{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T16:00:16Z","timestamp":1791820816276,"version":"4.3.4"},"reference-count":23,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T00:00:00Z","timestamp":1775520000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T00:00:00Z","timestamp":1775520000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJGX23B0"],"award-info":[{"award-number":["JPMJGX23B0"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP24K01897"],"award-info":[{"award-number":["JP24K01897"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J Eukaryotic Microbiology"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Wax esters (WEs) produced by\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Euglena gracilis<\/jats:italic>\n                    <\/jats:styled-content>\n                    under anaerobic conditions contain significant proportions of odd\u2010chain fatty acids and fatty alcohols (30%\u201340%). Propionyl\u2010CoA, the primer for odd\u2010chain fatty acid synthesis, is generally thought to be generated via the methylmalonyl\u2010CoA pathway; however, the enzymatic basis of this pathway in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>E. gracilis<\/jats:italic>\n                    <\/jats:styled-content>\n                    remains unclear. Here, we generated knockout (KO) mutants of candidate enzymes in the methylmalonyl\u2010CoA pathway\u2014succinyl\u2010CoA synthetase (SCS), methylmalonyl\u2010CoA mutase (MCM), methylmalonyl\u2010CoA epimerase (MCE), and propionyl\u2010CoA carboxylase (PCC)\u2014using the CRISPR\/Cas9 system and examined their roles in odd\u2010chain fatty acid synthesis and propionate assimilation. The methylmalonyl\u2010CoA pathway exhibited direction\u2010dependent functions. KO of MCE and PCC did not affect the proportion of odd\u2010chain WEs, indicating that these enzymes are not required for odd\u2010chain WE synthesis and suggesting an alternative route converting methylmalonyl\u2010CoA to propionyl\u2010CoA. In contrast, both enzymes were required for propionate assimilation. Functional differentiation was observed between SCS\u03b2 isozymes: SCS\u03b21 contributed to odd\u2010chain WE synthesis, whereas SCS\u03b22 primarily functioned in propionate assimilation. Partial impairment of propionate assimilation in SCS\u03b1 or SCS\u03b22 KO strains suggests involvement of additional routes. These findings improve our understanding of direction\u2010dependent roles in the methylmalonyl\u2010CoA pathway in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>E. gracilis<\/jats:italic>\n                    <\/jats:styled-content>\n                    and support further studies of odd\u2010chain lipid biosynthesis.\n                  <\/jats:p>","DOI":"10.1111\/jeu.70078","type":"journal-article","created":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T07:31:34Z","timestamp":1775547094000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Elucidation of Enzymatic Routes Underlying Odd\u2010Chain Fatty Acid Synthesis and Propionate Assimilation in\n                    <scp>\n                      <i>Euglena gracilis<\/i>\n                    <\/scp>"],"prefix":"10.1111","volume":"73","author":[{"given":"Ryunosuke","family":"Katayama","sequence":"first","affiliation":[{"name":"Department of Applied Biological Chemistry, Graduate School of Agriculture Osaka Metropolitan University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryuta","family":"Hayashi","sequence":"additional","affiliation":[{"name":"Department of Applied Biological Chemistry, Graduate School of Agriculture Osaka Metropolitan University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitsuhiro","family":"Ueda","sequence":"additional","affiliation":[{"name":"Department of Applied Biological Chemistry, Graduate School of Agriculture Osaka Metropolitan University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatsuji","family":"Sakamoto","sequence":"additional","affiliation":[{"name":"Department of Applied Biological Chemistry, Graduate School of Agriculture Osaka Metropolitan University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1639-0560","authenticated-orcid":false,"given":"Masami","family":"Nakazawa","sequence":"additional","affiliation":[{"name":"Department of Applied Biological Chemistry, Graduate School of Agriculture Osaka Metropolitan University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,7]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.68.1.271\u2010279.2002"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41597\u2010024\u201003404\u2010y"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00410835"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi992888d"},{"issue":"5","key":"e_1_2_10_6_1","first-page":"1097","article-title":"The Metabolic Pathway of Propionate in \n                     Euglena gracilis\n                   Z Grown Under Illumination","volume":"44","author":"Hosotani K.","year":"1980","journal-title":"Agricultural and Biological Chemistry"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014\u20105793(82)81310\u20100"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/pbi.13904"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006\u2010291X(88)90587\u20106"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.42.27573"},{"key":"e_1_2_10_11_1","first-page":"17","article-title":"High\u2010Yield Media for Photosynthesizing Euglena gracilis Z","volume":"14","author":"Koren L. 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