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Most\n                    <jats:italic>K. phaffii<\/jats:italic>\n                    bioprocesses for recombinant protein production rely on P\n                    <jats:sub>AOX1<\/jats:sub>\n                    to achieve dynamic control in two\u2010phase processes. Cells are first grown under conditions that repress P\n                    <jats:sub>AOX1<\/jats:sub>\n                    (growth phase), followed by methanol\u2010induced recombinant protein expression (production phase). In this study, we propose a methanol\u2010free approach for dynamic metabolic control in\n                    <jats:italic>K. phaffii<\/jats:italic>\n                    using optogenetics, which can help enhance input tunability and flexibility in process optimization and control. The light\u2010responsive transcription factor EL222 from\n                    <jats:italic>Erythrobacter litoralis<\/jats:italic>\n                    is used to regulate protein production from the P\n                    <jats:sub>C120<\/jats:sub>\n                    promoter in\n                    <jats:italic>K. phaffii<\/jats:italic>\n                    with blue light. We used two system designs to explore the advantages and disadvantages of coupling or decoupling EL222 integration with that of the gene of interest. We investigate the relationship between EL222 gene copy number and light dosage to improve production efficiency for intracellular and secreted proteins. Experiments in lab\u2010scale bioreactors demonstrate the feasibility of the outlined optogenetic systems as potential alternatives to conventional methanol\u2010inducible bioprocesses using\n                    <jats:italic>K. phaffii<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1002\/bit.29027","type":"journal-article","created":{"date-parts":[[2025,5,24]],"date-time":"2025-05-24T04:30:26Z","timestamp":1748061026000},"page":"2478-2498","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Balancing Doses of EL222 and Light Improves Optogenetic Induction of Protein Production in\n                    <i>Komagataella phaffii<\/i>"],"prefix":"10.1002","volume":"122","author":[{"given":"Shannon M.","family":"Hoffman","sequence":"first","affiliation":[{"name":"Department of Chemical and Biological Engineering, Hoyt Laboratory Princeton University Princeton New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebasti\u00e1n","family":"Espinel\u2010R\u00edos","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering, Hoyt Laboratory Princeton University Princeton New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah K.","family":"Kwartler","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology Princeton University Princeton New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makoto A.","family":"Lalwani","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering, Hoyt Laboratory Princeton University Princeton New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7209-4208","authenticated-orcid":false,"given":"Jos\u00e9 L.","family":"Avalos","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering, Hoyt Laboratory Princeton University Princeton New Jersey USA"},{"name":"Department of Molecular Biology Princeton University Princeton New Jersey USA"},{"name":"The Omenn\u2010Darling Bioengineering Institute, Princeton University Princeton New Jersey USA"},{"name":"The Andlinger Center for Energy and the Environment, Princeton University Princeton New Jersey USA"},{"name":"High Meadows Environmental Institute, Princeton University Princeton New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,5,24]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.200900233"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.gce.2023.10.004"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.3390\/biom13030441"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2023.1130583"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13568-022-01362-5"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.34133\/2021\/9857418"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2020.07.012"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/elsc.202100070"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-25777-z"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb1312s23"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bej.2005.04.005"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.27979"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.61268"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-33291-4"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.27095"},{"key":"e_1_2_11_17_1","doi-asserted-by":"crossref","unstructured":"Espinel\u2010R\u00edos S. andJ. 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