山竹壳多糖提取工艺优化及其生物活性研究

Optimization of the Extraction of Mangosteen(Garcinia mangostana)Pericarp Polysaccharides and Determination of Their Biological Activities

以山竹壳多糖提取率为指标,在单因素试验的基础上,选取料液比、提取温度、超声时间3个因素,采用正交试验优化山竹壳多糖的提取工艺;考察山竹壳多糖的抗氧化活性;通过微量肉汤稀释法评价山竹壳多糖的抑菌效果;采用细胞计数试剂盒(cell counting kit-8,CCK-8)法研究山竹壳多糖对肿瘤细胞的增殖抑制作用。结果表明,多糖最佳提取工艺为料液比1∶40(g/mL)、提取温度55℃、超声时间40 min,在此条件下山竹壳多糖的提取率为(12.99±0.48)%;山竹壳多糖清除DPPH自由基、羟基自由基以及ABTS+自由基的IC50值分别为69.25、462.50、18.39μg/mL,且其抗氧化性能随着多糖浓度的升高而增强;山竹壳多糖对金黄色葡萄球菌、大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、鲍曼不动杆菌最小抑菌浓度(minimum inhibitory concentration,MIC)依次分别为0.391、3.125、3.125、6.25、25.00 mg/mL;山竹壳多糖对人乳腺癌细胞(MCF-7)和小鼠黑色素瘤细胞(B16)无增殖抑制作用,对人结肠癌细胞(DLD-1)和人黑色素瘤细胞(A375)的增殖抑制效果与其浓度呈剂量依赖性,其作用于人结肠癌细胞(DLD-1)和人黑色素瘤细胞(A375)的IC50分别为76.66μg/mL和127.70μg/mL。

Mangosteen(Garcinia mangostana)pericarp was treated as the source material and the extracted yield of its polysaccharides was considered as the index.A series of orthogonal experiments,which were based on single-factor experimentation,were carried out to explore the effects of the ratio of material to liquid,extraction temperature,and ultrasound time on the extraction yield of polysaccharides.The antioxidant activity of the polysaccharides was investigated and their antibacterial effects were tested using the microbroth dilution method.The inhibitory effects of the polysaccharides on the proliferation of tumor cells was evaluated using a cell counting kit-8(CCK-8)assay.Based on the experimental results,the optimal conditions for polysaccharide extraction were determined to be:a solid-liquid ratio of 1∶40(g/mL),an extraction temperature of 55℃and an ultrasonic time of 40 min.Under the above conditions,the extraction rate of the polysaccharides was(12.99±0.48)%.The DPPH radical,OH radical and ABTS+radical scavenging assay IC50 values were 69.25,462.50,18.39μg/mL,respectively.The polysaccharides displayed antimicrobial activity for several bacterial species,including Staphylococcus aureus,Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa and Acinetobacter baumannii with MIC values of 0.391,3.125,3.125,6.25,25.00 mg/mL,respectively.The polysaccharides exhibited no cytotoxicity toward MCF-7 and B16 cells and exhibited lower cytotoxicity for DLD-1 and A375 cells with IC50 values of 76.66μg/mL and 127.70μg/mL,respectively.