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However, methanol is preferably avoided in industrial-scale fermentation given its toxicity, flammability, and explosiveness. To develop a protein production system under reduced methanol supply conditions, we attempted to characterize the AOX1 promoter induction activity by comparing derepression conditions with methanol induction conditions. This comparison is important because decreasing methanol consumption would enhance the industrial value of Ogataea minuta for heterologous protein production. For such a comparison, an alcohol oxidase-deficient (\u0394aox) strain was generated, with methanol only being used for AOX1 promoter induction. We also developed a culture process in a jar fermentor using the O. minuta \u0394aox strain under mixed feed conditions to achieve heterologous protein production comparable to that of the wild-type strain under low-methanol conditions.<\/jats:p>","DOI":"10.1093\/bbb\/zbae160","type":"journal-article","created":{"date-parts":[[2024,11,13]],"date-time":"2024-11-13T20:24:17Z","timestamp":1731529457000},"page":"102-109","source":"Crossref","is-referenced-by-count":2,"title":["Development of a protein production system using <i>Ogataea minuta<\/i> alcohol oxidase-deficient strain under reduced-methanol-consumption conditions"],"prefix":"10.1093","volume":"89","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2820-5334","authenticated-orcid":false,"given":"Masashi","family":"Tsuda","sequence":"first","affiliation":[{"name":"Bioprocess Technology Research Laboratories 1, Daiichi Sankyo Co., Ltd. , Chiyoda, Gunma ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuki","family":"Nakatani","sequence":"additional","affiliation":[{"name":"Bioprocess Technology Research Laboratories 1, Daiichi Sankyo Co., Ltd. , Chiyoda, Gunma ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Baba","family":"Satoshi","sequence":"additional","affiliation":[{"name":"Modality Research Laboratories II, Daiichi Sankyo Co., Ltd. , Shinagawa, Tokyo ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koichi","family":"Nonaka","sequence":"additional","affiliation":[{"name":"Technology Innovation Strategy and Intelligence, Daiichi Sankyo Co., Ltd. , Chiyoda, Gunma ,","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2024,11,15]]},"reference":[{"key":"2024122405070506800_bib1","doi-asserted-by":"publisher","first-page":"5301","DOI":"10.1007\/s00253-014-5732-5","article-title":"Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production","volume":"98","author":"Ahmad","year":"2014","journal-title":"Appl Microbiol Biotechnol"},{"key":"2024122405070506800_bib2","doi-asserted-by":"publisher","first-page":"365","DOI":"10.1007\/s12033-019-00164-8","article-title":"Yeast expression systems: overview and recent advances","volume":"61","author":"Baghban","year":"2019","journal-title":"Mol Biotechnol"},{"key":"2024122405070506800_bib3","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1186\/s12934-015-0292-7","article-title":"Development of a mixed feed strategy for a recombinant Pichia pastoris strain producing with a de-repression promoter","volume":"14","author":"Capone","year":"2015","journal-title":"Microb Cell Fact"},{"key":"2024122405070506800_bib4","doi-asserted-by":"publisher","first-page":"277","DOI":"10.1016\/S0141-0229(97)00042-2","article-title":"Recombinant protein production in an alcohol oxidase-defective strain of Pichia pastoris in fedbatch fermentations","volume":"21","author":"Chiruvolu","year":"1997","journal-title":"Enzyme Microb Technol"},{"key":"2024122405070506800_bib5","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1186\/1475-2859-5-17","article-title":"Operational strategies, monitoring and control of heterologous protein production in the methylotrophic yeast Pichia pastoris under different promoters: a review","volume":"5","author":"Cos","year":"2006","journal-title":"Microb Cell Fact"},{"key":"2024122405070506800_bib6","doi-asserted-by":"publisher","first-page":"2279","DOI":"10.1271\/bbb1961.44.2279","article-title":"Oxidation of methanol by the yeast, Pichia pastoris. 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