Preparation method of yak milk casein anticoagulant peptide

A technology of milk casein and anticoagulant peptide, which is applied in the biological field, can solve the problems of limited wide application, difficulty, and high content of hydrophobic amino acids in casein, and achieve the effects of improving nutritional value, reducing production costs, and high protein content

Active Publication Date: 2013-11-13
甘肃华羚生物技术研究中心 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Yak milk casein is a protein prepared by extracting and purifying the unique resource "Qula" in Northwest China. Due to the high content of hydrophobic amino acids in casein, it is not easily soluble in water and soluble in weak alkali, which limits its use in food. widely used

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: a kind of preparation method of yak milk casein anticoagulant peptide, comprises the following steps:

[0027] (1) Yak milk casein: purchased from Hualing Casein Co., Ltd., food grade.

[0028] (2) Dissolution: Weigh 10 g of yak casein, add 400 mL of water, adjust the reaction temperature to 30°C, pH 5.5, and stir to dissolve the yak casein.

[0029] (3) Enzymatic hydrolysis: Trypsin was purchased from Beijing Geyuan Tianrun Biotechnology Co., Ltd., 0.25 g of trypsin was weighed and added, and the reaction temperature was maintained at 30°C and pH 5.5 for 1 hour.

[0030] (4) To inactivate the enzyme, raise the temperature to 80°C and keep it for 15 minutes.

[0031] (5) Precipitate casein: adjust the pH to 4.6 with 5mol / L food grade hydrochloric acid to precipitate casein at its isoelectric point.

[0032] (6) Centrifugation: 4000g centrifugal force, centrifuge to remove the casein precipitate, and keep the supernatant.

[0033] (7) Spray drying: take ...

Embodiment 2

[0037] Embodiment 2: a preparation method of yak milk casein anticoagulant peptide, comprising the following steps:

[0038] (1) Yak milk casein: purchased from Hualing Casein Co., Ltd., food grade.

[0039] (2) Dissolution of yak casein: Weigh 25g of yak casein, add 500mL of water, adjust the reaction temperature to 40°C, pH6.5, and stir to dissolve the yak casein.

[0040] (3) Add trypsin: trypsin was purchased from Beijing Geyuan Tianrun Biotechnology Co., Ltd., and 0.25 g of trypsin was weighed and added, and the reaction temperature was maintained at 40°C and pH 6.5 for 2 hours.

[0041] (4) Inactivate trypsin, raise the temperature to 90°C and maintain for 10 minutes.

[0042] (5) Precipitate casein: adjust the pH to 4.6 with 5mol / L food grade hydrochloric acid to precipitate casein at its isoelectric point.

[0043] (6) Centrifugation: 8000g centrifugal force, centrifuge to remove the casein precipitate, and keep the supernatant.

[0044] (7) Spray drying: Take the c...

experiment example 3

[0048] Experimental example 3: a preparation method of yak milk casein anticoagulant peptide, comprising the following steps:

[0049] (1) Yak milk casein: purchased from Hualing Casein Co., Ltd., food grade.

[0050](2) Dissolving yak casein: Weigh 50 g of yak casein, add 500 mL of water, adjust the reaction temperature to 50°C, pH 7.5, and stir to dissolve the yak casein.

[0051] (3) Add trypsin: trypsin was purchased from Beijing Geyuan Tianrun Biotechnology Co., Ltd., and 0.25 g of trypsin was weighed and added, and the reaction temperature was maintained at 50° C., pH 7.5, and reacted for 4 hours.

[0052] (4) Inactivate trypsin, raise the temperature to 100°C and maintain for 5 minutes.

[0053] (5) Precipitate casein: adjust the pH to 4.6 with 5mol / L food grade hydrochloric acid to precipitate casein at its isoelectric point.

[0054] (6) Centrifugation: 10000g centrifugal force, remove the casein precipitate by centrifugation, and keep the supernatant.

[0055] (7)...

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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....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/16C07K7/06C07K7/08
Inventor 谢小冬高维东纪银莉宋礼文鹏程孙文静冯丹刘恭何潇葛静微
Owner 甘肃华羚生物技术研究中心
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