β-1,4-内切木聚糖酶的分离纯化及其性质  

Isolation and Characterization of β-1, 4-Endoxylanase from Trichoderma pseudokonigi

作　　者：邹永龙[1] 桑月婵 彭建新[1] 王国强[1] 

机构地区：[1]中国科学院上海植物生理研究所,上海200032

出　　处：《Acta Botanica Sinica》1999年第11期1212-1216,共5页Acta Botanica Sinica（植物学报：英文版）

基　　金：上海市自然科学基金;国家自然科学基金!39730040

摘　　要：通过硫酸铵分级盐析、DEAE-SephadexA50和DEAE－SepharoseCL-6B离子交换层析、FPLC等一系列分离纯化手段，从拟康氏木霉（TrichodermapseudokonigiRifai）固体培养基发酵抽提液中分离得到了Native－PAGE电泳纯的木聚糖酶，SDS-PAGE显示该酶为单肽链结构，分子量约为66kD。该酶的酶反应最适温度为55℃。酶反应的最适pH为4．5。该酶作用于山毛榉木聚糖（Beech－xylan）的Km为20mg/mL，Vmax为3．3μmol·min-1·mg-1。Hg2+、Cu2+对酶反应有较强的抑制作用，而Fe2+、Mn2+对该酶反应则有促进作用。β-1, 4-endoxylanase from Trichoderma pseudokonigi Rifai has been purified by anion-exchange chromatography on DEAE-Sephadex A50, DEAE-Sepharose CL-6B and mono Q. The endoxyanase was shown to be homogeneous by Native-PAGE and SDS-PAGE. This endoxlanase is a single-peptide chain protein with a molecular weight estimated as 66 kD. The endoxylanase was purified by 10-fold with a specific activity of 15. 87 U mg- 1. Optimum endoxyanase activity was obtained when the enzyme was incubated at PH 4. 5, 55 ℃ with a Km of 20 mg/mL and Vmax of 3.3 umol.min- 1 .mg-1. Hg2+ and Cu2+ have a strong inhibition while Fe2+ and Mn2+ have a increasing effect on the enzymatic reaction rate.

关 键 词：拟康氏木霉 β-1.4-内切木聚糖酶 分离 纯化 

分 类 号：Q949.323[生物学—植物学] TQ925[轻工技术与工程—发酵工程]