A novel genetic switch with an optimal ON/OFF ratio to preserve growth performance prior to Escherichia coli autolysis for enhanced plasmid release

当生物技术的主力生物，大肠杆菌（E。大肠杆菌）用于繁殖商业上有价值的质粒，例如全球扩展的细胞和基因治疗部门所需要的那些质粒，从大肠杆菌中有效回收质粒。coli细胞是关键。然而，所需质粒产物的这种回收可以证明是具有挑战性的，需要从复杂的工艺流中大规模地进行细胞破碎和质粒分离，本研究的目的是应用一种新型的基因开关，具有最佳的ON/OFF比率来控制大肠杆菌自溶，从而保持正常的生长性能，同时允许更简单的剪切诱导分离。已经报道了几种技术，其中E.大肠杆菌自溶被触发以提高重组蛋白或质粒DNA产物的释放。然而，到目前为止，这些E。由于控制自溶事件起始的遗传触发机制的“泄漏”，大肠杆菌菌株始终表现出受损的生长性能，并因此表现出受损的产量性能。这项技术如果成功，将为减少DNA制造步骤提供一种关键方法，使该过程更具成本效益和可持续性。

When the biotechnology workhorse organism, Escherichia coli (E. coli) is used to propagate commercially valuable plasmids, such as those demanded by the globally expanding cell and gene therapy sector, efficient recovery of plasmids from the E. coli cell is key. However, this recovery of the desired plasmid product can prove challenging, with requirements for cell disruption and plasmid isolation from a complex process stream at scale.The aim of this study is to apply a novel genetic switch, with an optimal ON/OFF ratio to control E.coli autolysis such that normal growth performance is maintained whilst allowing simpler less shear induced separation. Several technologies have been reported in which E. coli autolysis is triggered to improve liberation of recombinant protein or plasmid DNA products. However, to date these E. coli strains consistently exhibit compromised growth, and therefore yield, performance due to 'leakiness' of the genetic trigger mechanism that controls initiation of autolytic events. This technology if successful will provide a key approach in reducing the number of steps in DNA manufacture, making the process more cost effective and sustainable.