Preparation method, separation method and applications of forest frog oil antioxidant peptide component

A technology for antioxidant peptides and Rana oleifera oil, which is applied in the fields of peptide preparation methods, chemical instruments and methods, medical preparations of inactive ingredients, etc., can solve the problems of low content, difficult extraction, peptide library construction, screening and application. In the initial stage and other problems, to achieve the effect of benefiting the body's absorption, improving the bacteriostatic effect, and benefiting the stability

Active Publication Date: 2019-07-23
CHANGSHU INSTITUTE OF TECHNOLOGY
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Active peptides from living organisms are unlikely to meet large-scale demand due to low content and difficulty in extraction
Although gene combination and chemical combination peptide libraries have brought revolutionary progress to peptide drugs and the pharmaceutical industry, the construction, screening and application of peptide libraries are still in their infancy

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation method, separation method and applications of forest frog oil antioxidant peptide component
  • Preparation method, separation method and applications of forest frog oil antioxidant peptide component
  • Preparation method, separation method and applications of forest frog oil antioxidant peptide component

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of Rana Oil Antioxidant Peptide Component

[0031] Weigh a certain amount of Rana oil defatted powder, put it in 30mL distilled water and let it stand overnight. Adjust the pH to 1.0-1.5 with 1 mol / L hydrochloric acid aqueous solution, and then place it in a 37°C water bath for 5 min. Add a certain amount of pepsin, stir to make it evenly dispersed, and place it in a constant temperature shaker at 37°C for a certain period of time. After pepsin enzymatic hydrolysis, adjust the pH to 8.0-8.5 with 0.1mol / mL NaOH aqueous solution, put it in a 37°C water bath for 5 minutes, add a certain amount of trypsin, stir to make it evenly dispersed, and put it in a constant temperature oscillator. time. After trypsin enzymatic hydrolysis, place the enzymatic solution in a water bath at 95°C to 100°C to inactivate the enzyme for 10min to 15min, after cooling, centrifuge at 13000r / min for 30min, and freeze-dry the supernatant to obtain the antioxidant peptide co...

Embodiment 2

[0072] Example 2 Separation of Rana Oil Antioxidant Peptide Components

[0073] Using the PALL Minimate TM tangential flow ultrafiltration system, the ultrafiltration membranes with molecular weight cut-offs of 10kDa, 5kDa, 3kDa and 1kDa were used to fractionate the solution of antioxidant peptide components of Rana oil, and the components of different molecular weights were collected separately. (>10kDa), II (5kDa~10kDa), III(3kDa~5kDa), IV(1kDa~3kDa), V(<1kDa), after lyophilization, prepare solutions with a concentration of 5mg / mL. The salicylic acid method was used to compare the scavenging ability of hydroxyl radicals, determine the most active component and compare it with Rana oleoprotein.

[0074] As shown in Table 4, at the same concentration, component IV, that is, the peptide component with a molecular weight distribution of 1kDa to 3kDa, has the highest scavenging rate for hydroxyl radicals, which is 93.21%, while the Rana oleoprotein is only 10.12%, and the activit...

Embodiment 3

[0077] Example 3 In vivo antioxidant effect of Rana oil antioxidant peptide component IV

[0078] 60 male Kunming mice were selected as experimental animals, and were randomly divided into 6 groups, 10 in each group, that is, low-dose group (100mg / kg), middle-dose group (200mg / kg), high-dose group (100mg / kg) of Rana oil antioxidant peptide component IV group (400mg / kg), blank control group (distilled water), positive control group (Rana oleoprotein 200mg / kg), model control group (distilled water). Continuous feeding for 30 days, gavage once a day. On the last day, except for the blank control group, the other test groups were intragastrically given 50℅ethanol (12mL / kg) to establish the oxidative damage model. Six hours after modeling, 1 mL of blood was collected and centrifuged to determine the levels of malondialdehyde (MDA) and total superoxide dismutase (T-SOD). The experimental animals were sacrificed by neck dislocation, the liver was taken out, washed, homogenized and ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention provides a preparation method of an antioxidant peptide component and applications of the antioxidant peptide component in oral liquid preparations. The process conditions for preparinga forest frog oil antioxidant peptide component by pepsin/trypsin two-step hydrolysis mode are as follows: pepsin content is 2.0%, pepsin enzymolysis time is 3.5 h, trypsin content is 2.0%, and trypsin enzymolysis time is 3.0 h. Under the conditions, the forest frog oil antioxidant peptide component can remove 75.57% of hydroxyl free radicals, and has a yield of 63.45%. An ultrafiltration system intercepts the peptide component with the molecular weight of 1-3kDa so as to obtain the forest frog oil antioxidant peptide component IV with the highest activity. The peptide component has remarkableantioxidant effect in ethanol-induced oxidative damage model mouse body, can reduce the content of malondialdehyde and protein carbonyl, can increase the activity of total superoxide dismutase and the content of glutathione, and can be used for preparing a forest frog oil antioxidant peptide component oral liquid.

Description

technical field [0001] The invention belongs to the field of preparation and application of natural products, and in particular relates to a preparation method, separation method and application of an antioxidant peptide component of Rana oil. Background technique [0002] For the body, free radicals are a "double-edged sword". At physiological concentrations, free radicals are widely involved in cell signal transduction and life processes; in excess, they can cause mitochondrial oxidative stress, leading to aging and related diseases. Therefore, maintaining the balance of oxidation and antioxidants (redox balance) in cells is extremely important for maintaining health. The free radicals in the body are mainly reactive oxygen species (ROS), and some reactive nitrogen species (Reactive nitrogen species, RNS). ROS mainly include hydrogen peroxide, hydroxyl radicals, superoxide anions, hydroperoxides, etc.; RNS includes nitric oxide, pernitrate, nitrogen dioxide, etc. The bod...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34A61K38/01A61K9/08A61K47/46A61P39/06
CPCA61K9/0095A61K38/012A61K47/46A61P39/06C07K1/34C12P21/06
Inventor 张扬王逸飞李明珠刘舒悦于佳璐
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap