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PMID: 9023192 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Cloning, sequencing, and analysis of a gene cluster from Chelatobacter heintzii ATCC 29600 encoding nitrilotriacetate monooxygenase and NADH:flavin mononucleotide oxidoreductase.

Journal of bacteriology ·Vol. 179 ·No. 4 ·1997-02-00 ·Pages 1112-6

Xu Y, Mortimer MW, Fisher TS, Kahn ML, Brockman FJ, Xun L

Abstract

Nitrilotriacetate (NTA) is an important chelating agent in detergents and has also been used extensively in processing radionuclides. In Chelatobacter heintzii ATCC 29600, biodegradation of NTA is initiated by NTA monooxygenase that oxidizes NTA to iminodiacetate and glyoxylate. The NTA monooxygenase activity requires two component proteins, component A and component B, but the function of each component is unclear. We have cloned and sequenced a gene cluster encoding components A and B (nmoA and nmoB) and two additional open reading frames, nmoR and nmoT, downstream of nmoA. Based on sequence similarities, nmoR and nmoT probably encode a regulatory protein and a transposase, respectively. The NmoA sequence was similar to a monooxygenase that uses reduced flavin mononucleotide (FMNH2) as reductant; NmoB was similar to an NADH:flavin mononucleotide (FMN) oxidoreductase. On the basis of this information, we tested the function of each component. Purified component B was shown to be an NADH:FMN oxidoreductase, and its activity could be separated from that of component A. When the Photobacterium fischeri NADH:FMN oxidoreductase was substituted for component B in the complete reaction, NTA was oxidized, showing that the substrate specificity of the reaction resides in component A. Component A is therefore an NTA monooxygenase that uses FMNH2 and O2 to oxidize NTA, and component B is an NADH:FMN oxidoreductase that provides FMNH2 for NTA oxidation.

MeSH Terms
Amino Acid Sequence Base Sequence Biodegradation, Environmental Cloning, Molecular FMN Reductase Flavin Mononucleotide/metabolism Gene Expression Regulation, Bacterial Genes, Bacterial Gram-Negative Aerobic Bacteria/enzymology,genetics Mixed Function Oxygenases/chemistry,genetics,metabolism Molecular Sequence Data Molecular Weight Multigene Family NAD/metabolism NADH, NADPH Oxidoreductases/chemistry,genetics,metabolism Nitrilotriacetic Acid/metabolism Open Reading Frames Oxidation-Reduction Recombinant Proteins/metabolism Sequence Alignment
Chemicals
Recombinant Proteins NAD Flavin Mononucleotide Mixed Function Oxygenases nitrilotriacetate monooxygenase FMN Reductase NADH, NADPH Oxidoreductases Nitrilotriacetic Acid
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Xu Y
Department of Microbiology, Washington State University Tri-Cities, Richland 99352, USA.
Mortimer M W
Fisher T S
Kahn M L
Brockman F J
Xun L
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.4.1112-1116.1997
Published
1997-02-00
Pages
1112-6
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178806
Subset
IM
Databases
GENBANK
L49438
External Links
PubMed source DOI: 10.1128/jb.179.4.1112-1116.1997 PMC Full Text View journal details
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