食品科学 ›› 2012, Vol. 33 ›› Issue (11): 215-218.doi: 10.7506/spkx1002-6630-201211046

• 生物工程 • 上一篇    下一篇

大麦虫超氧化物歧化酶分离纯化及性质研究

张建新,刘 娜,何桂梅,郭 倩   

  1. 西北农林科技大学食品科学与工程学院
  • 出版日期:2012-06-15 发布日期:2012-07-27

Purification and Characterization of Superoxide Dismutase from Zophobas morio L. Larvae

ZHANG Jian-xin,LIU Na,HE Gui-mei,GUO Qian   

  1. (College of Food Science and Engineering, Northwest A & F University, Yangling 712100, China)
  • Online:2012-06-15 Published:2012-07-27

摘要: 目的:探讨大麦虫超氧化物歧化酶(SOD)分离纯化条件及其性质,为大麦虫利用提供科学依据。方法:以大麦虫为原料,经盐析沉淀、DEAE-Sepharose FF阴离子交换柱层析和Sephadex G-75凝胶过滤层析等纯化, 变性聚丙烯酰胺凝胶电泳(SDS-PAGE)方法对蛋白质的组分进行分离并确定其分子质量,原子吸收光谱仪鉴定酶的类型。结果:获得了一种酶比活力较高的大麦虫SOD,酶的纯化倍数为68.54倍,分子质量为37.30kD,属于含铜锌超氧化物歧化酶(Cu,Zn-SOD),且在278nm波长处有紫外吸收峰,稳定性较好,最适温度为40℃,最适pH值为6.0,对H2O2、β-巯基乙醇试剂十分敏感,受氯仿-乙醇混合液(3:5,V/V)的影响较小。结论:大麦虫SOD具有较高的酶比活力,稳定性好,具有较高的利用价值。

关键词: 大麦虫, 超氧化物歧化酶, 分离纯化, 性质

Abstract: Objective: To explore the purification and properties of superoxidae dismutase (SOD) from Zophobas morio L. larvae. Methods: SOD were obtained from Zophobas morio L. larvae by salting-out, DEAE-Sepharose FF column chromatography and Sephadex G-75 column chromatography. SDS-PAGE was used to determine its molecular weight and atomic absorption spectrometer was used to identify it types. Results: SOD with highly specific activity was obtained. The purification factor was 68.54. The purified SOD was identified as Cu, Zn-SOD with molecular weight of 37.30 kD. It had an absorption peak at 278 nm and good stability. Its optimal temperature and pH were 40 ℃ and 6.0, respectively. Its high sensitivity to H2O2 and β-mercaptoethanol was found , while a mixture of CHCl3 and CH3CH2OH (3:5, V/V) had little effect on its activity. Conclusion: The SOD extracted from Zophobas morio L.larvae has high enzymatic activity, stability and application potential.

Key words: Zophobas morio L., SOD, purification, characterization

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