食品科学 ›› 2009, Vol. 30 ›› Issue (19 ): 190-193.doi: 10.7506/spkx1002-6630-200919042

• 生物工程 • 上一篇    下一篇

亮菌漆酶的初步研究

凌庆枝1,2,董丽辉1,范三微1,高莉莉1,黄贝贝1,袁怀波2,魏兆军   

  1. 1.浙江医药高等专科学校 2.合肥工业大学生物与食品工程学院
  • 收稿日期:2009-07-05 出版日期:2009-10-01 发布日期:2010-12-29
  • 通讯作者: 凌庆枝
  • 基金资助:

    安徽省科技厅重点科研计划项目(07020303041);浙江医药高等专科学校科研基金项目(ZPCSR2008001)

Preliminary Study on Laccase from Armillariella tabescens

LING Qing-zhi1,2,DONG Li-hui1,FAN San-wei1,GAO Li-li1,HUANG Bei-bei1,YUAN Huai-bo2,WEI Zhao-jun2   

  1. 1. Zhejiang Pharmaceutical College, Ningbo 315100, China ;
    2. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei 230009, China
  • Received:2009-07-05 Online:2009-10-01 Published:2010-12-29
  • Contact: LING Qing-zhi

摘要:

对亮菌(Armillariella tabescens)固体发酵产生的漆酶进行研究,其粗酶液经过硫酸铵盐析、DEAE-cellulose柱纯化,活力回收率为42.3%,得到电泳纯的亮菌漆酶,SDS-PAGE法证实其分子量约为55.6kD。实验表明,亮菌漆酶最适反应温度为50℃、最适反应pH值为4.0,催化ABTS的米氏常数为0.018mmol/L,在pH4.0~5.0的偏酸性环境中,酶活力较高,活性较稳定。酸根阴离子和金属阳离子对酶活力都有不同程度的影响,其中SO42-、Na+对亮菌漆酶的活力有促进作用,CO32-、Cu2+、Mg2+对酶的活力有抑制作用。

关键词: 亮菌, 漆酶, 纯化, 性质

Abstract:

Laccase in Armillariella tabescens solid-state fermentation liquid was purified by ammonium sulfate salting-out and DEAE-cellulose column chromatography with an overall activity recovery of 42.3%. Electrophoretically pure laccase was obtained and its apparent molecular weight was approximately 55.6 kD, which was proved by SDS-PAGE analysis. The optimum reaction temperature and pH for this enzyme were 50 ℃ and 4.0, respectively, and the Km value was 0.018 mmol/L with 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) as a substrate. This enzyme maintained high and stable activity at pH 4.0-5.0. Various acid radical ions and metal ions had different effects on laccase activity, which was revealed an enhancement effect from SO42- and Na+, and an inhibition effect from CO32-, Cu2+ and Mg2+.

Key words: Armillariella tabescens, laccase, purification, characteristics

中图分类号: