The Preparation Of Enzyme On The Research Of Compounded Enzyme For Food Industry

Food enzyme which playing a very important role in the food processing industry is the largest part of enzyme preparation industry by making up30percent of the world zymin market. With the development of science and technology, the ongoing reseach on compound enzyme has made great progress. Enzyme preparation on food has been widely used for its high efficiency, energy saving, highly targeted and obvious effects, but new and highly reactive compound enzyme preparation still needs to be developed in our country because the limited variety of the current compound enzyme is far not enough to satisfy the huge demand of food industry. Research on the compound enzyme preparation needs high purity single-enzyme for basic experiment, according to the new characteristics of the single enzyme on substrate we can select the right single enzyme for compounding fast and accurately. In this thesis, four kinds of single enzymes have been separated and purified from the commecial pectinase preparation and laboratorial fermented broth by the technique of separation, purification and detection of protein. The four kinds of high purity single enzyme, which can be used for the reseach of food compound enzyme and laid the foundation for the further study of compound enzyme preparation. The main research contents and results were as follows:First of all, the preparation of aminopeptidase from bacillus subtilis fermentation liquor was done by salting out, chromatography and electrophoresis. The fermentation liquor was purified through ammonium sulfate fractionation, Phenyl Sepharose hydrophobic chromatography, Q Sepharose ion-exchange chromatography and Sephacryl S-200HR gel chromatography. The recovery rate was15.28%, specific activity reached to9066.67U/mg and78.26-fold purification was attained.Secondly, the preparation of laccase from lentinus tigrinus fermentation liquor was done by salting out, chromatography and electrophoresis. The fermentation liquor was purified through ammonium sulfate fractionation, DEAE Sepharose ion-exchange chromatography and Sephacryl S-200HR gel chromatography. The recovery rate was48.9%, specific activity reached to7827.33U/mg and40.41-fold purification was attained.Thirdly, the preparation of PG from commecial pectinase preparation was done by dialyse, salting out, chromatography and electrophoresis. The commecial pectinase preparation was purified through ammonium sulfate fractionation, DEAE Sepharose ion-exchange chromatography, Q Sepharose ion-exchange chromatography and Sephacryl S-200HR gel chromatography. The recovery rate was48.23%, specific activity reached to72.93U/mg and15.19-fold purification was attained.At last, the preparation of PE from commecial pectinase preparation was done by dialyse, salting out, chromatography and electrophoresis. The commecial pectinase preparation was purified through ammonium sulfate fractionation, DEAE Sepharose ion-exchange chromatography and Sephacryl S-200HR gel chromatography. The recovery rate was8.9%, specific activity reached to33.75U/mg and7.18-fold purification was attained.