{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T19:01:51Z","timestamp":1785438111997,"version":"3.56.0"},"reference-count":45,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T00:00:00Z","timestamp":1781654400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T00:00:00Z","timestamp":1781654400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100008982","name":"National Science Foundation","doi-asserted-by":"publisher","award":["CHE-2215854"],"award-info":[{"award-number":["CHE-2215854"]}],"id":[{"id":"10.13039\/501100008982","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R35GM147515"],"award-info":[{"award-number":["R35GM147515"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000006","name":"Office of Naval Research","doi-asserted-by":"publisher","award":["N00014-22-1-2266"],"award-info":[{"award-number":["N00014-22-1-2266"]}],"id":[{"id":"10.13039\/100000006","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100013316","name":"Strategic Environmental Research and Development Program","doi-asserted-by":"publisher","award":["WP2332"],"award-info":[{"award-number":["WP2332"]}],"id":[{"id":"10.13039\/100013316","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100013316","name":"Strategic Environmental Research and Development Program","doi-asserted-by":"publisher","award":["WP24-4206"],"award-info":[{"award-number":["WP24-4206"]}],"id":[{"id":"10.13039\/100013316","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2026,6,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Nitramines [R(R\u2032)N\u2013NO\n                      <jats:sub>2<\/jats:sub>\n                      ; R,R\u2032=H or alkyl] are valuable synthetic products, but knowledge of the biosynthetic processes that generate these compounds is limited. This work sought to elucidate the biosynthesis of a nitramine natural product,\n                      <jats:italic toggle=\"yes\">N<\/jats:italic>\n                      -nitroglycine (NNG) by\n                      <jats:italic toggle=\"yes\">Streptomyces noursei<\/jats:italic>\n                      . Stable isotope studies showed that\n                      <jats:italic toggle=\"yes\">S. noursei<\/jats:italic>\n                      cells supplemented with\n                      <jats:sc>l<\/jats:sc>\n                      -(\u03b5-\n                      <jats:sup>15<\/jats:sup>\n                      N)lysine, (\n                      <jats:sup>15<\/jats:sup>\n                      N)glycine, or (\n                      <jats:sup>13<\/jats:sup>\n                      C)hydrazinoacetic acid (HAA) incorporated 67%, 88%, and 67% of the isotope label into NNG, respectively, indicating that these compounds are biosynthetic precursors of NNG. Liquid chromatography coupled tandem mass spectrometry (LC-MS\/MS) of\n                      <jats:sup>15<\/jats:sup>\n                      N-Lys-labeled NNG confirmed that the nitro nitrogen of NNG originates from Lys. Bioinformatics analysis of the\n                      <jats:italic toggle=\"yes\">S. noursei<\/jats:italic>\n                      genome showed evidence for a biosynthetic gene cluster (BGC) that contained machinery for HAA biosynthesis (\n                      <jats:italic toggle=\"yes\">nngKLM<\/jats:italic>\n                      ), consistent with the results of the isotope labeling.\n                      <jats:italic toggle=\"yes\">In vitro<\/jats:italic>\n                      reconstitution of the gene products produced HAA. The borders of this BGC were defined by cross-referencing the predicted BGC with previously published differential proteomics data. Furthermore, we show that azaserine is produced alongside NNG in\n                      <jats:italic toggle=\"yes\">S. noursei<\/jats:italic>\n                      cultures, linking the two biosynthetic pathways via a proposed nitrosamine biosynthetic intermediate. Finally, the oxygen balance for NNG is \u221220.2% for the formation of carbon dioxide (CO\n                      <jats:sub>2<\/jats:sub>\n                      ), which is comparable to that of hexahydro-1,3,5-trinitro-1,3,5-triazene (common name: RDX; \u221221.6%). Crystal structure data of NNG indicate that the unit crystalizes as a pure material, not a hydrate, suggesting a favorable energetic crystallization phase. The combined results suggest a route that, with further development, could lead to sustainable production of energetic nitramines via synthetic biology or biocatalytic approaches.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      Chemical manufacture of nitramine energetics produces hazardous waste streams. Mitigation of these waste streams could be accomplished by replacing current production processes with enzymatic or synthetic biology approaches, enabling sustainable production of these compounds. This study identifies a\n                      <jats:italic toggle=\"yes\">Streptomyces noursei<\/jats:italic>\n                      biosynthetic gene cluster that produces a nitramine natural product,\n                      <jats:italic toggle=\"yes\">N<\/jats:italic>\n                      -nitroglycine, via the formation of a hydrazine precursor. The combined data, along with previously reported proteomics data, define the boundaries and genetic inventory of the biosynthetic gene cluster. Furthermore, structural and thermal degradation analyses of the natural product suggest favorable properties for the use of\n                      <jats:italic toggle=\"yes\">N<\/jats:italic>\n                      -nitroglycine, itself, as an energetic material. Further development of this pathway could enable sustainable approaches to produce energetic nitramines either by oxidation of non-native polyhydrazines to the corresponding polynitramines or by improving production and isolation of\n                      <jats:italic toggle=\"yes\">N<\/jats:italic>\n                      -nitroglycine from\n                      <jats:italic toggle=\"yes\">S. noursei<\/jats:italic>\n                      cultures.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aem.00631-26","type":"journal-article","created":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T14:09:17Z","timestamp":1779286157000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Hydrazinoacetic acid is a biosynthetic precursor of the bacterially produced nitramine,\n                    <i>N<\/i>\n                    -nitroglycine"],"prefix":"10.1128","volume":"92","author":[{"given":"Gabriel","family":"Padilla","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/036nfer12","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, University of Central Florida","place":["Orlando, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin M.","family":"Rathman","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/036nfer12","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, University of Central Florida","place":["Orlando, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shivaiah","family":"Vaddypally","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00kx1jb78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Temple University","place":["Philadelphia, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brenda","family":"Martinez Rodriguez","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/036nfer12","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, University of Central Florida","place":["Orlando, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0212-2557","authenticated-orcid":true,"given":"Michael J.","family":"Zdilla","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00kx1jb78","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, Temple University","place":["Philadelphia, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8968-7344","authenticated-orcid":true,"given":"David E.","family":"Graham","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01qz5mb56","id-type":"ROR","asserted-by":"publisher"}],"name":"Biosciences Division, Oak Ridge National Laboratory","place":["Oak Ridge, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9196-5275","authenticated-orcid":true,"given":"Jonathan D.","family":"Caranto","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/036nfer12","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, University of Central Florida","place":["Orlando, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/9783527628803"},{"key":"e_1_3_4_3_2","unstructured":"Abadin H Smith C Ingerman L Llados FT Barber LE Plewak D Diamond GL. 2013. Toxicological profile for RDX"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1039\/C8OB02750A"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.7b00203"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1039\/c0np00024h"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.21.279"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202507866"},{"key":"e_1_3_4_9_2","unstructured":"Graham DE Spain JC Parry RJ Hettich RL Mahan KM Klingeman DM Giannone RJ Gulvick CA Fida TT. 2018. Nitration enzyme toolkit for the biosynthesis of energetic materials (ORNL\/SPR-2017\/498). Oak Ridge National Lab Oak Ridge TN"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.1048"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2014.10.011"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(08)36001-7"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41557-024-01646-2"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.1991"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.joc.8b00367"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201800762"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-025-01898-0"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.8b05354"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1039\/d3sc01906c"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202304646"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.orglett.3c01229"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.3c12018"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-10079-x"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-021-00895-3"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2007.10.104"},{"key":"e_1_3_4_26_2","unstructured":"Bruker S SAINT S. 2002. Bruker AXS Inc. Madison Wisconsin USA."},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1107\/S2053273314026370"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889808042726"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.201900449"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-27523-x"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M114.629485"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1058092"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaf334"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.3c03419"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2210908119"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0812971106"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2015931118"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.201903500"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.8b13049"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-0894-z"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1021\/tx500371a"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201204116"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.1039\/D1OB01284C"},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.joc.0c02774"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1111\/1574-6976.12014"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01057588"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aem.00631-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aem.00631-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T15:23:01Z","timestamp":1783005781000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/aem.00631-26"}},"subtitle":[],"editor":[{"given":"Haruyuki","family":"Atomi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,6,17]]},"references-count":45,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,6,17]]}},"alternative-id":["10.1128\/aem.00631-26"],"URL":"https:\/\/doi.org\/10.1128\/aem.00631-26","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,17]]},"assertion":[{"value":"2026-03-28","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-20","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00631-26"}}