Immobilized Recombinant Cell Bioreactors

本研究项目主要研究了游离和固定化细胞生物反应器系统中重组细胞的动力学和蛋白质生产。在前人对lac基因表达研究的基础上，我们选择了一株枯草芽孢杆菌作为α-淀粉酶的组成型表达菌株。该质粒pBD214既含有结构基因，又含有酶分泌的启动子和信号序列。实验设计和数据收集程序与数学建模方面相结合，以揭示原型生物反应器中重组细胞中基因表达的统一图景。开发新生物技术的产品将需要高效的生物反应器和对这些反应器中的动力学的了解。细胞生长、质粒复制和脱落以及基因表达的动力学是一个复杂的、高度耦合的过程。鉴于目前的技术水平，有必要研究不同生物反应器系统中的重组细胞动力学，以确定给定过程中细胞生长和产物形成的优化。在许多情况下，准确、统一的模式将指导和加快进程的发展，但目前该领域缺乏这样的模式。这项工作解决了这些限制。

This research project is focused on the study of recombinant cell kinetics and protein production in both free and immobilized cell bioreactor systems. Building on previous work on lac gene expression, a Bacillus subtilis strain has been chosen with a plasmid for constitutive expression of a- amylase. The plasmid, pBD214, contains both the structural gene and the promoter and signal sequences for enzyme secretion. Experimental design and data gathering procedures are integrated with mathematical modeling aspects to uncover a unified picture of gene expression in recombinant cells in prototypical bioreactors. Development of products from the new biotechnology will require efficient bioreactors and an understanding of the kinetics in these reactors. The kinetics of cell growth, plasmid replication and shedding and gene expression are complex and highly coupled processes. Given the state-of- the-art, it is necessary to examine recombinant cell kinetics in different bioreactor systems to decide optimization of cell growth and product formation in a given process. Accurate, unified models would guide and accelerate process development in numerous cases but such models are currently lacking in the field. This work addresses such limitations.