{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T11:03:10Z","timestamp":1790593390427,"version":"4.1.0"},"reference-count":35,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000015","name":"U.S. Department of Energy","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000015","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000015","name":"U.S. Department of Energy","doi-asserted-by":"publisher","award":["DE\u2010AC36\u201008GO28308"],"award-info":[{"award-number":["DE\u2010AC36\u201008GO28308"]}],"id":[{"id":"10.13039\/100000015","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000015","name":"U.S. Department of Energy","doi-asserted-by":"publisher","award":["WBS 2.3.2.122"],"award-info":[{"award-number":["WBS 2.3.2.122"]}],"id":[{"id":"10.13039\/100000015","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2026,9]]},"abstract":"ABSTRACT<\/jats:title>\n \n Methylotrophic microbes are attractive alternatives to traditional heterotrophic production platforms, yet their efficiency is constrained by carbon loss through pyruvate decarboxylation and the oxidative branch of the RuMP cycle. The phosphoketolase (PKT) pathway provides a carbon\u2010conserving alternative by cleaving fructose\u20106\u2010phosphate and\/or xylulose\u20105\u2010phosphate into acetyl\u2010phosphate, which can subsequently be converted to acetyl\u2010coA without pyruvate decarboxylation. The remaining carbon intermediates are recycled through central metabolism to regenerate RuMP cycle intermediates without direct CO\n 2<\/jats:sub>\n release. Here, we engineered this strategy in\n \n Bacillus methanolicus<\/jats:italic>\n <\/jats:styled-content>\n , a thermophilic methylotroph with strong industrial potential. We first established a versatile expression toolkit comprising inducible and constitutive promoters, benchmarked using an sfGFP reporter. Leveraging this system, we heterologously expressed the phosphoketolase B (\n pktB<\/jats:italic>\n ) gene from\n Methylotuvimicrobium buryatense<\/jats:italic>\n 5GB1C which increased methanol\u2010to\u2010biomass yields by 18%\u201324% relative to controls and reduced biogenic CO\n 2<\/jats:sub>\n production by 9%\u201312%. Chromosomal integration of\n pktB<\/jats:italic>\n preserved these gains, demonstrating stability without reliance on plasmid\u2010based expression. Together, these results show that PKT\u2010driven metabolic rewiring enhances substrate yields in\n \n B. methanolicus<\/jats:italic>\n <\/jats:styled-content>\n and provides a scalable strategy to improve methylotrophic bioprocesses. This work expands the metabolic engineering toolbox for methylotrophs and highlights carbon\u2010conserving pathway design as a key lever for advancing single carbon (C1) biomanufacturing.\n <\/jats:p>","DOI":"10.1111\/1751-7915.70430","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T08:59:07Z","timestamp":1788253147000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rewiring Carbon Metabolism in\n \n Bacillus methanolicus<\/i>\n <\/scp>\n via Heterologous Phosphoketolase Expression Enhances Biomass Yield From Methanol and Reduces\n \n CO\n 2<\/sub>\n <\/scp>\n Loss"],"prefix":"10.1111","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3343-5804","authenticated-orcid":false,"given":"Yael J.","family":"Toporek","sequence":"first","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-4726-3121","authenticated-orcid":false,"given":"Jonathan R.","family":"Humphreys","sequence":"additional","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-4448-3948","authenticated-orcid":false,"given":"Thomas J.","family":"Musselwhite","sequence":"additional","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7965-8397","authenticated-orcid":false,"given":"Marcus S.","family":"Bray","sequence":"additional","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8740-3875","authenticated-orcid":false,"given":"Katie P.","family":"Michel","sequence":"additional","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2200-1210","authenticated-orcid":false,"given":"Kimberly A.","family":"Rosenbach","sequence":"additional","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1403-9689","authenticated-orcid":false,"given":"Michael T.","family":"Guarnieri","sequence":"additional","affiliation":[{"name":"Bioenergy & Sustainable Transportation Directorate, National Laboratory of the Rockies Golden Colorado USA"},{"name":"Department of Chemical and Biological Engineering Colorado School of Mines Golden Colorado USA"},{"name":"Renewable & Sustainable Energy Institute University of Colorado Boulder Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,1]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-20574-z"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2009.08.050"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12575"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.186.5.1229-1238.2004"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-006-0757-z"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.70322\/sbe.2025.10016"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.10.2970-2972.2000"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2019.10.002"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-015-0377-3"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/mSystems.00745-20"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.02380-14"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-41135-7"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00703-12"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2015.08.018"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymben.2017.03.007"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22094686"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.01481"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-015-1239-4"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/1751-7915.14067"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.188.8.3063-3072.2006"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms3785"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.71.2.728-733.2005"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymben.2025.02.013"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.01725"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.9.2929-2936.2001"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.13200"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00492-06"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41929-024-01137-0"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2024.103167"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.56.4.963-970.1990"},{"key":"e_1_2_10_32_1","unstructured":"Schendel F. J. R.Dillingham R. S.Hanson K.Sano andK.Matsui.2000.\u201cProduction of Glutamate Using Wild TypeBacillus methanolicus.\u201dGoogle Patents."},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymben.2004.02.001"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2024.06.013"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3002285"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2008.03.002"}],"container-title":["Microbial Biotechnology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1751-7915.70430","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/1751-7915.70430","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1751-7915.70430","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T10:23:37Z","timestamp":1790591017000},"score":1,"resource":{"primary":{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/10.1111\/1751-7915.70430"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9]]},"references-count":35,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1111\/1751-7915.70430"],"URL":"https:\/\/doi.org\/10.1111\/1751-7915.70430","archive":["Portico"],"relation":{},"ISSN":["1751-7915","1751-7915"],"issn-type":[{"value":"1751-7915","type":"print"},{"value":"1751-7915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9]]},"assertion":[{"value":"2026-04-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-29","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-01","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70430"}}