Department of Biochemistry, Jnana Sahyadri, Kuvempu University, Shankaraghatta, Shivamogga-577451, India.
Department of Biochemistry, Jnana Sahyadri, Kuvempu University, Shankaraghatta, Shivamogga-577451, India.
Int J Biol Macromol. 2024 Jun;269(Pt 1):132021. doi: 10.1016/j.ijbiomac.2024.132021. Epub 2024 Apr 30.
Challenges in enzyme and product recovery are currently intriguing in modern biotechnology. Coping enzyme stability, shelf life and efficiency, nanomaterials-based immobilization were epitomized of industrial practice. Herein, a α-amylase from Geobacillus thermoleovorans was purified and bound effectively on to a modified 3-Aminopropyltriethoxysilane (APTES)-FeO nanoparticle. It was revealed that the carrier-bound enzyme catalysis (pH 8 and 60 °C) was significant in contrast to the free enzyme (pH 7.5 and 55 °C). Furthermore, Zn and Cu were shown to cause inhibitory effects in both enzyme states. Unlike chloroform, toluene, benzene, and butanol, minimal effects were observed with ethanol, acetone, and hexane. The bound enzyme retained 27.4 % of its initial activity after being stored for 36 days. In addition, the reusability of the bound enzyme showed a gradual decline in activity after the first cycle; however, after 13 cycles, its residual activity at 53 % was observed. These data proved significant enough to use this enzyme for industrial starch and analogous substrate bio-processing.
目前，在现代生物技术中，酶和产物回收面临的挑战非常吸引人。为了应对酶的稳定性、保质期和效率问题，基于纳米材料的固定化技术已经成为工业实践的典范。本文从地芽孢杆菌中纯化得到了α-淀粉酶，并将其有效地固定在经过 3-氨丙基三乙氧基硅烷（APTES）-FeO 纳米粒子修饰的载体上。结果表明，与游离酶（pH7.5 和 55°C）相比，载体结合酶的催化作用（pH8 和 60°C）更为显著。此外，Zn 和 Cu 对两种酶状态都表现出抑制作用。与氯仿、甲苯、苯和丁醇不同，乙醇、丙酮和己烷对酶的影响较小。固定化酶在储存 36 天后仍保留初始活性的 27.4%。此外，固定化酶的重复使用性在第一个循环后表现出活性逐渐下降，但在 13 个循环后，观察到其残余活性为 53%。这些数据表明，该酶可用于工业淀粉和类似底物的生物加工，具有重要的应用价值。
