中华医学超声杂志

应用化学 ›› 2010, Vol. 27 ›› Issue (09): 1076-1082.DOI: 10.3724/SP.J.1095.2010.90742

• 研究论文 • 上一篇    下一篇

云芝漆酶在N,N'-亚甲基双丙烯酰胺（BIS）交联聚甲基丙烯酸基元上的固定及其修饰玻碳电极电化学行为

曾涵*,赵淑贤,龚兰新,许国强   

  1. (新疆师范大学生命科学与化学学院 乌鲁木齐 830054)
  • 收稿日期:2009-11-10 修回日期:2010-02-25 出版日期:2010-09-10 发布日期:2010-09-10
  • 通讯作者: 曾涵，男，硕士，讲师; E-mail:zenghan1289@163.com; 研究方向：天然高分子改性及应用

Immobilization of Laccase from Trametes Versicolor on the Matrix of N,N-ethylene bis(acrylamide) Cross-linked Poly(methyl acrylic acid) and the Electrochemical Behavior of its Modified Glassy Carbon Electrode

ZENG Han* , ZHAO Shu-Xian, GONG Lan-Xin, XU Guo-Qiang   

  1. (Institute of Life Science and Chemistry,Xinjiang Normal University,Urumuqi 830054)
  • Received:2009-11-10 Revised:2010-02-25 Published:2010-09-10 Online:2010-09-10
  • Contact: Han ZENG

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1912

被引次数

2

摘要：

以N,N′-亚甲基双丙烯酰胺（BIS）交联聚甲基丙烯酸作为固定漆酶的载体，以共价偶联法固定云芝漆酶并测定了固定基元的酶固定量和固定漆酶的比活力。 还研究了固定漆酶热稳定性、重复使用性以及固定漆酶催化2，6-二甲氧基苯酚（DMP）氧化的酶动力学参数。 实验结果表明，这种交联聚合物基元通过共价偶联法固定漆酶的量和固定漆酶的比活力分别可达26.37 mg/g和1.202 U/mg；在交联聚合物基元上固定的漆酶在50 ℃下放置2 h后仍然保持初始活力的83％，重复使用10次后仍保持初始活力的80％以上；交联聚合物固定漆酶催化DMP氧化的表观速率常数kcat可达1090 min-1，以固定漆酶的BIS交联聚甲基丙烯酸功能化碳纳米管修饰的玻碳电极在pH=4.4磷酸盐缓冲液中氧还原发生在+724 mV(vs.SCE)。

关键词: 漆酶, 酶固定, 热稳定性, 酶动力学, 氧还原

Abstract:

Laccase was immobilized on N,N′-methylene bis(acrylamide)(BIS) cross-linked poly(methyl acrylic acid) through covalent coupling strategy. The ratio of immobilization was determined and the specific activity of entrapped laccase was measured. Thermal stability, reusability of immobilized laccase and enzymatic parameters for 2,6-dimethoxyphenol(DMP) oxidation catalyzed by immobilized laccase were investigated. Results obtained from experiments indicate that the loading of laccase on this crosslinked copolymer matrix and the specific activity of immobilized laccase can reach up to 26.37 mg/g and 1.202 U/mg respectively. Laccase immobilization on this cross-linked copolymer matrix at 50 ℃ for 2 h can still retain about 83％ of its initial activity and can retain more than 80％ of its initial activity after 10 repeated uses. The apparent velocity constant kcat for DMP oxidation catalyzed by the entrapped enzyme can reach ca.1090 min-1. The reduction of oxygen at laccaseentrapped BIS cross-linked poly(methyl acrylic acid) modified glassy carbon electrode in phosphate buffer solution(pH=4.4) occurred near +724 mV(vs.SCE).

Key words: laccase, enzyme immobilization, thermal stability, enzymatic kinetics, oxygen reduction reaction

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