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›› 2007, Vol. 7 ›› Issue (2): 395-398.

• 生化工程专栏 • 上一篇    下一篇

溶胶-凝胶法固定化Candida sp.99-125脂肪酶

高阳,聂开立,王芳,谭天伟   

  1. 北京化工大学生命科学与技术学
  • 出版日期:2007-04-20 发布日期:2007-04-20

Immobilization of Lipase by Sol-Gel Entrapment

GAO Yang,NIE Kai-li,WANG Fang,TAN Tian-wei   

  1. College of Life Science and Technology, Beijing University of Chemical Technology
  • Online:2007-04-20 Published:2007-04-20

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被引次数

19

摘要： 利用正硅酸甲酯(TMOS)和丙基三甲氧基硅烷(PTMS)为复合硅源，以PEG(MW=20000)为稳定剂，以HCl为催化剂，经过溶胶-凝胶过程包埋假丝酵母99-125脂肪酶. 研究得到最适的固定化条件为：PTMS与TMOS的摩尔比4: 1, R值(水与硅源的摩尔比)20, 给酶量(酶占硅源的质量百分数)3.71%, PEG与酶的质量比(1~1.5):1, 硅源水解时间35 min. 在该条件下，固定化脂肪酶的最高酯化活力是游离酶最高酯化活力的2.02倍. 固定化脂肪酶在100℃保温2 h后酶活仍维持为59.1%，固定化酶催化特定酯化反应，经过8批连续反应96 h后酶活维持不变.

关键词: 溶胶-凝胶, 固定化, 脂肪酶

Abstract: Candida sp. 99-125 lipase was entrapped in organic-inorganic sol-gel powder prepared by polymerization of tetramethoxysilane (TMOS) and propyltrimethoxysilane (PTMS). PEG (MW=20000) and HCl were used as stabilizing additive and catalyst during the sol-gel process, respectively. The optimal preparation conditions were studied, which are PTMS/TMOS molar ratio at 4:1, water/silane molar ratio 20, enzyme loading 3.71% (w) of silane precursor and hydrolysis time of silane precursor for 35 min. The specific activity of immobilized lipase was 2.02 times higher than that of non-immobilized lipase under the optimal water content in esterification reaction system. The esterification activity of immobilized lipase retained 59.1% of the initial activity after 2 h incubation under 100℃, and the activity of immobilized lipase remained almost unchanged after 8 consecutive batches of each 12 h reaction.

Key words: sol-gel, entrapment, lipase