Identification of the Final Two Genes Functioning in Methanofuran Biosynthesis in Methanocaldococcus jannaschii

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dc.contributorVirginia Techen
dc.contributor.authorWang, Yuen
dc.contributor.authorXu, Huiminen
dc.contributor.authorJones, Michael K.en
dc.contributor.authorWhite, Robert H.en
dc.contributor.departmentBiochemistryen
dc.date.accessed2015-10-01en
dc.date.accessioned2015-11-28T21:53:48Zen
dc.date.available2015-11-28T21:53:48Zen
dc.date.issued2015-06-22en
dc.description.abstractAll methanofuran structural variants contain a basic core structure of 4-[N-(gamma -L-glutamyl)-p-(beta-aminoethyl) phenoxymethyl]( aminomethyl) furan (APMF-Glu) but have different side chains depending on the source organism. Recently, we identified four genes (MfnA, MfnB, MfnC, and MfnD) that are responsible for the biosynthesis of the methanofuran precursor gamma-glutamyltyramine and 5-(aminomethyl)-3-furanmethanol-phosphate (F1-P) from tyrosine, glutamate, glyceraldehyde-3-P, and alanine in Methanocaldococcus jannaschii. How gamma-glutamyltyramine and F1-P couple together to form the core structure of methanofuran was previously unknown. Here, we report the identification of two enzymes encoded by the genes mj0458 and mj0840 that catalyze the formation of F1-PP from ATP and F1-P and the condensation of F1-PP with gamma-glutamyltyramine, respectively, to form APMF-Glu. We have annotated these enzymes as MfnE and MfnF, respectively, representing the fifth and sixth enzymes in the methanofuran biosynthetic pathway to be identified. Although MfnE was previously reported as an archaeal adenylate kinase, our present results show that MfnE is a promiscuous enzyme and that its possible physiological role is to produce F1-PP. Unlike other enzymes catalyzing coupling reactions involving pyrophosphate as the leaving group, MfnF exhibits a distinctive alpha/beta two-layer sandwich structure. By comparing MfnF with thiamine synthase and dihydropteroate synthase, a substitution nucleophilic unimolecular (S-N-1) reaction mechanism is proposed for MfnF. With the identification of MfnE and MfnF, the biosynthetic pathway for the methanofuran core structure APMF-Glu is complete. IMPORTANCE This work describes the identification of the final two enzymes responsible for catalyzing the biosynthesis of the core structure of methanofuran. The gene products of mj0458 and mj0840 catalyze the formation of F1-PP and the coupling of F1-PP with gamma-glutamyltyramine, respectively, to form APMF-Glu. Although the chemistry of such a coupling reaction is widespread in biochemistry, we provide here the first evidence that such a mechanism is used in methanofuran biosynthesis. MfnF belongs to the hydantoinase A family (PF01968) and exhibits a unique alpha/beta two-layer sandwich structure that is different from the enzymes catalyzing similar reactions. Our results show that MfnF catalyzes the formation of an ether bond during methanofuran biosynthesis. Therefore, this work further expands the functionality of this enzyme family.en
dc.description.sponsorshipNational Science Foundationen
dc.description.sponsorshipMCB 1120346en
dc.description.sponsorshipVirginia Tech. Fralin Life Science Instituteen
dc.description.sponsorshipAgricultural Experiment Station Hatch Programen
dc.description.sponsorshipVA-135981en
dc.format.mimetypeapplication/pdfen
dc.identifier.citationWang, Yu, et al. (2015). Identification of the Final Two Genes Functioning in Methanofuran Biosynthesis in Methanocaldococcus jannaschii. Journal of Bacteriology, 197(17), 2850-2858. doi:10.1128/jb.00401-15en
dc.identifier.doihttps://doi.org/10.1128/jb.00401-15en
dc.identifier.issn0021-9193en
dc.identifier.urihttp://hdl.handle.net/10919/64214en
dc.identifier.urlhttp://jb.asm.org/content/197/17/2850en
dc.language.isoenen
dc.publisherAmerican Society for Microbiologyen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.titleIdentification of the Final Two Genes Functioning in Methanofuran Biosynthesis in Methanocaldococcus jannaschiien
dc.title.serialJournal of Bacteriologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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