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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.

Author information

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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