Genetic approaches to generate hyper-producing strains of goadsporin: the relationships between productivity and gene duplication in secondary metabolite biosynthesis

Biosci Biotechnol Biochem. 2014;78(3):394-9. doi: 10.1080/09168451.2014.885824. Epub 2014 Apr 16.

Abstract

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.

Keywords: Streptomyces; antibiotic biosynthesis; gene dosage; heterologous expression; secondary metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Gene Duplication
  • Gene Expression Regulation, Bacterial
  • Intercellular Signaling Peptides and Proteins
  • Peptide Biosynthesis / genetics*
  • Peptides / genetics*
  • Peptides / metabolism
  • Secondary Metabolism / genetics*
  • Streptomyces / genetics
  • Streptomyces / metabolism*

Substances

  • Intercellular Signaling Peptides and Proteins
  • Peptides
  • goadsporin