Flavin‐N5 Covalent Intermediate in a Nonredox Dehalogenation Reaction Catalyzed by an Atypical Flavoenzyme |
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Authors: | Dr Yumin Dai Dr Karina Kizjakina Ashley C Campbell Dr David A Korasick Prof Dr John J Tanner Prof Dr Pablo Sobrado |
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Affiliation: | 1. Department of Biochemistry, Virginia Tech, Blacksburg, Virginia, USA;2. Department of Biochemistry, University of Missouri, Columbia, Missouri, USA;3. Department of Chemistry, University of Missouri, Columbia, Missouri, USA |
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Abstract: | The flavin‐dependent enzyme 2‐haloacrylate hydratase (2‐HAH) catalyzes the conversion of 2‐chloroacrylate, a major component in the manufacture of acrylic polymers, to pyruvate. The enzyme was expressed in Escherichia coli, purified, and characterized. 2‐HAH was shown to be monomeric in solution and contained a non‐covalent, yet tightly bound, flavin adenine dinucleotide (FAD). Although the catalyzed reaction was redox‐neutral, 2‐HAH was active only in the reduced state. A covalent flavin‐substrate intermediate, consistent with the flavin‐acrylate iminium ion, was trapped with cyanoborohydride and characterized by mass spectrometry. Small‐angle X‐ray scattering was consistent with 2‐HAH belonging to the succinate dehydrogenase/fumarate reductase family of flavoproteins. These studies establish 2‐HAH as a novel noncanonical flavoenzyme. |
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Keywords: | covalent adduct dehalogenation flavin iminium adduct non-redox reactions |
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