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Bioactive peptide with dipeptidyl peptidase IV inhibitory activity

A bioactive peptide and dipeptidyl peptidase technology, applied in the field of bioactive peptide biology, can solve problems such as upper respiratory tract infection, gastrointestinal discomfort, allergic reaction, etc., achieve the effect of improving insulin resistance, simple preparation method, and stable activity

Pending Publication Date: 2022-07-29
XILINGOL VOCATIONAL COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long-term use of these chemically synthesized DPP-IV inhibitors may cause side effects such as allergic reactions, heart rate disorders, upper respiratory tract infections, and gastrointestinal discomfort

Method used

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  • Bioactive peptide with dipeptidyl peptidase IV inhibitory activity
  • Bioactive peptide with dipeptidyl peptidase IV inhibitory activity
  • Bioactive peptide with dipeptidyl peptidase IV inhibitory activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Acquisition of biologically active peptides with DPP-IV inhibitory activity

[0049] (1) Lactococcus lactis was inoculated into MRS liquid medium at an inoculum of 2% by volume, and cultured at 35°C for 12 hours; Yarrowia lipolytica was inoculated into YPD liquid culture at an inoculum of 2% by volume In the base, culture at 29°C and 200rpm for 12h; both bacteria proliferate continuously for 2 generations, and the number of viable bacteria of lactic acid bacteria and yeast after activation is not less than 10 respectively. 9 CFU / mL and 10 8 CFU / mL, spare;

[0050] (2) Fresh horse milk is purified by centrifugation and sterilized at 95°C for 10 minutes;

[0051] (3) When the horse milk was cooled to 30°C, the activated Yarrowia lipolytica and Lactococcus lactis were inoculated, and the number of viable bacteria in the horse milk after inoculation was 10 6 ~ 8 CFU / mL;

[0052] (4) Fermentation of horse milk, fermentation conditions are: 30 ℃, stirring spee...

Embodiment 2

[0059] Example 2: Enzyme Inhibition Kinetic Study of DPP-IV of Bioactive Peptide IAFP

[0060] (1) Use 0.1mol / L Tris-HCl buffer (pH 8.0) to prepare DPP-IV solution with enzyme activity of 80U / L, the solution concentration is 0.4mmol / L, 0.6mmol / L, 0.8mmol / L, 1.2 mmol / L, 1.6mmol / L, 2 mmol / L Gly-Pro-Pna substrate solution (GPP);

[0061] (2) Prepare a biologically active peptide IAFP solution with a concentration range of 0 to 2000 mmol / L;

[0062] (3) The DPP-IV inhibitory activity of the sample was determined by enzyme labeling method, and the DPP-IV inhibitory activity of different concentrations of bioactive peptide IAFP under different concentrations of GPP was measured to obtain the initial reaction speed;

[0063] (4) The Lineweaver-Burk double-reciprocal plot was used to determine the type of inhibitory effect of bioactive peptide IAFP on DPP-IV; the results are shown in Figure 9 , it can be seen from the figure that the biologically active peptide IAFP acts on DPP-IV ...

Embodiment 3

[0064] Example 3: Docking of small molecule peptide IAFP with dipeptidyl peptidase IV molecule

[0065] Download the corresponding DPP-IV (1N1M) model in the RCBS PDB protein database (https: / / www.rcsb.org / ). The small molecule peptide model was drawn using Chem Draw 19.0, and the NM2 force field was added to minimize its energy. Use Gold5.3.0 for flexible molecular docking, extract ligands for receptors, remove water molecules, add polar hydrogen, and use the Goldscore scoring system for docking model screening. Visual analysis of docked molecular complexes using Moe2019.10 software, see Figure 10 ,Depend on Figure 10 It can be seen that IAFP forms three hydrogen bonds with DPP-IV, and the binding site is close to the charge pocket of DPP-IVS'2.

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Abstract

The invention discloses a bioactive peptide which is derived from fermented horse milk, the amino acid sequence of the bioactive peptide is Ile-Ala-Phe-Pro, the molecular weight of the bioactive peptide is 500.6 Da, the bioactive peptide has dipeptidyl peptidase IV inhibitory activity, and the half inhibitory concentration (IC50) of the bioactive peptide to dipeptidyl peptidase IV is 0.001 mol / L; the traditional Chinese medicine composition can be used for preparing a high-glucose-induced Hepg2 cell model, can improve the glucose consumption capacity of the high-glucose-induced Hepg2 cell model, has the effect of inhibiting glucose uptake of the Hepg2 cell model, has an application prospect in the aspect of preparing health care products and medicines for treating or assisting in treating type 2 diabetes mellitus, and is simple to prepare and suitable for industrial production and market popularization and application.

Description

technical field [0001] The invention relates to a biologically active peptide with DPP-IV inhibitory activity in fermented horse milk, belonging to the field of biologically active peptide biotechnology. Background technique [0002] With the development of social economy and changes in people's lifestyles, the incidence of diabetes in the world is becoming more and more serious. The incidence of diabetes is getting younger and younger, and it will also cause many chronic complications. Diabetes mellitus is an endocrine and metabolic disease caused by the deficiency of insulin in the body or the reduction of its biological effect, or the coexistence of both, which leads to hyperglycemia and glucosuria, which leads to the disorder of fat and protein metabolism (WHO, 2006). Of all the types of diabetes, type 2 diabetes has the highest incidence, accounting for 90-95% of all diabetes. In its pathogenesis, DPP-IV can rapidly inactivate a variety of hormones such as incretin glu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K5/103C12P21/02C12R1/01C12R1/645
CPCC07K5/101C12P21/02C12R2001/01C12R2001/645
Inventor 郭元晟朱建军刘彦敏陈子衡伊日布斯郭梁雅梅
Owner XILINGOL VOCATIONAL COLLEGE
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