{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T03:13:37Z","timestamp":1775186017756,"version":"3.50.1"},"reference-count":37,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T00:00:00Z","timestamp":1733788800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["22-14-00291"],"award-info":[{"award-number":["22-14-00291"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["RM11916B861B9985"],"award-info":[{"award-number":["RM11916B861B9985"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["RM120172B6587496"],"award-info":[{"award-number":["RM120172B6587496"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004271","name":"Sapienza University of Rome","doi-asserted-by":"publisher","award":["22-14-00291"],"award-info":[{"award-number":["22-14-00291"]}],"id":[{"id":"10.13039\/501100004271","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004271","name":"Sapienza University of Rome","doi-asserted-by":"publisher","award":["RM11916B861B9985"],"award-info":[{"award-number":["RM11916B861B9985"]}],"id":[{"id":"10.13039\/501100004271","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004271","name":"Sapienza University of Rome","doi-asserted-by":"publisher","award":["RM120172B6587496"],"award-info":[{"award-number":["RM120172B6587496"]}],"id":[{"id":"10.13039\/501100004271","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Amino acid analogues with a phosphorus-containing moiety replacing the carboxylic group are promising sources of biologically active compounds. The H-phosphinic group, with hydrogen\u2013phosphorus\u2013carbon (H-P-C) bonds and a flattened tetrahedral configuration, is a bioisostere of the carboxylic group. Consequently, amino-H-phosphinic acids undergo substrate-like enzymatic transformations, leading to new biologically active metabolites. Previous studies employing NMR-based metabolomic and proteomic analyses show that in Escherichia coli, \u03b1-KG-\u03b3-PH (the distal H-phosphinic analogue of \u03b1-ketoglutarate) can be converted into L-Glu-\u03b3-PH. Notably, \u03b1-KG-\u03b3-PH and L-Glu-\u03b3-PH are antibacterial compounds, but their intracellular targets only partially overlap. L-Glu-\u03b3-PH is known to be a substrate of aspartate transaminase and glutamate decarboxylase, but its substrate properties with NAD+-dependent glutamate dehydrogenase (GDH) have never been investigated. Compounds containing P-H bonds are strong reducing agents; therefore, enzymatic NAD+-dependent oxidation is not self-evident. Herein, we demonstrate that L-Glu-\u03b3-PH is a substrate of eukaryotic GDH and that the pH optimum of L-Glu-\u03b3-PH NAD+-dependent oxidative deamination is shifted to a slightly alkaline pH range compared to L-glutamate. By 31P NMR, we observe that \u03b1-KG-\u03b3-PH exists in a pH-dependent equilibrium of keto and germinal diol forms. Furthermore, the stereospecific enzymatic synthesis of \u03b1-KG-\u03b3-PH from L-Glu-\u03b3-PH using GDH is a possible route for its bio-based synthesis.<\/jats:p>","DOI":"10.3390\/biom14121574","type":"journal-article","created":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T04:15:03Z","timestamp":1733804103000},"page":"1574","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Enzymatic Synthesis of Biologically Active H-Phosphinic Analogue of \u03b1-Ketoglutarate"],"prefix":"10.3390","volume":"14","author":[{"given":"Vsevolod L.","family":"Filonov","sequence":"first","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2833-9834","authenticated-orcid":false,"given":"Maxim A.","family":"Khomutov","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaroslav V.","family":"Tkachev","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Artem V.","family":"Udod","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4660-1322","authenticated-orcid":false,"given":"Dmitry V.","family":"Yanvarev","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0324-1398","authenticated-orcid":false,"given":"Fabio","family":"Giovannercole","sequence":"additional","affiliation":[{"name":"D\u00e9partement de Biologie, Universit\u00e9 de Namur, Rue de Bruxelles 61, 5000 Namur, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elena N.","family":"Khurs","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7443-6961","authenticated-orcid":false,"given":"Sergei N.","family":"Kochetkov","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3536-9927","authenticated-orcid":false,"given":"Daniela","family":"De Biase","sequence":"additional","affiliation":[{"name":"Department of Medico-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Corso della Repubblica 79, 04100 Latina, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6833-7855","authenticated-orcid":false,"given":"Alex R.","family":"Khomutov","sequence":"additional","affiliation":[{"name":"Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov St., 32, 119991 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3670","DOI":"10.1016\/j.bbagen.2013.02.008","article-title":"Natural products: A continuing source of novel drug leads","volume":"1830","author":"Cragg","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"5704","DOI":"10.1021\/acs.chemrev.6b00536","article-title":"Phosphonate biochemistry","volume":"117","author":"Horsman","year":"2017","journal-title":"Chem. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"20759","DOI":"10.1073\/pnas.1315107110","article-title":"Diversity and abundance of phosphonate biosynthetic genes in nature","volume":"110","author":"Yu","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1128\/CMR.00068-15","article-title":"Fosfomycin","volume":"29","author":"Falagas","year":"2016","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2824","DOI":"10.1021\/acssynbio.8b00219","article-title":"Genomic deoxyxylulose phosphate reductoisomerase (DXR) mutations conferring resistance to the antimalarial drug fosmidomycin in E. coli","volume":"7","author":"Pines","year":"2018","journal-title":"ACS Synth. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Jawaid, S., Seidle, H., Zhou, W., Abdirahman, H., Abadeer, M., Hix, J.H., van Hoek, M.L., and Couch, R.D. (2009). Kinetic characterization and phosphoregulation of the Francisella tularensis 1-deoxy-D-xylulose 5-phosphate reductoisomerase (MEP synthase). PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0008288"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.1111\/1462-2920.14534","article-title":"Identification of the first glyphosate transporter by genomic adaptation","volume":"21","author":"Wicke","year":"2019","journal-title":"Environ. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1903","DOI":"10.1021\/bi002438h","article-title":"The crystal structure of phosphinothricin in the active site of glutamine synthetase illuminates the mechanism of enzymatic inhibition","volume":"40","author":"Gill","year":"2001","journal-title":"Biochemistry"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3357","DOI":"10.1128\/AAC.01483-10","article-title":"The antibiotic dehydrophos is converted to a toxic pyruvate analogue by peptide bond cleavage in Salmonella enterica","volume":"55","author":"Circello","year":"2011","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"40036","DOI":"10.18632\/oncotarget.5486","article-title":"Specific inhibition by synthetic analogues of pyruvate reveals that the pyruvate dehydrogenase reaction is essential for metabolism and viability of glioblastoma cells","volume":"6","author":"Bunik","year":"2015","journal-title":"Oncotarget"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1021\/cr068444m","article-title":"Thiamin diphosphate catalysis: Enzymic and nonenzymic covalent intermediates","volume":"108","author":"Kluger","year":"2008","journal-title":"Chem. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1146\/annurev.biochem.78.091707.100215","article-title":"Biosynthesis of phosphonic and phosphinic acid natural products","volume":"78","author":"Metcalf","year":"2009","journal-title":"Annu. Rev. Biochem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1038\/nchembio.186","article-title":"Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli","volume":"5","author":"Bennett","year":"2009","journal-title":"Nat. Chem. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"96","DOI":"10.7164\/antibiotics.36.96","article-title":"Studies on the biosynthesis of bialaphos (SF-1293). 2. Isolation of the first natural products with a C-P-H bond and their involvement in the C-P-C bond formation","volume":"36","author":"Seto","year":"1983","journal-title":"J. Antibiot."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1038\/nchembio.2007.9","article-title":"Unusual transformations in the biosynthesis of the antibiotic phosphinothricin tripeptide","volume":"3","author":"Blodgett","year":"2007","journal-title":"Nat. Chem. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"687","DOI":"10.7164\/antibiotics.38.687","article-title":"Studies on the biosynthesis of bialaphos (SF-1293). 6. Production of N-acetyl-demethylphosphinothricin and N-acetylbialaphos by blocked mutants of Streptomyces hygroscopicus SF-1293 and their roles in the biosynthesis of bialaphos","volume":"38","author":"Imai","year":"1985","journal-title":"J. Antibiot."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1038\/s42004-020-00368-z","article-title":"Enzymatic kinetic resolution of desmethylphosphinothricin indicates that phosphinic group is a bioisostere of carboxyl group","volume":"3","author":"Cappadocio","year":"2020","journal-title":"Commun. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Khomutov, M.A., Giovannercole, F., Onillon, L., Demiankova, M.V., Vasilieva, B.F., Salikhov, A.I., Kochetkov, S.N., Efremenkova, O.V., Khomutov, A.R., and De Biase, D. (2023). A desmethylphosphinothricin dipeptide derivative effectively inhibits Escherichia coli and Bacillus subtilis growth. Biomolecules, 13.","DOI":"10.20944\/preprints202308.2199.v1"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Giovannercole, F., Gafeira Gon\u00e7alves, L., Armengaud, J., Varela Coelho, A., Khomutov, A., and De Biase, D. (2024). Integrated multi-omics unveil the impact of H-phosphinic analogues of glutamate and \u03b1-ketoglutarate on Escherichia coli metabolism. J. Biol. Chem., 300.","DOI":"10.1016\/j.jbc.2024.107803"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Plaitakis, A., Kalef-Ezra, E., Kotzamani, D., Zaganas, I., and Spanaki, C. (2017). The glutamate dehydrogenase pathway and its roles in cell and tissue biology in health and disease. Biology, 6.","DOI":"10.3390\/biology6010011"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6690","DOI":"10.1016\/S0021-9258(18)63461-1","article-title":"Sodium dodecyl sulfate inactivation of bovine liver glutamate dehydrogenase","volume":"244","author":"Rogers","year":"1969","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1006\/jmra.1995.1047","article-title":"Water suppression that works. Excitation sculpting using arbitrary wave-forms and pulsed-field gradients","volume":"112","author":"Hwang","year":"1995","journal-title":"J. Magn. Reson. Ser. A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1795","DOI":"10.1351\/pac200173111795","article-title":"NMR nomenclature. Nuclear spin properties and conventions for chemical shifts (IUPAC Recommendations 2001)","volume":"73","author":"Harris","year":"2001","journal-title":"Pure Appl. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"22988","DOI":"10.1074\/jbc.M109.020222","article-title":"Novel inhibitors complexed with glutamate dehydrogenase. Allosteric regulation by control of protein dynamics","volume":"284","author":"Li","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3089","DOI":"10.1016\/S0021-9258(19)44012-X","article-title":"Bovine glutamate dehydrogenase. The pH dependence of native and nitrated enzyme in the presence of allosteric modifiers","volume":"248","author":"Smith","year":"1973","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/S0076-6879(85)13006-5","article-title":"L-Glutamate dehydrogenase from bovine liver","volume":"113","author":"Fisher","year":"1985","journal-title":"Methods Enzymol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/aem.56.1.1-6.1990","article-title":"Stereospecific production of the herbicide phosphinothricin (glufosinate) by transamination: Isolation and characterization of a phosphinothricin-specific transaminase from Escherichia coli","volume":"56","author":"Schulz","year":"1990","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1007\/BF02007699","article-title":"Effect of phosphonic analogues of glutamic acid on glutamate decarboxylase","volume":"41","author":"Lacoste","year":"1985","journal-title":"Experientia"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4425","DOI":"10.1007\/s00253-018-8910-z","article-title":"Efficient reductive amination process for enantioselective synthesis of L-phosphinothricin applying engineered glutamate dehydrogenase","volume":"102","author":"Yin","year":"2018","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1021\/bi00780a006","article-title":"Nonlinear kinetics of glutamate dehydrogenase. Studies with substrates-glutamate and nicotinamide-adenine dinucleotide","volume":"10","author":"Barton","year":"1971","journal-title":"Biochemistry"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1042\/bj2480351","article-title":"Metabolism of 1-aminoethylphosphinate generates acetylphosphinate, a potent inhibitor of pyruvate dehydrogenase","volume":"248","author":"Laber","year":"1987","journal-title":"Biochem. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3257","DOI":"10.1002\/1521-3773(20020902)41:17<3257::AID-ANIE3257>3.0.CO;2-N","article-title":"Phosphite dehydrogenase: A versatile cofactor-regeneration enzyme","volume":"41","author":"Vrtis","year":"2002","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Aleshin, V.A., Bunik, V.I., Bruch, E.M., and Bellinzoni, M. (2022). Structural basis for the binding of allosteric activators leucine and ADP to mammalian glutamate dehydrogenase. Int. J. Mol. Sci., 23.","DOI":"10.20944\/preprints202208.0527.v1"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"25898","DOI":"10.1039\/D0RA04655H","article-title":"Phosphonopeptides containing free phosphonic groups: Recent advances","volume":"10","author":"Kafarski","year":"2020","journal-title":"RSC Adv."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Rudenko, A.Y., Mariasina, S.S., Bolikhova, A.K., Nikulin, M.V., Ozhiganov, R.M., Vasil\u2019ev, V.G., Ikhalaynen, Y.A., Khandazhinskaya, A.L., Khomutov, M.A., and Sergiev, P.V. (2024). Organophosphorus S-adenosyl-L-methionine mimetics: Synthesis, stability, and substrate properties. Front. Chem., 12.","DOI":"10.3389\/fchem.2024.1448747"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1134\/S0026893323040064","article-title":"Interaction of DNA methyltransferase Dnmt3a with phosphorus analogues of S-adenosylmethionine and S-adenosylhomocysteine","volume":"57","author":"Filonov","year":"2023","journal-title":"Mol. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.jbiotec.2020.03.001","article-title":"Tuning amino acid dehydrogenases with featured sequences for L-phosphinothricin synthesis by reductive amination","volume":"312","author":"Cheng","year":"2020","journal-title":"J. Biotechnol."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/14\/12\/1574\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:51:13Z","timestamp":1760115073000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/14\/12\/1574"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,10]]},"references-count":37,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["biom14121574"],"URL":"https:\/\/doi.org\/10.3390\/biom14121574","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,10]]}}}