CAS Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, People's Republic of China.
CAS Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, People's Republic of China.
Int J Biol Macromol. 2019 Feb 1;122:298-305. doi: 10.1016/j.ijbiomac.2018.10.177. Epub 2018 Oct 26.
In the present study, α-glucosidase (α-Glu) was immobilized on the chitosan-modified cellulose filter paper (CFP/CS). For α-Glu immobilization, glutaraldehyde (GA) was used as a coupling agent. CFP, environmentally friendly and commercially available with low cost, will avoid the tedious procedure for synthesizing immobilization carrier. In addition, the CFP/CS-immobilized α-Glu can be directly taken out from the reaction mixture after an enzymatic reaction. This makes the instantaneous separation of immobilized enzyme comes true and is convenient to the subsequent study. Combined with capillary electrophoresis (CE), the CFP/CS-immobilized α-Glu was then used for enzyme kinetic and inhibition study. The CFP/CS-immobilized α-Glu exhibited enhanced pH and temperature tolerance. In addition, the performance of the CFP/CS-immobilized α-Glu was studied. Immobilized α-Glu exhibited excellent batch-to-batch reproducibility (RSD = 6.7%, n = 5) and improved reusability (71.0% of its initial activity after 10 repeated cycles). Finally, immobilized α-Glu was used to screen enzyme inhibitors from 11 traditional Chinese medicines (TCMs). The results showed that CFP/CS has potential development prospects as a novel enzyme immobilization carrier.
在本研究中，α-葡萄糖苷酶（α-Glu）被固定在壳聚糖修饰的纤维素滤纸（CFP/CS）上。为了固定α-Glu，使用戊二醛（GA）作为偶联剂。CFP 环保且价格低廉，可避免合成固定载体的繁琐过程。此外，CFP/CS 固定的α-Glu 可以在酶反应后直接从反应混合物中取出。这使得瞬时分离固定酶成为可能，便于后续研究。结合毛细管电泳（CE），CFP/CS 固定的α-Glu 用于酶动力学和抑制研究。CFP/CS 固定的α-Glu 表现出增强的 pH 和温度耐受性。此外，研究了 CFP/CS 固定的α-Glu 的性能。固定的α-Glu 表现出出色的批间重现性（RSD=6.7%，n=5）和改进的可重复使用性（经过 10 次重复循环后，其初始活性的 71.0%）。最后，固定的α-Glu 用于从 11 种中药（TCM）中筛选酶抑制剂。结果表明，CFP/CS 作为一种新型酶固定载体具有潜在的发展前景。
