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Corn prolamin anti-inflammatory polypeptide and preparation method thereof

A technology of zein and peptides, which is applied in the direction of plant zein, chemical instruments and methods, specific peptides, etc., can solve the problem of non-absorption, ignore the influence of selective absorption by small intestinal epithelial cells, and ignore digestion and absorption in the gastrointestinal tract etc. to achieve good anti-inflammatory activity

Active Publication Date: 2018-04-03
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent "Anti-inflammatory peptide isolated from viscera of wrinkled abalone and its application" (201510594885.7), although the preparation of anti-inflammatory peptide by simulating gastrointestinal digestion method, ignores the influence of small intestinal epithelial cells on its selective absorption
In summary, the current methods for preparing functional polypeptides from zein ignore the digestion and absorption of the gastrointestinal tract, and cannot truly simulate the digestion of bioactive peptides in the gastrointestinal tract and the absorption of polypeptides by small intestinal endothelial cells

Method used

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  • Corn prolamin anti-inflammatory polypeptide and preparation method thereof
  • Corn prolamin anti-inflammatory polypeptide and preparation method thereof
  • Corn prolamin anti-inflammatory polypeptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Ultrasonic treatment of zein: quantitatively weigh zein, add distilled water to prepare a 1% (m / v) suspension, and perform single-frequency (40 kHz) ultrasonic treatment. The ultrasonic treatment time is 30min, the intermittent ratio is 10s / 3s, and the temperature is 25°C.

[0054] Preparation of zein enzymatic hydrolysis solution: adjust the pH of the above protein solution to 8, add alkaline protease, the ratio of enzyme to substrate is 1:20 (w / w), mix evenly, the enzymatic hydrolysis temperature is 50°C, and the enzymatic hydrolysis time is 2h. After the enzymolysis, adjust the pH of the mixture to 7.0, and inactivate the enzyme in a boiling water bath for 10 minutes, centrifuge to obtain a supernatant, desalt, concentrate, and freeze-dry to powder. The degree of hydrolysis of zein, the conversion rate of protein, and the anti-inflammatory activity of its hydrolyzate on EA.hy926 cells were measured.

[0055] Simulated gastrointestinal digestion: Prepare artificial g...

Embodiment 2

[0058] Ultrasonic treatment of zein: quantitatively weigh zein, add distilled water to prepare a 2% (m / v) suspension, and perform dual-frequency (20 / 28kHz) ultrasonic treatment. The ultrasonic treatment time is 20min, the intermittent ratio is 10s / 3s, and the temperature is 25°C.

[0059]Preparation of zein enzymatic hydrolysis solution: adjust the pH of the above protein solution to 7.5, add neutral protease, the ratio of enzyme to substrate is 1:30 (w / w), mix evenly, the enzymatic hydrolysis temperature is 55°C, and the enzymatic hydrolysis time is 4h. After enzymolysis, adjust the pH of the mixture to 7.0, inactivate the enzyme in a boiling water bath for 10 minutes, centrifuge to obtain the supernatant, desalt, concentrate, and freeze-dry to powder. The degree of hydrolysis of zein, the conversion rate of protein, and the anti-inflammatory activity of its hydrolyzate on EA.hy926 cells were measured.

[0060] Simulated gastrointestinal digestion: Prepare artificial gastric...

Embodiment 3

[0063] Ultrasonic treatment of zein: Quantitatively weigh zein, add distilled water to prepare a 5% (m / v) suspension, and perform three-frequency (20 / 28 / 40kHz) ultrasonic treatment. The ultrasonic treatment time is 10min, the intermittent ratio is 10s / 3s, and the temperature is 25°C.

[0064] Preparation of zein enzymatic hydrolysis solution: adjust the pH of the above protein solution to 8, add thermophilic protease, the ratio of enzyme to substrate is 1:50 (w / w), mix evenly, enzymolysis temperature is 70°C, and enzymolysis time is 2h. After enzymolysis, adjust the pH of the mixed solution to 7.0, inactivate the enzyme in a boiling water bath for 10 minutes, centrifuge to obtain the supernatant, desalt, concentrate, freeze-dry into powder, measure the degree of hydrolysis, protein conversion rate, and the effect on EA.hy926 cells anti-inflammatory activity.

[0065] Simulated gastrointestinal digestion: Prepare artificial gastric juice according to the United States Pharmaco...

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Abstract

The invention discloses corn prolamin anti-inflammatory polypeptide and a preparation method thereof, and belongs to the technical fields of deep processing of agricultural products and preparation offunctional food. The preparation method comprises the following steps: performing ultrasonic pretreatment on corn prolamin, performing enzymolysis by using protease to prepare corn anti-inflammatorypolypeptide, tracking the activity of the corn anti-inflammatory polypeptide by simulating gastrointestinal digestion, simulating intestinal epithelial cell absorption through Caco-2 cells and representing the corn pure peptide with high anti-inflammatory activity capable of resisting Caco-2 cell decomposition through gastrointestinal digestion and being absorbed by the inner wall of the small intestine simulated by the Caco-2 cells. The invention identifies the corn prolamin functional polypeptide with high anti-inflammatory activity for the first time, wherein the corn prolamin functional polypeptide can resist Caco-2 cell decomposition through gastrointestinal digestion and can be absorbed by the inner wall of the small intestine simulated by the Caco-2 cells. Through combination of thesimulated digestion system and the Caco-2 simulated intestine inner wall absorbing system, the change of the activity of the corn anti-inflammatory polypeptide is tracked; and the absorption rate ofthe corn polypeptide digestion peptide in the Caco-2 simulated intestine inner wall absorbing system is researched.

Description

technical field [0001] The invention provides a zein anti-inflammatory polypeptide and a preparation method thereof, which belong to the technical field of intensive processing of agricultural products and preparation of functional foods, and can be used as functional additives in functional foods or medicines. Background technique [0002] Chronic inflammation is involved in the occurrence and development of various diseases in the human body, and is the pathogenic basis of many diseases. Vascular endothelial cells, the single layer of squamous epithelial cells that line the inner surfaces of the heart, blood vessels, and lymphatic vessels, play crucial roles in vascular biology, such as regulation of vascular tone, hemostasis, neutrophil recruitment, hormone transport, and Liquid Filtration etc. Vascular inflammation is one of the key factors leading to endothelial dysfunction and is closely related to a variety of diseases, including atherosclerosis, diabetes, coronary a...

Claims

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Application Information

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IPC IPC(8): C07K14/425C12P21/06A23L33/18
CPCC12P21/06C07K14/425
Inventor 梁秋芳马海乐任晓锋
Owner JIANGSU UNIV
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