Novel tyrosinase inhibitor

A technology of tyrosinase and inhibitors, which is applied in the field of tyrosinase inhibitors, can solve the problems that the type and mechanism of inhibition have not been studied, and there is no report on the condensed tannin of golden osmanthus tea leaves, etc., and the price is low. , the effect of easy operation

Inactive Publication Date: 2017-02-22
HENAN UNIV OF URBAN CONSTR
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Some researchers also extracted potato tyrosinase inhibitors from four kinds of tea, Laoshan green tea, Fujian scented tea, Fujian Tieguanyin and Yunnan Pu'er, and studied the inhibitory effect, but did not study the type and mechanism of inhibit...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1. the acquisition of the condensed tannin extract of golden osmanthus tea leaves

[0025] Get the freeze-dried leaf powder of golden osmanthus (fresh leaves are collected in Anxi, Fujian, and the freeze-drying is carried out by vacuum freeze-drying machine (christ, ALPHA1-2LD Plus)) 10g, leaching with 200mL of 70% acetone at room temperature for 24h, centrifuged at 3500g for 15min , collect the supernatant. The above steps were repeated three times. The acetone extracts were combined, and the acetone was distilled off under reduced pressure at 38°C with a rotary evaporator (BUCHI, R-205V). The remaining aqueous phase was separated with n-hexane and dichloromethane successively to remove pigments, lipids and non-polar substances. The resulting aqueous solution was lyophilized and dissolved in 50% methanol, passed through a Sephadex LH-20 column (30×2.5 cm), and eluted successively with 50% methanol and 70% acetone. The acetone was distilled off under reduc...

Embodiment 2

[0026] Embodiment 2. Identification of the structural characteristics of the condensed tannin extract of the golden osmanthus tea leaves obtained in Example 1

[0027] By MALDI-TOF-MS (method reference Wei SD, Lin YM, Liao MM, Zhou HC, LiYY.Characterization and antioxidant properties of condensed tannins from the mangrove plant Aegiceras corniculatum.J Appl Polym Sci 2012,124:2463–2472.) , the gold osmanthus tea leaf condensed tannin extract that embodiment 1 obtains shows a series of mass spectrum peaks ( figure 1 ), HPLC-ESI-MS (method reference Wei SD, Lin YM, Liao MM, Zhou HC, LiYY. ) shows 11 corresponding peaks ( figure 2). It is finally determined that the condensed tannin extract obtained in Example 1 is a complex mixture consisting of homopolymers and heteropolymers formed by proanthocyanidins, protogeranidine and protodelphinidine, wherein proanthocyanidins are the main components. Proanthocyanidins are analogs of tyrosinase catalytic substrates, which can compet...

Embodiment 3

[0028] The condensed tannin extract of golden osmanthus tea leaves obtained in embodiment 1 inhibits tyrosinase activity of tyrosinase

[0029] The bioassay system is as follows: 0.3mL of 1mg / mL L-dopa (L-DOPA), 0.75mL of 0.2mM, phosphate buffer solution of pH=6.8, 1.8mL of distilled water, 0.1mL of different concentrations (respectively 0, 5, 15, 25, 35, 50, 65 μg / mL) of the condensed tannin extract obtained in Example 1, 0.05 mL of tyrosinase aqueous solution (sigma, mushroom tyrosinase). The constant temperature condition is 30° C., and the measurement wavelength is 475 nm.

[0030] like image 3 As shown, the half-inhibition rate of tyrosinase by the golden osmanthus condensed tannin extract obtained in Example 1 is 54.8±1.27 μg / mL.

[0031] Taking the concentration of L-DOPA as quantification and the amount of tyrosinase added as variable, the condensed tannin extracts (0, 6.67, 33.33, and 66.67 μg / mL) obtained in Example 1 of different concentrations were measured for ...

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Abstract

The application provides a novel tyrosinase inhibitor. Golden cassia tea is used as a raw material for extracting the tyrosinase inhibitor, and a condensed tannin extract is obtained by extracting, separating and purifying by the steps of acetone extraction, centrifugation, extraction, Sephadex LH-20 column chromatography, and the like. The extract is prepared from procyanidine, a homopolymer formed by propelargonidin and prodelphinidins, and a heteropolymer, wherein the procyanidine is taken as a main component. The extract has an obvious inhibitory influence effect on the activity of tyrosinase, and has the half inhibition rate of 54.8+/-1.27mu g/mL; the inhibition type of the extract is reversible inhibition, and the inhibition mechanism of the extract is in a hybrid type. The tyrosinase inhibitor provided by the application is convenient in material obtaining, low in price and simple and convenient to operate, thus having wide application prospect in the fields such as medical cosmetology, fruit and vegetable preservation and pest and disease control.

Description

technical field [0001] The application relates to the field of bioengineering, in particular to a novel tyrosinase inhibitor. Background technique [0002] Tea is one of the three major beverages in the world, the national drink of our country, and a natural green beverage in the 21st century. my country is a big tea-producing country. At present, the area of ​​tea gardens in my country is stable at 1.1 million hm 2 , with an annual output of more than 600,000 tons, ranking first in the world. However, as tea consumption becomes increasingly high-end, there are about 100,000 tons of middle and low-grade teas that are unsalable and backlogged. In addition, the by-products (tablets, powders, stems, etc.) of tea processing are at least 16,000 tons each year. The utilization rate of tea leaves is very low, which becomes a burden in the production of tea factories. Therefore, it is a research topic of great significance to develop new uses of tea and carry out comprehensive ut...

Claims

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Application Information

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IPC IPC(8): A61K31/353
CPCA61K31/353
Inventor 宋巍柳静佘秋生王珂杨海波魏淑东
Owner HENAN UNIV OF URBAN CONSTR
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