The Enzymes Which Initiating Honeysuckle Enzymatic Browning And The Research About Extraction And Purification Of Polyphenol Oxidase In Honeysuckle

Honeysuckle is a popular food in our daily life.But enzymatic browning will have a serious impact on their taste, flavor and color. In this paper, some enzymes of initiating honeysuckle browning and their mechanisation are introduced briefly; honeysuckle as an important medicinal and edible plant which has great medicinal and commercial value. Selecting honeysuckle as the research object, on which the extraction and purification of polyphenol oxidase to do the research. The main contents and conclusions are as follows:Enzymes involved in enzymatic browning of honeysuckle are: polyphenol oxidase(PPO), the main enzyme involved which participate in enzymatic browning on honeysuckle. Catalyzing the oxidation of phenolic compounds in honeysuckle, resulting in browning; peroxidase(POD), the POD bound to slow down the browning effect of honeysuckle, POD free state accelerated browning of honeysuckle; phenylalanine ammonia lyase(PAL) to promote the synthesis of phenolic compounds, resulting in browning of honeysuckle; catalase(CAT) can reduce the rate of oxidation of phenols, slowing the browning of honeysuckle to a certain extent.A comparative study of homogenization and acetone powder extracting polyphenol oxidase from honeysuckle found that homogenization could extract larger amount of enzyme, high activity, but low purity, the extraction effect is not ideal. Acetone powder method, while loss a large amount of enzyme, the overall activity is not high, but the purity is acceptable, and the acetone powder easy to save, after further purification to achieve a higher purity.Using acetone powder to extract polyphenol oxidase from honeysuckle, determining the optimal extraction process for pH 7.5, solid-liquid ratio was 1: 120, the standing time for 14.5 min, got average specific activity was 131.732 U / mg.The purification process of polyphenol oxidase from honeysuckle was preliminary studied, at 30% saturation of ammonium sulfate solution to remove hybrid proteins and at 80% saturation of ammonium sulfate solution initially separate target protein. the effect of elution as well Selecting DEAE-52 cellulose as ion exchangers because of its fast equilibrium and good elution, selecting 3.5 * 70 cm ion exchange column to purify polyphenol oxidaseenzyme solution at 20mL/h elution speed. interest protein was detected by electrophoresis,the results showed only one band and got electrophoretic homogeneity which relative molecular mass was 43000 D. molecular mass 43000 D.