Nadar Shamraja S, Rathod Virendra K
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.
Int J Biol Macromol. 2017 Feb;95:511-519. doi: 10.1016/j.ijbiomac.2016.11.084. Epub 2016 Nov 23.
The self-assembled glucoamylase metal-organic framework (glucoamylase-MOF) was synthesized by facile one-step method within 20min by simply mixing aqueous solution of 2-methylimidazole (160mM), glucoamylase (5mg/mL) and zinc acetate (40mM) at room temperature (28±2°C). The prepared glucoamylase-MOF was characterized by using FT-IR, confocal scanning laser microscopy, XRD and SEM. The robustness and thermal stability of glucoamylase embedded MOF was evaluated in terms of half-life (in the range of 60-80°C) which showed 6 folds increment as against free form. Further, in Michaelis-Menten kinetics studies, glucoamylase entrapped MOF exhibited higher K value and lower V value as compared to native enzyme. Moreover, the immobilized glucoamylase exhibited up to 57% of residual activity after six consecutive cycles of reuse, whereas it retained 91% of residual activity till 25days of storage. Finally, the conformational changes occurred after the encapsulation of glucoamylase in the interior of MOF, which was analyzed by using FT-IR data analysis tools.
通过在室温(28±2°C)下简单混合2-甲基咪唑水溶液(160mM)、糖化酶(5mg/mL)和醋酸锌(40mM),采用简便的一步法在20分钟内合成了自组装糖化酶金属有机框架(glucoamylase-MOF)。使用傅里叶变换红外光谱(FT-IR)、共聚焦扫描激光显微镜、X射线衍射(XRD)和扫描电子显微镜(SEM)对制备的糖化酶-MOF进行了表征。通过半衰期(在60-80°C范围内)评估了包埋糖化酶的MOF的稳定性和热稳定性,结果表明其半衰期相对于游离形式增加了6倍。此外,在米氏动力学研究中,与天然酶相比,包埋在MOF中的糖化酶表现出更高的K值和更低的V值。此外,固定化糖化酶在连续重复使用六个循环后仍表现出高达57%的残余活性,而在储存25天时仍保留91%的残余活性。最后,利用FT-IR数据分析工具分析了糖化酶包封在MOF内部后发生的构象变化。