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High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source

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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.

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Acknowledgments

We are grateful to Dr. Philip J. Shaw for critically editing the manuscript. Financial support (P-13-50136, P-14-50939) from the National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Thailand, is greatly appreciated. We also acknowledge Central Scientific Instrument Center, Faculty of Science and Technology, Thammasat University, Thailand, for kindly supporting the use of their HPLC.

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  1. Sompot Antimanon

    Present address: Food Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani, 12120, Thailand

Authors and Affiliations

  1. Department of Biotechnology, Faculty of Science and Technology, Rangsit Center, Thammasat University, Pathum Thani, 12120, Thailand

    Theppanya Charoenrat & Sompot Antimanon

  2. 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

    Kanokarn Kocharin, Sutipa Tanapongpipat & Niran Roongsawang

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  1. Theppanya Charoenrat
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Correspondence to Niran Roongsawang.

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Charoenrat, T., Antimanon, S., Kocharin, K. et al. High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source. Appl Biochem Biotechnol 180, 1618–1634 (2016). https://doi.org/10.1007/s12010-016-2191-8

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