Commercialisation of the Tat protein export pathway for biopharmaceutical production

超过30%的获得许可的治疗蛋白是在大肠杆菌中制造的，在这种情况下，从细胞质“出口”到周质是一种受欢迎的策略。目前的出口战略有严重的局限性，我们已经表明，‘TAT’出口路径比传统系统具有许多优势。这项研究将使我们能够完全商业化一套基于TAT的强大平台：1.我们将优化前面描述的系统，在该系统中，枯草芽孢杆菌TAT系统在大肠杆菌TAT缺失菌株中的表达导致输出到周质，然后释放到培养基中。我们将校准泄漏的外膜表型，以实现细胞稳健性与膜泄漏率之间的最佳平衡。我们将商业化一种新的平台，在该平台中，生物治疗药物通过TAT出口，然后在周质体内形成二硫键。我们将确定新的基于TAT的系统，它的运行效率比大肠杆菌系统更高。

Over 30% of licensed therapeutic proteins are made in the bacterium Escherichia coli, where 'export' out of the cytoplasm to the periplasm is a favoured strategy. Current export strategies have severe limitations and we have shown that the 'Tat' export pathway has a number of advantages over traditional systems. The research will enable us to fully commercialise a suite of powerful Tat-based platforms:1. We will optimise a previously-described system in which expression of a Bacillus subtilis Tat system in an E. coli tat deletion strain leads to export to the periplasm and then release to the medium. We will calibrate the leaky outer membrane phenotype to achieve the optimal balance of cell robustness vs membrane leakiness.2. We will commercialise a new platform in which biotherapeutics are exported by Tat with disulphide-bonds subsequently formed in the periplasm.3. We will identify new Tat-based systems that operate more efficiently than the E. coli system.