@article{5c2effae9466439e991f3856634684b5,
title = "Reshaping the 2-Pyrone Synthase Active Site for Chemoselective Biosynthesis of Polyketides",
abstract = "Engineering enzymes with novel reactivity and applying them in metabolic pathways to produce valuable products are quite challenging due to the intrinsic complexity of metabolic networks and the need for high in vivo catalytic efficiency. Triacetic acid lactone (TAL), naturally generated by 2-pyrone synthase (2PS), is a platform molecule that can be produced via microbial fermentation and further converted into value-added products. However, these conversions require extra synthetic steps under harsh conditions. We herein report a biocatalytic system for direct generation of TAL derivatives under mild conditions with controlled chemoselectivity by rationally engineering the 2PS active site and then rewiring the biocatalytic pathway in the metabolic network of E. coli to produce high-value products, such as kavalactone precursors, with yields up to 17 mg/L culture. Computer modeling indicates sterics and hydrogen-bond interactions play key roles in tuning the selectivity, efficiency and yield.",
keywords = "biocatalysis, metabolic engineering, polyketides, synthetic biology",
author = "Yu Zhou and Mirts, {Evan N.} and Sangdo Yook and Matthew Waugh and Rachel Martini and Jin, {Yong Su} and Yi Lu",
note = "Funding Information: We wish to thank Prof. Claudia Schmidt‐Dannert for providing pAC‐4CL1, Prof. Markus Jeschek for providing pCKmatBC, Prof. Jing‐Ke Weng for providing PmSPS1, Prof. Emily Que and Sky Price for HPLC analysis, Linggen Kong for the suggestions in molecular biology and synthetic biology work, Hirbod Heidari for the assistance in protein purification, Dr. Yunling Deng, Dr. Christopher Reed, Dr. Aaron Ledray, Mandira Banik, Whitney Lewis for their help with revising the manuscript. The work described in this report is supported by the DOE Center for Advanced Bioenergy and Bioproducts Innovation (U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research under Award Number DE‐SC0018420). Any opinions, findings, and conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the views of the U.S. Department of Energy. We also thank the Robert A. Welch Foundation (Grant F‐0020) for support of the Lu group research program at the University of Texas at Austin and NIH (1S10OD021508‐1) for purchasing the Bruker AVANCE III 500 NMR instrument. Publisher Copyright: {\textcopyright} 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.",
year = "2023",
month = jan,
day = "26",
doi = "10.1002/anie.202212440",
language = "English (US)",
volume = "62",
journal = "Angewandte Chemie - International Edition",
issn = "1433-7851",
publisher = "John Wiley & Sons, Ltd.",
number = "5",
}