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Biochemical Characterization of a Type III Polyketide Biosynthetic Gene Cluster from Streptomyces toxytricini

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Abstract

A type III polyketide biosynthetic gene cluster has been discovered in the industrially important strain Streptomyces toxytricini NRRL 15443, including four genes stp450-1, stts, stp450-2, and stmo. The stts gene encodes a putative type III polyketide synthase that is homologous to RppA, a 1,3,6,8-tetrahydroxynaphthalene (THN) synthase from Streptomyces griseus. The deduced protein product of stmo resembles the cupin-containing monooxygenase MomA from Streptomyces antibioticus that oxidizes THN into flaviolin. Two cytochrome P450s (CYPs), StP450-1 and StP450-2, are present in the gene cluster. StTS was overexpressed in Escherichia coli BL21(DE3) and identified as a THN synthase. The synthesized THN can be easily oxidized into flaviolin by air. Both CYPs were reconstituted in E. coli BL21(DE3) and can oxidize flaviolin to form oligomers. The k cat/K m values for StP450-1 and StP450-2 were 0.28 and 0.71 min−1 mM−1, respectively. UV irradiation test showed that expression of StTS in E. coli BL21(DE3) significantly protects the cells from UV radiation, and coexpression of StTS and StP450-1 provides even stronger protection.

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References

  1. Staunton, J., & Weissman, K. J. (2001). Natural Product Reports, 18, 380–416.

    Article  CAS  Google Scholar 

  2. Zhan, J. (2009). Current Topics in Medicinal Chemistry, 9, 1958–1610.

    Article  Google Scholar 

  3. Austin, M. B., & Noel, A. J. P. (2003). Natural Product Reports, 20, 79–110.

    Article  CAS  Google Scholar 

  4. Funa, N., Ohnishi, Y., Fujii, I., Shibuya, M., Ebizuka, Y., & Horinouchi, S. (1999). Nature, 400, 897–899.

    Article  CAS  Google Scholar 

  5. Funa, N., Ohnishi, Y., Ebizuka, Y., & Horinouchi, S. (2002). Journal of Biological Chemistry, 277, 4628–4635.

    Article  CAS  Google Scholar 

  6. Cortes, J., Velasco, J., Foster, G., Blackaby, A. P., Rudd, B. A. M., & Wilkinson, B. (2002). Molecular Microbiology, 44, 1213–1224.

    Article  CAS  Google Scholar 

  7. Li, S., Grüschow, S., Dordick, J. S., & Sherman, D. H. (2007). Journal of Biological Chemistry, 282, 12765–12772.

    Article  CAS  Google Scholar 

  8. Gross, F., Luniak, N., Perlova, O., Gaitatzis, N., Jenke-Kodama, H., Gerth, K., Gottschalk, D., Dittmann, E., & Muller, R. (2006). Archives of Microbiology, 185, 28–38.

    Article  CAS  Google Scholar 

  9. Harp, J. B. (1999). Drugs of Today, 35, 139–145.

    CAS  Google Scholar 

  10. Omura, T., & Sato, R. (1964). Journal of Biological Chemistry, 239, 2370–2378.

    CAS  Google Scholar 

  11. Davis, N. K., & Chater, K. F. (1990). Molecular Microbiology, 4, 1679–1691.

    Article  CAS  Google Scholar 

  12. Bergh, S., & Uhlén, M. (1992). Gene, 117, 131–136.

    Article  CAS  Google Scholar 

  13. Winter, J. M., Moffitt, M. C., Zazopoulos, E., McAlpine, J. B., Dorrestein, P. C., & Moore, B. S. (2007). Journal of Biological Chemistry, 282, 16362–16368.

    Article  CAS  Google Scholar 

  14. Snyder, S. A., Tang, Z. Y., & Gupta, R. (2009). Journal of the American Chemical Society, 131, 5744–5745.

    Article  CAS  Google Scholar 

  15. Zhao, B., Lamb, D. C., Lei, L., Kelly, S. L., Yuan, H., Hachey, D. L., & Waterman, M. R. (2007). Biochemistry, 46, 8725–8733.

    Article  CAS  Google Scholar 

  16. Zhao, B., Guengerich, F. P., Bellamine, A., Lamb, D. C., Izumikawa, M., Lei, L., Podust, L. M., Sundaramoorthy, M., Kalaitzis, J. A., Reddy, L. M., Kelly, S. L., Moore, B. S., Stec, D., Voehler, M., Falck, J. R., Shimada, T., & Waterman, M. R. (2005). Journal of Biological Chemistry, 280, 11599–11607.

    Article  CAS  Google Scholar 

  17. Funa, N., Funabashi, M., Ohnishi, Y., & Horinouchi, S. (2005). Journal of Bacteriology, 187, 8149–8155.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by a Utah State University Research Catalyst Grant. Richard Decker was supported by a Utah State University Engineering Undergraduate Research Program (EURP) Grant. We thank Prof. Lance Seefeldt and Dr. Zhiyong Yang in the Department of Chemistry and Biochemistry at Utah State University for their assistance in the CO difference spectrum assays.

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  1. Department of Biological Engineering, Utah State University, 4105 Old Main Hill, Logan, UT, 84322-4105, USA

    Jia Zeng, Richard Decker & Jixun Zhan

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  1. Jia Zeng
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  2. Richard Decker
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  3. Jixun Zhan
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Correspondence to Jixun Zhan.

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Zeng, J., Decker, R. & Zhan, J. Biochemical Characterization of a Type III Polyketide Biosynthetic Gene Cluster from Streptomyces toxytricini . Appl Biochem Biotechnol 166, 1020–1033 (2012). https://doi.org/10.1007/s12010-011-9490-x

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