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Preparation method of black-bone chicken oligopeptide and separation and identification method of active peptide fragment

An identification method and oligopeptide technology are applied in the field of biomedicine to achieve the effects of high yield, improved nutritional value and high degree of hydrolysis

Active Publication Date: 2012-11-21
CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the reported black-bone chicken research, only a preliminary exploration of black-bone chicken active peptides has been carried out, and the molecular weights are relatively large, most of which are concentrated in 2000-5000Da. Systematic research and development of specific active peptides

Method used

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  • Preparation method of black-bone chicken oligopeptide and separation and identification method of active peptide fragment
  • Preparation method of black-bone chicken oligopeptide and separation and identification method of active peptide fragment
  • Preparation method of black-bone chicken oligopeptide and separation and identification method of active peptide fragment

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Experimental program
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Effect test

Embodiment 1

[0065]1.83kg white striped black-bone chicken was made into meat puree with a chopper and a meat grinder, transferred to a hydrolysis pot, added 5L of deionized water, stirred evenly, heated and boiled for 0.5h, and cooled to room temperature. After the liquid material was treated with a colloid mill, it was heated to 90°C, and allowed to stand and cooled to room temperature. Centrifuge the liquid material at a speed of 3000r / min for 15 minutes, let it stand for 5 minutes, discard the upper layer of the liquid material, and keep the lower layer of liquid material. Return the liquid material to the hydrolysis kettle, heat to 50°C, adjust the liquid material to pH 7.5 with NaOH solution, add Protamex enzyme with a solid content of 3.0% in the liquid material, and enzymolyze the protein until the degree of protein hydrolysis of the liquid material reaches about 70%. The liquid material is passed through a high temperature instantaneous sterilization (UHT) system to kill enzymes. ...

Embodiment 2

[0067] 1902.3kg white striped black-bone chicken was made into meat puree with a chopping machine and a meat grinder, transferred to a hydrolysis tank, added 6000L of deionized water, stirred evenly, heated and boiled for 0.5h, and cooled to room temperature. After refining the liquid material with a colloid mill, heat it to 90°C, and let it stand to cool down to room temperature. Process the liquid material with a disc centrifuge to remove grease and retain the protein liquid material. Return the liquid material to the hydrolysis tank, heat to 55°C, adjust the liquid material to pH 8.5 with NaOH solution, add Protamex enzyme with a solid content of 5.0% in the liquid material, and perform enzymatic hydrolysis until the protein hydrolysis degree of the liquid material reaches about 70%. The liquid material is passed through a high temperature instantaneous sterilization (UHT) system to kill enzymes. Add ionized water to the liquid material to a solid content of 5%, and after ...

Embodiment 3

[0069] Use RP-HPLC to separate the components of black-bone chicken oligopeptides. The operation steps are: (1) Sample preparation: Weigh 1g of black-bone chicken oligopeptide dry powder, dilute it to a 100mL volumetric flask with mobile phase A, and oscillate ultrasonically for 10 minutes. The sample was fully dissolved and mixed to prepare a 0.1 mg / mL black-bone chicken oligopeptide solution. The sample solution was filtered with a polytetrafluoroethylene filter membrane with a pore size of 0.2 μm. (2) Sample loading: the sample loading volume is 50 μL. (3) Gradient elution: chromatographic column: XBridge BEH130 C18 column (4.6×250mm); mobile phase A: V (water): V (trifluoroacetic acid) = 100: 0.1; mobile phase B: V (acetonitrile): V (Water): V (trifluoroacetic acid) = 80: 20: 0.1; detection wavelength: UV220nm; flow rate: 0.6mL / min; column temperature: 32°C; gradient elution program: 0-20min, mobile phase B: 0% -5%; 20-30min, mobile phase B: 5%-5%; 30-80min, mobile phase...

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Abstract

A preparation method of black-bone chicken oligopeptides and a separation and identification method of active peptide fragments belong to the technical field of biological medicine. The preparation method particularly comprises the following steps: pretreating a black-bone chicken which is killed and has feather being stripped off by wringing, adding water, heating, and fat removal, adding Protamex for enzymatic hydrolysis, then performing procedures of centrifugation, ultrafiltration, concentration, spray drying, and the like to obtain the black-bone chicken oligopeptides. Detection shows that components in the black-bone chicken oligopeptides, which has a molecular weight of less than 1000 Da, account for more than 85%. The black-bone chicken oligopeptides have high DPPH free radical removing activity with an IC50 value of 4.52 mg / mL. Also, the black-bone chicken oligopeptides have strong ACE inhibition activity with an IC50 value of 2.86 mg / mL. A part of active peptide fragments are separated and identified by an RP-HPLC method and a Q-TOF2 method, wherein 2 antioxidation active peptide fragments are LWR and NM; and 5 blood pressure-reducing active peptide fragments are LER, GAGP, KPGV, LH, and NM.

Description

technical field [0001] The invention relates to the preparation of black-bone chicken oligopeptides and methods for separating and identifying active peptides, and belongs to the technical field of biomedicine. [0002] Background technique [0003] Black-bone chicken is a rare bird in my country's poultry breeds, which has special nutritional and medicinal value. Black-bone chicken is rich in nutritional value, and its main chemical components are protein, fat, calcium, phosphorus, iron, magnesium, vitamin B1, B2, etc. It contains more than 20 kinds of amino acids needed to form human protein, and has a special nourishing effect on the human body. It is recorded in "Compendium of Materia Medica" that black-bone chicken has the functions of tonifying deficiency, fatigue and weakness, curing thirst, benefiting pregnant women, treating women's collapse and some diseases of weakness. The efficacy of traditional black-bone chicken products in improving physiological functions, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K4/12C07K1/36C07K1/34C07K1/20G01N30/36G01N30/72C07K5/083C07K5/072C07K5/103A61P39/06A61P9/12
Inventor 蔡木易谷瑞增易维学马勇董哲潘兴昌林峰马永庆徐亚光鲁军金振涛陈亮刘文颖
Owner CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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