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Separation, identification and function analysis for tenebrio-molitor natural anti-oxidation octapeptide

A natural anti-oxidation and tenebrio molitor technology, applied in the field of biotechnology, can solve the problems of slow removal and excessive free radical generation, and achieve the effects of short cycle, purity, safety and environmental protection, and low production cost

Active Publication Date: 2016-11-09
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the body is in an unfavorable environment, and with age, the free radicals in the body will be produced too much or cleared too slowly

Method used

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  • Separation, identification and function analysis for tenebrio-molitor natural anti-oxidation octapeptide
  • Separation, identification and function analysis for tenebrio-molitor natural anti-oxidation octapeptide
  • Separation, identification and function analysis for tenebrio-molitor natural anti-oxidation octapeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The separation and identification of embodiment 1, Tenebrio molitor peptide:

[0042] (1) Sample processing, take 100g Tenebrio molitor larvae after ultrasonic treatment, repeatedly freeze-thaw, grind and mix through liquid nitrogen, obtain Tenebrio molitor homogenate sample;

[0043] (2) Protein extraction, add 50ml of protein extract to the Tenebrio molitor homogenate sample obtained above, place it at 4°C for overnight treatment, after sufficient extraction, centrifuge the obtained sample at 4°C, 12000g, for 30min Take the supernatant for later use, and the protein extract is 50mM HEPES pH7.0, 10mM b-ME, 1mM EDTA, 1.0mM Na3VO4 pH 7.0, 50mM NaF pH7.0, 1mM PMSF, 2mM benzamidine, 1mg / ml leupeptin, pepstain A, aprotinin;

[0044] (3) To remove macromolecular substances such as impurity proteins, place the supernatant obtained in step (2) in a water bath at 80°C for 1 hour, then centrifuge again, at 4°C, 12000g, for 60min, and take the middle supernatant (the upper layer...

Embodiment 2

[0052] Embodiment 2, the antioxidative experiment of Tenebrio molitor natural antioxidant octapeptide:

[0053] The present embodiment adopts improved FRAP method, the assay method of hydroxyl radical scavenging rate and the assay method of superoxide free radical (O2- ) scavenging rate to detect the antioxidant activity of Tenebrio molitor natural antioxidant octapeptide TM-1, And compared with the conventional antioxidant glutathione.

[0054] 1. Determination of reducing ability

[0055] The reducing ability was determined by the modified FRAP (Ferric Reducing Ability of Plasma) method. The detection principle is: antioxidants can reduce ferric ions to ferrous ions, and ferric ions can make phenanthroline substances form a stable complex, showing a clear blue color, and has a maximum at 593nm. Light absorption, according to the size of the absorbance to calculate the strength of the sample's antioxidant activity / reducing ability. This method does not reflect the scavengi...

Embodiment 3

[0067] Embodiment 3, Tenebrio molitor natural antioxidant octapeptide is used as a feed additive

[0068] 1. Product composition and use

[0069] 400 1-day-old white-feathered broilers were weighed separately, and the broilers were divided into 4 groups according to body weight, 100 in each group. Group 1 is the control group, group 2 is the 10mM / Kg Tenebrio molitor natural antioxidant octapeptide group, group 3 is the 50mM / Kg Tenebrio molitor natural antioxidant octapeptide group, and group 4 is the 250mM / Kg Tenebrio molitor natural antioxidant octapeptide group , drinking water use, the experimental period is 42d. To analyze the effect of Tenebrio molitor natural antioxidant octapeptide on chicken immunity and antioxidant capacity. Through this experiment, it was found that the natural antioxidant octapeptide can significantly increase the antibody level of mice, and has the function of regulating cellular immune response.

[0070] 2. Determination of immune organ index ...

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Abstract

The invention relates to a separation, identification and function analysis method for tenebrio-molitor peptide. The ultrasonic treatment technology, the technology screening technology, the ultrafiltration molecule flow intercepting technology, the molecular sieve technology, the reverse-phase high pressure liquid chromatography (HPLC) column chromatography technology, the mass spectrum technology, the amino acid analysis technology and the like are adopted, and octapeptide is separated from induced tenebrio molitor, wherein the amino acid sequence of the octapeptide is SEQ ID NO:1; the anti-oxidation activity of the octapeptide is further verified, it is verified that the anti-oxidation function of the octapeptide is higher than that of glutathione, and the basis is established for development and application of natural anti-oxidation octapeptide products.

Description

technical field [0001] The invention relates to a separation and identification method of Tenebrio molitor octapeptide and its application, belonging to the field of biotechnology. Background technique [0002] Tenebrio molitor Linne, also known as Tenebrio molitor Linne, is commonly known as mealworm, belongs to Insecta, Coleoptera, Pseudopedidae, Tenebrio genus. Tenebrio molitor was originally a warehouse pest, but in recent years, a large number of scientific and technological workers at home and abroad have conducted in-depth research on Tenebrio molitor in the fields of experimental material selection, feed production, food and health product research and development, and achieved a series of important breakthroughs, giving it Tenebrio molitor, which combines experimental insects and resource insects, is known as the "treasure house of protein feed". Tenebrio molitor is a kind of insect resource with various utilization values ​​such as feeding, food and medicine, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/14A23K20/147
CPCC07K7/06
Inventor 刘晓东王为栋张福波葛珍珍单虎王述柏秦志华
Owner QINGDAO AGRI UNIV
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