Pepe Alfonso, Tito Florencia Rocío, Daleo Gustavo Raúl, Guevara María Gabriela
Instituto de Investigaciones Biológicas, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Mar del Plata, 7600 Mar del Plata, Argentina.
Biotechnol Rep (Amst). 2019 Apr 1;22:e00330. doi: 10.1016/j.btre.2019.e00330. eCollection 2019 Jun.
The aim of this study was to optimize conditions to enhance fibrinogenolytic activity of subtilisin-like protease (SBTc-3). The effects of STBc-3 concentration (0.2-5 μM), pH value (6-10) and temperature (35-50 °C) on fibrinogenolytic activity were studied through response surface methodology (RSM). We obtained a model that predicts the response accurately. The relationship between enzyme concentration and fibrinogenolytic activity was linear, while the main effect from pH and temperature on the response was quadratic. From the RSM generated model the optimum pH was 8 and the optimum temperature was 43 °C, while higher concentrations of enzyme produce higher activities. Under optimum conditions there were no statistically significant differences between the experimental responses and the ones predicted from the model. This model also predicts the activity under physiological conditions. These results confirm that STBc-3 is a good candidate to be considered for therapeutic uses. The generated model will be useful for biotechnological purposes.
本研究的目的是优化条件以提高类枯草杆菌蛋白酶(SBTc-3)的纤维蛋白原溶解活性。通过响应面法(RSM)研究了SBTc-3浓度(0.2 - 5 μM)、pH值(6 - 10)和温度(35 - 50 °C)对纤维蛋白原溶解活性的影响。我们得到了一个能准确预测响应的模型。酶浓度与纤维蛋白原溶解活性之间的关系是线性的,而pH值和温度对响应的主要影响是二次的。从RSM生成的模型来看,最佳pH值为8,最佳温度为43 °C,而较高的酶浓度会产生更高的活性。在最佳条件下,实验响应与模型预测值之间没有统计学上的显著差异。该模型还能预测生理条件下的活性。这些结果证实SBTc-3是一种可考虑用于治疗用途的良好候选物。生成的模型将对生物技术目的有用。