Characterization and engineering of Streptomyces griseofuscus DSM 40191 as a potential host for heterologous expression of biosynthetic gene clusters.

Autor: Gren T; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark., Whitford CM; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark., Mohite OS; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark., Jørgensen TS; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark., Kontou EE; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark., Nielsen JB; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark., Lee SY; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark.; Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering, Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea., Weber T; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, bygning 220, 2800, Kgs. Lyngby, Denmark. tiwe@biosustain.dtu.dk.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2021 Sep 15; Vol. 11 (1), pp. 18301. Date of Electronic Publication: 2021 Sep 15.
DOI: 10.1038/s41598-021-97571-2
Abstrakt: Streptomyces griseofuscus DSM 40191 is a fast growing Streptomyces strain that remains largely underexplored as a heterologous host. Here, we report the genome mining of S. griseofuscus, followed by the detailed exploration of its phenotype, including the production of native secondary metabolites and ability to utilise carbon, nitrogen, sulphur and phosphorus sources. Furthermore, several routes for genetic engineering of S. griseofuscus were explored, including use of GusA-based vectors, CRISPR-Cas9 and CRISPR-cBEST-mediated knockouts. Two out of the three native plasmids were cured using CRISPR-Cas9 technology, leading to the generation of strain S. griseofuscus DEL1. DEL1 was further modified by the full deletion of a pentamycin BGC and an unknown NRPS BGC, leading to the generation of strain DEL2, lacking approx. 500 kbp of the genome, which corresponds to a 5.19% genome reduction. DEL2 can be characterized by faster growth and inability to produce three main native metabolites: lankacidin, lankamycin, pentamycin and their derivatives. To test the ability of DEL2 to heterologously produce secondary metabolites, the actinorhodin BGC was used. We were able to observe a formation of a blue halo, indicating a potential production of actinorhodin by both DEL2 and a wild type.
(© 2021. The Author(s).)
Databáze: MEDLINE
Externí odkaz: