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Wheat germ protein polypeptide with alpha-glucosidase inhibitory activity and preparation thereof

A glucosidase and activity-inhibiting technology, which is applied in the field of wheat germ protein polypeptide and its preparation and application, can solve the problems of high price, aggravation, and scarce source, and achieve the effect of low price, wide range of sources, and small molecular weight

Pending Publication Date: 2021-12-17
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these drugs have some obvious side effects, such as abdominal pain, bloating, increased flatulence, diarrhea, cramps, etc.
And these drugs are in great demand, but the sources are scarce and expensive

Method used

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  • Wheat germ protein polypeptide with alpha-glucosidase inhibitory activity and preparation thereof
  • Wheat germ protein polypeptide with alpha-glucosidase inhibitory activity and preparation thereof
  • Wheat germ protein polypeptide with alpha-glucosidase inhibitory activity and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1. Preparation of α-glucosidase activity inhibitory polypeptide

[0057] (1) Extraction of wheat germ protein: Dissolve 10g of defatted wheat germ powder in 100mL of distilled water, adjust the pH to 11.0 with 1M NaOH, stir and extract at 50°C for 2h, centrifuge at 4000rpm for 15min, take the supernatant, adjust the pH to 4.5 with 1M HCl, and centrifuge at 4000rpm After 15 minutes, the precipitation was wheat germ protein, which was freeze-dried for later use.

[0058] (2) Directed enzymatic hydrolysis to produce peptides: Bioinformatics combined with bioenzymatic hydrolysis technology is used to directional enzymatically hydrolyze wheat germ protein. First, bioinformatics methods were used to analyze the protein structure, and the wheat germ protein amino acid / gene sequence was downloaded from the UniProtKB protein data / NCBI gene bank, and the amino acid sequence was analyzed using the "Profiles of potential biological activity" module in the Biopep database...

Embodiment 2

[0062] Example 2. Detection of α-glucosidase inhibitory activity of biologically active polypeptides

[0063] Detection method: Add 50 μL 0.1mol / L PBS (0.01M, pH6.8), 50 μL 0.5U / mL α-glucosidase and 50 μL peptide solution in sequence to the wells of a 96-well microtiter plate (use PBS to prepare different concentrations of peptide solutions ), mixed at 37°C for 15 minutes, then added 100 μL of 1.5mmol / L 4-nitrophenyl-α-D-glucopyranoside (PNPG), mixed evenly and incubated at 37°C for 15 minutes before detection The absorbance value at a wavelength of 405nm was repeated three times, and the average value was taken.

[0064] The formula of enzyme activity inhibition rate: I%=[1-(A1-A2) / (A3-A4)] * 100%; In the formula: A1 is the absorbance value of sample solution group, and A2 is to replace enzyme solution with PBS and record A3 is the absorbance value of the blank control group measured by replacing the sample solution with PBS solution, and A4 is the background absorbance valu...

Embodiment 3

[0066] Example 3. Intestinal sac reversal experiment in rats

[0067] Male SD rats were fasted overnight before the experiment, and the jejunum part was taken, rinsed with normal saline to remove the contents, cut into 3 cm long segments, and the jejunum segment was turned over with a gavage needle, with brush-like ciliates facing outwards, and both ends were knotted and sealed, and placed Add 5mL containing 5M sucrose, in 37 ℃ Ringer's solution, add or not add the bioactive polypeptide prepared in the above-mentioned embodiment 1, incubate 90min, detect the glucose content in the solution every 15min, calculate the glucose production rate, evaluate the decline of biologically active polypeptide. blood sugar effect. image 3 The results showed that compared with no bioactive peptide, 1mg / mL wheat germ peptide (<1KDa) could significantly reduce the rate of glucose production and exert an excellent hypoglycemic effect.

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Abstract

The invention provides a wheat germ protein polypeptide with alpha-glucosidase inhibitory activity and preparation and application thereof, and belongs to the field of biological medicine. The bioactive polypeptide provided by the invention has alpha-glucosidase inhibitory activity, has the effect of reducing blood sugar, has small molecular weight and high stability, and has wide application prospects in the fields of medical health, food health care and the like. The bioactive polypeptide can be used for preparing a drug for treating and / or preventing diabetes, or can be used as a functional food additive for long-term treatment and health care of diabetic patients. The bioactive polypeptide provided by the invention is derived from natural vegetable protein, has the characteristics of stability, safety, easy absorption by human body and the like, and can be used for large-scale industrial production.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a wheat germ protein polypeptide with alpha-glucosidase inhibitory activity and its preparation and application. Background technique [0002] Diabetes mellitus is a group of chronic, multifactorial metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by defective insulin secretion or impaired biological action, or both. Long-term high blood sugar leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves, such as foot ulcers, diabetic retinopathy, kidney disease, stroke, cardiovascular disease, neuropathy, etc. . Diabetes can be divided into type 1 or type 2 diabetes, and both type 1 and type 2 diabetes have obvious genetic heterogeneity. Diabetes tends to occur in families, and 1 / 4-1 / 2 patients have a family history of diabetes, and there are at least 60 genetic syndromes clinically a...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K1/36C07K1/34C07K1/30C12P21/06A61K38/08A61P3/10A23L33/18
CPCC07K7/06C12P21/06A61P3/10A23L33/18A61K38/00A23V2002/00A23V2200/328A23V2250/55
Inventor 李洪岩王静刘维维孙宝国温洋洋范浩然路士熠
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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