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壳聚糖水凝胶微球:一种用于可溶性葡聚糖酶交联以提高稳定性和循环利用效率的有效共价基质。

Chitosan hydrogel microspheres: an effective covalent matrix for crosslinking of soluble dextranase to increase stability and recycling efficiency.

作者信息

Shahid Faiza, Aman Afsheen, Nawaz Muhammad Asif, Karim Asad, Ul Qader Shah Ali

机构信息

The Karachi Institute of Biotechnology and Genetic Engineering (KIBGE), University of Karachi, Karachi, 75270, Pakistan.

Department of Biochemistry, University of Karachi, Karachi, 75270, Pakistan.

出版信息

Bioprocess Biosyst Eng. 2017 Mar;40(3):451-461. doi: 10.1007/s00449-016-1713-7. Epub 2016 Nov 30.

Abstract

Dextranase is a unique biocatalyst that has high specificity and stereo-selectivity towards a complex biopolymer known as dextran. Dextranase has wide industrial application, but most of the time harsh environmental conditions adversely affect the functionality and stability of the enzyme. To overcome this issue, a covalent cross-linking immobilization method was adapted in the current study utilizing a nontoxic and biocompatible matrix known as chitosan. Chitosan hydrogel microspheres were synthesized using chitosan which exhibited noteworthy physical and mechanical strength. After treatment with glutaraldehyde, chitosan hydrogel microspheres were used for immobilization of dextranase. The kinetic characteristics of immobilized dextranase were compared with that of the soluble enzyme. A shift in optimum pH and temperature from 7.0 to 7.5 and 50 to 60 °C was observed after immobilization, respectively. Recycling efficiency, thermal stability, and activation energy distinctly improved after immobilization, whereas anchoring of substrate at the active site of the soluble dextranase exhibited an increase in K with no change in V after crosslinking. This technique involves the reduction in the size of carrier molecules (microspheres) that provide a larger surface area for improved immobilization efficiency. Therefore, it is concluded that increased stability and reusability of this immobilized biocatalyst makes it a promising aspirant for the utilization at commercial level.

摘要

葡聚糖酶是一种独特的生物催化剂,对一种名为葡聚糖的复杂生物聚合物具有高特异性和立体选择性。葡聚糖酶具有广泛的工业应用,但大多数情况下,恶劣的环境条件会对该酶的功能和稳定性产生不利影响。为了克服这一问题,本研究采用了一种共价交联固定化方法,利用一种名为壳聚糖的无毒且生物相容性良好的基质。使用壳聚糖合成了壳聚糖水凝胶微球,其表现出显著的物理和机械强度。用戊二醛处理后,壳聚糖水凝胶微球用于固定葡聚糖酶。将固定化葡聚糖酶的动力学特性与可溶性酶的动力学特性进行了比较。固定化后分别观察到最适pH从7.0变为7.5,最适温度从50℃变为60℃。固定化后回收效率、热稳定性和活化能明显提高,而交联后可溶性葡聚糖酶活性位点处底物的锚定导致K增加而V不变。该技术涉及减小载体分子(微球)的尺寸,从而提供更大的表面积以提高固定化效率。因此,可以得出结论,这种固定化生物催化剂稳定性和可重复使用性的提高使其成为商业应用的有前途的候选者。

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