Ultrasonic sound assistant method for simulated digestion of casein active peptides and health food applications

A casein and protease technology, which is applied in the fields of intensive processing of dairy products and functional food preparation, can solve the problems of inability to absorb peptides, ignoring the digestion and absorption of gastrointestinal tract, and ignoring the influence of selective absorption of small intestinal epithelial cells.

Active Publication Date: 2018-04-27
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 peptides from casein 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 peptides by small intestinal endothelial cells

Method used

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  • Ultrasonic sound assistant method for simulated digestion of casein active peptides and health food applications
  • Ultrasonic sound assistant method for simulated digestion of casein active peptides and health food applications
  • Ultrasonic sound assistant method for simulated digestion of casein active peptides and health food applications

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Ultrasonic treatment of casein: Weigh casein quantitatively, dissolve in phosphate buffer solution with pH=7.8, and the concentration of casein is 1g / 100mL. Single frequency (40kHz) sonication. The ultrasonic treatment time is 30min, the intermittent ratio is 10s / 3s, and the temperature is 25°C.

[0056] Preparation of casein 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, enzymolysis temperature is 50°C, and enzymolysis 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 casein, the conversion rate of protein, and the ACE inhibitory activity of its hydrolyzate were measured.

[0057] Simulated gastrointestinal digestion: Prepare artificial gastric juice accord...

Embodiment 2

[0060] Ultrasonic treatment of casein: Weigh casein quantitatively, dissolve in phosphate buffer solution with pH=7.8, and the concentration of casein is 2g / 100mL. Dual frequency (20 / 40kHz) ultrasonic processing. The ultrasonic treatment time is 20min, the intermittent ratio is 10s / 3s, and the temperature is 30°C.

[0061] Preparation of casein hydrolyzate: 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, enzymolysis temperature is 55°C, and enzymolysis 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 casein, the conversion rate of protein, and the ACE inhibitory activity of its hydrolyzate were measured.

[0062] Simulated gastrointestinal digestion: Prepare artificial gastric juice according to the ...

Embodiment 3

[0065]Ultrasonic treatment of casein: Weigh casein quantitatively, dissolve in phosphate buffer solution with pH=7.8, and the concentration of casein is 5g / 100mL. Tri-frequency (20 / 40 / 60kHz) sonication. The ultrasonic treatment time is 10min, the intermittent ratio is 10s / 3s, and the temperature is 40°C.

[0066] Preparation of casein hydrolyzate: adjust the pH of the above protein solution to 8, add papain, 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, the pH of the mixture was adjusted to 7.0, and the enzyme was inactivated in a boiling water bath for 10 minutes. The supernatant was obtained by centrifugation, desalted, concentrated, and lyophilized into powder. The degree of hydrolysis, protein conversion rate, and ACE inhibitory activity were measured.

[0067] Simulated gastrointestinal digestion: Prepare artificial gastric juice according to the United States Pharmacopoeia (USP...

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Abstract

The invention discloses an ultrasonic sound assistant method for simulated digestion of casein active peptides and health food applications, and belongs to the technical fields of dairy product deep processing and health food preparation. The method comprises the following steps: ultrasonic pretreatment of casein is firstly employed; enzymatic hydrolysis is carried out with protease in order to prepare casein active peptides with ACE inhibitory activity; ACE inhibitory activity of the casein active peptides is traced by means of simulation of gastrointestinal tract digestion, and after Caco-2cells simulate absorption of small intestine epithelial cells, the casein functional polypeptides with high ACE inhibitory activity which are digested by gastrointestinal tracts and absorbed by smallintestine inner walls which are simulated by Caco-2 cells are represented. The simulated digestive system and the small intestine inner wall absorption system simulated by Caco-2 cells are firstly combined in order to trace activity change of casein polypeptides with ACE inhibition; three kinds of casein functional polypeptides with high ACE inhibitory activity which are digested by stomach and intestine and absorbed by small intestine inner walls which are simulated by Caco-2 cells are firstly identified.

Description

technical field [0001] The invention provides an ultrasonic-assisted simulated digestion method of casein active peptide and its application in functional food, which belongs to the technical field of deep processing of dairy products and preparation of functional food, and can be used as a functional product for preparing health food or medicine. Background technique [0002] Hypertension (hypertension) refers to an increase in systemic arterial blood pressure (systolic and / or diastolic) as the main feature (systolic blood pressure ≥ 140 mm Hg, diastolic blood pressure ≥ 90 mm Hg), which may be accompanied by heart, brain, kidney, etc. Clinical syndrome of functional or organic damage to organs. Hypertension is the most common chronic disease and the main risk factor for cardiovascular and cerebrovascular diseases. At present, there are more than 200 million hypertensive patients in China, so the current situation is very serious. Angiotensin converting enzyme (ACE EC 3.4...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/103C07K5/083C12P21/06A23L33/18
Inventor 任晓锋梁秋芳张熙陆峰侯婷马海乐
Owner JIANGSU UNIV
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