Submerged fermentation of Streptomyces uncialis providing a biotechnology platform for uncialamycin biosynthesis, engineering, and production

Author:

Hindra 1ORCID,Yang Dong12ORCID,Luo Jun1ORCID,Huang Tingting1,Yan Xiaohui1ORCID,Adhikari Ajeeth1,Teijaro Christiana N1ORCID,Ge Huiming1,Shen Ben123ORCID

Affiliation:

1. Department of Chemistry, The Scripps Research Institute, Jupiter, FL 33458, USA

2. Natural Products Discovery Center at Scripps Research, The Scripps Research Institute, Jupiter, FL 33458, USA

3. Department of Molecular Medicine, The Scripps Research Institute, Jupiter, FL 33458, USA

Abstract

Abstract Uncialamycin (UCM) belongs to the anthraquinone-fused subfamily of 10-membered enediyne natural products that exhibits an extraordinary cytotoxicity against a wide spectrum of human cancer cell lines. Antibody-drug conjugates, utilizing synthetic analogues of UCM as payloads, are in preclinical development. UCM is exclusively produced by Streptomyces uncialis DCA2648 on solid agar medium with low titers (∼0.019 mg/l), limiting its supply by microbial fermentation and hampering its biosynthetic and engineering studies by in vivo pathway manipulation. Here, we report cultivation conditions that enable genetic manipulation of UCM biosynthesis in vivo and allow UCM production, with improved titers, by submerged fermentation of the engineered S. uncialis strains. Specifically, the titer of UCM was improved nearly 58-fold to ∼1.1 mg/l through the combination of deletion of biosynthetic gene clusters encoding unrelated metabolites from the S. uncialis wild-type, chemical mutagenesis and manipulation of pathway-specific regulators to generate the engineered S. uncialis strains, and finally medium optimization of the latter for UCM production. Genetic manipulation of UCM biosynthesis was demonstrated by inactivating selected genes in the engineered S. uncialis strains, one of which afforded a mutant strain accumulating tiancimycin B, a common biosynthetic intermediate known for the anthraquinone-fused subfamily of enediyne natural products. These findings highlight a biotechnology platform for UCM biosynthesis, engineering, and production that should facilitate both its fundamental studies and translational applications.

Funder

National Institutes of Health

China Scholarship Council

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,Biotechnology,Bioengineering

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