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Effects of temperature shift strategies on human preproinsulin production in the fed-batch fermentation of recombinantEscherichia coli

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Abstract

Preproinsulin is a well-known precursor of human insulin for the regulation of blood glucose levels. In this study, fed-batch fermentations of recombinantEscherichia coli JM109/pPT-MRpi were carried out for the overexpression of human preproinsulin. The expression of human preproinsulin was controlled by the temperature inducibleP2 promoter. The time-course profiles of fed-batch fermentation and SDS-PAGE analysis showed that human insulin expression was triggered by a culture temperature change from 30 to 37°C. Fermentation shift strategies, including the multi-step increase of temperature and the modulation of initiation time, were optimized to obtain high titers of cell mass and preproinsulin. The optimized fed-batch fermentation, consisting of a three-step shift of culture temperature from 30 to 37°C for 2 h, gave the best results of 43.1 g/L of dry cell weight and 33.3% preproinsulin content, which corresponded to 2.0- and 1.2-fold increases, respectively, as compared to those of fed-batch culture at a constant temperature of 37°C.

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Authors and Affiliations

  1. Interdisciplinary Program of Bioengineering, Seoul National University, 151-742, Seoul, Korea

    Young-Jin Son & Jin-Ho Seo

  2. Biotechnology Laboratory, CKDBiO Research Institute, CKDBiO, 425-100, Ansan, Korea

    Young-Jin Son, Sang-Yong Lee, Sung-Jin Oh, Chang-Kyu Kim & Byoung-Taek Choi

  3. Health Industry Center, Chungbuk TechnoPark, 363-883, Chungbuk, Korea

    Kyong-Hee Park

  4. Center for Agricultural Biomaterials, Seoul National University, 151-921, Seoul, Korea

    Yong-Cheol Park

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  1. Young-Jin Son
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Correspondence to Jin-Ho Seo.

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Son, YJ., Park, KH., Lee, SY. et al. Effects of temperature shift strategies on human preproinsulin production in the fed-batch fermentation of recombinantEscherichia coli . Biotechnol. Bioprocess Eng. 12, 556–561 (2007). https://doi.org/10.1007/BF02931354

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  • DOI: https://doi.org/10.1007/BF02931354

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