). (2015) XoxFencoding an alternative methanol dehydrogenase is widespread in coastal marine environments. Environmental Microbiology, 17 (10). pp. 3937-3948. Permanent WRAP URL:http://wrap.warwick.ac.uk/67990 Copyright and reuse:The Warwick Research Archive Portal (WRAP) makes this work by researchers of the University of Warwick available open access under the following conditions. Copyright © and all moral rights to the version of the paper presented here belong to the individual author(s) and/or other copyright owners. To the extent reasonable and practicable the material made available in WRAP has been checked for eligibility before being made available.Copies of full items can be used for personal research or study, educational, or not-for profit purposes without prior permission or charge. Provided that the authors, title and full bibliographic details are credited, a hyperlink and/or URL is given for the original metadata page and the content is not changed in any way. Publisher's statement:"This is the peer reviewed version of the following article: which has been published in final form at http://dx.doi.org/10.1111/1462-2920.12896 This article may be used for noncommercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving." A note on versions:The version presented here may differ from the published version or, version of record, if you wish to cite this item you are advised to consult the publisher's version. Please see the 'permanent WRAP url' above for details on accessing the published version and note that access may require a subscription. Abstract 22The xoxF gene, encoding a pyrroloquinoline quinone-dependent methanol dehydrogenase, is found in all 23 known proteobacterial methylotrophs. In several newly discovered methylotrophs, XoxF is the active methanol 24 dehydrogenase, catalysing the oxidation of methanol to formaldehyde. Apart from that, its potential role in 25 methylotrophy and carbon cycling is unknown. So far, the diversity of xoxF in the environment has received 26 little attention. We designed PCR primer sets targeting clades of the xoxF gene, and used 454 pyrosequencing 27 of PCR amplicons obtained from DNA of four coastal marine environments for a unique assessment of the 28 diversity of xoxF in these habitats. Phylogenetic analysis of the data obtained revealed a high diversity of xoxF 29 genes from two of the investigated clades, and substantial differences in sequence composition between 30 environments. Sequences were classified as being related to a wide range of both methylotrophs and non-31 methylotrophs from Alpha-, Beta-and Gammaproteobacteria. The most prominent sequences detected were 32 related to the family Rhodobacteraceae, the genus Methylotenera and the OM43 clade of Methylophilales, 33 and are thus related to organisms that employ XoxF for methanol oxidation. Furthermore, our analyses 34 revealed a high degree of so far undescribed sequences, suggesting a high number of unknown species in 35 these habitats. 36

Nitrogen is a key limiting resource for biomass production in the marine environment. Methylated amines, released from the degradation of osmolytes, could provide a nitrogen source for marine microbes. Thus far, studies in aquatic habitats on the utilization of methylamine, the simplest methylated amine, have mainly focussed on the fate of the carbon from this compound. Various groups of methylotrophs, microorganisms that can grow on one-carbon compounds, use methylamine as a carbon source. Non-methylotrophic microorganisms may also utilize methylamine as a nitrogen source, but little is known about their diversity, especially in the marine environment. In this proof-of-concept study, stable isotope probing (SIP) was used to identify microorganisms from a coastal environment that assimilate nitrogen from methylamine. SIP experiments using N methylamine combined with metagenomics and metaproteomics facilitated identification of active methylamine-utilizing Alpha- and Gammaproteobacteria. The draft genomes of two methylamine utilizers were obtained and their metabolism with respect to methylamine was examined. Both bacteria identified in these SIP experiments used the γ-glutamyl-methylamide pathway, found in both methylotrophs and non-methylotrophs, to metabolize methylamine. The utilization of N methylamine also led to the release of N ammonium that was used as nitrogen source by other microorganisms not directly using methylamine.

Copies of full items can be used for personal research or study, educational, or not-for profit purposes without prior permission or charge. Provided that the authors, title and full bibliographic details are credited, a hyperlink and/or URL is given for the original metadata page and the content is not changed in any way. Publisher's statement:"This is the peer reviewed version of the following article: which has been published in final form at http://dx.doi.org/10.1111/1462-2920.12935 This article may be used for noncommercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving." A note on versions:The version presented here may differ from the published version or, version of record, if you wish to cite this item you are advised to consult the publisher's version. Please see the 'permanent WRAP url' above for details on accessing the published version and note that access may require a subscription.