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Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides

Author

Listed:
  • Wan-Qiu Liu

    (ShanghaiTech University)

  • Xiangyang Ji

    (ShanghaiTech University)

  • Fang Ba

    (ShanghaiTech University)

  • Yufei Zhang

    (ShanghaiTech University)

  • Huiling Xu

    (ShanghaiTech University)

  • Shuhui Huang

    (ShanghaiTech University)

  • Xiao Zheng

    (ShanghaiTech University)

  • Yifan Liu

    (ShanghaiTech University
    ShanghaiTech University
    Shanghai Clinical Research and Trial Center)

  • Shengjie Ling

    (ShanghaiTech University
    ShanghaiTech University
    Shanghai Clinical Research and Trial Center)

  • Michael C. Jewett

    (Stanford University)

  • Jian Li

    (ShanghaiTech University
    ShanghaiTech University
    Shanghai Clinical Research and Trial Center)

Abstract

Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a major class of natural products with diverse chemical structures and potent biological activities. A vast majority of RiPP gene clusters remain unexplored in microbial genomes, which is partially due to the lack of rapid and efficient heterologous expression systems for RiPP characterization and biosynthesis. Here, we report a unified biocatalysis (UniBioCat) system based on cell-free gene expression for rapid biosynthesis and engineering of RiPPs. We demonstrate UniBioCat by reconstituting a full biosynthetic pathway for de novo biosynthesis of salivaricin B, a lanthipeptide RiPP. Next, we delete several protease/peptidase genes from the source strain to enhance the performance of UniBioCat, which then can synthesize and screen salivaricin B variants with enhanced antimicrobial activity. Finally, we show that UniBioCat is generalizable by synthesizing and evaluating the bioactivity of ten uncharacterized lanthipeptides. We expect UniBioCat to accelerate the discovery, characterization, and synthesis of RiPPs.

Suggested Citation

  • Wan-Qiu Liu & Xiangyang Ji & Fang Ba & Yufei Zhang & Huiling Xu & Shuhui Huang & Xiao Zheng & Yifan Liu & Shengjie Ling & Michael C. Jewett & Jian Li, 2024. "Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-48726-y
    DOI: 10.1038/s41467-024-48726-y
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