木材蓝变生物控制菌康氏木霉胞外几丁质酶的纯化及特性

Purification and Properties of Extracellular Chitinase from Trichoderma koningii Used for Biological Control of Wood Blue-stain

本研究以木材蓝变生物控制菌康氏木霉(6111-ET)为供试菌株进行胞外几丁质酶纯化和特性的研究。将液态PDA培养基中的康氏木霉(Trichoderma koningii)置于恒温摇床(200 r/min,28℃)培养10天。培养滤液经过硫酸铵分级沉淀、Q-HP强阴离子交换柱层析、Phenyl-Sepharose疏水层析、CM-Sepharose弱阳离子交换柱层析、Phenyl-Sepharose疏水柱层析后得到凝胶电泳(SDS-PAGE)谱带单一的几丁质酶,其分子量为36kDa,最适温度为40℃,最适pH为4.0。多种金属阳离子对该酶具有影响,Ca2+、Fe2+对酶活力有比较明显的激活作用,其中Fe2+的作用最强;K+、Na+、Cu2+有明显的抑制作用。

In the present study,purifiying and properties analyzing of an extracellular chitinase from Trichoderma koningii were carried out with T.koningii（6111-ET） as testing fungus.T.koningii was cultured in PDA broth at 28℃ on a rotary shaker at 200r/min for 10 days.The extracellular chitinase was purified to SDS-PAGE homogeneous through ammonium sulfate fraction,Q-HP chromatography,Phenyl-Sepharose chromatogram,CM-Sepharose chromatogram and Phenyl-Sepharose chromatogram.The molecular weight of the chitinase was about 36kD.Its best activity was obtained at 40℃ and pH4.0.Many kinds of metal ions had effects on the chitinase activities.Ca2＋ and Fe2＋ showed positive effects,whereas K＋、Na＋ and Cu2＋ showed negative effects.The effect of Fe2＋ was the strongest.