Use of black mulberry extract in preparing tyrosinase inhibitor
A technology of tyrosinase and compound, applied in the field of natural compounds, can solve the problems of lack of fresh-keeping method, economic loss and the like
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Embodiment 1
[0018] Example 1: Determination of tyrosinase inhibitory activity.
[0019] The determination of tyrosinase inhibitory activity takes kojic acid as a reference substance, and the method is as follows.
[0020] The substrate dopa was first dissolved in phosphate buffer (25 mM, pH 6.8) to a concentration of 8 mM. To a 96-well plate, 20 μL of the above substrate solution, 90 μL of phosphate buffer, and 20 μL of compounds of different concentrations were added. Add 70 μL of tyrosinase-containing phosphate buffer (100 U / mL, pH 6.8) to start the reaction, and react at 37° C. for 10 minutes. Before and after the reaction, the absorbance of each well was measured with a Multiscan MK3 automatic multifunctional microplate reader (Thermo) at 492 nm.
[0021] According to the absorbance at 492nm, the inhibitory rate (%) of the compound to tyrosinase is calculated, and the concentration of the inhibitor when the enzyme activity inhibitory rate (%) reaches 50% is determined as IC 50 valu...
Embodiment 2
[0031] Example 2: Time Analysis of Tyrosinase Inhibition.
[0032] The substrate dopa was first dissolved in phosphate buffer (25 mM, pH 6.8) to a concentration of 8 mM. To a 96-well plate, add 20 μL of the above-mentioned different concentrations of the substrate solution, 90 μl of phosphate buffer, and 20 μL of the compound with a final concentration of 60 μM. Add 70 μL of tyrosinase-containing phosphate buffer (100 U / mL, pH 6.8) to start the reaction, and measure the absorbance of each well at different times. Calculate enzyme inhibition rate, method is the same as "embodiment 1". Results: See Table 2.
[0033] The tyrosinase inhibition activity of table 2 different time
[0034] time (min)
[0035] 30
[0036] Conclusion: the enzyme inhibition rate was 63.49% when reacting for 15 minutes, and as time went on, the enzyme inhibition rate decreased, but the inhibition rate to tyrosinase remained above 50% in 180 minutes, so the formula ( 1) The comp...
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