Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase

Se-Young Jun; Kevin M Lewis; Buhyun Youn; Luying Xun; ChulHee Kang

doi:10.1111/mmi.13363

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Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase

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Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase

Se-Young Jun, Kevin M Lewis, Buhyun Youn, Luying Xun and ChulHee Kang

Molecular microbiology, Vol.100(6), pp.989-1003

06/2016

DOI: https://doi.org/10.1111/mmi.13363

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https://hdl.handle.net/2376/113496

PMCID: PMC4899218

PMID: 26928990

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Abstract

Amino Acid Sequence

Catalytic Domain

Oxidoreductases - metabolism

Oxidoreductases Acting on CH-NH Group Donors - chemistry

Models, Molecular

Edetic Acid - metabolism

Mixed Function Oxygenases - metabolism

Oxidoreductases Acting on CH-NH Group Donors - metabolism

Phyllobacteriaceae - metabolism

Structure-Activity Relationship

Hydroquinones - metabolism

FMN Reductase - metabolism

FMN Reductase - chemistry

Molecular Docking Simulation

Catalysis

NADH, NADPH Oxidoreductases - metabolism

Phyllobacteriaceae - enzymology

Flavin Mononucleotide - metabolism

Flavins - metabolism

Ethylenediaminetetraacetate (EDTA) is currently the most abundant organic pollutant due to its recalcitrance and extensive use. Only a few bacteria can degrade it, using EDTA monooxygenase (EmoA) to initiate the degradation. EmoA is an FMNH2 -dependent monooxygenase that requires an NADH:FMN oxidoreductase (EmoB) to provide FMNH2 as a cosubstrate. Although EmoA has been identified from Chelativorans (ex. Mesorhizobium) sp. BNC1, its catalytic mechanism is unknown. Crystal structures of EmoA revealed a domain-like insertion into a TIM-barrel, which might serve as a flexible lid for the active site. Docking of MgEDTA(2-) into EmoA identified an intricate hydrogen bond network connected to Tyr(71) , which should potentially lower its pKa. Tyr(71) , along with nearby Glu(70) and a peroxy flavin, facilitates a keto-enol transition of the leaving acetyl group of EDTA. Further, for the first time, the physical interaction between EmoA and EmoB was observed by ITC, molecular docking and enzyme kinetic assay, which enhanced both EmoA and EmoB activities probably through coupled channelling of FMNH2 .

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Title: Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase
Creators: Se-Young Jun - Department of Chemistry, Washington State University, Pullman, WA, 99164-4630, USA
Kevin M Lewis - Department of Chemistry, Washington State University, Pullman, WA, 99164-4630, USA
Buhyun Youn - Department of Biological Sciences, Pusan National University, Busan, South Korea
Luying Xun - School of Molecular Biosciences, Washington State University, Pullman, WA, 99164-4660, USA
ChulHee Kang - School of Molecular Biosciences, Washington State University, Pullman, WA, 99164-4660, USA
Publication Details: Molecular microbiology, Vol.100(6), pp.989-1003
Academic Unit: Chemistry, Department of; Molecular Biosciences, School of
Publisher: England
Grant note: R01 GM111254 / NIGMS NIH HHS
Identifiers: 99900547716901842
Language: English
Resource Type: Journal article