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嗜热子囊菌光孢变种Thermoascus aurantiacus var.levisporusβ-葡萄糖苷酶的分离纯化及特性研究

Keywords: 嗜热子囊菌光孢变种,Thermoascus,aurantiacus,var.levisporus,β-葡萄糖苷酶,纯化,特性

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Abstract:

采用室内培养、测定方法,对嗜热子囊菌光孢变种Thermoascusaurantiacusvar.levisporus产生的β-葡萄糖苷酶进行了分离纯化及特性研究。粗酶液经硫酸铵沉淀、DEAE-SepharoseFastFlow阴离子层析、Phenyl-Sepharose疏水层析等步骤获得了凝胶电泳均一的β-葡萄糖苷酶。结果表明,经12%SDS-PAGE测得酶的单亚基分子量约为118kDa,凝胶过滤层析测得酶的分子量约为350kDa。该酶反应的最适温度和最适pH分别为80℃和4.5 ̄5.0,在pH5.0条件下,该酶在70℃条件下基本稳定,80℃保温30min,剩余酶活为15%。金属离子对β-葡萄糖苷酶活性影响较大,其中Ca2+、Ba2+对酶有激活作用;Ag+、Fe3+、Cu2+对酶有显著的抑制作用。该酶对水杨苷具有很强的底物特异性。

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