食品科学

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壳聚糖固定化羊血超氧化物歧化酶工艺条件优化及酶学性质比较研究

付文力1,杨学山1,*,杨孝朴1,祝 霞2,韩舜愈2,杜 娜2,李蔚蔚1,白 钰1   

  1. 1.甘肃农业大学生命科学技术学院,甘肃 兰州 730070;2.甘肃农业大学食品科学与工程学院,甘肃 兰州 730070
  • 出版日期:2013-10-15 发布日期:2013-09-27
  • 通讯作者: 杨学山
  • 基金资助:

    甘肃省农业生物技术研究与应用开发项目(GNSW-2010-03)

Optimization of Process Conditions for Chitosan-immobilized SOD from Sheep Blood and Its Enzymatic Characteristics

FU Wen-li1,YANG Xue-shan1,*,YANG Xiao-pu1,ZHU Xia2,HAN Shun-yu2,DU Na2,LI Wei-wei1,BAI Yu1   

  1. 1. College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
    2. College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China
  • Online:2013-10-15 Published:2013-09-27
  • Contact: YANG Xue-shan1

摘要:

以羊血为材料获得超氧化物歧化酶(SOD)酶液,采用壳聚糖进行SOD固定化,对壳聚糖用量、戊二醛体积分数、反应时间及pH值对SOD固定化的影响进行研究,通过正交试验优化工艺参数,并比较固定化酶与天然酶的酶学性质。结果表明:10mL比活力为5219.3U/mg的SOD样液与0.25g用体积分数0.7%戊二醛活化的壳聚糖在pH5.6的条件下充分反应2h,所得的固定化酶的活力最强;固定化SOD比天然SOD热稳定性提高了65.2%,耐酸碱性提高了48.4%,Km值为0.18mmol/L。因此,壳聚糖可用于制备性能较优的固定化羊血超氧化物歧化酶。

关键词: 羊血SOD, 壳聚糖, 固定化, 酶活性

Abstract:

The process conditions for immobilizing SOD from sheep blood by chitosan were explored. Chitosan dosage,
glutaraldehyde concentration, reaction time and pH were optimized by orthogonal array design and a comparative evaluation
of enzymatic properties was made between free and immobilized SOD. The immobilization conditions were optimized as
following: enzyme activity of 5219.3 U/mg, chitosan dosage of 0.25 g, glutaraldehyde concentration of 0.7%, reaction time
of 2 h and pH 5.6. Under the optimal process conditions, the highest activity of immobilized enzyme was obtained, and the
thermal stability was increased by 65.2% when compared with natural SOD. Meanwhile, the resistance to acid and alkali
demonstrated an increase of 48.4%, and Km was 0.18 mmol/L. Chitosan is useful to prepare high-performance immobilized
SOD from sheep blood.

Key words: sheep blood SOD, chitosan, immobilization, enzyme activity

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