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首页> 外文期刊>Biotechnology and bioprocess engineering >Effects of Temperature Shift Strategies on Human Preproinsulin Production in the Fed-batch Fermentation of Recombinant Escherichia coli
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Effects of Temperature Shift Strategies on Human Preproinsulin Production in the Fed-batch Fermentation of Recombinant Escherichia coli

机译：温度变化策略对重组大肠杆菌补料分批发酵中人胰岛素原生产的影响

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摘要

Preproinsulin is a well-known precursor of human insulin for the regulation of blood glucose levels. In this study, fed-batch fermentations of recombinant Escherichia coli JM109/pPT-MRpi were carried out for the overexpression of human preproinsulin. The expression of human preproinsulin was controlled by the temperature inducible P2 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℃. 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℃ 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℃.
机译：前胰岛素原是人胰岛素的众所周知的前体，用于调节血糖水平。在这项研究中，重组人大肠杆菌JM109 / pPT-MRpi的分批补料发酵用于人类前胰岛素原的过表达。人前胰岛素原的表达受温度诱导的P2启动子控制。分批补料发酵和SDS-PAGE分析的时程图表明，培养温度从30℃变化到37℃会触发人胰岛素的表达。优化了发酵转移策略，包括温度的多步升高和起始时间的调节，以获得高滴度的细胞质量和胰岛素原。优化的分批补料发酵，包括将培养温度从30℃逐步转变为37℃2个小时，可得到43.1 g / L的干细胞重量和33.3％的胰岛素原含量，相当于2.0-与在37℃恒温下分批补料培养相比，分别增加了1.2倍和1.2倍。

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