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

Kentaro HaginakaDepartment of Biotechnology, Toyama Prefectural University, Imizu, Toyama, Japan

Shumpei AsamizuDepartment of Biotechnology, The University of Tokyo, Tokyo, Japan

Taro OzakiDepartment of Biotechnology, The University of Tokyo, Tokyo, Japan

Yasuhiro IgarashiDepartment of Biotechnology, Toyama Prefectural University, Imizu, Toyama, Japan

Tamotsu FurumaiDepartment of Biotechnology, Toyama Prefectural University, Imizu, Toyama, Japan

Hiroyasu OnakaDepartment of Biotechnology, Toyama Prefectural University, Imizu, Toyama, Japan;Department of Biotechnology, The University of Tokyo, Tokyo, JapanCorrespondence[email protected]

Figures & data

Fig. 1. Chemical structure and biosynthetic gene organization of goadsporin.

Note: (A) Chemical structure of goadsporin. (B) Gene organization of goadsporin biosynthetic Gene cluster in Streptomyces sp. TP-A0584. The expression vectors of godI and godR were constructed in this study.

Table 1. Plasmids used in this study.

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Fig. 2. HPLC profiles of extracts from the culture broth of goadsporin-producing strains.

Note: (A) Streptomyces sp. TP-A0584 wild-type strain. (B) Strain TP-A0584 (pGSBC1), harboring a duplicate set of goadsporin biosynthetic gene clusters in the genome. C, Strain TP-A0584 (pGSBC1 and pTYM1-GSBC) harboring a triplicate set of goadsporin biosynthetic gene clusters in the genome. The arrow in A corresponds to goadsporin. Sample preparation and HPLC conditions are described in “Materials and Methods.” Peaks were detected at 254 nm, the chromatographs were drawn to the same scale, and the gradient diagram is shown on the right-hand scale.

Fig. 3. Amounts of goadsporin produced by Streptomyces sp. TP-A0584 by the duplicate and triplicate gene cluster containing strains.

Note: Circles, Streptomyces sp. TP-A0584; triangles, strain TP-A0584 (pGSBC1); squares, strain TP-A0584 (pGSBC1 and pTYM1-GSBC). The concentrations of goadsporin are average values for two independent experiments, TP-A0584 and TP-A0584 (pGSBC1) and three independent experiments, TP-A0584 (pGSBC1 and pTYM1-GSBC).

Fig. 4. Amount of goadsporin produced by the surrogate hosts.

Note: Circles, S. lividans (pGSBC1); squares, S. lividans (pGSBC1 and pTYM1-GSBC). The concentrations of goadsporin are average values for two independent experiments.

Fig. 5. Transcriptional and self-resistance effects on goadsporin production.

Note: Circles, Streptomyces sp. TP-A0584 (control); triangles, godI constitutive expression strain of Streptomyces sp. TP-A0584; squares, godR constitutive expression strain of Streptomyces sp. TP-A0584. The concentrations of goadsporin are average values for two independent experiments.

Supplemental material

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