@article{79a96b92896c4a969a5fac6e46b06a6a,
title = "Site-selective chlorination of pyrrolic heterocycles by flavin dependent enzyme PrnC",
abstract = "Halogenation of pyrrole requires strong electrophilic reagents and often leads to undesired polyhalogenated products. Biocatalytic halogenation is a highly attractive approach given its chemoselectivity and benign reaction conditions. While there are several reports of enzymatic phenol and indole halogenation in organic synthesis, corresponding reports on enzymatic pyrrole halogenation have been lacking. Here we describe the in vitro functional and structural characterization of PrnC, a flavin-dependent halogenase that can act on free-standing pyrroles. Computational modeling and site mutagenesis studies identified three key residues in the catalytic pocket. A moderate resolution map using single-particle cryogenic electron microscopy reveals PrnC to be a dimer. This native PrnC can halogenate a library of structurally diverse pyrrolic heterocycles in a site-selective manner and be applied in the chemoenzymatic synthesis of a chlorinated analog of the agrochemical fungicide Fludioxonil.",
author = "Peh, {Guang Rong} and Terence Tay and Tan, {Lee Ling} and Elaine Tiong and Jiawu Bi and Goh, {Yi Ling} and Suming Ye and Fu Lin and Tan, {Cheryl Jia Xin} and Tan, {Yong Zi} and Joel Wong and Huimin Zhao and Wong, {Fong Tian} and Ang, {Ee Lui} and Lim, {Yee Hwee}",
note = "We (Y.H.L., E.L.A., and F.T.W.) gratefully acknowledge financial support from the National Research Foundation, Singapore (NRF-CRP19-2017-05-00) and Agency for Science, Technology and Research (A*STAR), Singapore (C211917006, C211917010 and C233017004) for this work. F.L. thanks A*STAR BII and NSCC (13002658) for the generous and strong support of computational resources. Y.Z.T. and C.J.X.T. gratefully acknowledge funding from the National Research Foundation, Singapore (NRF Fellowship A-8001346-00-00), National University of Singapore (PYP Fellowship A-0008405-00-00, A-0008405-01-00), Ministry of Education, Singapore (MOE AcRF Tier 1 A-8000037-00-00, MOE AcRF Tier 2 A-6100427-01-00); Agency for Science, Technology and Research, Singapore. J.B. gratefully acknowledges A*STAR Graduate Academy (A*GA) for his PhD scholarship. Lastly, the authors would also like to thank Prashant Deshmukh for help with preparing the protein for cryo-EM, Shi Jian for cryo-EM data collection at NUS Centre for Bioimaging Sciences (CBIS), and Dr Choon Boon Cheong (ISCE2) for some of the NMR analysis. The authors acknowledge the insightful suggestions and control experiments provided by the anonymous reviewers and Professor Jared Lewis (Indiana University), which enhanced the quality of this manuscript. We (Y.H.L., E.L.A., and F.T.W.) gratefully acknowledge financial support from the National Research Foundation, Singapore (NRF-CRP19-2017-05-00) and Agency for Science, Technology and Research (A*STAR), Singapore (C211917006, C211917010 and C233017004) for this work. F.L. thanks A*STAR BII and NSCC (13002658) for the generous and strong support of computational resources. Y.Z.T. and C.J.X.T. gratefully acknowledge funding from the National Research Foundation, Singapore (NRF Fellowship A-8001346-00-00), National University of Singapore (PYP Fellowship A-0008405-00-00, A-0008405-01-00), Ministry of Education, Singapore (MOE AcRF Tier 1 A-8000037-00-00, MOE AcRF Tier 2 A-6100427-01-00); Agency for Science, Technology and Research, Singapore. J.B. gratefully acknowledges A*STAR Graduate Academy (A*GA) for his PhD scholarship. Lastly, the authors would also like to thank Prashant Deshmukh for help with preparing the protein for cryo-EM, Shi Jian for cryo-EM data collection at NUS Centre for Bioimaging Sciences (CBIS), and Dr Choon Boon Cheong (ISCE) for some of the NMR analysis. The authors acknowledge the insightful suggestions and control experiments provided by the anonymous reviewers and Professor Jared Lewis (Indiana University), which enhanced the quality of this manuscript. 2",
year = "2024",
month = dec,
doi = "10.1038/s42004-023-01083-1",
language = "English (US)",
volume = "7",
journal = "Communications Chemistry",
issn = "2399-3669",
publisher = "Springer Nature",
number = "1",
}