Preparation method of yak milk casein anticoagulant peptide

Abstract

The invention relates to a method for preparing, separating and purifying anticoagulant peptide by hydrolyzing yak milk casein with trypsin. The method comprises the following steps: (1), adjusting the reaction temperature at 30-50 DEG C and pH at 5.5-7.5, and stirring to dissolve yak milk casein; (2) hydrolyzing by adding trypsin, maintaining the reaction temperature at 30 DEG C and pH at 5.5-7.5, and hydrolyzing for 1-4 hours; (3) heating to 80-100 DEG C, maintaining for 5-15 minutes, and inactivating trypsin; (4) adjusting pH to 4.6 with 5mol / L edible grade hydrochloric acid to separate out casein at an isoelectric point thereof; (5) centrifugalizing for 5-10 minutes by a centrifugal force at 4000-10000g to remove precipitates and reserving supernate; (6) spraying and drying the supernate; (7) separating and purifying hydrolysates by a glucan column; (8) detecting the activities of the components by an agarose-fibrous protein flat band method; and (9) detecting an amino acid sequence: detecting the amino acid sequence of the yak milk casein anticoagulant peptide by an N-terminal sequencing method.

Description

technical field

[0001] The invention uses trypsin to hydrolyze yak milk casein to prepare, separate and purify anticoagulant peptide, and belongs to the field of biotechnology. Background technique

[0002] Blood coagulation is an indispensable physiological function of the human body, but blood coagulation in diseases can induce thrombus, cause pulmonary infarction, cerebral thrombosis, retinal arteriovenous occlusion, myocardial infarction and limb and peripheral vascular embolism. At present, cardiovascular and cerebrovascular diseases are the diseases with the highest morbidity, disability and mortality in my country. According to the survey, in recent years, 1-3 people in every 1,000 people suffer from thrombosis, and one person suffers from stroke or myocardial infarction every 12s. And fell down.

[0003] Cardiovascular and cerebrovascular diseases have become the diseases with the highest morbidity and mortality in my country, most of which are related to thrombosis....