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Novel intestinal mucosa anti-oxidation active peptide and preparation method thereof

An antioxidant active peptide and antioxidant activity technology, applied in the field of new intestinal mucosal antioxidant active peptide and its preparation, to achieve the effects of easy structure regulation, strong stability, and good application potential

Pending Publication Date: 2020-02-28
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few reports on the optimization of the preparation process of intestinal mucosal polypeptides by enzymatic hydrolysis and the existence of bioactive peptides in the enzymatic hydrolysis solution

Method used

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  • Novel intestinal mucosa anti-oxidation active peptide and preparation method thereof
  • Novel intestinal mucosa anti-oxidation active peptide and preparation method thereof
  • Novel intestinal mucosa anti-oxidation active peptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The preparation method of antioxidant peptide of the present invention is as follows:

[0025] Extraction of intestinal mucosal protein: fresh intestinal mucosa was filtered to remove impurities, dried in an oven, and stored at 4°C after cooling at room temperature; 5 g of intestinal mucosa was dissolved in 25 mL of phosphate buffer (pH 6.5); the solution was homogenized Mix well, conditions: 10000 r / min, 10 s each time, repeat 1-2 times; prepare intestinal mucosa solution with a ratio of material to liquid 1:4, homogenate at 10000 rpm for 10 s, add papain, the enzyme dosage is 5000 U / g protein, enzymolysis temperature 50°C, enzymolysis time 6 h, inactivation in boiling water bath for 5 min, centrifugation at 4000 r / min for 15 min, supernatant was collected and freeze-dried for later use.

[0026] Separation of intestinal mucosal antioxidant peptides by ultrafiltration: the lyophilized powder of intestinal mucosal proteolyzate was obtained according to the above method...

Embodiment 2

[0029] Embodiment 2 studies the natural antioxidant peptide activity that obtains in embodiment 1:

[0030] 1. Peptide artificial synthesis steps:

[0031] (1) Hanging resin: Add 2g of CTC resin to a 125mL reaction column, then add 50ml of DCM to swell for 30min, add Fmoc-Lys (BOC)-OH (2eq) and DIPEA (4eq), react for 2.5h, then add 2ml of MeOH to block After 30 minutes, drain the solvent, wash with DCM (50ml*3) and DMF (50ml*3) successively.

[0032] (2) Deprotection: add 50ml 20% piperidine / DMF, react for 30min, wash with DMF (50ml*5).

[0033] (3) Condensed amino acids: add Fmoc-Leu (2eq), HBTU (2eq), DIPEA (4eq), then add 50ml DMF, react for 1.5h, wash with DMF (50ml*5).

[0034] (4) Peptide chain growth: Repeat step 2 and step 3 in sequence until the last amino acid Fmoc-Asp(OtBu)-OH is connected, and then wash with DMF (50ml*5), DCM (50ml*2), and MeOH (50ml*2), drained.

[0035] (5) Cleavage: Add 5 times the volume of TFA:TIS:H2O (95:2.5:2.5) to the drained resin, and...

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Abstract

The invention relates to the technical field of biology and particularly discloses a novel intestinal mucosa anti-oxidation active peptide and a preparation method thereof. Amino acid sequences of theanti-oxidation active peptide are shown as Asp-Gly-Leu-Lys-His-Leu-Asp-Asn-Leu-Lys and Asn-Asp-Trp-Glu-Asp-His-Leu-Ala-Val-Lys. The anti-oxidation active peptide is derived from intestinal mucosa protein. The peptide has more hydrophobic amino acids and strong anti-oxidation activity and is strong in DPPH free radical scavenging action and high in reducing power. The preparation method of the peptide has advantages of simplicity, easiness in large-scale production and the like. The novel intestinal mucosa anti-oxidation active peptide and the preparation method thereof have significant socialbenefits, economical benefits or ecological benefits in increasing of industrial added values of natural casings, abatement of environmental pollution and promotion of comprehensive utilization of casing processing byproduct resources.

Description

technical field [0001] The invention relates to a novel intestinal mucosal antioxidant active peptide and a preparation method thereof, belonging to the field of comprehensive utilization of animal by-product resources. Background technique [0002] As a kind of biologically active peptides, antioxidant peptides are the most studied biologically active peptides, which have the functions of scavenging free radicals and inhibiting the formation of active oxygen. Antioxidant peptides can affect other biological activities, and have a certain relationship with some physiological regulatory functions, such as immune regulation, lowering cholesterol, and anti-tumor. Many studies have found that there are peptide sequences with antioxidant activity in food source proteins, which are more active than proteins and amino acids, and can be released by enzyme engineering technology. Food-derived antioxidant peptides have become a potential base for developing natural antioxidants becau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/36C07K1/34C07K1/16C12P21/06
CPCC07K7/06C12P21/06
Inventor 葛庆丰刘瑞丁琪琪陈磊于海
Owner YANGZHOU UNIV
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