A kind of bioactive polypeptide fspankkltpkky and its preparation method and application

A technology of biologically active peptides and biologically active peptides, applied in the field of protein, to achieve the effect of promoting the increase of induction amount, improving the ability and improving the quality

Active Publication Date: 2022-03-29
ZHEJIANG PANDA DAIRY GRP COMPANY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research on the related functions of the protein disulfide-isomerase A3 protein fragment in the prior art

Method used

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  • A kind of bioactive polypeptide fspankkltpkky and its preparation method and application
  • A kind of bioactive polypeptide fspankkltpkky and its preparation method and application
  • A kind of bioactive polypeptide fspankkltpkky and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 The artificial synthesis of active peptide FSPANKKLTPKKY

[0033] 1. Synthesis of Bioactive Peptides

[0034] Synthetic bioactive peptide FSPANKKLTPKKY.

[0035] 2. Confirmation of Bioactive Peptides

[0036] 1) UPLC analysis

[0037] The UPLC conditions are as follows:

[0038] Instruments: Waters ACQUITY UPLC ultra-high performance liquid phase, electrospray, quadrupole, time-of-flight mass spectrometer

[0039] Column specification: BEH C18 column

[0040] Flow rate: 0.4mL / min

[0041] Temperature: 50°C

[0042] UV detection wavelength: 210nm

[0043] Injection volume: 2μL

[0044] Gradient conditions: liquid A: water containing 0.1% formic acid (v / v), liquid B: acetonitrile containing 0.1% formic acid (v / v)

[0045] Time(min) %A %B

0 95.0 5.0

1.50 80.0 20.0

3.50 60.0 40.0

5.00 40.0 60.0

7.00 15.0 85.0

8.00 0.0 100.0

11.00 0.0 100.0

11.50 95.0 5.0

13.00 95.0 5.0

[0046] 2) Mass spectrometry analysis

[0047] The mass spectro...

Embodiment 2

[0061] Example 2 Immune Activity Experiment of Bioactive Peptides

[0062] 1. Determination of the amount of bioactive polypeptide FSPANKKLTPKKY that promotes the induction of nitric oxide in macrophages (Griess method)

[0063] 1. Experimental reagents and instruments:

[0064] Reagents: experimental animals balb / c mice (male 6-8 weeks old) spleen lymphocyte-derived biologically active peptide FSPANKKLTPKKY; LPS, purchased from Sigma; neutral red staining solution, produced by Beyond Biotechnology Research Institute.

[0065] Equipment: LRH-250F biochemical incubator Shanghai Heng Technology Co., Ltd.; GL-22M high-speed refrigerated centrifuge Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.; Hera cell 150 CO 2 Incubator Heraeus; DragonWellscan MK3 microplate reader Labsystems.

[0066] 2. Test method:

[0067] The number of cells added is 2×10 6 / ml cell suspension 100μl / well, after adherent purification, add 200μl / well of RPMI1640 complete culture solution (10% FBS) ...

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Abstract

The present invention relates to the field of proteins, in particular to a biologically active polypeptide FSPANKKLTPKKY and its preparation method and application. The amino acid sequence of the biologically active polypeptide FSPANKKLTPKKY is Phe-Ser-Pro-Ala-Asn-Lys-Lys-Leu-Thr-Pro-Lys ‑Lys‑Tyr. Through the in vitro immune function regulation experiment, it is verified that the polypeptide FSPANKKLTPKKY has a good immune regulation function. The biologically active peptide FSPANKKLTPKKY of the present invention can significantly enhance the in vitro proliferation ability of lymphocytes, and has the ability to increase the induction of macrophage nitric oxide under certain concentration conditions, improve the body's ability to resist external pathogen infection, and reduce the body's morbidity , improving the quality of life is of great significance to the development of food, health products and medicines with immune regulation function.

Description

technical field [0001] The invention relates to the field of proteins, in particular to a biologically active polypeptide FSPANKKLTPKKY and its preparation method and application. Background technique [0002] In recent years, bioactive peptides have become a familiar term. Because of its many potential biological functions, it has attracted more and more attention and has become one of the hot spots of scientific research. At present, the beneficial effects of many bioactive peptides have also been well proven, such as anti-cancer, lowering blood pressure, antibacterial, lowering cholesterol, anti-diabetic and other properties. More than 3000 different bioactive peptides have been reported in the current most authoritative bioactive peptide database BIOPEP-UMW. [0003] At present, most studies on bioactive peptides focus on food-derived peptides, while there are few studies and reports on non-food-derived peptides. And it has been confirmed by research results that, com...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/90C12N15/61A61K38/52A61K8/66A61Q19/00A61P29/00A61P37/02A23L33/10
CPCC12N9/90A61K8/66A61Q19/00A61P29/00A61P37/02A23L33/10C12Y503/04001A61K38/00A23V2002/00A23V2200/324
Inventor 张少辉陈承余肖珊珊石毅洪志骏王娟李锡安占文静梁世排梁世赞
Owner ZHEJIANG PANDA DAIRY GRP COMPANY
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