高等学校化学学报 ›› 2001, Vol. 22 ›› Issue (4): 678.

• 研究论文 • 上一篇    下一篇

聚碳酸乙烯撑酯与双端氨基聚乙二醇交联体系固定化酶载体的合成与表征

黄家贤, 丁伦汉, 曹喆, 李悦, 蒋艳   

  1. 南开大学化学系离子交换与吸附国家重点实验室, 天津 300071
  • 收稿日期:1999-12-12 出版日期:2001-04-24 发布日期:2001-04-24
  • 通讯作者: 黄家贤(1936年出生),男,博士,教授,博士生导师,主要从事大环高分子、感光高分子、高分子酶载体、离子型聚合、原子转移活性自由基聚合和高分子中间体研究.
  • 基金资助:

    国家自然科学基金(批准号:59973009)资助

Synthesis and Characterizations of New Supports for Enzymes Immobilization from Poly(vinylene carbonate) and α-(2-Aminoethyleneamino)-ω-(2-aminoethyleneamino)-poly(ethylene oxide)

HUANG Jia-Xian, DING Lun-Han, CAO Zhe, LI Yue, JIANG Yan   

  1. Department of Chemistry, The State Key Labortory of Functional Polymer Materials for Adsorption and Seperation, Nankai University, Tianjin 300071, China
  • Received:1999-12-12 Online:2001-04-24 Published:2001-04-24

摘要: 将聚碳酸乙烯撑酯(PVCA)与α,ω-双端氨基聚乙二醇(H2N-PEG-NH2)溶于DMF,于液蜡中进行交联反应制得亲水性固定化酶载体,将其与胰蛋白酶进行偶联反应制备了固定化胰蛋白酶.酶蛋白的比活力及其于载体上的结合量与反应条件有关,当w(PVCA)/w(H2N-PEG-NH2)为0.5时,二者均处于最高值.此固定化酶酶促反应的最适pH值和Km值均较之溶液酶有显著提高,但二者的最适酶促反应温度却相当一致.

关键词: 聚乙二醇, 聚碳酸乙烯撑酯, 固定化酶, 载体, 胰蛋白酶

Abstract: In this paper, poly(ethylene oxide) end capped with amine groups(PEO) was prepared and dissolved with poly(vinylene carbonate)(PVCA) in DMFsolution. The solution was suspended in paraffin oil to carry out the crosslinking reaction and produce beadlike hydrophilic supports containing reactive cyclic carbonate groups for enzyme immobilization. In above reaction, PEOwas acted as both crosslinking agent and hydrophilic modifier of poly(vinylene carbonate). Evaluation of the supports using trypsin as a model enzyme shows that the specific activity of immobilized trypsin and amount of enzymes coupled to supports were related to PVCA/PEOratio, reaction time, concentration and pHvalue of enzyme solution, etc.. The amount of trypsin coupled and the specific activity reach their maximum at w (PVCA)/ w (PEG)=1/2. The optimal pHvalue for trypsin immobilized increases as compared with the optimal pHvalue for native trypsin. The Km value of immobilized and native trypsin was determined to be 325 mmol/Land 152 mmol/L, respectively.

Key words: Poly(ethylene glycol), Poly(vinylene carbonate), Fixed enzyme, Support, Trysin

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