仿刺参水溶性海参皂苷的分离制备及抗真菌活性的研究

Isolation of Water-Soluble Holothurian Glycoside from Apostichopus joponicus and Studies of Its Antifungal Activity

利用海洋生物资源进行天然抗真菌新药的开发，从冻干海参加工废液中提取了水溶性海参皂苷，并对其抗真菌活性进行了研究。采用大孔吸附树脂法提取水溶性海参皂苷，经Libermann Burchard反应等对提取物的性质进行鉴定后，利用管碟法对所得提取物的抗菌活性进行了测定，并利用琼脂稀释法测定了所得水溶性海参皂苷对6株供试真菌的最低抑菌浓度（MIC）。提取物在Molish反应、沉淀反应和泡沫实验中均为阳性，说明提取物中确含有皂苷类成分；在Libermann Burchard反应中，液体的颜色处在红色与紫色之间，显示提取物可能是1种三萜类皂苷。水溶性海参皂苷对6株供试真菌均具有显著的抑制作用，且在浓度为0．5～4mg／mL的范围内，其抑菌活性与所用浓度之间呈明显的正相关性；在浓度为4mg／mL时，水溶性海参皂苷对6株供试真菌抑制活性的大小依次为裂殖酵母菌、啤酒酵母菌、白色念珠菌、葡萄炭疽病原菌、黄瓜枯萎病原菌和黑曲霉，海参皂苷对6株真菌的MIC值依次分别为0．002，0．016，0．063，0．063，0．125．0．250mg／mL。

To exploit new natural antifungal medicaments from marine echinoderm resources, the water-soluble holothurian glycoside was isolated from the liquid residue of freeze-drying sea cucumber Apostichopus joponicus and its antifungal activity was examined. The water-soluble holothurian glycoside was extracted by column chromatography of AB-8 macroporous resin, and the Libermann-Burchard reaction, Molish reaction, Deposit reaction, and Foaming test were utilized to characterize its properties. The antifungal activities of the water-soluble holothurian glycoside were examined with six trains of fungi, i. e., Candida albicans, Colletotrichum gloeosporioides , Fusarium oxysporum f. sp. cucumerinum , Schizosaccharomyces pombe , Saccharomyes cerevisiae and Aspergillus niger, by agar diffusion methods, and their minimum inhibitory concentration （MIC） of the water-soluble holothurian glycoside was determined by agar dilution methods, respectively. The isolated fractions were all positive in Molish reaction, Deposit reaction and Foaming test, which indicated that the obtained fractions did contain the water-soluble holothurian glycoside. The fractions underwent color changes between red and purple in Libermann-Burchard reaction, and it indicated that the water-soluble holothurian glycoside isolated in this study was probably a kind of triterpenoid glycoside. The water-soluble holothurian glycoside exhibited remarkable antifungal activities on the 6 strains of fungi, and its antifungal activities increased with concentrations in a dose-dependent manner within the concentrations of 0.5 and 4 mg/mL. The effect of the antifungal activities of the water-soluble holothurian glycoside at a concentration of 4 mg/mL on the 6 strains of fungi was as follows, S. pombe 〉 S. cerevisiae 〉 C. albicans 〉 C. gloeosporioides 〉 F. oxysporum 〉 A. niger, and the MIC value of the water-soluble holothurian glycoside on them was 0. 002, 0. 016, 0. 063, 0.063, 0. 125 and 0. 250 mg/mL, respectively. The water-soluble holothurian glycoside, probably a kind of triterpenoid glycoside, was isolated by AB-8 macroporous resin successfully and exhibited an obvious antifungal activity on different strains of fungi. The water-soluble holothurian glycoside obtained in this study may have great potentials and values in the development of natural marine antifungal medicaments.