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Various heterologous protein production systems have been developed using prokaryotic and eukaryotic hosts. Especially methylotrophic yeast in eukaryotic hosts is suggested to be particularly valuable because such systems have the following advantages: protein secretion into culture broth, eukaryotic quality control systems, a post-translational modification system, rapid growth, and established recombinant DNA tools and technologies such as strong promoters, effective selection markers, and gene knock-in and -out systems. Many methylotrophic yeasts such as the genera <jats:italic>Candida<\/jats:italic>, <jats:italic>Ogataea<\/jats:italic>, and <jats:italic>Komagataella<\/jats:italic> have been studied since methylotrophic yeast was first isolated in 1969. The methanol-consumption-related genes in methylotrophic yeast are strongly and strictly regulated under methanol-containing conditions. The well-regulated gene expression systems under the methanol-inducible gene promoter lead to the potential application of heterologous protein production in methylotrophic yeast. In this review, we describe the recent progress of heterologous protein production technology in methylotrophic yeast and introduce <jats:italic>Ogataea minuta<\/jats:italic> as an alternative production host as a substitute for <jats:italic>K. phaffii<\/jats:italic> and <jats:italic>O. polymorpha<\/jats:italic>.<\/jats:p>","DOI":"10.1007\/s11274-024-04008-9","type":"journal-article","created":{"date-parts":[[2024,5,11]],"date-time":"2024-05-11T02:01:17Z","timestamp":1715392877000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Recent progress on heterologous protein production in methylotrophic yeast systems"],"prefix":"10.1007","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2820-5334","authenticated-orcid":false,"given":"Masashi","family":"Tsuda","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koichi","family":"Nonaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,5,11]]},"reference":[{"key":"4008_CR1","doi-asserted-by":"publisher","first-page":"5301","DOI":"10.1007\/s00253-014-5732-5","volume":"98","author":"M Ahmad","year":"2014","unstructured":"Ahmad M, Hirz M, Pichler H, Schwab H (2014) Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. 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