食品科学 ›› 2011, Vol. 32 ›› Issue (21): 171-176.doi: 10.7506/spkx1002-6630-201121035

• 生物工程 • 上一篇    下一篇

醋酸纤维素-聚四氟乙烯复合膜固定化脂肪酶的研究

梁单琼1,周晓丹2,时 敏2,王 雪2,于殿宇2,*   

  1. 1.黑龙江省造纸工业研究所 2.东北农业大学食品学院
  • 出版日期:2011-11-15 发布日期:2011-11-11
  • 基金资助:
    国家“863”计划项目(2010AA101503)

Immobilization and Characterization of Lipase Immobilized onto Cellulose Acetate-Polytetrafluoroethylene Composite Membrane

LIANG Dan-qiong1,ZHOU Xiao-dan2,SHI Min2,WANG Xue2,YU Dian-yu2,*   

  1. (1. Heilongjiang Research Institute of Pulp and Paper Industry, Mudanjiang 157013, China; 2. College of Food Science, Northeast Agricultural University, Harbin 150030, China)
  • Online:2011-11-15 Published:2011-11-11

摘要: 以醋酸纤维素(CA)和聚四氟乙烯(PTFE)为材料制备醋酸纤维素-聚四氟乙烯复合膜,采用吸附-交联相结合的固定化方法,用该复合膜固定化脂肪酶。研究温度、吸附时间、酶液质量浓度、交联时间和交联剂体积分数对脂肪酶固定化效率和催化效果的影响,并对固定化酶膜的酶学性质进行研究。得到最佳的固定化条件为:温度25℃、吸附时间2h、酶液质量浓度0.02g/mL、交联时间3h、交联剂(戊二醛)体积分数0.2%,固定化酶最大酶活力为17.2U/cm2。固定化酶膜的酶学性质为:最适温度35℃,比游离酶降低了5℃;最适pH8.5,与游离酶相比pH值向碱性偏移1.0;经10次(10h /次)重复使用后,固定化酶相对酶活力为55.5%。SEM结果显示CA-PTFE复合膜能较好的固定化脂肪酶。

关键词: 醋酸纤维素/聚四氟乙烯复合膜, 脂肪酶, 固定化, 酶活力, SEM

Abstract: In the present work, we immobilized lipase onto cellulose acetate (CA)-polytetrafluoroethylene (PTFE) composite membrane by a combined method of adsorption and cross-linking. The effects of different immobilization conditions such as immobilization temperature, adsorption time, lipase concentration, cross-linking time and cross-linking agent concentration on lipase immobilization efficiency and catalytic activity were explored. Immobilized lipase was also characterized. The optimal immobilization conditions were reaction temperature of 25 ℃, adsorption time of 2 h, lipase concentration of 0.02 g/mL, crosslinking time of 3 h, and cross-linking agent concentration of 0.2%. Under the optimal immobilization conditions, the maximum activity of immobilized lipase reached up to 17.2 U/cm2. The optimal temperature and pH of immobilized lipase were 35 ℃ and 8.5, respectively, which exhibited a decrease by 5 ℃ and shift to alkaline side by 1.0 when compared with the free enzyme. After repeated use for 10 times for 10 h each time, the residual activity was still up to 55.5%. The SEM micrographs revealed better immobilization of the enzyme on CA-PTFE composite membrane.

Key words: CA-PTFE composite membrane, lipase, immobilization, lipase activity

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