Engineering Salinispora tropica for heterologous expression of natural product biosynthetic gene clusters

TitleEngineering Salinispora tropica for heterologous expression of natural product biosynthetic gene clusters
Publication TypeJournal Article
Year of Publication2018
AuthorsZhang J.J, Moore BS, Tang X.Y
JournalApplied Microbiology and Biotechnology
Volume102
Pagination8437-8446
Date Published2018/10
Type of ArticleArticle
ISBN Number0175-7598
Accession NumberWOS:000444761100024
KeywordsBiotechnology & Applied Microbiology; direct cloning; diversity; Engineering; escherichia-coli; genetic; heterologous expression; integrase; marine actinomycete salinispora; natural products; phi-c31; reveals; Salinispora; salinosporamide; secondary metabolism; site-specific recombination; streptomyces-coelicolor
Abstract

The marine actinomycete genus Salinispora is a remarkably prolific source of structurally diverse and biologically active secondary metabolites. Herein, we select the model organism Salinispora tropica CNB-440 for development as a heterologous host for the expression of biosynthetic gene clusters (BGCs) to complement well-established Streptomyces host strains. In order to create an integratable host with a clean background of secondary metabolism, we replaced three genes (salA-C) essential for salinosporamide biosynthesis with a cassette containing the Streptomyces coelicolor Phi C31 phage attachment site attB to generate the mutant S. tropica CNB-4401 via double-crossover recombination. This mutagenesis not only knocks-in the attachment site attB in the genome of S. tropica CNB-440 but also abolishes production of the salinosporamides, thereby simplifying the strain's chemical background. We validated this new heterologous host with the successful integration and expression of the thiolactomycin BGC that we recently identified in several S. pacifica strains. When compared to the extensively engineered superhost S. coelicolor M1152, the production of thiolactomycins from S. tropica CNB-4401 was approximately 3-fold higher. To the best of our knowledge, this is the first example of using a marine actinomycete as a heterologous host for natural product BGC expression. The established heterologous host may provide a useful platform to accelerate the discovery of novel natural products and engineer biosynthetic pathways.

DOI10.1007/s00253-018-9283-z
Short TitleAppl. Microbiol. Biotechnol.
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