Determination Of Activityand High Activity Feed Phytase Extraction, Purification

In recent years, domestic and foreign research on phytase is a focus topic. Thispaper explores the characterization of plant phytase, using the unique feedingdual-purpose triticale, according to the national standard of crude enzyme liquid activitymeasurement of the new standard, then purified triticale phytase, in order to testHISTEP activities in plants of Triticale phytase activity, explore its enzymaticproperties.Methods this study summarizes the previous09years, according to new GBstandard, determination of phytase activity using vanadium ammonium molybdatemethod. To determine the extraction buffer solution pH5.0, solid to liquid ratio is10, atroom temperature200rpm shock1H, without the addition of active agent under thecondition of extracting the highest phytase activity, the optimum extraction conditions.According to the optimal extraction conditions of extraction of crude enzyme solution,concentrated liquid of phytase by salting out, dialysis desalting by preliminary. ThenDE-52anion chromatography column on the salt removing enzyme purification,purified by SDS-PAGE electrophoresis purity of purified set purification, triticalephytase characterization, found that the enzyme in pH3.0~7is relatively stable;phytase reaction of Triticale optimum temperature of37℃;at80℃for1h, triticale theresidual activity of phytase in20%above, phytase Km=0.373mmol/L triticale,Vmax=243.9mmol/min.The experimental materials for the special feeding dual-purpose triticale,experimental method is easy to operate, the conclusion of Triticale Phytase activity ishigh, can get the extraction and purification effect, and belongs to the pure plant phytase,use safety and environmental protection in the actual production, the content of thisexperiment for wheat phytase by promising.