title: Genetic approaches to generate hyper-producing strains of goadsporin: the relationships between productivity and gene duplication in secondary metabolite biosynthesis.
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.
authors.affiliation: a Department of Biotechnology , Toyama Prefectural University , Imizu, Toyama , Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Haginaka
authors.forename: Shumpei
authors.initials: S
authors.lastname: Asamizu
authors.forename: Taro
authors.initials: T
authors.lastname: Ozaki
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Igarashi
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Furumai
authors.forename: Hiroyasu
authors.initials: H
authors.lastname: Onaka
mesh_terms: D000595:Amino Acid Sequence; D020440:Gene Duplication; D015964:Gene Expression Regulation, Bacterial; D036341:Intercellular Signaling Peptides and Proteins; D010452:Peptide Biosynthesis / Q000235:genetics*; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D064210:Secondary Metabolism / Q000235:genetics*; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C449027:goadsporin
keywords: Streptomyces; antibiotic biosynthesis; gene dosage; heterologous expression; secondary metabolism