Three cholesterol-lowering peptides with cholesterol esterase inhibitory activity and application thereof

A cholesterol esterase and activity-inhibiting technology, applied in the protein field, can solve the problems of high price, hepatotoxicity, myotoxicity, etc., and achieve the effect of clear target, high stability and clear mechanism of action

Active Publication Date: 2022-07-22
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]Cholesterol-lowering drugs currently on the market mainly include statins, fibrates, niacin, etc. Among them, statins have significant curative effect, but are expensive and have liver Potential side effects such as toxicity and myotoxicity

Method used

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  • Three cholesterol-lowering peptides with cholesterol esterase inhibitory activity and application thereof
  • Three cholesterol-lowering peptides with cholesterol esterase inhibitory activity and application thereof
  • Three cholesterol-lowering peptides with cholesterol esterase inhibitory activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Preparation of adzuki bean protein

[0023] Defatted adzuki bean powder and distilled water were mixed in a ratio of 1:10 (w / v), and the pH of the solution was adjusted to 8.5. After continuous stirring at 40° C. for 1 h, the supernatant was collected, and after the pH of the supernatant was adjusted to 4.5, it was allowed to stand at room temperature for 1 h to promote protein precipitation. 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

[0024] Example 2 Preparation of proteolytic solution

[0025] A 5% adzuki bean protein solution was prepared for two-stage enzymatic hydrolysis with pepsin and trypsin. First, 4% (w / w) pepsin was added, the pH of the enzymatic hydrolysis was 2.0, the enzymatic hydrolysis temperature was 37 °C, and the enzymatic hydrolysis time was 2 h; after the pepsin hydrolysis was completed, the pH of the enzymatic hydrolysis solution was adjusted to 5.3, The pH value was maintained at 7.5, then 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 was completed, the enzyme was heated in boiling water to inactivate the enzyme, and the enzymatic hydrolysis solution was centrifuged to take the supernatant.

Embodiment 3

[0026] Example 3 Preparation of cholesterol-lowering crude peptide

[0027] The supernatant collected in step (2) was ultrafiltered by using 10kDa and 3kD ultrafiltration devices. Briefly, 12 mL of the proteolysis supernatant was transferred to a centrifugal ultrafiltration tube, and centrifuged at 5000 × g for 30 min to obtain an ultrafiltrate. The effect of different fractions (>10 kDa, 3-10 kDa and figure 1 ). The specific process of the cholesterol esterase activity inhibition experiment is as follows:

[0028] In a 96-well microtiter plate, 50 μL of sample, 50 μL of 25 μg / mL cholesterol esterase solution and 50 μL of 10 mM p-nitrophenyl butyrate as substrates were prepared in phosphate buffer (containing 100 mM NaCl, 5.16 mM) at pH 7.0 mM sodium taurocholate) for 5 min at 25°C. The microplate reader records the absorbance at 405 nm. Simvastatin was used as a positive control and calculated according to formula (1).

[0029]

[0030] In formula (1): A: the absorban...

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PUM

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Abstract

The invention provides a peptide with cholesterol esterase inhibitory activity as well as application and a screening method thereof. The sequence of the peptide is selected from SEQ ID NO.1-3. The peptide disclosed by the invention has a good cholesterol lowering effect and human intestinal absorbability, is high in stability, non-toxic, clear in action mechanism and definite in target spot, meets the requirements of pharmaceutical preparation development, is also easily added into various health foods as a functional factor, and has a good market prospect in the medicine and food industries.

Description

technical field [0001] The present application belongs to the field of proteins, and specifically, the present application provides three cholesterol-lowering peptides with cholesterol esterase inhibitory activity and their application and screening methods. Background technique [0002] The control of cholesterol levels has received much attention as plasma cholesterol levels are associated with various diseases including coronary artery disease, cancer, obesity and diabetes. Over the past few decades, cholesterol esterase has been extensively studied as a potential target for controlling cholesterol levels. Cholesterol esterase is an alpha / beta hydrolase primarily present in the lumen of the small intestine and secreted from the pancreas into the gut. Under normal circumstances, dietary cholesterol esters are difficult to absorb directly in the gut. Cholesterol esterase can hydrolyze cholesterol esters to produce cholesterol and free fatty acids, and then these unesterif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K7/08C12P21/06A61K38/08A61K38/10A61P3/06G01N33/68A23L33/18
CPCC07K7/06C07K7/08C12P21/06A61P3/06G01N33/6848A23L33/18A61K38/00A23V2002/00A23V2200/3262A23V2250/55
Inventor 沈群赵卿宇付永霞薛勇
Owner CHINA AGRI UNIV
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