Abstract：
In the present study, generation of prebiotic fructooligosaccharides (FOS) using Aspergillus tamarii FTase was optimized by applying response surface methodology. Optimal FOS (251 g L-1 ) was generated at 28.4°C, pH 7.0 and 50% (w/v) sucrose leading to 1.97-fold yield enhancement. The m-FTase was purified using ultrafiltration followed by HiTrap Q HP anion exchange chromatography resulting in 2.15-fold purified FTase with 12.76 U mg-1 specific activity. Purified FTase (75 kDa) had Km and Vmax values of 1049.717 mM and 2.094 µmol min-1 mg-1 , respectively. FOS incorporation led to upregulation of caspase 3, caspase 9, and Bax genes suggesting mitochondrial apoptosis activation in cancer cells. The study describes characteristics of purified FTase from A. tamarii, production optimization of FOS and unravels the role of FOS in anticancer activity against HT-29 cells. PRACTICAL APPLICATION: This study provides detailed insights of kinetic and thermodynamic characteristics of purified FTase, a prebiotic FOS-generating enzyme. Moreover, the role of the apoptotic genes involved in anticancer activity, and the prebiotic potential of FOS is also investigated. These findings are important in the context of FOS applications, and the optimized production strategies make it useful for industrial application.
摘要：
在本研究中,通过应用响应面法优化了使用tamarii曲霉FTase的益生元低聚果糖（FOS）的生成。最佳FOS（251gL-1）在28.4°C下产生，pH7.0和50％（w/v）蔗糖导致产量提高1.97倍。使用超滤纯化m-FTase，然后进行HiTrapQHP阴离子交换色谱，得到2.15倍纯化的FTase，比活性为12.76Umg-1。纯化的FTase（75kDa）的Km和Vmax值为1049.717mM和2.094µmolmin-1mg-1，分别。FOS掺入导致caspase3，caspase9和Bax基因上调，表明癌细胞中线粒体凋亡激活。该研究描述了来自A.tamarii的纯化FTase的特性，FOS的生产优化和揭示FOS在抗HT-29细胞抗癌活性中的作用。实际应用：本研究提供了纯化FTase的动力学和热力学特性的详细见解，一种产生FOS的益生元酶。此外，参与抗癌活性的凋亡基因的作用，还研究了FOS的益生元潜力。这些发现在FOS应用的背景下很重要，优化的生产策略使其对工业应用有用。
