摘要
本研究应用Box-Behnken设计-响应面法优化菲牛蛭多肽的提取工艺,并考察其美白抗衰作用.在单因素实验的基础上,以NaCl溶液浓度、提取时间、提取次数为影响因素,多肽得率为响应值,设计并利用响应面模型,得到菲牛蛭多肽提取的最佳条件:NaCl溶液浓度4.3％,超声时间4 h,超声次数2次(2+2 h).该模型显著,与实际试验拟合良好.通过酪氨酸酶催化左旋多巴氧化速率法考察菲牛蛭多肽的酪氨酸酶抑制活性;以秀丽隐杆线虫为模式生物,考察菲牛蛭多肽对线虫的身体长度、运动能力、生殖能力、活性氧(reactive oxygen species,ROS)和脂褐素的影响.结果显示,菲牛蛭多肽对酪氨酸酶有显著的抑制作用,其IC50为0.58 mg·mL-1,强于阳性对照熊果苷(2.24 mg·mL-1).与空白组相比,0.1、0.5和1.0mg·mL-1的菲牛蛭多肽对线虫体长、产卵量和头尾摆动无显著性差异;0.5 mg·mL-1的菲牛蛭多肽显著降低线虫体内ROS水平;0.5和1.0 mg·mL-1的菲牛蛭多肽显著降低线虫体内脂褐素水平.菲牛蛭多肽具有良好的美白抗衰作用,其作用机制可能与抑制酪氨酸酶活性和抗氧化作用有关.
Abstract
The objective of this study was to optimise the extraction process of peptide of Poecilobdella manillensis by the Box-Behnken design-response surface methodology,and to investigate its whitening and anti-aging effects.Based on single factor experiments,NaCl solution concentration,extracting time and extracting times were taken as influencing factors,and peptide yield was used as the response value.The response surface model was designed and used to obtain the optimal extraction conditions of peptides of Poecilobdella manillensis:NaCl solution concentration of 4.3％,ultrasonic time of 4 h,and ultrasonic times of 2 times(2+2 h),which was significant and well-fitted to the actual experiment.The tyrosinase inhibition activity of peptide of Poecilobdella manillensis was evaluated by measuring the oxidation rate of levodopa(L-DOPA)catalyzed by tyrosinase.Moreover,using Caenorhabditis elegans as a model organism,the effects of peptide of Poecilobdella manillensis on body length,locomotion,reproductive capacity,reactive oxygen species(ROS),and lipofuscin levels were determined.The results showed that peptide of Poecilobdella manillensis exhibited a significant inhibitory effect on tyrosinase,with an IC50 of 0.58 mg·mL-1,which was stronger than that of the positive control arbutin(2.24 mg·mL-1).Compared to the control group,0.1,0.5 and 1.0 mg·mL-1 peptide of Poecilobdella manillensis showed no significant differences in the body length,eggs,and body bending of Caenorhabditis elegans.However,0.5 mg·mL-1 peptide of Poecilobdella manillensis significantly reduced ROS levels in Caenorhabditis elegans;0.5 and 1.0 mg·mL-1 peptide of Poecilobdella manillensis significantly reduced lipofuscin levels in Caenorhabditis elegans.Peptide of Poecilobdella manillensis exhibits effective whitening and anti-aging activities,potentially mediated by its inhibition of tyrosinase activity and antioxidant effects.
维普
万方数据