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Xylose oxidative pathways that oxidize xylose into xylonate can be more advantageous than conventional xylose assimilation pathways because of fewer reaction steps without loss of carbon and ATP. Moreover, commodity chemicals like 3,4\u2010dihydroxybutyrate and 3\u2010hydroxybutyrolactone can be produced from the intermediates of xylose oxidative pathway. However, successful implementations of xylose oxidative pathway in yeast have been hindered because of the secretion and accumulation of xylonate which is a key intermediate of the pathway, leading to low yield of target product. Here, high\u2010yield production of 3,4\u2010dihydroxybutyrate from xylose by engineered yeast was achieved through genetic and environmental perturbations. Specifically, 3,4\u2010dihydroxybutyrate biosynthetic pathway was established in yeast through deletion of <jats:italic>ADH6<\/jats:italic> and overexpression of <jats:italic>yneI<\/jats:italic>. Also, inspired by the mismatch of pH between host strain and key enzyme of XylD, alkaline fermentations (pH\u2009\u2265\u20097.0) were performed to minimize xylonate accumulation. Under the alkaline conditions, xylonate was re\u2010assimilated by engineered yeast and combined product yields of 3,4\u2010dihydroxybutyrate and 3\u2010hydroxybutyrolactone resulted in 0.791\u2009mol\/mol\u2010xylose, which is highest compared with previous study. These results shed light on the utility of the xylose oxidative pathway in yeast.<\/jats:p>","DOI":"10.1002\/bit.28278","type":"journal-article","created":{"date-parts":[[2022,11,2]],"date-time":"2022-11-02T03:29:17Z","timestamp":1667359757000},"page":"511-523","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Enhanced production of 3,4\u2010dihydroxybutyrate from xylose by engineered yeast via xylonate re\u2010assimilation under alkaline condition"],"prefix":"10.1002","volume":"120","author":[{"given":"Takahiro","family":"Yukawa","sequence":"first","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiro","family":"Bamba","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mami","family":"Matsuda","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"},{"name":"Engineering Biology Research Center Kobe University Kobe Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takanobu","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kentaro","family":"Inokuma","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Technology and Innovation Kobe University Kobe Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jungyeon","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Food Science and Human Nutrition University of Illinois at Urbana\u2010Champaign Urbana Illinois USA"},{"name":"Carl R. 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