DFRKN-1碱性磷酸酶的提取纯化和主要理化性质研究

目的]提取纯化根结线虫天敌真菌-1(Destroying fungi of root-knot nematode-1,DFRKN-1)的碱性磷酸酶,并对其主要理化性质进行研究。方法]以DFRKN-1为提酶材料,经正丁醇提取,硫酸铵分级沉淀分离,透析获得酶的粗提取液,用SephadexG-150凝胶过滤柱层析纯化,获得纯化碱性磷酸酶,并测定纯化碱性磷酸酶的理化性质。结果]酶纯度达到电泳纯,提纯倍数达到15.7倍,比活力达5333 U/mg;该酶的最适pH值为8.9,最适温度为43℃,米氏常数为6.82 mmol/L;K+、Ba2+、Mg2+对该碱性磷酸酶的活性均有激活作用,甲醛、草酸、酒石酸对该碱性磷酸酶均有抑制作用。结论]试验结果可为进一步深入研究DFRKN-1提供参考。

Study on Isolation, Purification and Some Properties of Alkaline Phosphatase from DFRKN-1

Objective] The research aimed to isolate the alkaline phosphatase from destroying fungi of root-knot nematode-1(DFRKN-1),and studied some properties of the enzyme.Method] By n-butanol extraction,ammonium sulfate fractionation and sodium chloride dialyzation,the partially purified enzyme was gained by means of gel filtration on sephadex G-150 column,and some properties of the enzyme were determined.Result] The purification attained to 15.7 times and the specific activity was 5 333 U/mg.The optimum pH value was 8.9.The optimum temperature was 43 ℃,The Michaelis-Menten contant(Km) was 6.82 mmol/L.The AKP was active by K+、Ba2+、Mg2+ and inhibited by formaldehyde,tartrate and oxalate.Conclusion] The results can be provided the scientific basis for the research of DFRKN-1.