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绿色木霉β-葡萄糖苷酶的分离纯化及酶学性质

作  者:
刘颖;韩春然;张帅;张玮玮;窦博鑫
单  位:
哈尔滨商业大学食品工程学院
关键词:
绿色木霉;β-葡萄糖苷酶;分离纯化;酶学性质
摘  要:
针对绿色木霉AS3.3711产生的β-葡萄糖苷酶组分,先后运用包括乙酸铵沉淀、透析、Sephadex G-150葡聚糖凝胶柱层析在内的一系列分离纯化技术对该纤维素酶进行纯化,得到β-葡萄糖苷酶纯化组分,并对该酶的酶学性质进行研究。纯化后酶液的蛋白质量浓度为8.12 mg/mL、酶活力为4.08 U/mL,纯化倍数达到18.48,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecylsulfate polyacrylamide gel electrophoresis,SDS-PAGE)测定分子质量为66.0 kD。绿色木霉β-葡萄糖苷酶在酸性条件下稳定性良好,最适pH值为5.0;在温度60~70℃能长时间保持较高酶活力,最适反应温度为60℃。金属离子中,Ca2+、Mg2+、K+对绿色木霉AS3.3711β-葡萄糖苷酶活力起到促进作用,Ca2+促进作用最强;而Zn2+、Fe3+对该酶有抑制作用,Ag+、Cu2+、Hg2+重金属离子使β-葡萄糖苷酶几乎丧失了全部活性。
译  名:
Separation,Purification and Enzymatic Properties of β-Glucosidase from Tridchoderma viride
作  者:
LIU Ying;HAN Chun-ran;ZHANG Shuai;ZHANG Wei-wei;DOU Bo-xin;College of Food Engineering,Harbin University of Commerce;
关键词:
Tridchoderma viride;;β-glucosidase;;separation and purification;;enzymatic properties
摘  要:
An extracellular β-glucosidase produced by Trichoderma viride AS3.3711 was separated and purified by ammonium acetate precipitation, dialysis and Sephadex G-150 column chromatography. The results of enzyme characterization showed that the protein concentration of the purified enzyme solution was 8.12 mg/mL, and the β-glucosidase activity was 4.08 U/mL, with a purification factor of 18.48. Its molecular weight was 66.0 kD. The β-glucosidase fromTrichoderma viride displayed a strong stability under acidic conditions and its optimum pH value was 5.0. In the range of 60–70 ℃, it could still maintain a high enzyme activity for a long time with an optimum reaction temperature of 60 ℃. Some metal ions including Ca2+, Mg2+ and K+ improved its activity with Ca2+ having the strongest effect. However, Zn2+ and Fe3+ inhibited the enzyme. Heavy metal ions such as Ag+, Cu2+ and Hg2+ almost totally inactivated its activity.

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