Department of Microbiology, Dr. Harisingh Gour Vishwavidyalaya (A Central University), Sagar, India.
J Food Sci. 2022 Jul;87(7):3294-3306. doi: 10.1111/1750-3841.16173. Epub 2022 May 31.
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 ) 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 specific activity. Purified FTase (75 kDa) had K and V values of 1049.717 mM and 2.094 µmol min mg , 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.
在本研究中，应用响应面法优化了米曲霉 FTase 生成益生元果寡糖 (FOS)。在 28.4°C、pH7.0 和 50%（w/v）蔗糖条件下生成最优 FOS（251 g L），产率提高了 1.97 倍。m-FTase 经超滤和 HiTrap Q HP 阴离子交换层析纯化，FTase 的纯化倍数为 2.15 倍，比酶活为 12.76 U mg。纯化的 FTase（75 kDa）的 K 和 V 值分别为 1049.717 mM 和 2.094 µmol min mg。FOS 的掺入导致 caspase 3、caspase 9 和 Bax 基因上调，提示癌细胞中线粒体凋亡的激活。该研究描述了米曲霉中纯化 FTase 的特性、FOS 的生产优化，并揭示了 FOS 在 HT-29 细胞抗癌活性中的作用。实用程序：本研究提供了有关纯化 FTase 的动力学和热力学特性的详细信息，FTase 是一种生成益生元 FOS 的酶。此外，还研究了参与抗癌活性的凋亡基因以及 FOS 的益生元潜力。这些发现对于 FOS 的应用非常重要，优化的生产策略使其在工业应用中具有实用性。
