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Para-position derivatives of fungal anthelmintic cyclodepsipeptides engineered with Streptomyces venezuelae antibiotic biosynthetic genes

Nature Biotechnology volume 22, pages 848–855 (2004)Cite this article

Abstract

PF1022A, a cyclooctadepsipeptide possessing strong anthelmintic properties and produced by the filamentous fungus Rosellinia sp. PF1022, consists of four alternating residues of N-methyl-L-leucine and four residues of D-lactate or D-phenyllactate. PF1022A derivatives obtained through modification of their benzene ring at the para-position with nitro or amino groups act as valuable starting materials for the synthesis of compounds with improved anthelmintic activities. Here we describe the production of such derivatives by fermentation through metabolic engineering of the PF1022A biosynthetic pathway in Rosellinia sp. PF1022. Three genes cloned from Streptomyces venezuelae, and required for the biosynthesis of p-aminophenylpyruvate from chorismate in the chloramphenicol biosynthetic pathway, were expressed in a chorismate mutase–deficient strain derived from Rosellinia sp. PF1022. Liquid chromatography–mass spectrometry and NMR analyses confirmed that this approach facilitated the production of PF1022A derivatives specifically modified at the para-position. This fermentation method is environmentally safe and can be used for the industrial scale production of PF1022A derivatives.

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Figure 1
Figure 2: Biosynthetic pathway of PF1022A and strategy for fermentation of unnatural PF1022A derivatives.
Figure 3: Restriction maps and Southern blot analysis.
Figure 4: Schematic representation of the cluster genes and plasmids.
Figure 5: Identification of PF1022A derivatives in the culture broth of the TF-11 strain.

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Acknowledgements

We thank M. Ohyama and O. Sakanaka for the synthesis of PF1022A derivatives and p-substituted phenyllactates, N. Sano for the LC-MS analysis, S. Miki for the HRMS analysis, S. Gomi for the NMR analysis, H. Nakazato for help in the isolation of the tef1 gene, T. Matsunobu and S. Amano for the purification of PF1022-220 and C.J. Thompson for helpful discussions and manuscript reading.

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  1. Microbiological Resources and Technology Laboratories, Meiji Seika Kaisha, Ltd., 788 Kayama, Odawara-shi, Kanagawa, 250-0852, Japan

    Koji Yanai, Naomi Sumida, Kaoru Okakura, Tatsuki Moriya, Manabu Watanabe & Takeshi Murakami

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Correspondence to Koji Yanai.

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Yanai, K., Sumida, N., Okakura, K. et al. Para-position derivatives of fungal anthelmintic cyclodepsipeptides engineered with Streptomyces venezuelae antibiotic biosynthetic genes. Nat Biotechnol 22, 848–855 (2004). https://doi.org/10.1038/nbt978

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