title: High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source.
abstract: The yeast Ogataea thermomethanolica has recently emerged as a potential host for heterologous protein expression at elevated temperature. To evaluate the feasibility of O. thermomethanolica as heterologous host in large-scale fermentation, constitutive production of fungal phytase was investigated in fed-batch fermentation. The effect of different temperatures, substrate feeding strategies, and carbon sources on phytase production was investigated. It was found that O. thermomethanolica can grow in the temperature up to 40 °C and optimal at 34 °C. However, the maximum phytase production was observed at 30 °C and slightly decreased at 34 °C. The DOT stat control was the most efficient feeding strategy to obtain high cell density and avoid by-product formation. The table sugar can be used as an alternative substrate for phytase production in O. thermomethanolica. The highest phytase activity (134 U/mL) was obtained from table sugar at 34 °C which was 20-fold higher than batch culture (5.7 U/mL). At a higher cultivation temperature of 38 °C, table sugar can be used as a low-cost substrate for the production of phytase which was expressed with an acceptable yield (85 U/mL). Lastly, the results from this study reveal the industrial favorable benefits of employing O. thermomethanolica as a host for heterologous protein production.
authors.affiliation: Department of Biotechnology, Faculty of Science and Technology, Rangsit Center, Thammasat University, Pathum Thani, 12120, Thailand.
authors.forename: Theppanya
authors.initials: T
authors.lastname: Charoenrat
authors.affiliation: Department of Biotechnology, Faculty of Science and Technology, Rangsit Center, Thammasat University, Pathum Thani, 12120, Thailand.|Food Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani, 12120, Thailand.
authors.forename: Sompot
authors.initials: S
authors.lastname: Antimanon
authors.affiliation: Microbial Cell Factory Laboratory, Bioresources Technology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand.
authors.forename: Kanokarn
authors.initials: K
authors.lastname: Kocharin
authors.affiliation: Microbial Cell Factory Laboratory, Bioresources Technology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand.
authors.forename: Sutipa
authors.initials: S
authors.lastname: Tanapongpipat
authors.affiliation: Microbial Cell Factory Laboratory, Bioresources Technology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand. niran.roo@biotec.or.th.
authors.forename: Niran
authors.initials: N
authors.lastname: Roongsawang
mesh_terms: D010832:6-Phytase / Q000096:biosynthesis*; D000222:Adaptation, Physiological; D061249:Batch Cell Culture Techniques; D001709:Biotechnology / Q000379:methods*; D002244:Carbon / Q000494:pharmacology*; D002452:Cell Count; D019422:Dietary Sucrose / Q000494:pharmacology*; D005240:Feasibility Studies; D005285:Fermentation / Q000187:drug effects; D010100:Oxygen / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis*; D004718:Saccharomycetales / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D013696:Temperature
chemical_list: D019422:Dietary Sucrose; D011994:Recombinant Proteins; D002244:Carbon; D010832:6-Phytase; D010100:Oxygen
keywords: Fed-batch fermentation; High-temperature process; Ogataea thermomethanolica; Phytase; Table sugar; Thermotolerant methylotrophic yeast