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

Abstract:

: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 .

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Jun SY,Lewis KM,Youn B,Xun L,Kang C

doi

10.1111/mmi.13363

subject

Has Abstract

pub_date

2016-06-01 00:00:00

pages

989-1003

issue

6

eissn

0950-382X

issn

1365-2958

journal_volume

100

pub_type

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