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Assessing and harnessing updated polyketide synthase modules through combinatorial engineering

Author

Listed:
  • Katherine A. Ray

    (The University of Texas at Austin)

  • Joshua D. Lutgens

    (The University of Texas at Austin)

  • Ramesh Bista

    (The University of Texas at Austin)

  • Jie Zhang

    (The University of Texas at Austin)

  • Ronak R. Desai

    (The University of Texas at Austin)

  • Melissa Hirsch

    (The University of Texas at Austin)

  • Takeshi Miyazawa

    (The University of Texas at Austin)

  • Antonio Cordova

    (The University of Texas at Austin)

  • Adrian T. Keatinge-Clay

    (The University of Texas at Austin)

Abstract

The modular nature of polyketide assembly lines and the significance of their products make them prime targets for combinatorial engineering. The recently updated module boundary has been successful for engineering short synthases, yet larger synthases constructed using the updated boundary have not been investigated. Here we describe our design and implementation of a BioBricks-like platform to rapidly construct 5 triketide, 25 tetraketide, and 125 pentaketide synthases to test every module combination of the pikromycin synthase. Anticipated products are detected from 60% of the triketide synthases, 32% of the tetraketide synthases, and 6.4% of the pentaketide synthases. We determine ketosynthase gatekeeping and module-skipping are the principal impediments to obtaining functional synthases. The platform is also employed to construct active hybrid synthases by incorporating modules from the erythromycin, spinosyn, and rapamycin assembly lines. The relaxed gatekeeping of a ketosynthase in the rapamycin synthase is especially encouraging in the quest to produce designer polyketides.

Suggested Citation

  • Katherine A. Ray & Joshua D. Lutgens & Ramesh Bista & Jie Zhang & Ronak R. Desai & Melissa Hirsch & Takeshi Miyazawa & Antonio Cordova & Adrian T. Keatinge-Clay, 2024. "Assessing and harnessing updated polyketide synthase modules through combinatorial engineering," Nature Communications, Nature, vol. 15(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-50844-6
    DOI: 10.1038/s41467-024-50844-6
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