Use of cashew nut shell liquid active substances as tyrosinase inhibitors and plant-derived antibacterial agents
A technology of cashew nut shell oil and active substances, applied in the direction of plant growth regulators, plant growth regulators, chemicals for biological control, etc., can solve the problem of no further detection of plant pathogenic fungi, high cost, no further detection and extraction Purification of antibacterial agents and other issues to achieve the effect of inhibiting mushroom tyrosinase diphenolase activity, simple steps, and simple and fast processing
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Embodiment 1
[0021] Embodiment 1: Cashew nut shell liquid active material is used as tyrosinase inhibitor and antibacterial agent preparation method
[0022] The active ingredient of the extracted and purified cashew nut shell oil accounts for 95%, and the emulsifier is 5%, and it is stirred for 1 hour to fully mix. That is, the tyrosinase inhibitor and the plant-derived antibacterial agent were obtained and stored at 4°C for future use. When used, dilute with organic solvent methanol or DMSO to prepare the required concentration.
Embodiment 2
[0023] Embodiment 2: Determination of the inhibitory effect of active substances of cashew nut shell oil on tyrosinase diphenolase activity
[0024]The active substance prepared in Example 1 was tested for inhibition of mushroom tyrosinase diphenolase, and the active substance prepared in Example 1 was dissolved in deionized double-distilled water to prepare solutions containing different concentrations of effectors. With 0.5mM L-DOPA as substrate, in 3mL assay system of 0.05M phosphate buffer (pH 6.8), first add 0.1mL of different concentrations of effectors to the cuvette, then add 2.8mL in advance at 30°C Add 0.05mL of mushroom tyrosinase aqueous solution to the substrate solution kept in a constant temperature water bath, mix immediately, measure the optical density value at a wavelength of 475nm with time at a constant temperature of 30°C, and obtain the enzyme enzyme from the slope of the line. Vitality, extinction coefficient according to ε = 3700 (mM cm) -1 calculate....
Embodiment 3
[0026] Example 3: Measuring Analysis of the Mechanism and Type of Cashew Nut Shell Oil Active Substances Inhibiting Acidase
[0027] The cashew nut shell oil active substance prepared in Example 1 was respectively subjected to molecular inhibition mechanism and type analysis tests with tyrosinase. In the assay activity system, the enzyme concentration was 3.33 μg / mL, and the substrate L-DOPA concentration was changed to measure Effects of different concentrations of effectors on the initial velocity of enzyme catalysis, plotting the initial velocity of the enzyme reaction against the substrate concentration is a set of hyperbolas, indicating that the enzymatic reaction follows the Michaelis-Menten kinetic equation. Use the Lineweaver-Burk double reciprocal plot to determine the type of inhibition of the effector. The Michaelis constant K of the enzyme catalyzing the oxidation of L-DOPA was obtained from the intercept of the straight line m and the maximum speed V m . The re...
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