Korpys-Woźniak Paulina, Kubiak Piotr, Celińska Ewelina
Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, ul. Wojska Polskiego 48, 60-637 Poznań, Poland.
Biotechnol Rep (Amst). 2021 Sep 11;32:e00669. doi: 10.1016/j.btre.2021.e00669. eCollection 2021 Dec.
Depending on the suboptimal factor, the target protein secretion can be over 1000-fold below the theoretical maximum. The bottlenecks may be alleviated by co-overexpression of "secretory helpers" (). Here we proposed twelve functionally spanning the whole transcription-translation-translocation-folding-maturation-excretion pipeline. The genes were co-transformed with an easy-to-track reporter, and tested less than two temperatures. Our results indicated a clear distinction in the effects triggered by involved in either synthesis or trafficking of the heterologous polypeptides. For superior operation of synthesis-related , namely and , the secretory pathway's capacity must be released by applying decreased temperature (25 °C). The other considered (e.g. ) did not give such spectacular results in the amounts of the target heterologous polypeptide, but allowed to maintain secretory capacity under unfavorable thermal conditions. This study provides generalizable guidelines for cloning/culturing strategies aiming at enhancement of heterologous protein secretion in .
根据次优因素，目标蛋白分泌量可能比理论最大值低1000倍以上。通过共过量表达“分泌辅助蛋白”（），瓶颈可能会得到缓解。在此，我们提出了十二个功能涵盖整个转录-翻译-转运-折叠-成熟-分泌流程的（辅助蛋白）。这些基因与一个易于追踪的报告基因共转化，并在低于两个温度下进行测试。我们的结果表明，参与异源多肽合成或转运的（辅助蛋白）所引发的效应存在明显差异。为了使与合成相关的（辅助蛋白），即（某两个辅助蛋白）和（某两个辅助蛋白）更好地发挥作用，必须通过降低温度（25°C）来释放分泌途径的能力。其他被考虑的（辅助蛋白）（例如）在目标异源多肽的产量方面没有给出如此显著的结果，但能在不利的热条件下维持分泌能力。本研究为旨在提高（某生物）中异源蛋白分泌的克隆/培养策略提供了可推广的指导方针。
