Study On The Extration,Isolation,Purification,Biologic Active Of Earthworm Fibrinolytic Enzymes And Compound Amino Acid From Earthworm

The earthworm is a commonly used animal ingredients, the body is rich in protein andactive substances.Earthworm and human relations are very close as a traditional Chinesemedicine, but also the active ingredient the extraction lumbrokinase, a variety of amino acids.Lumbrokinase is extracted from the earthworm fibrinolytic activity of the protein, with therole of thrombolytic, is one of the fibrinolytic drugs is the most development prospects.Earthworms contain a variety of amino acids, using a variety of enzymatic hydrolysis ofearthworm extract protein slurry hydrolysis, compound amino acids in the earthworm. Thisarticle reviews the current research status quo, carried out basic research on the the earthwormpart of the active ingredient.This paper uses a hierarchical analysis of salt than pure love earthworm or kinase in theextraction process for research, by fiber protein dissolved ring size and protein content of theearthworm ratio to the kinase than economic vitality. And designed through the single factorexperiment has studied extraction buffer solution pH, the first salting out ammonium sulfatedensity, the second salting out ammonium sulfate density to the earthworm activating enzymeextraction influence, and has determined the enzyme molecular weight with SDS-PAGE, andhas discussed the metal ion pair enzyme vigor influence. In the preparation of compoundamino acids, the earthworm has been skimmed, bleaching, deodorization experimentconducted a study, select extract of hydrolysis time, pH, temperature, substrate concentrationon effect of hydrolysis conditions. And uses the orthogonal experiment to design theinspection best extraction temperature, the best extraction time, best pH, the best substrateconcentration, optimized the extraction technological conditions.Lumbrokinase extraction process results showed that the optimum extraction conditions forlumbrokinase: buffer pH7.5, for the first time the optimal concentration of saturatedammonium sulfate salting40%80%ammonium sulfate concentration of the secondsalting,under these conditions lumbrokinase highest specific activity. Improve the enzymeactivity of ion K⁺, Na⁺, Mg²⁺, Ca²⁺and Fe³⁺, Pb²⁺inhibitors. By electrophoresis experimentshows that the molecular weight of earthworms extracted protein is widely distributed and aredistributed from15kDa to90kDa, mainly in the15kDa,50kDa,70kDa and90kDa nearby.Through experimental research on earthworm skimmed, and bleaching, deodorization,came to the following conclusion:â‘ skim, deodorization, bleaching methods, extractionmethod of cable-better than soaking.â‘¡properties for solvent degreasing, deodorization,bleaching, the Institute selected best solvent to acetone.â‘¢Dang to acetone for solvent Shi,cable type extraction method, and liquid solid than for2:1of conditions Xia, return time ofextended on decolorization, and skim, and de stink no effect, are can get white, and no stink of earthworm protein, skim rate for up100%; and no water ethanol in same conditions Xia（cable type extraction method, and liquid solid than for2:1）, return time of extended ondecolorization no effect, on skim effect micro-has growth, but except stink effect verydifference, cannot eradication smell. This experiment using a variety of proteases hydrolysisof earthworm liquid, as different protease enzyme cutting sites of different hydrolysis morethoroughly so that you can make.Neutral protease hydrolysis of20G protein slurry, the substrate concentration for3%,adjust the pH is about7, maintaining temperature of40℃, after hydrolysis of2H, measuredby amino nitrogen content of33.62mg/100ml;Slurry of alkaline protease hydrolysis of20G protein, the substrate concentration for2%,adjust the pH to8per cent, maintaining temperature of50℃, after hydrolysis of2H,measured by amino nitrogen content of41.32mg/100ml;Tryptic hydrolysis of20G protein slurry, the substrate concentration for3%, adjust the pHto8per cent, maintaining temperature of50℃, after hydrolysis of3h, measured by aminonitrogen content of57.43mg/100ml...