Studies On Enzymatic Reduction Of β-lactoglobulin

In this study, self-made as the raw materialβ-lactoglobulin, From the modified protein and lower content ofβ-lactoglobulin the point of view, optimization of a degradation process ofβ-lactoglobulin, and the establishment of a small peptide for the detection of substances in whey gel electrophoresis detection methods, and Through this method,β-lactoglobulin degradation of the molecular weight of 10kD and below the qualitative detection of small peptides, studies were divided into three parts.Test 1, as polyacrylamide gel electrophoresis protein analysis, samples of small molecular weight peptide analysis results are poor, for the whey peptide small molecules in terms of practical application, this study concentrated on one of plastic, separating gel concentration and crosslinking degree were discussed on the electrophoresis process voltage and current control, and different staining methods were compared.Finally optimized for the establishment of a rapid detection of 10kD and below the small peptide Tricine-SDS-PAGE method.Test 2, forβ-lactoglobulin hydrolysis-resistant, single-enzyme hydrolysis of its limited capacity, the problem of low degree of hydrolysis, respectively, of the alkaline protease and papain, neutral protease, pepsin combination ofβ-milk globulin by double digestion, to the degree of hydrolysis (DH) and soluble nitrogen (TCA-SN) the degree of response was characterized to determine the alkaline protease enzyme papain is the best combination. Orthogonal experiment to determine the best combination of reaction conditions were: activity dosage of 5310U/g of alkaline protease hydrolysis 150min first, then join the activity dosage of 4500 U/g of papain, the reaction time to 180min, up to 36.21% degree of hydrolysis. Use the conclusion of test 1 analyzed the result ofβ-lactoglobulin hydrolysis, to understand their degradation.Test 3, little modification of existing proteins in combination, the researcher adopted the heat treatment and test II explored by the combination of two-enzyme reaction, combined use of two modified techniques. First heat treatment by single factor experiments the best reaction time and reaction temperature, further double-enzyme reaction.To the degree of hydrolysis (DH) and soluble nitrogen (TCA-SN) as an index, by Tricine-SDS-PAGE analysis of dual-treated hydrolyzate peptide distribution. The results showed that in 85℃, 20min after the dual enzyme treatment,β-lactoglobulin and the residual rate of less than 1%.