Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 78, Issue 3, Pages 394-399Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09168451.2014.885824
Keywords
antibiotic biosynthesis; gene dosage; heterologous expression; Streptomyces; secondary metabolism
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Funding
- New Energy and Industrial Technology Development Organization (NEDO) of Japan
- Institution of Fermentation, Osaka
- JSPS KAKENHI [25108707]
- Grants-in-Aid for Scientific Research [26850044, 25108707] Funding Source: KAKEN
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Improving the productivity of secondary metabolites is highly beneficial for the utilization of natural products. Here, we found that gene duplication of the goadsporin biosynthetic gene locus resulted in hyper-production. Goadsporin is a linear azole containing peptide that is biosynthesized via a ribosome-mediated pathway in Streptomyces sp. TP-A0584. Recombinant strains containing duplicated or triplicated goadsporin biosynthetic gene clusters produced 1.46- and 2.25-fold more goadsporin than the wild-type strain. In a surrogate host, Streptomyces lividans, chromosomal integration of one or two copies of the gene cluster led to 342.7 and 593.5 mg/L of goadsporin production. Expression of godI, a self-resistance gene, and of godR, a pathway-specific transcriptional regulator, under a constitutive promoter gave 0.79- and 2.12-fold higher goadsporin production than the wild-type strain. Our experiments indicated that a proportional relationship exists between goadsporin production per culture volume and the copy number of the biosynthetic gene cluster.
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