Latypova, Ekaterina, Yang, Song, Wang, Yi-Shun, Wang, Tiansong, Chavkin, Theodore A., Hackett, Murray, Schaefer, Hendrik and Kalyuzhnaya, Marina G.. (2010) Genetics of the glutamate-mediated methylamine utilization pathway in the facultative methylotrophic beta-proteobacterium Methyloversatilis universalis FAM5. Molecular Microbiology, Vol.75 (No.2). pp. 426-439. ISSN 0950-382X

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Abstract

The ability of some microbial species to oxidize monomethylamine via glutamate-mediated pathways was proposed in the 1960s; however, genetic determinants of the pathways have never been described. In the present study we describe a gene cluster essential for operation of the N-methylglutamate pathway in the methylotrophic beta-proteobacterium Methyloversatilis universalis FAM5. Four major polypeptides from protein fractions displaying high activities of N-methylglutamate synthetase, N-methylglutamate dehydrogenase and g-glutamylmethylamide synthetase were selected for mass spectrometry-based identification. The activities of enzymes were associated with the presence of peptides identified as ferredoxin-dependent glutamate synthase (GltB2), large subunit of putative heterotetrameric sarcosine oxidase (SoxA) and glutamine synthetase type III (GSIII) respectively. A gene cluster (8.3 kb) harbouring gltB2, soxA and gsIII-like genes was amplified from M. universalis FAM5, sequenced and assembled. Two partial and six complete open reading frames arranged in the order soxBDAG-gsIII-gltB132 were identified and subjected to mutational analysis, functional and metabolic profiling. We demonstrated that gltB-like and sox-like genes play a key role in methylamine utilization and encode N-methylglutamate synthetase and N-methylglutamate dehydrogenase respectively. Metabolic, enzymatic and mutational analyses showed that the gsIII-like gene encodes g-glutamylmethylamide synthetase; however, this enzyme is not essential for oxidation of methylamine.

Item Type:	Journal Article
Subjects:	Q Science > QD Chemistry Q Science > QR Microbiology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)
Journal or Publication Title:	Molecular Microbiology
Publisher:	Wiley-Blackwell Publishing Ltd.
ISSN:	0950-382X
Date:	January 2010
Volume:	Vol.75
Number:	No.2
Number of Pages:	14
Page Range:	pp. 426-439
Identification Number:	10.1111/j.1365-2958.2009.06989.x
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	Royalty Research Fund, National Science Foundation, NSF, NIGMS
Grant number:	65-1818, MCB-0842686, MCB-06044269, GM-58933
URI:	http://wrap.warwick.ac.uk/id/eprint/16584

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