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Precursor-directed biosynthesis of 6-deoxyerythronolide B analogues is improved by removal of the initial catalytic sites of the polyketide synthase

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Journal of Industrial Microbiology & Biotechnology

Abstract

Precursor-directed biosynthesis has been shown to be a powerful tool for the production of polyketide analogues that would be difficult or cost prohibitive to produce from medicinal chemistry efforts alone. It has been most extensively demonstrated using a KS1 null mutation (KS10) to block the first round of condensation in the biosynthesis of the erythromycin polyketide synthase (DEBS) for the production of analogues of its aglycone, 6-deoxyerythronolide B (6-dEB). Here we show that removing the DEBS loading domain and first module (mod1Δ), rather than using the KS10 system, can lead to an increase in the utilization of some chemical precursors and production of 6-dEB analogues (R-6dEB) in both Streptomyces coelicolor and Saccharopolyspora erythraea. While the difference in utilization of the precursor was diketide specific, in strains fed (2R*, 3S*)-5-fluoro-3-hydroxy-2-methylpentanoate N-propionylcysteamine thioester, twofold increases in both utilization of the diketide and 15-fluoro-6dEB (15F-6dEB) production were observed in S. coelicolor, and S. erythraea exhibited a tenfold increase in production of 15-fluoro-erythromycin when utilizing the mod1Δ rather than the KS10 system.

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Acknowledgments

We would like to thank John Carney and Chau Tran for assistance with compound analysis, Chaitan Khosla for the gift of DEBS1+TE polyclonal antibody, and Sumati Murli for her helpful comments during the preparation of this manuscript.

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Authors and Affiliations

  1. Kosan Biosciences Inc., 3832 Bay Center Place, Hayward, CA, 94545, USA

    Shannon L. Ward, Ruchir P. Desai, Zhihao Hu & Leonard Katz

  2. Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad de Rosario, Suipacha 531, 2000, Rosario, Argentina

    Hugo Gramajo

Authors
  1. Shannon L. Ward
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  2. Ruchir P. Desai
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  3. Zhihao Hu
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  4. Hugo Gramajo
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  5. Leonard Katz
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Correspondence to Shannon L. Ward.

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Ward, S.L., Desai, R.P., Hu, Z. et al. Precursor-directed biosynthesis of 6-deoxyerythronolide B analogues is improved by removal of the initial catalytic sites of the polyketide synthase. J Ind Microbiol Biotechnol 34, 9–15 (2007). https://doi.org/10.1007/s10295-006-0156-6

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  • DOI: https://doi.org/10.1007/s10295-006-0156-6

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