华南理工大学学报(自然科学版) ›› 2011, Vol. 39 ›› Issue (9): 152-157,164.doi: 10.3969/j.issn.1000-565X.2011.09.026

• 生物工程 • 上一篇    下一篇

亚热带木腐真菌产漆酶及其酶学性质

傅恺 付时雨 张丽 周攀登 詹怀宇   

  1. 华南理工大学 制浆造纸工程国家重点实验室,广东 广州 510640
  • 收稿日期:2010-12-27 修回日期:2011-03-11 出版日期:2011-09-25 发布日期:2011-08-02
  • 通信作者: 付时雨(1963-) ,男,教授,博士生导师,主要从事制浆漂白生物技术以及木质纤维素降解与转化的微生物及酶系研究. E-mail: shyfu@scut.edu.cn E-mail:su27s@126.com
  • 作者简介:傅恺(1984-) ,男,博士生,主要从事制浆造纸生物技术研究.
  • 基金资助:

    国家自然科学基金资助项目( 30771689) ; 广东省科技计划项目( 2007B031600002)

Production and Enzymatic Properties of Laccase from a Subtropical Wood-Rot Fungus

Fu Kai Fu  Shi-yu Zhang  Li Zhou  Pan-deng Zhan  Huai-yu   

  1. State Key Laboratory of Pulp and Paper Engineering,South China University of Technology,Guangzhou 510640,Guangdong,China
  • Received:2010-12-27 Revised:2011-03-11 Online:2011-09-25 Published:2011-08-02
  • Contact: 付时雨(1963-) ,男,教授,博士生导师,主要从事制浆漂白生物技术以及木质纤维素降解与转化的微生物及酶系研究. E-mail: shyfu@scut.edu.cn E-mail:su27s@126.com
  • About author:傅恺(1984-) ,男,博士生,主要从事制浆造纸生物技术研究.
  • Supported by:

    国家自然科学基金资助项目( 30771689) ; 广东省科技计划项目( 2007B031600002)

摘要: 对选育的一株木腐真菌Psathyrella candolleana 进行液体深层发酵生产漆酶,通过硫酸铵盐析和离子交换色谱等纯化技术分离发酵酶液后,得到纯度较高的漆酶,并对其酶学性质进行研究. 结果表明: P. candolleana 在发酵罐中的漆酶产量最高可达9100 U/L; 分离纯化后得到漆酶活性组分,其比活力达到438. 0 U/mg,纯化倍数为13. 2,得率为50. 1%; 该漆酶蛋白分子质量约为61. 2 ku,米氏常数为16 μmol /L; 该漆酶催化反应最适温度和pH 值分别为45℃和5. 0,在4 ~ 10 ℃和pH 9. 0 的条件下漆酶活力稳定性较好,Mg2+、Mn2+、Zn2+ 等金属离子对其活力的影响较小.

关键词: 白黄小脆柄菇, 白腐菌, 漆酶, 纯化, 漆酶活力, 稳定性

Abstract:

Laccase from a newly isolated wood-rot fungus Psathyrella Candolleana,was produced via the submerged fermentation,and high-purity laccase was obtained by means of the ammonium sulfate precipitation and the ion-exchange chromatography. Then,the enzymatic properties of the laccase were also analyzed. The results indicate that the maximal activity of the laccase from P. candolleana in the fermentor reaches 9100U/L,that the active
component of the laccase achieved after 13. 2-fold purification is of a specific activity of 438. 0U/mg with a yield of 50. 1%,that the molecular mass and Michaelis constant of the laccase are respectively about 61. 2ku and 16μmol /L,and that the optimal pH value and temperature for the laccase activity are 5. 0 and 45℃,respectively. Moreover,it is also found that the laccase activity is stable under 4 ~10℃ at pH 9. 0 and is not sensitive to Mg2 + ,Mn2 + and Zn2 + .

Key words: Psathyrella Candolleana, white-rot fungus, laccase, purification, laccase activity, stability

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