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Production of recombinant human epidermal growth factor in Pichia pastoris

机译:巴斯德毕赤酵母中重组人表皮生长因子的生产

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摘要

This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27 μg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60 h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.
机译:进行该研究以在巴斯德毕赤酵母GS115中表达人表皮生长因子(hEGF)。为了这个目的,将hEGF基因克隆到pPIC9K表达载体中,然后通过电穿孔整合到巴斯德毕赤酵母中。基于ELISA的分析表明,分泌到培养基中的hEGF的量会受到发酵条件的影响,特别是受培养基,pH和温度的影响。最佳hEGF产生的最佳培养基是在6.0和7.0的pH范围内缓冲的BMMY。通过用0.5%(v / v)甲醇诱导培养60小时可获得最高量的hEGF,平均产量为2.27μg/ mL。人工神经网络(ANN)分析显示,pH和温度的变化均显着影响hEGF的产生,pH的变化对hEGF的产生的影响比温度的变化稍高。

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