Whey protein peptide and high-F-value oligopeptide with liver injury protection effect as well as preparation method and application of whey protein peptide and high-F-value oligopeptide

A whey protein peptide and protease technology, applied in the field of active peptides, can solve the problems of poor reproducibility, difficulty in large-scale production, and low efficiency, and achieve the effect of increasing the content

Active Publication Date: 2022-08-02
JIANGSU QIRUI MEDICINE TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although high F value oligopeptides have a very wide range of biological activities, due to the high standards of their products, large-scale production is not easy, such as poor control of the directional enzymatic hydrolysis process, poor reproducibility, and the removal effect of aromatic amino acids poor, low efficiency

Method used

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  • Whey protein peptide and high-F-value oligopeptide with liver injury protection effect as well as preparation method and application of whey protein peptide and high-F-value oligopeptide
  • Whey protein peptide and high-F-value oligopeptide with liver injury protection effect as well as preparation method and application of whey protein peptide and high-F-value oligopeptide
  • Whey protein peptide and high-F-value oligopeptide with liver injury protection effect as well as preparation method and application of whey protein peptide and high-F-value oligopeptide

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Experimental program
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preparation example Construction

[0035] The present invention provides a method for preparing a whey protein peptide, comprising the following steps: after pre-treating whey protein with alkaline protease, exposing the aromatic amino acid of the peptide chain at the C-terminus or N-terminus with endoprotease, and then Exoprotease is used to hydrolyze the exposed aromatic amino acids to obtain an enzymatic hydrolysis solution; the enzymatic hydrolysis solution is subjected to ion exchange column chromatography to collect a permeate, wherein the permeate contains the whey protein peptide.

[0036] The present invention utilizes alkaline protease to pretreat whey protein, and the pretreatment includes performing first enzymatic hydrolysis on whey protein with alkaline protease under alkaline conditions to obtain a first enzymatic hydrolysis solution. The pH value of the first enzymatic hydrolysis system of the present invention is preferably about 8.5, and the temperature of the first enzymatic hydrolysis is pref...

Embodiment 1

[0047] Step 1. Alkaline protease pretreatment

[0048] Measure 6L of deionized water, adjust its pH to about 8.5 with 1M NaOH, raise the temperature to 55°C, weigh 0.5g of alkaline protease, and slowly add 500g of whey protein after stirring evenly, stirring continuously at 45 r / min. , to obtain the first enzymatic hydrolyzate;

[0049] Step 2. Further treatment with endonuclease

[0050] During the reaction of the first enzymatic hydrolysis solution, the pH was continuously detected during the period. When the pH dropped to about 7.0 to 7.5, and the temperature dropped to 37 to 42 °C at the same time, 6.5 g of trypsin was weighed, and after enzymatic hydrolysis for 2 hours, the second enzymatic hydrolysis was obtained. liquid;

[0051] Step 3. Exonuclease treatment

[0052] Weigh 1.5 g of flavor protease, add it to the second enzymatic hydrolysis solution, continue enzymatic hydrolysis for 4 hours, heat up to 100 °C, and kill the enzyme for 10 min to obtain the third enzym...

Embodiment 2

[0111] Activation rate assay for alcohol dehydrogenase (ADH)

[0112] Alcohol dehydrogenase (ADH) catalyzes the dehydrogenation of ethanol to form acetaldehyde in the weak base range in the presence of oxidative aldehyde coenzyme I (NAD+) as follows:

[0113]

[0114] The present invention adopts the Waller-Hooch method with slight modifications to determine the inhibitory activity of high F-value whey protein peptides to alcohol dehydrogenase; prepare a 5mL test tube, add 1.5mL of pH8.8 sodium pyrophosphate buffer, 27mmol / L-oxidative coenzyme I (NAD+) 1.0 mL, 11.5% ethanol solution 0.5 mL, test sample 0.1 mL (50 μg / mL sample solution or ultrapure water), the above samples are mixed evenly and placed in a 25°C water bath The pot was warmed for 5 min, and finally 0.1 mL of alcohol dehydrogenase (ADH, 0.25 U / mL) in a 25°C warm bath was added to the test tube. Use a multi-function microplate reader to measure the absorbance at a wavelength of 340nm, measure the absorbance ev...

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Abstract

The invention provides a group of whey protein peptides and high-F-value oligopeptides with a liver injury protection effect as well as a preparation method and application thereof, and relates to the technical field of active peptides. According to the method, the high-F-value oligopeptide is prepared by hydrolyzing whey protein through three enzymes, firstly, a small amount of alkaline protease is used for pretreating the high-F-value oligopeptide, the solubility of the high-F-value oligopeptide is improved, the peptide chain of the high-F-value oligopeptide is damaged, the subsequent enzymolysis efficiency is improved, then endonuclease treatment and exonuclease treatment are sequentially adopted, and finally the high-F-value oligopeptide exists in enzymatic hydrolysate in a free state, so that the high-F-value oligopeptide is obtained. And finally adsorbing the aromatic amino acid by using specific resin to obtain whey protein peptide with F value greater than 20, and finally separating and screening to obtain two oligopeptides with high F value. The whey protein peptide and the high-F-value oligopeptide can reduce the content of malondialdehyde and triglyceride in the liver and increase the content of reduced glutathione, and have an auxiliary protection effect on chemical liver injury.

Description

technical field [0001] The invention belongs to the technical field of active peptides, and in particular relates to a group of whey protein peptides and high F-value oligopeptides with liver injury protection functions, and a preparation method and application thereof. Background technique [0002] F value (Fischer ratio) is the molar ratio of branched-chain amino acids (BCAA: Val, Ile, Leu) and aromatic amino acids (AAA: Trp, Tyr, Phe), in memory of the famous German scholar Fischer.J.E in the 20th century. In the 1970s, it was named after the "pseudo-neurotransmitter hypothesis". High-F oligopeptides are bioactive peptides with a molar ratio of branched-chain amino acids and aromatic amino acids obtained by using protease to act on plant protein or animal protein, which is much higher than the ratio of these two types of amino acids in the human body. [0003] Due to its unique amino acid composition and physiological function, high F-value oligopeptides have attracted g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K7/06C07K1/18A61K38/01A61P1/16A23L33/18
CPCC12P21/06C07K7/06A61K38/018A61P1/16A23L33/18A61K38/00A23V2002/00A23V2200/32A23V2250/54252A23V2250/55
Inventor 吴明辉朱敏支晓娜冯小深朱安琪
Owner JIANGSU QIRUI MEDICINE TECH CO LTD
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