该文研究了黑糙米花青素的抗氧化能力并探究黑糙米花青素对酪氨酸酶的抑制机理。以DPPH自由基、ABTS阳离子自由基和羟自由基清除能力为指标通过实验评价其抗氧化能力,其半抑制浓度IC50值分别为40.34、20.3、287.63 μg/mL,表明黑糙米花青素具有良好的抗氧化能力。通过体外模拟试验及酶促动力学模型研究花青素对酪氨酸酶的抑制效果,黑糙米花青素对酪氨酸酶的半抑制浓度IC50值为94.2 μg/mL,其抑制效果强于阳性对照物熊果苷,其IC50值为126.95 μg/mL,且其对酪氨酸酶的抑制类型为可逆竞争性抑制。通过分子对接和动力学模拟探究矢车菊素-3-O-葡萄糖苷(cyanidin-3-O-glucoside,C3G)、熊果苷对酪氨酸酶活性的抑制机制,C3G与酪氨酸酶的结合位点主要是TYR140、PRO366、TRP386和HIS390这4个氨基酸残基,通过离子键和氢键相互作用,熊果苷、C3G与酪氨酸酶的结合能分别为-11.82、-16.9 kcal/mol。根据以上结果得出,C3G与酪氨酸酶结合的更稳定,能够通过竞争性结合酪氨酸酶的结合位点,进而降低该酶的催化效率,从而抑制黑色素的形成。花色苷具有良好的抗氧化活性,对酪氨酸酶有良好的抑制效果,可用于抗衰老和美白产品的应用。
The antioxidant capacity of black brown rice anthocyanins was studied and the inhibition mechanism of black brown rice anthocyanins on tyrosinase was investigated.The antioxidant capacity of black brown rice anthocyanins was evaluated by DPPH free radical, ABTS cationic radical and hydroxyl radical scavenging experiments and the half inhibitory concentration IC50 values were 40.34, 20.3, and 287.63 μg/mL, respectively, indicating that black brown rice anthocyanins have good antioxidant capacity.The inhibitory effect of anthocyanins on tyrosinase was studied by simulation tests and enzymatic kinetic models in vitro.The IC50 value of the half inhibitory concentration of anthocyanins on tyrosinase was 94.2 μg/mL, which was stronger than that of the positive control arbutin (IC50=126.95 μg/mL), and its inhibition type of tyrosinase was reversible competitive inhibition.The inhibitory mechanism of C3G and arbutin on tyrosinase activity was analyzed by molecular docking and kinetic simulation, the binding sites of cyanidin-3-O-glucoside(C3G) and tyrosinase were mainly four amino acid residues, including TYR140, PRO366, TRP386, and HIS390, through the interaction of ionic bonds and hydrogen bonds, the binding energies of arbutin, C3G and tyrosinase were -11.82 and -16.9 kcal/mol, respectively.So C3G binded to tyrosinase more stably.C3G could competitively bind to the binding site of tyrosinase, thereby reducing the catalytic efficiency of the enzyme and inhibiting the formation of melanin.Therefore, anthocyanins have good antioxidant activity and have a good inhibitory effect on tyrosinase, which can be used in the application of anti-aging and whitening products.
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