Department of Chemical Engineering Institute of Chemical Technology, Matunga (E) Mumbai, 400019, India.
Department of Chemical Engineering Institute of Chemical Technology, Matunga (E) Mumbai, 400019, India.
Enzyme Microb Technol. 2018 Jan;108:11-20. doi: 10.1016/j.enzmictec.2017.08.008. Epub 2017 Aug 30.
The enzyme under lower-intensity ultrasonic irradiation leads to favorable conformational changes, thereby enhancing its activity. In this study, lipase activity was augmented upto 1.6-folds after ultrasonic treatment at 22kHz and 11.38Wcm for 25min. This highly activated lipase was encapsulated within zeolite imidazolate framework-8 (ZIF-8) as a metal-organic framework (MOF) material via facile one-step biomineralization method by simply mixing aqueous solution of 2-methylimidazole (13.3mmol) and zinc acetate (1.33mmol) along with sonicated lipase within 10min at room temperature (28±2°C). The prepared lipase-MOF was characterized by using FT-IR, FT-Raman, XRD, BET, confocal scanning laser microscopy, TGA and SEM. Further, the thermal stability of lipase embedded MOF was evaluated in the range of 55-75°C on the basis of half-life which showed 3.2 folds increment as against free lipase. In Michaelis-Menten kinetics studies, sonicated lipase entrapped MOF showed nearly same K and V values as that of sonicated free lipase. Moreover, the immobilized lipase exhibited up to 54% of residual activity after seven successive cycles of reuse, whereas it retained 90% of residual activity till twenty-five days of storage. Finally, the conformational changes occurred in lipase after sonication treatment and encapsulation within MOF were analyzed by using FT-IR data analysis tools and fluorescent spectroscopy.
在低强度超声辐射下,酶会导致有利的构象变化,从而提高其活性。在这项研究中,经过 22kHz 和 11.38Wcm 超声处理 25min 后,脂肪酶活性提高了 1.6 倍。通过简单混合 2-甲基咪唑(13.3mmol)和乙酸锌(1.33mmol)的水溶液以及在室温(28±2°C)下超声处理的脂肪酶 10min,这种高度激活的脂肪酶可以作为金属有机骨架(MOF)材料被封装在沸石咪唑酯骨架-8(ZIF-8)内通过简便的一步生物矿化方法。所制备的脂肪酶-MOF 通过傅里叶变换红外光谱(FT-IR)、傅里叶变换拉曼光谱(FT-Raman)、X 射线衍射(XRD)、BET、共聚焦扫描激光显微镜、热重分析(TGA)和扫描电子显微镜(SEM)进行了表征。此外,在 55-75°C 的范围内根据半衰期评估了包埋在 MOF 中的脂肪酶的热稳定性,其半衰期比游离脂肪酶增加了 3.2 倍。在米氏动力学研究中,超声处理的脂肪酶包埋的 MOF 显示出与超声处理的游离脂肪酶几乎相同的 K 和 V 值。此外,固定化脂肪酶在连续七次重复使用后仍保持 54%的残余活性,而在 25 天的储存期间仍保持 90%的残余活性。最后,通过傅里叶变换红外光谱数据分析工具和荧光光谱分析了超声处理后脂肪酶的构象变化和包埋在 MOF 内的构象变化。