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A new pathway for polyketide synthesis in microorganisms

Nature volume 400, pages 897–899 (1999)Cite this article

Abstract

Chalcone synthases, which biosynthesize chalcones (the starting materials for many flavonoids1,2), have been believed to be specific to plants. However, the rppA gene from the Gram-positive, soil-living filamentous bacterium Streptomyces griseus encodes a 372-amino-acid protein that shows significant similarity to chalcone synthases3. Several rppA-like genes are known, but their functions and catalytic properties have not been described. Here we show that a homodimer of RppA catalyses polyketide synthesis: it selects malonyl-coenzyme-A as the starter, carries out four successive extensions and releases the resulting pentaketide to cyclize to 1,3,6,8-tetrahydroxynaphthalene (THN). Site-directed mutagenesis revealed that, as in other chalcone synthases4,5, a cysteine residue is essential for enzyme activity. Disruption of the chromosomal rppA gene in S. griseus abolished melanin production in hyphae, resulting in ‘albino’ mycelium. THN was readily oxidized to form 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin), which then randomly polymerized to form various coloured compounds. THN formed by RppA appears to be an intermediate in the biosynthetic pathways for not only melanins but also various secondary metabolites containing a naphthoquinone ring. Therefore, RppA is a chalcone-synthase-related synthase that synthesizes polyketides and is found in the Streptomyces and other bacteria.

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Figure 1: SDS–PAGE analysis of RppA.
Figure 2: Reverse-phase HPLC analysis of products synthesized by RppA in vitro, showing absorbance at 254 nm and radioactivity of each fraction.
Figure 3: Biosynthesis of THN by RppA.
Figure 4: Phenotypes of the rppA-disrupted strain of S. griseus.

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Acknowledgements

This work was supported by the Nissan Science Foundation, by the ‘Research for the Future’ Program of JSPS, and by the Bio Design Program from the Ministry of Agriculture, Forestry and Fisheries of Japan.

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Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Bunkyo-ku, 113-8657, Tokyo, Japan

    Nobutaka Funa, Yasuo Ohnishi & Sueharu Horinouchi

  2. Department of Natural Product Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, 113-0033, Tokyo, Japan

    Isao Fujii, Masaaki Shibuya & Yutaka Ebizuka

Authors
  1. Nobutaka Funa
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  2. Yasuo Ohnishi
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  3. Isao Fujii
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  4. Masaaki Shibuya
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  6. Sueharu Horinouchi
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Correspondence to Sueharu Horinouchi.

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Funa, N., Ohnishi, Y., Fujii, I. et al. A new pathway for polyketide synthesis in microorganisms. Nature 400, 897–899 (1999). https://doi.org/10.1038/23748

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