响应面法优化川芎蛋白提取工艺及抗氧化活性研究

Optimization of Extraction Process and Antioxidant Activity ofLigusticum chuanxiong Protein by Response Surface Methodology

为优化川芎蛋白(Ligusticum chuanxiong protein,LCP)的提取工艺,并考察其抗氧化活性。基于单因素实验,采用Box-Behnken响应面法,以料液比、提取时间、提取溶剂pH为考察因素,LCP得率为指标,优化LCP提取工艺;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)测定LCP的分子量范围;测定LCP的等电点(pI)及溶解度,并考察LCP的抗氧化活性。结果表明:最佳提取条件为料液比1:15(g/mL)、提取时间1.5 h、pH6,在优化条件下LCP得率为(2.36%±0.13%),实测值与理论值较为接近,表明该数学模型可用于优化LCP提取工艺。最优条件下提取的LCP分子量在17~48 kDa,等电点为3.88,pH8时溶解度为96%,羟自由基清除能力IC_(50)为1.18 mg/mL、超氧阴离子自由基清除能力IC_(50)为0.57 mg/mL、1,1-二苯基-2-苦基肼基(DPPH)自由基清除能力IC_(50)为1.31 mg/mL。该法提取的LCP具有较好的抗氧化活性,可为LCP抗氧化活性的进一步研发提供实验思路。

The aim of this study was to optimize the extraction process of Ligusticum chuanxiong protein(LCP)and to investigate its antioxidant activity.Based on single factor experiment,Box-Behnken response surface methodology was used to optimize the extraction process of LCP,with solid-liquid ratio,extraction time,extraction solvent pH as the factors,and extraction rate of LCP as the index.The molecular weight range of LCP was determined by sodium dodecyl sulphatepolyacrylamide gel electrophoresis(SDS-PAGE).The isoelectric point(pI)and solubility of LCP were determined,and the antioxidant activity of LCP was investigated.The results showed that the optimal extraction conditions were solid-liquid ratio of 1:15(g/mL),extraction time of 1.5 h and pH of 6.Under the optimized conditions,the extraction yield of LCP was(2.36%±0.13%),and the measured value was close to the theoretical value,indicating that the mathematical model could be used to optimize the extraction process of LCP.Under the optimal conditions,the molecular weight of the extracted LCP was 17~48 kDa,the isoelectric point was 3.88,the solubility was 96%at pH8.The IC_(50)of hydroxyl radical scavenging activity was 1.18 mg/mL,the IC_(50)of superoxide anion radical scavenging activity was 0.57 mg/mL,and the IC_(50)of 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical scavenging activity was 1.31 mg/mL.LCP extracted by this method has good antioxidant activity,which can provide experimental ideas for further research and development of LCP antioxidant activity.