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A kind of dandelion seed antioxidant tetrapeptide and preparation method thereof

An antioxidant, dandelion technology, applied in the field of food biology, can solve the problem of little research on dandelion seeds

Inactive Publication Date: 2019-03-12
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many studies on dandelion whole herb at home and abroad, and mainly on dandelion flavonoids, polysaccharides and terpenoids, but little research on dandelion seed proteins and peptides

Method used

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  • A kind of dandelion seed antioxidant tetrapeptide and preparation method thereof
  • A kind of dandelion seed antioxidant tetrapeptide and preparation method thereof
  • A kind of dandelion seed antioxidant tetrapeptide and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0021] The preparation method of antioxidant tetrapeptide of the present invention is as follows:

[0022] Extraction of dandelion seed protein: dandelion seeds were pulverized by a high-speed traditional Chinese medicine grinder, extracted with 0.1mol / L NaOH solution, stirred at 40°C for 140min, centrifuged at 10000 rpm for 20min, the supernatant was taken, adjusted with 1mol / L hydrochloric acid The pH value is 4.0, centrifuged at 12000 rpm for 20 min, and the precipitate is freeze-dried to obtain dandelion seed protein powder.

[0023] Enzymolysis of dandelion seed protein: use alkaline protease to enzymolyze dandelion seed protein, the protein concentration is 2% (w / v), the enzyme-to-substrate ratio is 7.8%, the enzymatic hydrolysis time is 4.90h, the pH is 8.5, the temperature is 45°C, the enzyme After the solution was completed, the enzyme was inactivated in a boiling water bath for 10 minutes, then rapidly cooled to room temperature, centrifuged at 10,000 rpm for 20 minu...

Embodiment 2

[0029] The natural antioxidant tetrapeptide activity obtained in embodiment 1 is studied:

[0030] DPPH free radical scavenging activity: Take 1mL of samples with different concentrations and 1mL of DPPH solution (0.1mM, prepared in 95% ethanol) and mix them well, let stand in the dark at room temperature for 30min, and measure the absorbance at 517nm; use 1mL of 95% ethanol solution instead of DPPH solution for Sample reference group; blank group is 1mL DPPH solution and 1mL 95% ethanol solution. The DPPH free radical scavenging activity of the sample was calculated according to the following formula (1):

[0031] (1)

[0032] In the formula: A i —Absorbance value of sample group; A j — Absorbance value of sample reference group; A 0 — Absorbance value of the blank group

[0033] ABTS free radical scavenging activity: prepare 7mM ABTS storage stock solution and 2.45mM potassium persulfate solution, mix them at a ratio of 1:1 before use, place at room temperature for 1...

Embodiment 3

[0044] The antioxidant tetrapeptide of the present invention has inhibitory effect on linoleic acid peroxidation system:

[0045] Take 1mL of samples of different concentrations in a stoppered colorimetric tube, add 2mL of 95% ethanol, 26μL of linoleic acid and 2mL of phosphate buffer (50mM, pH 7.0), mix well, seal it in a dark place and keep it for 40 ℃ constant temperature. In the blank group, 1 mL of distilled water was used instead of the sample. The peroxidation degree of the system was measured every 24 hours. The level of lipid peroxidation was determined by ferric thiocyanate (FTC) method. Take 100 μL of the mixture, mix with 4.7mL 75% ethanol solution, 0.1mL 30% ammonium thiocyanate, add 0.1mL FeCl 2 Solution (20mM, 3.5% hydrochloric acid solution preparation), after fully mixing, accurately time 3min, and measure the absorbance value at a wavelength of 500nm.

[0046] From the measurement results, it can be seen that ( Figure 8 ), the antioxidant tetrapeptide s...

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Abstract

The invention discloses dandelion seed antioxidant tetrapeptide and a preparation method thereof. The sequence of dandelion seed antioxidant tetrapeptide is Trp-Thr-Leu-Asn (WTLN). In-vitro experiments show that dandelion seed antioxidant tetrapeptide is capable of effectively eliminating DPPH, ABTS, hydroxyl free radicals, superoxide anion free radicals and the like and simultaneously inhibiting linoleic acid peroxidation. Dandelion seed antioxidant tetrapeptide has the characteristics of simple structure, strong antioxidant activity and the like, can be used as an excellent substitute of existing artificially synthesized antioxidants and is of an important value in the aspects of development and application of novel food additives.

Description

technical field [0001] The invention relates to a dandelion seed antioxidant tetrapeptide, a preparation method thereof and an application in the food field, belonging to the field of food biotechnology. Background technique [0002] Active oxygen radicals are derivatives of oxygen, which are extremely unstable due to unpaired orbital electrons, including hydroxyl radicals, superoxide radicals, hydrogen peroxide and singlet oxygen. Some reactive oxygen species and other free radical products are produced during human metabolism, and the excessive accumulation of these substances will cause oxidative damage to biological macromolecules such as DNA, protein, lipid and carbohydrate. Many evidences show that active oxygen free radicals play a decisive role in causing many serious diseases such as cancer, cardiovascular disease, diabetes, rheumatoid arthritis, and senile dementia. Reactive oxygen radicals are thought to be involved in the process of lipid oxidation, which produc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/117C07K1/36C07K1/20C07K1/16C12P21/06A23L33/18
CPCC07K5/1024
Inventor 洪晶张杨胡磊孟春
Owner FUZHOU UNIV
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