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A kind of houttuynia glycoprotein and its preparation and application

A technology of Houttuynia cordata and glycoprotein, which is applied in the field of Houttuynia cordata glycoprotein, and can solve the problems that norovirus inhibitory effect has not been reported.

Active Publication Date: 2021-04-30
JIANGXI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the inhibitory effect of Houttuynia cordata extract on norovirus

Method used

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  • A kind of houttuynia glycoprotein and its preparation and application
  • A kind of houttuynia glycoprotein and its preparation and application
  • A kind of houttuynia glycoprotein and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Houttuynia glycoprotein

[0027] (1) With the fresh Houttuynia cordata purchased from the market, remove impurities and dry in an oven at 50°C until powder can be formed, pulverize, pass through an 80-mesh sieve, seal and place in a desiccator for subsequent use;

[0028] (2) Take 5 g of Houttuynia cordata powder in step (1) in a 2L beaker, add 275 mL of distilled water (Houttuynia cordata (g): water (ml) = 1:55), and then add 5 mg of cellulase (enzyme activity 25U / mg), stirred at 55°C for 2 hours to carry out the enzymolysis reaction, centrifuged the enzymolysis solution once at 8000×g for 10 minutes, obtained a supernatant and a precipitate, collected the supernatant once, added activated carbon to make the concentration 5g / L, and stirred at room temperature Discoloration for 30 minutes, 8000×g secondary centrifugation for 10 minutes to obtain the secondary supernatant and secondary precipitation; take the secondary supernatant and add trichloroacetic a...

Embodiment 2

[0031] Example 2: Influence of Houttuynia cordata glycoprotein on Norovirus activity inhibition and protein shell integrity

[0032] 1. Test material

[0033] Viruses: human norovirus GI (NoV GI), human norovirus GII (NoV GII) (provided by the Zhejiang Provincial Center for Disease Control and Prevention), mouse norovirus MNV-1 (purchased from the Microbial Culture Collection Center of the Chinese Academy of Sciences ).

[0034] Test drug: the pure Houttuynia glycoprotein product prepared in Example 1 was dissolved in water to prepare a 100 mg / mL Houttuynia glycoprotein mother solution.

[0035] Detection kits: QIAGEN Virus RNA Extraction Kit, Norovirus ELISA Kit (Bayfar R-Biopharm, Germany); One Step PrimescriptTM RT-PCR Kit was purchased from TaKaRa Company.

[0036] Main instruments: ABI7500 fluorescent quantitative PCR instrument, Hitachi scanning electron microscope TM4000, microplate reader.

[0037] 2. Test method

[0038] (1) Time-dependent test of antiviral effect...

Embodiment 3

[0065] Example 3: Houttuynia cordata glycoprotein inhibits the replication of murine norovirus in its host cell

[0066] 1. Test material

[0067] Viruses and cells: Murine Norovirus MNV-1 and RAW264.7 cells (purchased from the Culture Collection Center of Microorganisms, Chinese Academy of Sciences).

[0068] Test drug: the pure Houttuynia glycoprotein product prepared in Example 1 was dissolved in water to prepare a 100 mg / mL Houttuynia glycoprotein mother solution.

[0069] Detection kits: QIAGEN Virus RNA Extraction Kit, Norovirus ELISA Kit (Bayfar R-Biopharm, Germany); One Step PrimescriptTM RT-PCR Kit was purchased from TaKaRa Company.

[0070] Main instruments: ABI7500 fluorescent quantitative PCR instrument.

[0071] 2. Test method

[0072] A certain amount of RAW 264.7 cells were inoculated in a 12-well plate containing basal medium (DMEM+10%PBS+1%PS) (cell density 1×10 6 / well), add different final concentrations (1, 10, 25, 50mg / mL) houttuynia glycoprotein and 1×1...

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Abstract

The invention discloses a Houttuynia cordata glycoprotein and its preparation and application. The pure product yield of the Houttuynia cordata glycoprotein prepared by the invention is 1.245-1.827%, the total sugar concentration in the glycoprotein is 55.6-62.9%, and the uronic acid The mass concentration is 15.3-19.5%, and the protein mass concentration is 9.52-10.87%. The houttuynia cordata glycoprotein of the present invention is greater than or equal to 50 mg / mL, and the inhibition rate of human norovirus GI, GII and murine norovirus MNV‑1 for 120 minutes is greater than 85%, and can be used to prepare norovirus activity inhibitors .

Description

[0001] (1) Technical field [0002] The invention relates to a houttuynia glycoprotein, in particular to a houttuynia glycoprotein and its preparation and application in anti-norovirus. [0003] (2) Background technology [0004] Norovirus (NoV) belongs to the Norovirus genus of the family Caliciviruses, and was first discovered by Kapikian and other scholars using electron microscopy in 1972. Usually, according to the differences in the nucleotide sequence of the RNA polymerase coding region of the NoV virus or the amino acid sequence of the coat protein region, it can be divided into five different genomes (GGI, GGII, GGIII, GGIV and GGV). Among them, types GI, GII and GIV mainly infect humans, while type GV mainly infects mice. Norovirus is highly infectious and is one of the most common pathogenic microorganisms that cause acute gastroenteritis in humans. Among them, GII type is the dominant strain that has been reported in my country, and can usually be transmitted throug...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K1/14C07K1/18C07K1/30A61K38/16A61P31/14
CPCA61K38/00A61P31/14C07K14/415
Inventor 廖宁波章荣华张严峻龚黎明陈江
Owner JIANGXI AGRICULTURAL UNIVERSITY
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