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α-淀粉酶在活化的缩二脲基丙烯腈纤维上的固定化:提高酶稳定性和可重复使用性的新方法。

Immobilisation of α-amylase on activated amidrazone acrylic fabric: a new approach for the enhancement of enzyme stability and reusability.

机构信息

King Abdulaziz City for Science and Technology, P.O. Box 6086, 11442, Riyadh, Saudi Arabia.

Chemistry Department, Faculty of Sciences and Arts, University of Jeddah, Khulais, P.O. Box 355, Khulais, 21921, Saudi Arabia.

出版信息

Sci Rep. 2019 Sep 3;9(1):12672. doi: 10.1038/s41598-019-49206-w.

Abstract

In this study, amidrazone acrylic fabric was applied as an immobilising support for α-amylase. The immobilised α-amylase was characterised by Fourier transform infrared spectroscopy and scanning electron microscopy. Furthermore, the optimum conditions for immobilisation efficiency, immobilisation time, reusability, kinetic parameters and pH, for the immobilisation process were examined. The study demonstrated that with 4% cyanuric chloride, and a pH of 7.0, the highest immobilization efficiency of 81% was obtained. Around 65% of the initial activity was maintained after storage at 4 °C for 8 weeks. The immobilised enzyme retained 53% of its original activity after being reused 15 times and exhibited improved stability compared with the free enzyme in relation to heavy metal ions, pH, temperature and inhibitors. The immobilised enzyme presented kinetic parameters of 2.6 mg starch and 0.65 µmol maltose/mL for K and V respectively, compared with 3.7 mg starch and 0.83 µmol maltose/ mL for the free enzyme. The improvements in the enzyme's catalytic properties, stability and reusability obtained from immobilisation make amidrazone acrylic fabric support a good promising candidate for bio-industrial applications.

摘要

在这项研究中,脒基嗪丙烯腈纤维被用作α-淀粉酶的固定化载体。固定化α-淀粉酶的特征通过傅里叶变换红外光谱和扫描电子显微镜进行了研究。此外,还研究了固定化效率、固定化时间、重复使用性、动力学参数和 pH 值等固定化过程的最佳条件。研究表明,在 4%的三聚氯氰和 pH 值为 7.0 的条件下,固定化效率最高可达 81%。在 4°C 下储存 8 周后,仍保持约 65%的初始活性。固定化酶重复使用 15 次后,保留了其原始活性的 53%,与游离酶相比,在重金属离子、pH 值、温度和抑制剂方面表现出更好的稳定性。固定化酶的动力学参数分别为 2.6mg 淀粉和 0.65µmol 麦芽糖/mL 的 K 和 V,而游离酶的相应参数为 3.7mg 淀粉和 0.83µmol 麦芽糖/mL。固定化后,酶的催化性能、稳定性和可重复使用性得到了提高,脒基嗪丙烯腈纤维支持物成为生物工业应用的一个有前途的候选材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2158/6722121/e35c3a5e82c2/41598_2019_49206_Fig1_HTML.jpg

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