Genetic approaches to generate hyper-producing strains of goadsporin: the relationships between productivity and gene duplication in secondary metabolite biosynthesis. Issue 3 (4th March 2014)

• Record Type: Journal Article
• Authors: Haginaka, Kentaro; Asamizu, Shumpei; Ozaki, Taro; Igarashi, Yasuhiro; Furumai, Tamotsu; Onaka, Hiroyasu
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 78:Issue 3(2014)
• Page Start: 394
• Page End: 399
• Publication Date: 2014-03-04
• Subjects: antibiotic biosynthesis -- gene dosage -- heterologous expression -- Streptomyces -- secondary metabolism; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2014.885824
• Languages: English
• ISSNs: 0916-8451

Abstract

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. Graphical abstract: : Proportional relationship exists between Streptomyces antibiotic production and the copy number of the biosynthetic gene cluster.