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Preparation method of yak milk casein antibacterial peptide

A technology of milk casein and bacteriostatic peptides, which is applied in the field of preparation of milk casein bacteriostatic peptides, can solve the problems of few research reports on antimicrobial peptides, and achieve the effects of reducing production costs, low prices, and low production costs

Inactive Publication Date: 2013-01-16
甘肃华羚生物技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Antimicrobial peptides have attracted much attention due to their good stability and harmlessness to normal cells of higher animals, but there are few research reports on antimicrobial peptides derived from food protein

Method used

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  • Preparation method of yak milk casein antibacterial peptide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: a kind of preparation method of yak milk casein bacteriostatic peptide is characterized in that comprising the following steps:

[0025] (1) Yak milk casein: purchased from Hualing Casein Co., Ltd., food grade.

[0026] (2) Dissolution of yak casein: Weigh 25g of yak casein, add 500mL of water, adjust the reaction temperature to 55°C, pH 8.0, and stir to dissolve the yak casein.

[0027] (3) Add trypsin: trypsin was purchased from Beijing Geyuan Tianrun Biotechnology Co., Ltd., the titer was determined to be no less than 2500 units, calculated on the basis of 25g of yak milk casein, 0.25g of enzyme was weighed, and the reaction temperature was maintained at 55°C. pH8.0, react for 2 hours.

[0028] (4) Inactivate trypsin, raise the temperature to 80°C and keep it for 15 minutes.

[0029] (5) Precipitate casein: adjust the pH to 4.6 with 5mol / L food grade hydrochloric acid to precipitate casein at its isoelectric point.

[0030] (6) Centrifugation: 4000g ...

Embodiment 2

[0034] Embodiment 2: a kind of preparation method of yak milk casein bacteriostatic peptide is characterized in that comprising the following steps:

[0035] (1) Yak milk casein: purchased from Hualing Casein Co., Ltd., food grade.

[0036] (2) Dissolution of yak casein: Weigh 20 g of yak casein, add 500 mL of water, adjust the reaction temperature to 55°C, pH 8.0, and stir to dissolve the yak casein.

[0037] (3) Add trypsin: add trypsin: trypsin was purchased from Beijing Geyuan Tianrun Biotechnology Co., Ltd., the titer was determined to be no less than 2500 units, calculated on the basis of 20g of yak casein, and 0.5g of enzyme was weighed to maintain the reaction Temperature 55°C, pH 8.0, react for 2 hours.

[0038] (4) Inactivate trypsin, raise the temperature to 80°C and keep it for 15 minutes.

[0039] (5) Precipitate casein: adjust the pH to 4.6 with 5mol / L food grade hydrochloric acid to precipitate casein at its isoelectric point.

[0040] (6) Centrifugation: 120...

Embodiment 3

[0044] Embodiment 3: a kind of preparation method of yak milk casein bacteriostatic peptide is characterized in that comprising the following steps:

[0045] (1) Yak milk casein: purchased from Hualing Casein Co., Ltd., food grade.

[0046] (2) Dissolution of yak casein: Weigh 40 g of yak casein, add 500 mL of water, adjust the reaction temperature to 55°C, pH 8.0, and stir to dissolve the yak casein.

[0047] (3) Add trypsin: add trypsin: trypsin was purchased from Beijing Geyuan Tianrun Biotechnology Co., Ltd., the titer was determined to be no less than 2500 units, calculated based on 40g of yak casein, and 0.5g of enzyme was weighed to maintain the reaction Temperature 55°C, pH 8.0, react for 2 hours.

[0048] (4) Inactivate trypsin, raise the temperature to 80°C and keep it for 15 minutes.

[0049] (5) Precipitate casein: adjust the pH to 4.6 with 5mol / L food grade hydrochloric acid to precipitate casein at its isoelectric point.

[0050] (6) Centrifugation: 12000g cen...

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Abstract

The invention relates to a preparation method of a yak milk casein antibacterial peptide and belongs to the technical field of biology. The preparation method includes steps of dissolving yak milk casein, adding trypsin for hydrolysis, inactivating the trypsin, precipitating casein, performing centrifugation to remove precipitation, subjecting yak milk casein to sephadex column chromatography seperation, determining molecular weight by mass spectra, performing ultrafiltration and performing spray drying, wherein ultrafiltration membranes with interception of 1000Da and 2500Da are used for collecting peptide fragments with the molecular weight range of 1000Da to 2500Da. According to the yak milk casein antibacterial peptide prepared by the method, peptide molecular weight distribution is concentrated, bacteriostatic activity is high, the minimum inhibition concentration of staphylococcus aureus is 270 mu g / mL, and the minimum inhibition concentration of Escherichia coli is 980 mu g / mL.

Description

technical field [0001] The invention relates to a preparation method of milk casein bacteriostatic peptide and belongs to the field of biotechnology. Background technique [0002] Milk is the main food source for newborn mammals. The newborn baby has not yet completed the development of the immune system, and its disease resistance and immune function mainly come from breast milk, which is due to the presence of a variety of biologically active peptides. Milk contains a variety of bioactive factors, including antibacterial and antiviral bioactive peptides such as lactoferrin. Studies have shown that these biologically active peptides can be released by hydrolyzing milk proteins with appropriate proteases under in vitro conditions. [0003] Casein accounts for 78.8% of the protein content of milk and is a complete protein. In recent years, studies have shown that in addition to nutritional functions, casein also has very important physiological functions and is an importan...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/16
Inventor 谢小冬宋礼纪银莉高维东何潇葛静微苏艳萍
Owner 甘肃华羚生物技术研究中心
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