机构地区: 华南理工大学轻工与食品学院轻化工研究所
出 处: 《华南理工大学学报(自然科学版)》 2006年第3期111-116,共6页
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2h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Ba^2+,Pb^2+,Fe^2+,Fe^2+,Al^3+和高浓度(12mmol/L)的Cu^2+对木聚糖酶的活性有抑制作用,而Ca^2+,Zn^2+和4mmol/L的Cu^2+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5.37g/L,最大反应速率为0.94μmol/min. 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, Ba^2+ , Pb^2+ , Fe^2+ , Fe^2+ , Al^3+ and 12mmol/L Cu^2+ are strong inhibitors to xylanase, while Ca^2+ , Zn^2+ and 4mmol/L Cu^2+ 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 0.94 μ mol/min.
领 域: [生物学]