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首页> 外文期刊>Bioprocess and Biosystems Engineering >Using a kinetic model that considers cell segregation to optimize hEGF expression in fed-batch cultures of recombinant Escherichia coli
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Using a kinetic model that considers cell segregation to optimize hEGF expression in fed-batch cultures of recombinant Escherichia coli

机译：使用考虑细胞分离的动力学模型优化重组大肠杆菌分批补料培养中的hEGF表达

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Growth inhibition of recombinant Escherichia coli during the expression of human epidermal growth factor was observed. The recombinant cells could be segregated into three populations based on their cell division and plasmid maintenance abilities: dividing and plasmid-bearing cells, dividing and plasmid-free cells, and viable-but-non-culturable (VBNC) cells. Fed-batch fermentations were performed to investigate the effect of cell segregation on the kinetics of growth and foreign protein production. The results showed that a low concentration of inducer caused weak induction, whereas high levels cause strong induction, resulting in cells segregating into VBNC bacteria and producing a low foreign protein yield. A kinetic model for cell segregation was proposed and its predictions correlated well with experimental data for cell growth and protein expression. The optimal induction strategy could then be predicted by the model, and this prediction was then verified by experimentally deriving the conditions necessary for maximum expression of recombinant protein.
机译：观察到在人表皮生长因子表达过程中重组大肠杆菌的生长抑制。根据重组细胞的细胞分裂和质粒维持能力，可以将其分为三个种群：分裂的和携带质粒的细胞，分裂的和不含质粒的细胞以及存活但不可培养的（VBNC）细胞。进行分批补料发酵以研究细胞分离对生长动力学和外源蛋白质产生的影响。结果表明，低浓度的诱导剂诱导弱诱导，而高水平的诱导剂强烈诱导，导致细胞分离成VBNC细菌并产生低的外源蛋白产量。提出了细胞分离的动力学模型，其预测与细胞生长和蛋白质表达的实验数据很好地相关。然后可以通过模型预测最佳诱导策略，然后通过实验推导重组蛋白最大表达所必需的条件来验证该预测。

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