Purification And Some Characterization Of A Group Of Glycolysis Enzymes And Alkaline Phosphatase From Pig Muscle Tissue

Nucleotide that present a taste is the main material that form the flavor characteristics in meat. Among them inosine-5’-monophosphate (IMP), as one of the important indexes that measure the fresh degree of pork.And the formation and biodegradable of it have a close realation between some enzymes. This paper firstly have a corresponding understanding for the changes of the inosine-5’-monophosphate (IMP) conent from pork tissue, and through the related purification and some characterization of a group of glycolysis enzymes and alkaline phosphatase, we discussed the effects on the formation and degradation of inosine-5’-monophosphate (IMP), Aims to explore mechanism of some enzymes in flavour of induction nucleotide the of substance.(1) Changes of inosine-5’-monophosphate (IMP) conents from pork tissue Changes of inosine-5’-monophosphate (IMP) contents in longissimus dorsi at4℃were analyzed by HPLC, to understand the changes of inosine-5’-monophosphate (IMP) after the pig is killed, discussed the relationship between it with the quality preliminary. The result showed that storage time affected the concentration of IMP,concentration of IMP increased until reaching top on the next day, then decreased gradually. Therefore, the fresh degree of pork could be forecasted by determining the change of IMP, and it was one of the major quality variables during storage and process.(2) Exploration of the purification of glycolysis enzyme and the degradation characterization from pork tissue. To purify glycolysis enzyme, we adopted following procedures:Acetone precipitation, ion-exchange chromatography, SDS-PAGE electrophoresis, LC-MS and so on. And to study the further degradation function through the high performance liquid chromatography (HPLC) method. In order to discuss the relationship between glycolysis enzyme from fresh pork with the quality and the flavor respectively. It has been proven the effect of this kind of purification plan is remarkable. Get a set of glycolysis enzyme. After appraisal,they are phosphoric acid oleic acid kinase gump,3,3heterogeneous pyruvate kinase,1,3heterogeneous pyruvate kinase, enolase enzymes respectively. This group of glycolysis enzymes ensure the best degradation of the concentration is lOμg/ml through Single factor experiment, it can degrade ATP effectively, but it has no degradation effection to IMP. The decrease of ATP is9.2395μg in the processes, the degradation products of ATP include ADP, IMP; and3minutes degradation products contains2.1451μg ADP,3minutes degradation products contains IMP and ADP,the contents are1.0687μg and2.0093μg. This group of glycolysis enzymes are the key enzymes in the glycolysis processes, they not only related to pork quality closely, but aslo have certain effect to the flavor substances metabolic processes.(3)Purification of alkaline phosphataseand some characterization research from pork tissue. An alkaline phosphatase purified from raw meat by following procedures:ammonium sulfate precipitation, ion-exchange chromatography on DEAE-Sepharose Fast Flow column, fellow by gel filtration through Sephacryl S-200and SDS-PAGE electrophoresis. The purification multiples is57.34, and the specific activity is28.77U/mg. The preparation was formed a single band on SDS-PAGE and its molecular weight was determined to be about66kD. The optimum pH and optimum temperature for the enzyme to catalyze the hydrolysis of phenylphosphonic acid disodium salt(p-NPP)were pH9.7and30℃, The enzyme is stable in the range of pH from7.0to10.5and at the temperature below40℃. Mg2+and Ca2+activated the enzyme while Zn2+and EDTA inhibited the enzyme.