{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T05:09:14Z","timestamp":1778908154139,"version":"3.51.4"},"reference-count":64,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T00:00:00Z","timestamp":1776470400000},"content-version":"vor","delay-in-days":17,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T00:00:00Z","timestamp":1775001600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP21am0101118"],"award-info":[{"award-number":["JP21am0101118"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP22ama121006"],"award-info":[{"award-number":["JP22ama121006"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP25ama121004"],"award-info":[{"award-number":["JP25ama121004"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["18K19875"],"award-info":[{"award-number":["18K19875"]}],"id":[{"id":"10.13039\/501100001691","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":[[2026,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Ogataea methanolica<\/jats:italic>\n                    is a methylotrophic yeast that can produce diverse recombinant proteins using methanol as the sole carbon and energy source. Unlike most yeast species, which possess a single alcohol oxidase,\n                    <jats:italic>O.\u2009methanolica<\/jats:italic>\n                    encodes two isoenzymes, Mod1p and Mod2p. This study examines the structural and functional differences between Mod1p and Mod2p homooctamers. Both enzymes were purified from\n                    <jats:italic>MOD<\/jats:italic>\n                    \u2010disrupted strains and analysed using cryogenic electron microscopy, achieving resolutions of 1.9 and 2.7\u2009\u00c5 for Mod1p and Mod2p, respectively. The two isozymes assemble as tetramers of dimers stabilized by extensive intersubunit interactions, largely mediated by protruding loop regions and C\u2010terminal extensions. Despite overall structural similarities, Mod1p and Mod2p exhibit subtle differences in surface charge distribution and sequence composition within the FAD\u2010binding domain. These variations correlate with distinct cofactor preferences, with Mod1p binding arabityl FAD and Mod2p binding canonical FAD. Thin\u2010section electron microscopy further revealed that Mod1p and Mod2p form both homomeric and hybrid octamers that assemble into peroxisomal crystalloids essential for methanol metabolism. Collectively, our findings provide mechanistic insight into alcohol oxidase diversity in methylotrophic yeasts, advancing our understanding of methanol utilization and its applications in biotechnology.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.70355","type":"journal-article","created":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T09:54:59Z","timestamp":1776506099000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Cryo\u2010\n                    <scp>EM<\/scp>\n                    Structures of Alcohol Oxidase Isozymes Reveal Structural Determinants of Cofactor Variation and Enzymatic Activity in\n                    <i>Ogataea methanolica<\/i>"],"prefix":"10.1111","volume":"19","author":[{"given":"Hao\u2010Liang","family":"Cai","sequence":"first","affiliation":[{"name":"The United Graduate School of Agricultural Sciences Gifu University  Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atsuhiro","family":"Shimada","sequence":"additional","affiliation":[{"name":"The United Graduate School of Agricultural Sciences Gifu University  Gifu Japan"},{"name":"The Graduate School of Natural Sciences and Technologies Gifu University  Gifu Japan"},{"name":"Faculty of Applied Biological Sciences Gifu University  Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tasuku","family":"Hamaguchi","sequence":"additional","affiliation":[{"name":"RIKEN SPring\u20108 Center  Sayo Hyogo Japan"},{"name":"Institute of Multidisciplinary Research for Advanced Materials Tohoku University  Sendai Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akira","family":"Mizoguchi","sequence":"additional","affiliation":[{"name":"Graduate School of Medicine Mie University  Tsu Mie Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5520-4391","authenticated-orcid":false,"given":"Koji","family":"Yonekura","sequence":"additional","affiliation":[{"name":"RIKEN SPring\u20108 Center  Sayo Hyogo Japan"},{"name":"Institute of Multidisciplinary Research for Advanced Materials Tohoku University  Sendai Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kyohei","family":"Tsuchiyama","sequence":"additional","affiliation":[{"name":"The Graduate School of Natural Sciences and Technologies Gifu University  Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masaya","family":"Shimada","sequence":"additional","affiliation":[{"name":"The United Graduate School of Agricultural Sciences Gifu University  Gifu Japan"},{"name":"The Graduate School of Natural Sciences and Technologies Gifu University  Gifu Japan"},{"name":"Faculty of Applied Biological Sciences Gifu University  Gifu Japan"},{"name":"Preemptive Food Research Center Gifu University  Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akio","family":"Ebihara","sequence":"additional","affiliation":[{"name":"The United Graduate School of Agricultural Sciences Gifu University  Gifu Japan"},{"name":"The Graduate School of Natural Sciences and Technologies Gifu University  Gifu Japan"},{"name":"Faculty of Applied Biological Sciences Gifu University  Gifu Japan"},{"name":"Preemptive Food Research Center Gifu University  Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4835-154X","authenticated-orcid":false,"given":"Kazutoshi","family":"Tani","sequence":"additional","affiliation":[{"name":"Graduate School of Medicine Mie University  Tsu Mie Japan"},{"name":"Center for Computational Sciences University of Tsukuba  Ibaraki Japan"},{"name":"Center for Quantum and Information Life Sciences University of Tsukuba  Tsukuba Ibaraki Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomoyuki","family":"Nakagawa","sequence":"additional","affiliation":[{"name":"The United Graduate School of Agricultural Sciences Gifu University  Gifu Japan"},{"name":"The Graduate School of Natural Sciences and Technologies Gifu University  Gifu Japan"},{"name":"Faculty of Applied Biological Sciences Gifu University  Gifu Japan"},{"name":"Preemptive Food Research Center Gifu University  Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,4,18]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010024\u201007487\u2010w"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"e_1_2_10_4_1","first-page":"1407","article-title":"Formation and Distribution of Modified FAD Between Isozymes of Alcohol Oxidase in the Methylotrophic Yeast Pichia methanolica","volume":"63","author":"Ashin V. V.","year":"1998","journal-title":"Biochemistry (Moscow)"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.181342398"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.energy.2019.05.171"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1099\/00221287\u2010137\u201010\u20102381"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/1751\u20107915.13811"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/yea.3807"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2201711119"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909042073"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475\u20102859\u20105\u201017"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1128\/mcb.9.3.1316\u20101323.1989"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444910007493"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/bs.enz.2020.05.003"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2020.08.008"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiosc.2019.01.008"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002530000464"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkg556"},{"key":"e_1_2_10_20_1","first-page":"1537","article-title":"Multiple Molecular Forms of Alcohol Oxidase From the Methylotrophic Yeast Pichia methanolica","volume":"61","author":"Gruzman M. B.","year":"1996","journal-title":"Biochemistry (Moscow)"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475\u20102859\u20105\u201039"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcou.2018.06.019"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/0263\u20107855(96)00018\u20105"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/yea.1490"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0135651100"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0149846"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1099\/ijs.0.63491\u20100"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1023\/a:1001761008817"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2005.07.007"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253\u2010014\u20106224\u20103"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/yea.896"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0061(19990915)15:12<1223::AID-YEA450>3.0.CO;2-X"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1263\/jbb.91.225"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/0922-338X(96)81469-3"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2007.321"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42003\u2010018\u20100157\u20109"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253\u2010014\u20106240\u20103"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1080\/00021369.1969.10859497"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200462121"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.151.1.376\u2010383.1982"},{"key":"e_1_2_10_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsyr.2005.06.005"},{"key":"e_1_2_10_43_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20084"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.1002\/bbb.1773"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097\u20100061(19980115)14"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2015.08.008"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2003.07.013"},{"key":"e_1_2_10_48_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12915\u2010015\u20100186\u20105"},{"key":"e_1_2_10_49_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1399004713029283"},{"key":"e_1_2_10_50_1","doi-asserted-by":"publisher","DOI":"10.1039\/d5sc02960k"},{"key":"e_1_2_10_51_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.114.15.2863"},{"key":"e_1_2_10_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0969\u20102126(00)00133\u20107"},{"key":"e_1_2_10_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2006.05.009"},{"key":"e_1_2_10_54_1","first-page":"443","article-title":"Production of Formaldehyde by a Mutant of Methanol Yeast, \n                     Candida boidinii\n                   S2","volume":"63","author":"Tani Y.","year":"1985","journal-title":"Journal of Fermentation Technology"},{"key":"e_1_2_10_55_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/22.22.4673"},{"key":"e_1_2_10_56_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2006.07.016"},{"key":"e_1_2_10_57_1","doi-asserted-by":"publisher","DOI":"10.1128\/mcb.1.10.949\u2010957.1981"},{"key":"e_1_2_10_58_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00402303"},{"key":"e_1_2_10_59_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0159476"},{"key":"e_1_2_10_60_1","doi-asserted-by":"publisher","DOI":"10.1016\/1047\u20108477(92)90070\u2010Q"},{"key":"e_1_2_10_61_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.58.1245"},{"key":"e_1_2_10_62_1","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/101298"},{"key":"e_1_2_10_63_1","doi-asserted-by":"publisher","DOI":"10.1042\/BA20090030"},{"key":"e_1_2_10_64_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4193"},{"key":"e_1_2_10_65_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.42166"}],"container-title":["Microbial Biotechnology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1751-7915.70355","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/1751-7915.70355","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/1751-7915.70355","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T04:27:22Z","timestamp":1778905642000},"score":1,"resource":{"primary":{"URL":"https:\/\/sfamjournals.onlinelibrary.wiley.com\/doi\/10.1111\/1751-7915.70355"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4]]},"references-count":64,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2026,4]]}},"alternative-id":["10.1111\/1751-7915.70355"],"URL":"https:\/\/doi.org\/10.1111\/1751-7915.70355","archive":["Portico"],"relation":{},"ISSN":["1751-7915","1751-7915"],"issn-type":[{"value":"1751-7915","type":"print"},{"value":"1751-7915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4]]},"assertion":[{"value":"2026-01-21","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-06","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-18","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70355"}}