{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T17:05:35Z","timestamp":1784567135017,"version":"3.55.0"},"reference-count":30,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2015,1,1]],"date-time":"2015-01-01T00:00:00Z","timestamp":1420070400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2015,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            With a completely reengineered and humanized glycosylation pathway, glycoengineered\n            <jats:named-content content-type=\"genus-species\">Pichia pastoris<\/jats:named-content>\n            has emerged as a promising production host for the manufacture of therapeutic glycoproteins. However, the extensive genetic modifications have also negatively affected the overall fitness levels of the glycoengineered host cells. To make glycoengineered\n            <jats:named-content content-type=\"genus-species\">Pichia<\/jats:named-content>\n            strains more compatible with a scalable industrial fermentation process, we sought to identify genetic solutions to broadly improve cell robustness during fermentation. In this study, we report that mutations within the\n            <jats:named-content content-type=\"genus-species\">Pichia pastoris<\/jats:named-content>\n            <jats:italic>ATT1<\/jats:italic>\n            (\n            <jats:italic>PpATT1<\/jats:italic>\n            ) gene (a homolog of the\n            <jats:named-content content-type=\"genus-species\">Saccharomyces cerevisiae<\/jats:named-content>\n            <jats:italic>GAL4<\/jats:italic>\n            [\n            <jats:italic>ScGAL4<\/jats:italic>\n            ] transcriptional activator) dramatically increased the cellular fitness levels of glycoengineered\n            <jats:named-content content-type=\"genus-species\">Pichia<\/jats:named-content>\n            strains. We demonstrate that deletion of the\n            <jats:italic>PpATT1<\/jats:italic>\n            gene enabled glycoengineered\n            <jats:named-content content-type=\"genus-species\">Pichia<\/jats:named-content>\n            strains to improve their thermal tolerance levels, reduce their cell lysis defects, and greatly improve fermentation robustness. The extension of the duration of fermentation enabled the\n            <jats:italic>PpATT1<\/jats:italic>\n            -modified glycoengineered\n            <jats:named-content content-type=\"genus-species\">Pichia<\/jats:named-content>\n            strains to increase their product yields significantly without any sacrifice in product quality. Because the\n            <jats:italic>ATT1<\/jats:italic>\n            gene could be deleted from any\n            <jats:named-content content-type=\"genus-species\">Pichia<\/jats:named-content>\n            strains, including empty hosts and protein-expressing production strains alike, we suggest that the findings described in this study are broadly applicable to any\n            <jats:named-content content-type=\"genus-species\">Pichia<\/jats:named-content>\n            strains used for the production of therapeutic proteins, including monoclonal antibodies, Fc fusions, peptides, hormones, and growth factors.\n          <\/jats:p>","DOI":"10.1128\/aem.02619-14","type":"journal-article","created":{"date-parts":[[2014,10,25]],"date-time":"2014-10-25T04:07:55Z","timestamp":1414210075000},"page":"260-271","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Inactivation of a\n            <i>GAL4<\/i>\n            -Like Transcription Factor Improves Cell Fitness and Product Yield in Glycoengineered Pichia pastoris Strains"],"prefix":"10.1128","volume":"81","author":[{"given":"Bo","family":"Jiang","sequence":"first","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca","family":"Argyros","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Bukowski","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanie","family":"Nelson","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathan","family":"Sharkey","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sehoon","family":"Kim","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Copeland","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert C.","family":"Davidson","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronghua","family":"Chen","sequence":"additional","affiliation":[{"name":"Informatics and IT, Merck &amp; Co., Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Zhuang","sequence":"additional","affiliation":[{"name":"Informatics and IT, Merck &amp; Co., Boston, Massachusetts, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natarajan","family":"Sethuraman","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terrance A.","family":"Stadheim","sequence":"additional","affiliation":[{"name":"GlycoFi Inc., Discovery Biologics, Merck &amp; Co., Lebanon, New Hampshire, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0910-917"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10295-013-1235-0"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1517\/17460440903413504"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2007.09.001"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2011.11.002"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2008.12.015"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.4161\/mabs.3.3.15532"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.biologicals.2011.06.002"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-9-93"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0931263100"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10295-010-0746-1"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.embor.7400679"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(85)90336-8"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.X114.547323"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1099\/ijs.0.63491-0"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1544"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2011.04.014"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb1303bs82"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1486"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp324"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp352"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1754"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2013.05.009"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2010.11.004"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1007\/10_2013_259"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2007.05.017"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000612"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1026"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1093\/glycob\/cwh104"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1130256"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02619-14","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02619-14","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T20:15:40Z","timestamp":1645647340000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.02619-14"}},"subtitle":[],"editor":[{"given":"J. L.","family":"Schottel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2015,1]]},"references-count":30,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2015,1]]}},"alternative-id":["10.1128\/AEM.02619-14"],"URL":"https:\/\/doi.org\/10.1128\/aem.02619-14","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,1]]},"assertion":[{"value":"2014-08-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-10-15","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-12-11","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}