以高压静电纺PVA纳米纤维膜为载体化学合成法固定葡萄糖淀粉酶

Immobilization of glucoamylase onto electrospinning PVA nanofiber membrane via chemical process

利用静电纺丝纳米纤维具有高比表面积和多孔疏松结构的优势,固定葡萄糖淀粉酶(GA)以提高酶的重复使用性。采用活化酯法,以甲苯基磺酰氟(PMSF)为基团活化剂在乙醇溶液中活化聚乙烯醇(PVA)纳米纤维膜,再在水溶液以共价键连接固定GA。通过对红外光谱的分析,表明GA可成功固定在静电纺丝形成的PVA纳米纤维膜表面。与游离的GA相比较,固定化的GA体现出较高的酶活性,更宽泛的适用温度和pH,以及更好的热稳定性和存贮稳定性,具有较好的重复使用性。此法制备生物活性酶膜操作简单,具有一定的应用前景。

Immobilized glucoamylase that was able to improve the reusability of the enzyme was synthesized on elec- trospinning nanofibers with high surface area and porous structure. P-toluenesulfonyl fluoride（PMSF）regarded as an activa- tor, polyvinyl alcohol（PVA）nanofiber membrane was activated in alcohol by using an activated ester method, and then the glucoamylase was fixed to PVA nanofibers by covalent connection in an aqueous solution. Fourier transform infrared spec- troscopy（FT-IR ）indicated that glucoamylase can be successfully fixed on the surface of the membrane. Compared with free glucoamylase,the immobilized glucoamylase had enhanced the enzyme stability against changed in pH and temperature, as well as had higher activity and storage stability. The immobilized glucoamylase had also the better reusability. Based on the electrospinning technology and covalent connection, the enzyme memhranes with biological activity can be successfully pre- pared, moreover, which will provide a certain application.