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Horse milk-derived small molecule peptide and application thereof

A small molecule peptide, dipeptidyl peptidase technology, applied in applications, medical preparations containing active ingredients, peptides, etc., can solve problems such as upper respiratory tract infection, gastrointestinal discomfort, heart rate disorder, etc., to improve glucose uptake, Improve cell viability, simple preparation effect

Active Publication Date: 2022-03-04
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used DPP-IV inhibitors are mostly chemically synthesized, and long-term use is likely to cause side effects such as allergic reactions, heart rate disorders, upper respiratory tract infections, and gastrointestinal discomfort.

Method used

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  • Horse milk-derived small molecule peptide and application thereof
  • Horse milk-derived small molecule peptide and application thereof
  • Horse milk-derived small molecule peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Acquisition of Small Molecular Peptides

[0033] 1. Fermentation of horse milk: take 400 mL of fresh horse milk and sterilize it at 60 °C for 30 minutes, wash and resuspend Lactococcus lactis ( Lactococcus lactis ) and Yarrowia lipolytica ( Yarrowia lipolytica ) Y7, each inoculated 2 mL into sterilized horse milk, and then fermented at 32 ℃, 225 rpm for 68 h; among them, Yarrowia lipolytica ( Yarrowia lipolytica ) Y7 was disclosed in "Tang Rong et al. Research on ACE inhibitory peptides produced by fermenting horse milk with yeast and lactic acid bacteria, Food Science, 2021", Lactococcus lactis ( Lactococcus lactis ) purchased from Chr. Hansen;

[0034] 2. Fermentation broth pretreatment: the fermented horse milk was adjusted to pH 4.3 with 0.1 mol / L HCL, centrifuged at 10000 g, 4°C for 10 min, and the supernatant was taken; the pH was adjusted to 7.0 with 0.1 mol / L NaOH, Centrifuge at 10,000 g, 4 °C for 10 min to take the supernatant and freeze-dry...

experiment example 2

[0040] Experimental example 2: Enzyme inhibition kinetics experiment of small molecule peptide FPGGP on dipeptidyl peptidase Ⅳ (DPP-Ⅳ)

[0041] Prepare the following solutions with 0.1mol / L Tris-HCL buffer (pH8.0) as solvent: DPP-Ⅳ solution with enzyme activity 80U / L; Pro-pNA substrate solution; small molecular peptide FPGGP solution with a concentration range of 0-2000 μm, using the above-mentioned dipeptidyl peptidase IV inhibitory activity detection test method, to obtain the absorbance value changes of different concentrations of small molecular peptides on different concentrations of substrates ( That is, the initial reaction velocity), and the Lineweaver-Burk double reciprocal plot was used to determine the type of inhibitory effect of the small molecular peptide FPGGP on DPP-Ⅳ; the results are shown in Figure 9 , it can be seen from the figure that the small molecular peptide FPGGP acts on dipeptidyl peptidase IV through anti-competitive inhibition.

experiment example 3

[0042] Experimental Example 3: Molecular docking of small molecule peptide FPGGP with dipeptidyl peptidase IV

[0043] Download the corresponding DPP-IV (1N1M) model in the RCBS PDB protein database (https: / / www.rcsb.org / ). Use Chem Draw 19.0 to draw the small molecule peptide model, add NM2 force field to minimize its energy. Use Gold5.3.0 for flexible molecular docking, ligand extraction for receptors, removal of water molecules, addition of polar hydrogen, and use of the Goldscore scoring system for docking model screening. Using Moe2019.10 software for visual analysis of docking molecular complexes, see the results Figure 10 ,Depend on Figure 10 It can be seen that FPGGP forms two hydrogen bonds with dipeptidyl peptidase IV, and the binding site is close to the charge pocket of dipeptidyl peptidase IVS'2.

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Abstract

The invention discloses a small molecule peptide, the amino acid sequence of the small molecule peptide is Phe-Pro-Gly-Gly-Pro, and the molecular weight of the small molecule peptide is 545.6 Da. The small molecule peptide has dipeptidyl peptidase IV inhibitory activity, and the half inhibitory concentration (IC50) of the small molecule peptide to dipeptidyl peptidase IV is 0.664 mg / mL; in addition, the insulin resistance of high-glucose-induced (human liver cancer) Hepg2 cells can be improved, the activity of the Hepg2 cells is remarkably improved, and the glucose consumption and glucose uptake of the Hepg2 cells are improved; the small molecule peptide disclosed by the invention can be applied to preparation of special medical foods, health care products and medicines for treating or assisting in treating the type 2 diabetes mellitus, is simple to prepare and is suitable for industrial production and market popularization and application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a small molecule peptide derived from fermented horse milk and its application. Background technique [0002] Diabetes has become a global public health problem. The International Diabetes Federation (IDF) predicts that by 2045, the total number of people suffering from diabetes worldwide will exceed 700 million. In recent years, with the change of population and lifestyle in our country, the incidence of diabetes is also increasing year by year, and the age is getting younger day by day. In addition, diabetic patients are in a state of high blood sugar for a long time, which can easily induce various complications such as peripheral neuropathy, vascular disease, abnormal visual system, endocrine and metabolic disorders, etc., seriously affecting the health and quality of life of patients. [0003] Diabetes mellitus is a metabolic disease that is caused by the deficiency of insulin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/07A61P3/10A23L33/18
CPCC07K7/06A61P3/10A23L33/18A61K38/00A23V2002/00A23V2200/328A23V2250/55Y02P20/55
Inventor 伊日布斯陈子衡陈亚辉成超王嘉鑫严金平
Owner KUNMING UNIV OF SCI & TECH
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