biostudies-literatureUnknown110(1)Gao WMethanol has emerged as a promising substrate in biomanufacturing, where high tolerance and efficient assimilation are critical to overcome the cytotoxic effects of methanol and its intermediate metabolites. In this study, a Pichia kudriavzevii P22 strain (genome size: 17.3 Mb) was isolated from fermented camel milk. Remarkably, this strain tolerated up to 15% (v/v) methanol and survived under multiple stress conditions, including pH 2, 47 °C, 5 g/L acetic acid, and 100 g/L NaCl. In a 5-L fed-batch fermentation system operated under open, non-sterile conditions with 7.5% (v/v) methanol as the sole carbon source, P22 remained viable for 78 h. The maximum cell dry weight (CDW) reached 27.56 g/L, with proteins accounting for 54%. Furthermore, a preliminary investigation of the methanol metaboApplied microbiology and biotechnology77https://www.ebi.ac.uk/biostudies/studies/S-EPMC12935861biostudies-literatureRobust Pichia kudriavzevii for open production of methylotrophic protein.PMC12935861Yin GWang RGao WZhang MChen HWang YYue HXu JKuang ZXu ZfalseRobust Pichia kudriavzevii for open production of methylotrophic protein.Methanol has emerged as a promising substrate in biomanufacturing, where high tolerance and efficient assimilation are critical to overcome the cytotoxic effects of methanol and its intermediate metabolites. In this study, a Pichia kudriavzevii P22 strain (genome size: 17.3 Mb) was isolated from fermented camel milk. Remarkably, this strain tolerated up to 15% (v/v) methanol and survived under multiple stress conditions, including pH 2, 47 °C, 5 g/L acetic acid, and 100 g/L NaCl. In a 5-L fed-batch fermentation system operated under open, non-sterile conditions with 7.5% (v/v) methanol as the sole carbon source, P22 remained viable for 78 h. The maximum cell dry weight (CDW) reached 27.56 g/L, with proteins accounting for 54%. Furthermore, a preliminary investigation of the methanol metabo2026-01-01T00:00:00Z2026 Feb2026-07-16T22:04:34.812Z2026-07-11T03:11:24.708ZS-EPMC129358614173556610.1007/s00253-026-13745-w