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A casein-derived cholesterol-lowering peptide and its preparation method and application

A technology for lowering cholesterol and casein peptide is applied in the field of cholesterol lowering peptide derived from casein and its preparation, and achieves the effects of improving efficiency, high enzymatic cleavage efficiency and inhibiting absorption

Active Publication Date: 2020-04-21
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved: In order to overcome the defects of the existing technology, make up the gap in the study of cholesterol-lowering peptides derived from casein in milk, obtain a new source of cholesterol-lowering peptides, and prepare them by artificial synthesis to inhibit exogenous cholesterol Absorbed cholesterol-lowering peptide, the invention provides a casein-derived cholesterol-lowering peptide and its preparation method and application

Method used

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  • A casein-derived cholesterol-lowering peptide and its preparation method and application
  • A casein-derived cholesterol-lowering peptide and its preparation method and application
  • A casein-derived cholesterol-lowering peptide and its preparation method and application

Examples

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preparation example Construction

[0038] Preparation of biuret reagent: Weigh 1.5g of copper sulfate and 6.0g of potassium sodium tartrate, dissolve them in 500mL of distilled water, add 300mL of 10% (W / V) NaOH solution during dissolution and stirring, and dilute to 1L with distilled water.

[0039] Preparation of peptide content standard curve: Take ten 10mL volumetric flasks and prepare 0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6 in sequence with 5% (W / V) TCA (trichloroacetic acid) aqueous solution and 1.8mg / mL Gly-Gly-Tyr-Arg tetrapeptide standard solution, then take 6.0mL standard solution, add 4.0mL biuret reagent, mix well, let stand for 10min, centrifuge at 2000r / min for 10min, take the supernatant Measure the OD value at 540nm (use the first tube as blank control). The concentration of the peptide was taken as X (mg / mL) on the abscissa, and the absorbance (OD value) was taken as Y on the ordinate to create a standard curve.

[0040] Determination of sample polypeptide content: Take 2.5mL sample solution...

Embodiment 1

[0046] A casein-derived cholesterol-lowering peptide, the cholesterol-lowering peptide comprising the following amino acid sequences: SEQ ID NO:1 and SEQ ID NO:2.

[0047] A method for preparing a casein-derived cholesterol-lowering peptide, comprising the following steps:

[0048] (1) Preparation of casein hydrolysis solution: prepare 50mg / mL casein solution, add 50U / mg neutral protease, the final concentration of protease is 60mg / mL; use 1mol / L NaOH and 0.5mol / L HCl solution to adjust the enzymolysis The pH is 7.0, the enzymatic hydrolysis temperature is 47.5°C, 1mol / L NaOH is continuously added during the enzymatic hydrolysis to keep the pH of the solution constant, and the enzymatic hydrolysis time is 3.5h; supernatant;

[0049] (2) Preparation of casein peptide ultrafiltrate: the supernatant collected in step (1) is subjected to ultrafiltration, the ultrafiltration membrane pressure is 0.25MPa, and the flow rate is 7.5mL / min; the ultrafiltration membrane used in the ultr...

Embodiment 2

[0059] The difference between this example and Example 1 lies in the step (1) preparation of casein hydrolysis solution: prepare 40 mg / mL casein solution, add neutral protease to a final concentration of 50 mg / mL, enzymolysis pH 6.0, enzymolysis temperature at 45°C, enzymatic hydrolysis for 4 hours, inactivated by heat, and centrifuged to obtain the supernatant.

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Abstract

The invention discloses a cholesterol lowering peptide with a casein source as well as a preparation method and application of the cholesterol lowering peptide. The cholesterol lowering peptide comprises the following amino acid sequences: SEQ ID NO:1 and SEQ ID NO:2. The peptide is prepared by the steps: hydrolyzing casein through protease so as to obtain enzymatic hydrolysate; performing ultrafiltration, chromatographic separation and ultra-high performance liquid chromatography mass spectrometry on the enzymatic hydrolysate so as to obtain a polypeptide sequence, synthesizing the polypeptide, and verifying the cholesterol lowering activity, thereby obtaining the product. The cholesterol lowering peptide with the casein source comes from cow's milk; a crude cholesterol lowering peptide obtained through gel filtration chromatography in the method disclosed by the invention does not need to be subjected to liquid chromatography separation, the amino acid sequences of the cholesterol lowering peptide are directly analyzed by adopting ultra-high performance liquid chromatography mass spectrometry (UPLC-MS) analysis, the cholesterol lowering peptide is synthesized so as to further verify the cholesterol lowering activity, the sequence of the cholesterol lowering peptide is determined, and the efficiency is improved; moreover, the cholesterol lowering peptide with the casein source disclosed by the invention can be used for preparing functional foods, so that the absorption of exogenous cholesterol is inhibited.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a functional factor and a preparation method thereof, in particular to a cholesterol-lowering peptide derived from casein, a preparation method and application thereof. Background technique [0002] Excessive blood cholesterol levels, especially low-density lipoprotein levels, may increase the risk of cardiovascular disease, and the accumulation of cholesterol on arterial walls can lead to diseases such as atherosclerosis. At present, most of the cholesterol-lowering drugs used in clinical treatment are chemical drugs, which will bring different degrees of toxic and side effects to patients. For example, bile acid integrators such as cholestyramine and colestipol, as well as statins, have certain risks to the health of patients and are not suitable for long-term use. The food-derived active peptide is a functional food with few clinical side effects and a good market prospect. [000...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C07K5/103C12P21/06A23L33/18
CPCA23V2002/00C07K5/101C07K14/4732C12P21/06A23V2200/3262A23V2250/55
Inventor 郭宇星潘道东高彬熊彬屹姜潇潇
Owner NANJING NORMAL UNIVERSITY