Five pancreatic lipase inhibitors derived from small red bean protein and application thereof

A pancrelipase and inhibitor technology, applied in peptide/protein components, medical preparations containing active ingredients, metabolic diseases, etc., can solve problems such as sudden death, achieve good lipid-lowering effect, clear target, and clear mechanism of action Effect

Active Publication Date: 2022-07-29
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Orlistat is a pancreatic lipase inhibitor drug that is widely recognized and purchased in the market, but it has side effects such as insomnia, fatigue, nausea, vomiting, flatulence, and even arrhythmia, heart failure, and sudden death

Method used

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  • Five pancreatic lipase inhibitors derived from small red bean protein and application thereof
  • Five pancreatic lipase inhibitors derived from small red bean protein and application thereof
  • Five pancreatic lipase inhibitors derived from small red bean protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Extraction of adzuki bean protein

[0022] The adzuki bean protein was extracted by the method of alkali dissolution and acid precipitation. Specifically, defatted adzuki bean powder and distilled water were mixed at a solid-liquid ratio of 1:10 (w / v), and the pH of the solution was adjusted to 8.5. After shaking in a water bath for 1 h at 40°C, the supernatant was collected, and after the pH value of the supernatant was adjusted to 4.5, the protein was allowed to stand for 1 h at room temperature to precipitate the protein. The precipitate was collected, washed 3 times with distilled water, and the pH of the protein was adjusted to 7.0. Finally, freeze-dry and store at -20°C.

Embodiment 2

[0023] Example 2 In vitro simulated digestion

[0024] Adzuki bean protein was hydrolyzed by double-enzyme (pepsin and trypsin) method to obtain protein hydrolyzate. Specifically, 4% (w / w) pepsin was first added to 5% adzuki bean protein solution, the pH of the enzymatic hydrolysis was 2.0, the enzymatic hydrolysis temperature was 37 °C, and the enzymatic hydrolysis time was 2 h; then the pH of the enzymatic hydrolysis solution was The pH value was adjusted to 5.3, the pH value was maintained at 7.5, 4% trypsin (w / w) was added, the enzymatic hydrolysis temperature was 37°C, and the enzymatic hydrolysis time was 2 h. After the enzymatic hydrolysis, the enzyme was inactivated in a boiling water bath for 10 min, and the enzymatic hydrolysis solution was centrifuged to take the supernatant.

Embodiment 3

[0025] Example 3 Screening of lipid-lowering peptides

[0026] The supernatant collected in step (2) was ultrafiltered by using 10 kDa and 3 kDa centrifugal filters to obtain different fractions (>10 kDa, 3-10 kDa and figure 1 ). The specific process of the pancreatic lipase activity inhibition experiment is as follows:

[0027] In a 96-well microtiter plate, 50 μL of sample, 40 μL of 2.5 mg / mL pancrelipase solution and 50 μL of 10 mM p-nitrophenyl butyrate were used as substrates and incubated in phosphate buffer pH 7.3 for 30 min at 37 °C . The microplate reader records the absorbance at 405 nm. Orlistat was used as a positive control and calculated according to formula (1).

[0028]

[0029] In formula (1): A: the absorbance of the control; B: the absorbance of the control blank; C: the absorbance of the sample; D: the absorbance of the sample blank.

[0030] The <3kDa fractions were sequenced by mass spectrometry using liquid chromatography-tandem mass spectrometry,...

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PUM

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Abstract

The invention provides a peptide with a pancreatic lipase inhibitor effect. The sequence of the peptide is selected from SEQ ID NO.1-5; the invention also provides application of the peptide in preparation of medicines, foods and health-care products for treating hyperlipemia. The peptide is a plant-derived, non-toxic and harmless natural substance, and experiments prove that the peptide has remarkable pancreatic lipase inhibitory activity and lipid-lowering effect; the compound is clear in action mechanism and definite in target spot, is expected to become a substitute medicine for preventing and treating hyperlipemia, and has a good market prospect in the industries of medicine and functional food.

Description

technical field [0001] The present application belongs to the protein field, and specifically, the present application provides five pancreatic lipase inhibitors derived from adzuki bean protein and applications thereof. Background technique [0002] Dietary fats exist primarily in the form of triglycerides, but such molecules cannot be absorbed in the gut unless hydrolyzed into smaller molecules such as monoglycerides and free fatty acids. Pancreatic lipase is a water-soluble lipolytic enzyme secreted by the pancreas, which plays an important role in the digestion of triglycerides. It can digest 50%-70% of dietary fat in food and is the main lipolytic enzyme in the human body. Dietary lipids are first hydrolyzed by pancreatic lipase, decomposed into chylomicrons, and then form micelles with cholesterol and bile salts, which are finally absorbed by cells in the intestinal tract. However, excessive hydrolysis can lead to the development of diseases such as hyperlipidemia. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K7/08C12P21/06A61K38/08A61K38/10A61P3/06A23L33/18
CPCC07K7/06C07K7/08C12P21/06A61P3/06A23L33/18A61K38/00
Inventor 沈群赵卿宇付永霞赵亮星
Owner CHINA AGRI UNIV
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