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Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis

Author

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  • Allwin D. McDonald

    (University of Wisconsin−Madison)

  • Peyton M. Higgins

    (University of Wisconsin−Madison)

  • Andrew R. Buller

    (University of Wisconsin−Madison)

Abstract

Enzymes with high activity are readily produced through protein engineering, but intentionally and efficiently engineering enzymes for an expanded substrate scope is a contemporary challenge. One approach to address this challenge is Substrate Multiplexed Screening (SUMS), where enzyme activity is measured on competing substrates. SUMS has long been used to rigorously quantitate native enzyme specificity, primarily for in vivo settings. SUMS has more recently found sporadic use as a protein engineering approach but has not been widely adopted by the field, despite its potential utility. Here, we develop principles of how to design and interpret SUMS assays to guide protein engineering. This rich information enables improving activity with multiple substrates simultaneously, identifies enzyme variants with altered scope, and indicates potential mutational hot-spots as sites for further engineering. These advances leverage common laboratory equipment and represent a highly accessible and customizable method for enzyme engineering.

Suggested Citation

  • Allwin D. McDonald & Peyton M. Higgins & Andrew R. Buller, 2022. "Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis," Nature Communications, Nature, vol. 13(1), pages 1-11, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-32789-w
    DOI: 10.1038/s41467-022-32789-w
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    References listed on IDEAS

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    1. Emily E. Wrenbeck & Laura R. Azouz & Timothy A. Whitehead, 2017. "Single-mutation fitness landscapes for an enzyme on multiple substrates reveal specificity is globally encoded," Nature Communications, Nature, vol. 8(1), pages 1-10, August.
    2. Corin C. Wagen & Spencer E. McMinn & Eugene E. Kwan & Eric N. Jacobsen, 2022. "Screening for generality in asymmetric catalysis," Nature, Nature, vol. 610(7933), pages 680-686, October.
    3. Linfeng Xu & Kai-Chun Chang & Emory M. Payne & Cyrus Modavi & Leqian Liu & Claire M. Palmer & Nannan Tao & Hal S. Alper & Robert T. Kennedy & Dale S. Cornett & Adam R. Abate, 2021. "Mapping enzyme catalysis with metabolic biosensing," Nature Communications, Nature, vol. 12(1), pages 1-7, December.
    4. Fuqiang Ma & Meng Ting Chung & Yuan Yao & Robert Nidetz & Lap Man Lee & Allen P. Liu & Yan Feng & Katsuo Kurabayashi & Guang-Yu Yang, 2018. "Efficient molecular evolution to generate enantioselective enzymes using a dual-channel microfluidic droplet screening platform," Nature Communications, Nature, vol. 9(1), pages 1-8, December.
    5. Edward Kalkreuter & Kyle S. Bingham & Aaron M. Keeler & Andrew N. Lowell & Jennifer J. Schmidt & David H. Sherman & Gavin J. Williams, 2021. "Computationally-guided exchange of substrate selectivity motifs in a modular polyketide synthase acyltransferase," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
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