Decoupling cell formation from recombinant protein synthesis is a potent strategy to intensify bioprocesses. Escherichia coli strains with mutations in the glucose uptake components lack catabolite repression, display low growth rate, no overflow metabolism, and high recombinant protein yields. Fast growth rates were promoted by the simultaneous consumption of glucose and glycerol, and this was followed by a phase of slow growth, when only glucose remained in the medium. A glycerol‐repressible genetic circuit was designed to autonomously induce recombinant protein expression. The engineered strain bearing the genetic circuit was cultured in 3.9 g L−1 glycerol + 18 g L−1 glucose in microbioreactors with online oxygen transfer rate monitoring. The growth was fast during the simultaneous consumption of both carbon sources (C‐sources), while expression of the recombinant protein was low. When glycerol was depleted, the growth rate decreased, and the specific fluorescence reached values 17% higher than those obtained with a strong constitutive promoter. Despite the relatively high amount of C‐source used, no oxygen limitation was observed. The proposed approach eliminates the need for the substrate feeding or inducers addition and is set as a simple batch culture while mimicking fed‐batch performance.

中文翻译：

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通过甘油抑制遗传回路实现自诱导蛋白表达的运输控制生长解偶联

将细胞形成与重组蛋白合成分离是强化生物过程的有效策略。大肠杆菌葡萄糖摄取成分发生突变的菌株缺乏分解代谢物抑制，表现出低生长速率、无溢出代谢和高重组蛋白产量。同时消耗葡萄糖和甘油促进了快速生长，随后是缓慢生长阶段，此时培养基中仅保留葡萄糖。设计了甘油抑制遗传电路来自主诱导重组蛋白表达。携带遗传电路的工程菌株在 3.9 g L-1 中培养−1甘油+18克/升−1微生物反应器中的葡萄糖，具有在线氧转移率监测。在同时消耗两种碳源（C源）的过程中，生长速度很快，但重组蛋白的表达量较低。当甘油耗尽时，生长速率降低，并且特异性荧光达到比用强组成型启动子获得的值高17%。尽管使用的碳源量相对较高，但没有观察到氧限制。所提出的方法消除了对底物补料或诱导剂添加的需要，并设置为简单的分批培养，同时模仿分批补料性能。