华南理工大学学报(自然科学版) ›› 2006, Vol. 34 ›› Issue (3): 111-116.

• 生物工程 • 上一篇    下一篇

康氏木霉木聚糖酶的分离纯化及特性

陆登俊 肖凯军 郭祀远 郑必胜 蔡妙颜   

  1. 华南理工大学 轻化工研究所,广东 广州 510640
  • 收稿日期:2005-07-13 出版日期:2006-03-25 发布日期:2006-03-25
  • 通信作者: 陆登俊(1978-),女,博士生,主要从事天然产物分离纯化新方法新技术的研究. E-mail:dj6688@126.com
  • 作者简介:陆登俊(1978-),女,博士生,主要从事天然产物分离纯化新方法新技术的研究.
  • 基金资助:

    广东省科技攻关项目(2004B10201001);广东省科技计划项目(2004B20401001,2003B31701);广州市科技攻关项目(2002Z3-E0321)

Separation,Purification and Characteristics of Xylanase Separated from Trichoderma Koningii

Lu Deng-jun  Xiao Kai-jun  Guo Si-yuan  Zheng Bi-sheng  Cai Miao-yan   

  1. Research Institute of Light Industry and Chemical Engineering,South China Univ.of Tech.,Guangzhou 510640,Guangdong,China
  • Received:2005-07-13 Online:2006-03-25 Published:2006-03-25
  • Contact: 陆登俊(1978-),女,博士生,主要从事天然产物分离纯化新方法新技术的研究. E-mail:dj6688@126.com
  • About author:陆登俊(1978-),女,博士生,主要从事天然产物分离纯化新方法新技术的研究.
  • Supported by:

    广东省科技攻关项目(2004B10201001);广东省科技计划项目(2004B20401001,2003B31701);广州市科技攻关项目(2002Z3-E0321)

摘要: 使用硫酸铵分级沉淀、Sephadex G-25凝胶色谱脱盐和Sephadex G-100凝胶色谱等分离纯化技术,从康氏木霉(Trichoderma Koningii)发酵液中分离出木聚糖酶,纯化后的木聚糖酶经十二烷基硫酸钠一聚丙烯酰胺(SDS-PAGE)凝胶电泳鉴定为单一组分,其相对分子质量为55208.所得的木聚糖酶的最适反应温度为65 ℃,pH值为6.0.该酶稳定性较好,在30~60℃下放置2 h能保持83%以上的酶活;在3.0~10.0的pH值范围内能保持85%以上的酶活.研究表明:金属离子Ba2+,Pb2+,Fe2+,Fe3+,Al3+¨和高浓度(12 mmoL/L)的Cu2+对木聚糖酶的活性有抑制作用,而Ca2+,Zn2+ 和4mmol/L的Cu2+对该酶反应有促进作用.该木聚糖酶作用于Birchwood木聚糖的米氏常数为5.37 g/L,最大反应速率为0.94μmol/min.

关键词: 木聚糖酶, 康氏木霉, 分离, 纯化, 酶活

Abstract:

Xylanase was separated and purified from a culture filtrate of Trichoderma Koningii through ammonium sulfate precipitation.Sephadex G-25 and Sephadex G-100 column chromatography.The purified xylanase was then proved by SDS.PAGE to be a single composition with a relative molecular mass of 55 208.Good thermostability of xylanase with an optimal activity at 65 ℃ and pH 6.0 was also revealed.The results show that more than 83% or 85% of the relative activity of xylanase can be remained when respectively stored at 30~60 ℃ for 2 h or in the pH range of 3.0 ~10.0. Moreover,Ba2+,Pb2+,Fe2+,Fe3+,,Al3+ and 12mmol/L Cu2+ are strong inhibitors to xyla-nase.while Ca2+ ,Zn2+ and 4 mmol/L Cu2+ are stimulators.The Michaelis.Menten constant of xylanases in the enzymatic hydrolysis of birchwood xylan iS 5.37 g/L.and the maximal reaction velocity iS 0.94μmol/min.

Key words: xylanase, Trichoderma Koningii, separation, purification, enzyme activity