{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T11:43:45Z","timestamp":1792496625136,"version":"4.5.0"},"reference-count":56,"publisher":"American Chemical Society (ACS)","issue":"20","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006151","name":"Basic Energy Sciences","doi-asserted-by":"publisher","award":["DE-FG02-95ER20175"],"award-info":[{"award-number":["DE-FG02-95ER20175"]}],"id":[{"id":"10.13039\/100006151","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,20]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Glycerol is abundant in natural environments and is utilized by many microbes. In aerobes, it is typically converted, via glycerol-3-phosphate (G3P), to dihydroxyacetone phosphate (DHAP) by a quinone-linked dehydrogenase thereby coupling glycolysis and respiration. Under anaerobic conditions, an NAD+-dependent dehydrogenase has been proposed to oxidize G3P to DHAP in several bacteria but this reaction is thermodynamically unfavorable. Herein, we have purified a heterotrimeric electron bifurcating enzyme from the archaeon Pyrococcus furiosus where the simultaneous oxidation of both G3P (Eo\u2032 \u2212190 mV) and reduced ferredoxin (Em \u223c \u2212450 mV) enables reduction of NAD+(Eo\u2032 \u2212320 mV) and production of DHAP. Surprisingly, this ferredoxin-dependent NAD+G3P oxidoreductase (FngABC) is unrelated evolutionarily to the four known families of bifurcating enzyme. Analysis by UV\u2013visible spectroscopy and of its predicted structure suggest FngABC has a nonclassical bifurcation mechanism. Phylogenetic and genome context analyses of 4,273 P. furiosus FngA homologues revealed they are widespread with \u223c35% found mainly in Clostridia as FngABC and \u223c10% found mainly in Actinomyces as FngAB, most of which utilize polyols in addition to glycerol. The remaining \u223c40% are present in Gammaproteobacteria and halophilic archaea but here FngA is associated with homologues of E. coli GlpBC, which are unrelated to FngBC, and these enzymes typically only use glycerol. It is therefore proposed that P. furiosus FngABC and the previously characterized E. coli GlpABC are disparate members of a new electron bifurcating family widespread in the microbial world that are involved in the metabolism of a range of sugars as well as glycerol.<\/jats:p>","DOI":"10.1021\/acs.biochem.6c00512","type":"journal-article","created":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T11:01:35Z","timestamp":1791370895000},"page":"3247-3257","source":"Crossref","is-referenced-by-count":0,"title":["A New Family of Electron-Bifurcating Enzymes Represented\nby Ferredoxin- and NAD-Dependent Glycerol-3-Phosphate Oxidoreductase"],"prefix":"10.1021","volume":"65","author":[{"given":"Gerrit J.","family":"Schut","sequence":"first","affiliation":[{"name":"University of Georgia , , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farris L.","family":"Poole","sequence":"additional","affiliation":[{"name":"University of Georgia , , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0881-8870","authenticated-orcid":true,"given":"Michael W. W.","family":"Adams","sequence":"additional","affiliation":[{"name":"University of Georgia , , , ,","place":["Athens, Georgia, United States, 30602"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,7]]},"reference":[{"issue":"3","key":"2026102007161356000_cit1","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1128\/JB.01417-07","article-title":"Coupled\nferredoxin and crotonyl coenzyme A (CoA) reduction\nwith NADH catalyzed by the butyryl-CoA dehydrogenase\/Etf complex from Clostridium kluyveri","volume":"190","author":"Li","year":"2008","journal-title":"J. Bacteriol."},{"issue":"1","key":"2026102007161356000_cit2","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1186\/s40709-014-0023-y","article-title":"The ecology\nof Dunaliella in high-salt environments","volume":"21","author":"Oren","year":"2014","journal-title":"J. Biol. Res."},{"issue":"1","key":"2026102007161356000_cit3","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1038\/s42003-025-07953-9","article-title":"An unusual glycerol-3-phosphate\ndehydrogenase\nin Sulfolobus acidocaldarius elucidates the diversity\nof glycerol metabolism across Archaea","volume":"8","author":"Schmerling","year":"2025","journal-title":"Commun.\nBiol."},{"issue":"8","key":"2026102007161356000_cit4","doi-asserted-by":"publisher","DOI":"10.1128\/aem.00886-25","article-title":"Biochemical\nproperties of glycerol kinase from the\nhypersaline-adapted archaeon Haloferax volcanii","volume":"91","author":"Sanchez","year":"2025","journal-title":"Appl. Environ. Microbiol."},{"issue":"1","key":"2026102007161356000_cit5","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1038\/sj.jim.7000155","article-title":"Fermentation\nof glycerol by Clostridium pasteurianum: batch and\ncontinuous culture studies","volume":"27","author":"Biebl","year":"2001","journal-title":"J. Ind. Microbiol.\nBiot."},{"issue":"9","key":"2026102007161356000_cit6","doi-asserted-by":"publisher","first-page":"6080","DOI":"10.1074\/jbc.M113.525535","article-title":"Glycerol dehydrogenase plays a dual role in glycerol\nmetabolism and 2,3-butanediol formation in Klebsiella pneumoniae","volume":"289","author":"Wang","year":"2014","journal-title":"J. Biol. Chem."},{"issue":"1","key":"2026102007161356000_cit7","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1111\/1751-7915.13938","article-title":"In silico and in vivo analyses reveal\nkey metabolic pathways enabling the fermentative utilization of glycerol\nin Escherichia coli","volume":"15","author":"Clomburg","year":"2022","journal-title":"Microb. Biotechnol."},{"issue":"6","key":"2026102007161356000_cit8","doi-asserted-by":"publisher","first-page":"831","DOI":"10.1007\/s10529-013-1240-4","article-title":"Fermentation of\nglycerol and production of valuable chemical and biofuel molecules","volume":"35","author":"Mattam","year":"2013","journal-title":"Biotechnol. Lett."},{"issue":"3","key":"2026102007161356000_cit9","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.copbio.2007.05.002","article-title":"Anaerobic fermentation\nof glycerol: a path to economic\nviability for the biofuels industry","volume":"18","author":"Yazdani","year":"2007","journal-title":"Curr. Opin.\nBiotechnol."},{"issue":"3","key":"2026102007161356000_cit10","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1007\/s002530051523","article-title":"Microbial\nproduction\nof 1,3-propanediol","volume":"52","author":"Biebl","year":"1999","journal-title":"Appl. Microbiol. 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Evol."},{"issue":"W1","key":"2026102007161356000_cit34","doi-asserted-by":"publisher","first-page":"W78","DOI":"10.1093\/nar\/gkae268","article-title":"Interactive Tree of\nLife (iTOL) v6: recent updates to the phylogenetic\ntree display and annotation tool","volume":"52","author":"Letunic","year":"2024","journal-title":"Nucleic Acids\nRes."},{"issue":"14","key":"2026102007161356000_cit35","doi-asserted-by":"publisher","first-page":"168018","DOI":"10.1016\/j.jmb.2023.168018","article-title":"EFI-GNT, and EFI-CGFP: Enzyme Function\nInitiative (EFI) web resource for genomic enzymology tools","volume":"435","author":"Oberg","year":"2023","journal-title":"J. Mol. 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Microbiol."},{"issue":"3","key":"2026102007161356000_cit44","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1007\/s00792-022-01277-w","article-title":"The catalytic and structural basis\nof archaeal glycerophospholipid biosynthesis","volume":"26","author":"de Kok","year":"2022","journal-title":"Extremophiles"},{"key":"2026102007161356000_cit45","doi-asserted-by":"publisher","first-page":"1357","DOI":"10.3389\/fmicb.2018.01357","article-title":"On the natural\nhistory\nof flavin-based electron bifurcation","volume":"9","author":"Baymann","year":"2018","journal-title":"Front.\nMicrobiol."},{"issue":"1","key":"2026102007161356000_cit46","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1021\/acsbiomedchemau.5c00232","article-title":"Photoinduced electron\ntransfer informs on pathway coupling\nin flavin-based electron bifurcation","volume":"6","author":"Wiley","year":"2026","journal-title":"ACS Bio.\nMed. Chem. Au"},{"key":"2026102007161356000_cit47","doi-asserted-by":"publisher","first-page":"108796","DOI":"10.1016\/j.abb.2021.108796","article-title":"Flavins in the electron\nbifurcation process","volume":"701","author":"Kayastha","year":"2021","journal-title":"Arch. Biochem. 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Bacteriol"},{"issue":"19","key":"2026102007161356000_cit51","doi-asserted-by":"publisher","first-page":"9959","DOI":"10.1016\/S0021-9258(19)68723-5","article-title":"The anaerobic sn-glycerol-3-phosphate\ndehydrogenase\nof Escherichia coli. Purification and\ncharacterization","volume":"256","author":"Schryvers","year":"1981","journal-title":"J. Biol. Chem."},{"issue":"3\u20134","key":"2026102007161356000_cit52","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1139\/o95-018","article-title":"Physiological role of GlpB of anaerobic glycerol-3-phosphate\ndehydrogenase of Escherichia coli","volume":"73","author":"Varga","year":"1995","journal-title":"Biochem. 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