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过程工程学报 ›› 2016, Vol. 16 ›› Issue (3): 494-499.DOI: 10.12034/j.issn.1009-606X.215389

• 生化工程专栏 • 上一篇    下一篇

以静电纺PAA/PVA纤维膜为载体活化酯法固定葡萄糖淀粉酶

隋春红 王昭懿 韦雨清 王程   

  1. 吉林医药学院化学教研室 武汉大学信息学部印刷与包装系 吉林医药学院化学教研室 吉林医药学院化学教研室
  • 收稿日期:2015-11-18 修回日期:2016-01-04 出版日期:2016-06-20 发布日期:2017-04-28
  • 通讯作者: 王程

Immobilization of Glucoamylase onto Electrospun PAA/PVA Microfibrous Membrane by Active Ester Method

SUI Chun-hong WANG Zhao-yi WEI Yu-qing WANG Cheng   

  1. Jilin Medical University School of Printing and Packaging, Wuhan Universit Jilin Medical University Jilin Medical University
  • Received:2015-11-18 Revised:2016-01-04 Online:2016-06-20 Published:2017-04-28
  • Contact: WANG Cheng

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摘要： 以高压静电纺聚丙烯酸(PAA)/聚乙烯醇(PVA)纤维膜为载体，以羟基琥珀酰亚胺和二环己基碳二亚胺为基团偶联剂和活化剂，采用活化酯法固定葡萄糖淀粉酶. 结果表明，葡萄糖淀粉酶可固定于高压静电纺PAA/PVA纤维膜表面，固载后纤维直径约为435 nm，固定化酶的最大反应速率为5.77 g/(L×h)，米氏常数Km=10.45 g/L；游离酶与固定化酶的最适pH值和温度基本相同，贮存12 d固定酶活力为新鲜酶的62%，游离酶活力约为新鲜酶的40%；固定化酶重复使用10次仍能保持40.5%活性.

关键词: 静电纺丝, 固定化酶, 活化酯, 聚丙烯酸

Abstract: The carboxyl groups of poly(acrylic acid) (PAA)/polyvinyl alcohol (PVA) microfibrous membranes were activated by using dicyclohexyl-carbodiimide and N-hydroxysuccinimide. The free amines of glucoamylase were reacted with the activated carboxyls to form ester bond, which immobilized the enzyme onto the electrospun PAA/PVA microfiber by covalent bonds. The enzymatic reaction of the immobilized glucoamylase was investigated. The results showed that glucoamylase was attached on the surface of electrospun PAA/PVA composite microfibrous membrane, and the diameter of the fiber was about 435 nm. The maximum reaction velocity (Vmax) and michaelis constant (Km) of the immobilized glucoamylase were 5.77 g/(L×h) and 10.45 g/L, respectively. The optimum pH range and temperature of the immobilized enzyme were similar to those of the free enzyme. Immobilized glucoamylase had higher storage stability. It retained 62% of the original activity in 12 d, as compared to about 40% for the free enzyme. The immobilized glucoamylase could be used for 10 times while retaining 40.5% of its activity.

Key words: electrospinning, immobilized enzyme, active ester, poly(acrylic acid)

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