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Functional studies have established essential roles for individual ALDH isoforms in tumor initiation, progression, and metastasis; however, the mechanisms by which these enzymes promote malignancy remain poorly understood. Here we show that aldehyde dehydrogenase 1B1 (ALDH1B1), a mitochondrial enzyme highly expressed in colorectal cancer (CRC) and pancreatic ductal adenocarcinoma (PDAC), generates \u03b3-butyrobetaine (GBB) and \u03b3-aminobutyric acid (GABA) in CRC cells. The biosynthesis of both aminocarboxylic acids has been attributed to the cytosolic enzyme ALDH9A1, and we demonstrate that mitochondrial GBB and GABA are functionally distinct from the cytosolic pools of these metabolites. We further demonstrate that mitochondrial GBB can function as an antiport substrate of carnitine-acylcarnitine translocase (CACT), the transporter that mediates fatty acid uptake into the mitochondrial inner matrix. This activity complements the role of cytosolic GBB as the biosynthetic precursor to carnitine. Accordingly, ALDH1B1 can markedly enhance mitochondrial fatty acid oxidation (FAO), a catabolic process that has been linked to CRC and PDAC stemness, progression, and metastasis. Our findings reveal an unexpected role for ALDH1B1 in carnitine metabolism, GABA biosynthesis, and FAO and illustrate how the compartmental reprogramming of metabolic pathways can promote tumor growth.<\/jats:p>","DOI":"10.64898\/2026.09.24.753323","type":"posted-content","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T13:00:39Z","timestamp":1790341239000},"source":"Crossref","is-referenced-by-count":0,"title":["ALDH1B1 promotes mitochondrial \u03b3-butyrobetaine biosynthesis and fatty acid oxidation in colorectal cancer"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9337-1454","authenticated-orcid":false,"given":"Zhiping","family":"Feng","sequence":"first","affiliation":[{"name":"Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7032-8747","authenticated-orcid":false,"given":"Thomas E.","family":"Bearrood","sequence":"additional","affiliation":[{"name":"Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7861-5609","authenticated-orcid":false,"given":"Sydney L.","family":"Campbell","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7599-1294","authenticated-orcid":false,"given":"Adrianne M.","family":"Kinsey","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Stanford University, Stanford, CA, US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7624-3090","authenticated-orcid":false,"given":"Nedas","family":"Matulionis","sequence":"additional","affiliation":[{"name":"Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0765-0893","authenticated-orcid":false,"given":"Che-Hong","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4787-6910","authenticated-orcid":false,"given":"Takuya","family":"Seike","sequence":"additional","affiliation":[{"name":"Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7000-9612","authenticated-orcid":false,"given":"Logan","family":"Leak","sequence":"additional","affiliation":[{"name":"Department of Biology, Stanford University, Stanford, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3480-1837","authenticated-orcid":false,"given":"A. 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