Application of a Fed-Batch Bioprocess for the Heterologous Production of hSCOMT in Escherichia coli
Application of a Fed-Batch Bioprocess for the Heterologous Production of hSCOMT in Escherichia coli
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DOI：
10.4014/jmb.0812.658
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发表时间：
2009-09-01
影响因子：
2.8
通讯作者：
Queiroz, J. A.
中科院分区：
文献类型：
--
作者：
Passarinha, L. A.;Bonifacio, M. J.;Queiroz, J. A.
本文介绍了一种在重组大肠杆菌BL 21（DE 3）中分批补料培养生产人可溶性儿茶酚-O-甲基转移酶（hSCOMT）的方法。第一次，一个简单的模型应用于重组hSCOMT表达系统，并区分诱导后的蛋白质生产阶段的初始细胞生长阶段。具体而言，动力学模型预测随着时间的推移培养物中的生物质、底物和产物浓度，并且从通过测试几种进料曲线设计的一系列分批进料实验中鉴定。该模型的主要优点是其参数可以更可靠地从不同的补料分批策略中识别，例如甘油脉冲和指数，然后是恒定的底物添加。有趣的是，有限的数据量，所提出的模型圆满地完成了三个状态变量的实验结果，并没有详尽的过程知识是必需的。通过验证实验获得的测量数据与模型预测值的比较，表明该模型具有很强的外推能力，可为COMT生产系统提供新的补充信息。
in this paper, a fed-batch cultivation process in recombinant Escherichia coli BL21(DE3) bacteria, for the production of human soluble catechol-O-methyltransferase (hSCOMT), is presented. For the first time, a straightforward model is applied in a recombinant hSCOMT expression system and distinguishes an initial cell growth phase from a protein production phase upon induction. Specifically, the kinetic model predicts biomass, substrate, and product concentrations in the culture over time and was identified from a series of fed-batch experiments designed by testing several feed profiles. The main advantage of this model is that its parameters can be identified more reliably from distinct fed-batch strategies, such as glycerol pulses and exponential followed by constant substrate additions. Interestingly, with the limited amount of data available, the proposed model accomplishes satisfactorily the experimental results obtained for the three state variables, and no exhaustive process knowledge is required. The comparison of the measurement data obtained in a validation experiment with the model predictions showed the great extrapolation capability of the model presented, which could provide new complementary information for the COMT production system.