y-Glutamylmethylamide Is an Essential Intermediate in the Metabolism of Methylamine by Methylocella silvestris

Chen, Y, Scanlan, J, Song, L, Crombie, A, Rahman, MT, Schafer, H and Murrell, JC (2010) y-Glutamylmethylamide Is an Essential Intermediate in the Metabolism of Methylamine by Methylocella silvestris. Applied and Environmental Microbiology, 76 (13). pp. 4530-4537. ISSN 0099-2240

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Abstract

Methylocella silvestris BL2, a facultative methane utilizer, can grow on monomethylamine (MMA) as a sole carbon and nitrogen source. No activity of MMA dehydrogenase was detectable. Instead, this bacterium utilizes a methylated amino acid pathway (?-glutamylmethylamide [GMA] and N-methylglutamate [NMG]) for MMA metabolism. The activities of the two key enzymes in this pathway, GMA synthetase and NMG dehydrogenase, were found when the bacterium was grown on MMA. GMA was detected by high-performance liquid chromatography-mass spectrometry only when the bacterium was grown on MMA but not when it was grown on methanol. Proteomic analysis of soluble and membrane fractions of the proteome from MMA- and methanol-grown cultures revealed that an eight-gene cluster (Msil2632 to Msil2639) was induced by MMA and cotranscribed as an operon, as shown by reverse transcription-PCR. GMA-dissimilating enzyme activity was also detected when it was grown on MMA. Formaldehyde and ammonium production from GMA was dependent on glutamate but not on a-ketoglutarate. Marker exchange mutagenesis of a putative GMAS gene homologue (gmas, Msil2635) within this eight-gene cluster, with a kanamycin gene cassette, abolished growth of M. silvestris on MMA as either a sole carbon or a sole nitrogen source. Overall, our results suggest that gmas is essential in MMA metabolism by M. silvestris.

Item Type: Article
Faculty \ School: Faculty of Science > School of Environmental Sciences
University of East Anglia > Faculty of Science > Research Centres > Centre for Ecology, Evolution and Conservation
University of East Anglia > Faculty of Science > Research Groups > Environmental Biology
University of East Anglia > Faculty of Science > Research Groups > Resources, Sustainability and Governance
University of East Anglia > Faculty of Science > Research Groups > Marine and Atmospheric Sciences
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Depositing User: Rhiannon Harvey
Date Deposited: 27 Mar 2012 12:07
Last Modified: 25 Jul 2018 04:11
URI: https://ueaeprints.uea.ac.uk/id/eprint/38491
DOI: 10.1128/AEM.00739-10

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