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Capability of a large bacterial artificial chromosome clone harboring multiple biosynthetic gene clusters for the production of diverse compounds

The Journal of Antibiotics volume 77, pages 288–298 (2024)Cite this article

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Abstract

The biosynthetic gene clusters (BGCs) for the macrocyclic lactone-based polyketide compounds are extremely large-sized because the polyketide synthases that generate the polyketide chains of the basic backbone are of very high molecular weight. In developing a heterologous expression system for the large BGCs amenable to the production of such natural products, we selected concanamycin as an appropriate target. We obtained a bacterial artificial chromosome (BAC) clone with a 211-kb insert harboring the entire BGC responsible for the biosynthesis of concanamycin. Heterologous expression of this clone in a host strain, Streptomyces avermitilis SUKA32, permitted the production of concanamycin, as well as that of two additional aromatic polyketides. Structural elucidation identified these additional products as ent-gephyromycin and a novel compound that was designated JBIR-157. We describe herein sequencing and expression studies performed on these BGCs, demonstrating the utility of large BAC clones for the heterologous expression of cryptic or near-silent loci.

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Acknowledgements

MI deceased on 23 December 2015. This work was supported by AMED under Grant JP19ae0101045 to KS and JSPS KAKENHI Grant Number JP23H04569 to KK.

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Author notes

  1. Takehiro Nishimura

    Present address: Faculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Minato-ku, Tokyo, 105-8512, Japan

  2. Haruo Ikeda

    Present address: Technology Research Association for Next Generation Natural Products Chemistry, 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan

  3. Deceased: Miho Izumikawa.

Authors and Affiliations

  1. Department of Life Science and Biotechnology, National Institute of Advanced Industrial Science and Technology (AIST), 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan

    Kei Kudo, Hikaru Suenaga & Kazuo Shin-ya

  2. Technology Research Association for Next Generation Natural Products Chemistry, 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan

    Takehiro Nishimura & Kazuo Shin-ya

  3. Japan Biological Informatics Consortium (JBIC), 2-4-32 Aomi, Koto-ku, Tokyo, 135-0064, Japan

    Miho Izumikawa, Ikuko Kozone & Junko Hashimoto

  4. Okinawa Institute of Science and Technology Graduate University, 1919-1 Tancha, Onna-son, Kunigami-gun, Okinawa, 904-0495, Japan

    Manabu Fujie & Nori Satoh

  5. Kitasato Institute for Life Sciences, Kitasato University, 1-15-1 Kitasato, Minami-ku, Sagamihara, Kanagawa, 252-0373, Japan

    Haruo Ikeda

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Correspondence to Kazuo Shin-ya.

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Kudo, K., Nishimura, T., Izumikawa, M. et al. Capability of a large bacterial artificial chromosome clone harboring multiple biosynthetic gene clusters for the production of diverse compounds. J Antibiot 77, 288–298 (2024). https://doi.org/10.1038/s41429-024-00711-9

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