鸡蛋花多糖提取工艺优化及生物活性研究

孙宁云; 姚欣; 张英慧; 杨安平; 刘辉

doi:10.13386/j.issn1002-0306.2021050198

鸡蛋花多糖提取工艺优化及生物活性研究

Optimization of Extraction Process of Polysaccharides from Plumeria rubra L.cv. Acutifolia and Evaluation of Biological Activity

摘要

摘要: 目的：优化鸡蛋花（Plumeria rubra L. cv. Acutifolia）多糖的提取工艺，并对鸡蛋花多糖（PRLAP）体外抗氧化、抗菌、抗肿瘤活性进行评价。方法：通过单因素实验设计正交试验，确定PRLAP的最佳提取条件；通过测PRLAP对DPPH、ABTS和OH自由基清除能力来评价PRLAP的抗氧化活性；采用微量肉汤稀释法测PRLAP对大肠杆菌（Escherichia coli）、金黄色葡萄球菌（Staphylococcus aureus）、铜绿假单胞菌（Pseudomonas aeruginosa）、鲍曼不动杆菌（Acinetobacter baumannii）、肺炎克雷伯菌（Klebsiella pneumoniae）的最小抑菌浓度（minimal inhibit concentration，MIC）来评价PRLAP的抑菌活性；此外，通过CCK-8法检测PRLAP对人黑色素瘤细胞（A375）、小鼠黑色素瘤细胞（B16）、人结直肠癌上皮细胞（DLD-1）、人乳腺癌细胞（MCF-7）细胞活力的影响来评价PRLAP的抗肿瘤能力。结果：PRLAP最佳提取条件：超声功率264 W、料液比1:50（g/mL）、提取时间40 min、提取温度35 ℃。此条件下，PRLAP的得率为14.01%±0.22%。PRLAP清除DPPH、ABTS、OH自由基的半抑制浓度（IC₅₀）分别为0.1934、0.2315、2.469 mg/mL。PRLAP对大肠杆菌和金黄色葡萄球菌的MIC分别为0.391和0.195 mg/mL，对铜绿假单胞菌、肺炎克雷伯菌和鲍曼不动杆菌的MIC均为1.562 mg/mL。细胞活力实验结果表明，在一定浓度范围内，除DLD-1细胞外，PRLAP对A357、B16和MCF-7细胞活力有一定的抑制作用且与浓度均呈剂量依赖性，IC₅₀分别为101.3、285.6、423.1 μg/mL。结论：本文首次对鸡蛋花多糖提取工艺进行了优化，通过一系列实验证明PRLAP具有良好的体外抗氧化、抑菌以及一定的抗肿瘤活性，为进一步研究开发鸡蛋花提供科学理论支撑。

Abstract: Objective: To optimize the extraction process of polysaccharides from Plumeria rubra L. cv. Acutifolia, and make acomment about antioxidant, antibacterial and anti-tumor activities of Plumeria polysaccharide. Methods: The orthogonal experiment was designed by single factor experiment and the optimal extraction conditions were determined. The antioxidant activity was evaluated by measuring its scavenging ability to DPPH, ABTS, and OH free radical assays. The minimum inhibitory concentration of Plumeria polysaccharide against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae was measured by the micro broth dilution method to evaluate the antibacterial activity of Plumeria polysaccharide. In addition, CCK-8 assay was used to evaluate the inhibitory effect of Plumeria polysaccharide on human melanoma cells (A375), mouse melanoma cells (B16), human colorectal cancer epithelial cells (DLD-1) and human breast cancer cells (MCF-7). Results: The optimum conditions of Plumeria polysaccharide were as follows:Ultrasonic power was 264 W, solid-liquid ratio was 1:50 (g/mL), extraction time was 40 min, extraction temperature was 35 ℃. Under these conditions, the yield of Plumeria polysaccharide was 14.01%±0.22%. The half-inhibitory concentrations of Plumeria polysaccharide in scavenging DPPH, ABTS and OH free radicals were 0.1934, 0.2315 and 2.469 mg/mL, respectively. The minimum inhibitory concentrations of Plumeria polysaccharide against Escherichia coli and Staphylococcus aureus was 0.391 and 0.195 mg/mL, respectively, and the minimum inhibitory concentrations against Pseudomonas aerugino, Klebsiella pneumoniae and Acinetobacter baumannii were all 1.562 mg/mL. The results of cell viability experiments showed that within a certain concentration range, except for DLD-1 cells, Plumeria polysaccharide had a certain inhibitory effect on the viability of A357, B16 and MCF-7 cells and was dose-dependent with the concentration, with IC₅₀ of 101.3, 285.6 and 423.1 μg/mL. Conclusion: This article optimizes the extraction process of Plumeria polysaccharides for the first time. A series of experiments prove that Plumeria polysaccharide has good antioxidant, antibacterial and anti-tumor activities in vitro. This research provides a certaindata theoretical support for the further development and utilization of Plumeria.