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羧甲基纤维素-明胶-二氧化硅纳米杂化物:一种用于α淀粉酶的高效载体基质。

Carboxymethyl cellulose-gelatin-silica nanohybrid: an efficient carrier matrix for alpha amylase.

作者信息

Singh Vandana, Ahmad Shakeel

机构信息

Department of Chemistry, University of Allahabad, Allahabad 211002, India.

Department of Chemistry, University of Allahabad, Allahabad 211002, India.

出版信息

Int J Biol Macromol. 2014 Jun;67:439-45. doi: 10.1016/j.ijbiomac.2014.03.051. Epub 2014 Apr 4.

Abstract

Carboxymethyl cellulose (CMC)-gelatin (G) dual templated polymerization of tetramethoxysilane (TMOS) furnished an efficient hybrid carrier support for alpha amylase. The material has been characterized using FTIR, XRD SEM, TGA and BET studies. The amylase was immobilized at the presynthesized hybrid support by adsorption and the immobilized enzyme was used to optimize the conditions for soluble starch hydrolysis. The immobilization did not change the optimum working pH (pH 5) and temperature (40°C) of the enzymatic reaction. The kinetic parameters of the immobilized (Km=9.970mgmL(-1); Vmax=66.23mgmL(-1)min(-1)) and free amylase (KM=4.0509mgmL(-1), Vmax=4.2909mgmL(-1)min(-1)) indicated that the immobilization has enhanced the catalytic function of diastase alpha amylase. The immobilized enzyme showed higher shelf life as compared to the free enzyme in solution and it could be reused for seven consecutive cycles where 85% of the initial activity was exhibited even in the last cycle. The present material is as efficient as our previously reported material CMC-AgNps-Si.

摘要

羧甲基纤维素(CMC)-明胶(G)双模板法聚合四甲氧基硅烷(TMOS)为α淀粉酶提供了一种高效的杂化载体支持物。通过傅里叶变换红外光谱(FTIR)、X射线衍射(XRD)、扫描电子显微镜(SEM)、热重分析(TGA)和比表面积分析(BET)等研究对该材料进行了表征。通过吸附作用将淀粉酶固定在预先合成的杂化载体上,并使用固定化酶优化可溶性淀粉水解的条件。固定化过程并未改变酶促反应的最佳工作pH(pH 5)和温度(40°C)。固定化淀粉酶(Km = 9.970mg/mL;Vmax = 66.23mg/mL·min-1)和游离淀粉酶(KM = 4.0509mg/mL,Vmax = 4.2909mg/mL·min-1)的动力学参数表明,固定化增强了淀粉酶α淀粉酶的催化功能。与溶液中的游离酶相比,固定化酶表现出更长的保质期,并且可以连续重复使用七个循环,即使在最后一个循环中仍能保持85%的初始活性。本材料与我们之前报道的材料CMC-AgNps-Si一样高效。

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