表达重组猪α干扰素的大肠埃希工程菌分批发酵动力学模型的建立

Building α fermentation kinetic model for the expression of recombinant porcine interferon-α in bioengineered Escherichia coli

关键词:  大肠埃希工程菌  重组猪ɑ干扰素  分批发酵  发酵动力学模型。

Key words:  Escherichia coli engineering bacterium  recombinant porcine interferon-ɑ  batch fermentation  fermentation kinetic model.

摘要:

为探究表达重组猪α干扰素的大肠埃希工程菌的分批发酵动力学基本规律，以表达猪α干扰素的基因工程重组大肠埃希菌BL21(DE3）为实验对象，观察并研究了重组大肠埃希工程菌在30 L发酵罐分批发酵过程中菌体生长、重组猪α干扰素生成和基质消耗的变化规律，根据经典的Logistic方程、Luedeking-Piret方程和类似Luedeking-Piret方程对其实验结果进行非线性回归分析。结果表明：菌体生长呈典型的S型曲线，蛋白生成和菌体生长无明显的相关性，属于非生长偶联型；重组猪ɑ干扰素的产生和大肠埃希工程菌的生长速率及菌体积累量相关，菌体生长、基质消耗和产物形成模型的拟合度R2分别为0.99030、0.91095和0.96683，能较好地描述发酵中的动力学特征；重组猪ɑ干扰素蛋白得率提高了40％，由此建立的模型将为后续的连续补料、高密度发酵和工业化放大生产奠定了基础。

Abstract:

To explore the Anhui jiuchuan biotechnology co. LTD. Technical development department''s E. coli expressing recombinant pig interferon alpha(α) batch fermentation kinetics of the basic law of engineering bacteria, as a token of pig interferon α genetic engineering recombinant E. coli BL21 (DE3) as the experimental object, observe and study the recombinant E. coli and engineering bacteria in 30 L fermentor batch fermentation process of bacteria growth, restructuring the pig interferon α generation and the change rule. According to the classical Logistic equation, luedeking-piret equation and similar luedeking-piret equation, nonlinear regression analysis was carried out on the test results. The results showed that the growth of bacteria showed a typical s-shaped curve, and there was no obvious correlation between protein production and growth of bacteria. Restructuring pig interferon α production and the growth rate of E. coli and engineering bacteria and bacteria accumulation, bacteria growth, substrate consumption and product form model fitting R^2 is 0.99030, 0.91095 and 0.96683 respectively,which can well describe the dynamics characteristic in the fermentation; Restructuring the pig interferon α protein yield increased by 40%. Therefore, the established model will lay a foundation for the subsequent high-density fermentation and industrial amplification.