Preparation and Performance Analysis of Nanoflower-immobilized Glucose Isomerase
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Abstract:
In this paper, glucose isomerase (GI) nanoflowers were prepared using the co-precipitation method, and the immobilization conditions were optimized. The morphological characteristics and enzymatic properties of nanoflower-immobilized enzymes were also explored. The results showed that by adding 9 mL of PBS buffer at pH value 7.4 to 40 μL of enzyme solution and mixing with 30 μL CuSO4, and then allowing the reaction to proceed at 35 ℃ for 18 h, Glucose Isomerase @ Nanoflowers (GI@NFs) were obtained, with a recovery rate of enzyme activity at 183.06%. Scanning electron microscopy characterization showed that GI@NFs had a complete nanoflower structure, while Fourier transform infrared spectrometry showed that GI@NFs had characteristic absorption of enzymes and PO43-, and X-ray diffraction results further proved that its carrier was Cu3(PO4)2. In the study of enzymatic properties, the optimum reaction temperature for GI@NFs was 60 ℃, which was 10 ℃ higher than that for the free enzyme; the optimal reaction pH was 8, which was higher than that for the free enzyme. The temperature stability and pH stability of GI@NFs were significantly higher than those for the free enzyme. The immobilized enzymes were reused eight times, and the remaining enzyme activity was 60.32% of the original activity. The results showed that the nanoflower structure improved the activity of glucose isomerase, exhibited better reusability and stability and had certain potential for application.
