Tyrosinase inhibitory activity of bunge auriculate root polysaccharide and application of bunge auriculate root polysaccharide

A technology of tyrosinase diphenolase and Baishouwu, which is applied in the direction of organic active ingredients, skin care preparations, medical preparations containing active ingredients, etc., to achieve low production costs, obvious effects, and the effect of preventing browning

Active Publication Date: 2013-09-25
YANTAI BAOFULING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report that the polysaccharide extracted from Baishouwu me

Method used

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  • Tyrosinase inhibitory activity of bunge auriculate root polysaccharide and application of bunge auriculate root polysaccharide
  • Tyrosinase inhibitory activity of bunge auriculate root polysaccharide and application of bunge auriculate root polysaccharide
  • Tyrosinase inhibitory activity of bunge auriculate root polysaccharide and application of bunge auriculate root polysaccharide

Examples

Experimental program
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Effect test

Embodiment 1

[0019] The preparation of the total polysaccharides of Baishouwu: The roots of Baishouwu are crushed with a pulverizer. After 20g of Baishouwu powder is extracted with multiple times of ethanol, the residue is air-dried, and the material-to-liquid ratio is 1:20 for boiling water extraction. The extract is α- Amylase and glucoamylase are used to remove starch, papain and Sevag reagent (chloroform: n-butanol is 5:1) are used to remove protein, the supernatant is concentrated to a certain volume, and ethanol is added for precipitation. Washing with ethanol, acetone, ether, and vacuum drying to obtain 1.39 g of Polysaccharides from Radix Polygoni Multiflori.

Embodiment 2

[0021] The preparation of the total polysaccharides of Baishouwu: The roots of Baishouwu are crushed with a pulverizer and passed through a 20-mesh sieve. The powder of Baishouwu is extracted with ethanol for several times, and the residue is air-dried until there is no alcohol. The ratio is 1:20 for boiling water. Extraction, water bath 2h, 4000r / min centrifugation for 5min, take the supernatant and suction filtration. Add amylase to the suction filtered supernatant, adjust to PH=6, 85℃ water bath for 1.5h, use I 2 / KI detected to I 2 Does not change color. Add glucoamylase (45°C in advance), adjust the pH to 6, and place in a water bath at 60°C for 1 hour. Suction filtration to remove the residue, collect the extract, and concentrate to a small volume. Add the mixture of chloroform and methanol (5:1) in a ratio of 1:3 to the above small volume concentrate, centrifuge at 3500r / min for 3min, the precipitate between the mixture and the supernatant is the protein, collect the sup...

Embodiment 3

[0023] Experimental method of inhibiting tyrosinase activity by total polysaccharides of Baishouwu:

[0024] Reagents: mushroom tyrosinase (EC.1.14.18.1, specific activity of 5370U / mg) and levodopa (1-dopa) are all products of Sigma, and hydroquinone (AR grade) is a product of Beijing Zizhu Pharmaceutical Co., Ltd. , Other reagents (AR grade) are all products of Sinopharm Shanghai Chemical Reagent Company.

[0025] Instrument: AL104 electronic analytical balance, manufactured by METTLER TOLEDO (Shanghai) Instrument Co., Ltd.; Muitiskan MK3 multifunctional microplate reader, product of Shanghai Regal Analytical Instrument Co., Ltd.

[0026] The tyrosinase was dissolved in phosphate buffer to prepare a 500U / mL solution, aliquoted and frozen, and diluted with phosphate buffer to 25U / mL before the experiment. Before the experiment, weigh an appropriate amount of levodopa, dissolve it with phosphate buffer to prepare a 15mmol / L solution, shake it, and fully dissolve it for later use. Ta...

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Abstract

The invention relates to tyrosinase inhibitory activity of total polysaccharide which is extracted and separated from traditional Chinese medicine bunge auriculate root and clarifies a new application of the total polysaccharide in the fields of medicines, food and daily chemicals. Influence of the total polysaccharide on tyrosinase diphenol enzyme activity is detected by virtue of an enzymatic reaction, experimental results prove that the total polysaccharide of bunge auriculate root has strong inhibiting effect on mushroom tyrosinase diphenol enzyme activity, IC50 is 31.8mug/ml, and an inhibition mechanism of the total polysaccharide belongs to reversible competitive inhibition. The total polysaccharide of bunge auriculate root can be used for preparing a medicine used for treating hyperpigmentation diseases, can be used for preparing health care food and cosmetics which are used for preventing skin from being sunburnt, whitening and removing freckles and can be also used for preparing a fruit and vegetable colour retention agent which can prevent brown stain of food. The application of the total polysaccharide of bunge auriculate root based on tyrosinase inhibition activity has the characteristics of nature, high efficiency, safety and no toxicity.

Description

Technical field [0001] The invention relates to the tyrosinase inhibitory activity of Polysaccharides of Radix Polysaccharides and its application. The invention relates to the total polysaccharide extracted and separated from the traditional Chinese medicine Baishouwu having the activity of inhibiting tyrosinase, and clarifies its preparation method and its new application in the fields of medicine, food and daily chemical industry. Background technique [0002] Tyrosinase is a copper-containing polyphenol oxidase. It is the rate-limiting enzyme in the biosynthesis of melanin. It catalyzes the hydroxylation of tyrosine to dopa, the oxidation of dopa to dopaquinone, and 5, 6 -Dihydroxyindole (DHI) is oxidized to 5,6-indolequinone in three steps. Its expression and activity determine the amount and speed of melanin production. Excessive reaction can cause hypermelanism, such as freckles, chloasma, etc. Excessive melanin production is the root cause of the formation of chloasma; ...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A23L1/29A23L1/272A61K8/73A61Q19/02A61Q19/00A61Q17/04A61P17/00A23L5/41A23L33/00
Inventor 王晓岚姚文杰苑亮
Owner YANTAI BAOFULING BIOTECH CO LTD
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