食品科学 ›› 2009, Vol. 30 ›› Issue (11): 207-210.doi: 10.7506/spkx1002-6630-200911046

• 生物工程 • 上一篇    下一篇

Aspergillus ficuum菊粉酶的化学修饰

王 静1,金征宇2,江 波2,孙宝国1,曹雁平1   

  1. 1. 北京工商大学化学与环境工程学院 2. 江南大学食品学院
  • 收稿日期:2008-09-05 修回日期:2009-01-14 出版日期:2009-06-01 发布日期:2010-12-29
  • 通讯作者: 王静 E-mail:jwang810@yahoo.com.cn
  • 基金资助:

    国家自然科学基金项目(20376029);北京市优秀人才培养项目(20071D0500300141)

Chemical Modification of Aspergillus ficuum Inulinases

WANG Jing1,JIN Zheng-yu2,JIANG Bo2,SUN Bao-guo1,CAO Yan-ping1   

  1. (1. College of Chemistry and Environmental Engineering, Beijing Technology and Business University, Beijing 100037, China;
    2. School of Food Science and Technology, Jiangnan University, Wuxi 214122, China)
  • Received:2008-09-05 Revised:2009-01-14 Online:2009-06-01 Published:2010-12-29
  • Contact: WANG Jing1, E-mail:jwang810@yahoo.com.cn

摘要:

应用NBS、DEPC、EDC、DIC、Ch-T、PMSF 和DTT 化学修饰剂对Aspergillus ficuum 产内切菊粉酶和外切菊粉酶进行化学修饰,测定与其活性相关的氨基酸残基,结果表明,构成内切菊粉酶和外切菊粉酶活性中心的必需氨基酸残基均含有色氨酸和羧基氨基酸(谷氮酸或天门冬氨酸),组氨酸可能是酶活性中心的组成氨基酸。邹氏作图法进一步确认外切菊粉酶活性中心必需色氨酸残基数目为2,内切菊粉酶活性中心必需色氨酸残基数目为1 。

关键词: Aspergillus ficuum, 菊粉酶, 化学修饰, 活性中心

Abstract:

Exo-inulinase and endo-inulinase from Aspergillus ficuum were subject to chemical modification of seven protein modification reagents and to identification of amino acid residues essential for enzyme activity. The results suggested that Nbromosuccinimide (NBS), carbodiimide (EDC) and diethyl pyrocarbonate (DEPC) resulted in a great loss of endo- and exoinulinase activities, indicating that Trp, carboxyl amino acid (Glu or Asp) and His residues may be parts of the enzyme active sites, respectively. Two Trp residues essential for exo-inulinase active site and one for endo-inulinase were confirmed by Tsou’s method.

Key words: Aspergillus ficuum, inulinase, chemical modification, active site

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