酶法制取五谷虫脂肪酸及其抗肿瘤活性成分的研究.pdf

酶法制取五谷虫脂肪酸及其抗肿瘤活性成分的研究 麦波 钱俊青 周文武 （浙江工业大学药学院，杭州 310014 ） 摘要：许多对中药及天然药物的研究表明，油脂及脂肪酸类物质对各种癌症及肿瘤细胞有着不同程度的抑 制以及杀灭活性。五谷虫粉中含有大量的油脂和脂肪酸成分，具有一定的抗癌作用，因此通过酶解处理五 谷虫粉的石油醚提取物，来获得具有更高抗癌活性的脂肪酸药效成分。 五谷虫粉石油醚提取物作为酶解的原料，在有机相中溶解后，以磷酸盐缓冲液作为水相，通过脂肪酶 的水解作用得到高含量游离脂肪酸的提取物。首先通过单因素条件实验得到合适的脂肪酶加入量，酶解的 时间和温度，缓冲溶液的ph ，缓冲液和有机相加入比例以及料液比。然后，使用plackett-burman （PB ）和 中心复合设计（Central Composite Design ）对上述的6 个影响酶解效率的因素进行筛选，并对其中影响最为 明显的3 个因素料液比，酶解的时间和温度优化，得到的五谷虫脂肪酸含量达到95.36 ％的工艺条件为：酶 解时间 16h40m，酶解温度为39.5，料液比为1:7.65，缓冲溶液pH 为8.0，缓冲液与有机相体积比为3:2， 酶量为 10mg·g-1 。对脂肪酸含量为58.6 ％的样品进行了抗癌活性的检测，五谷虫脂肪酸(0.063 mg ·mL-1)时 经过72h ，对HL-60 人白血病细胞株和A-549 人肺癌细胞株的抑制效应分别为94.3 ％和91.4 ％，表明经过 酶解得到的五谷虫脂肪酸相较于五谷虫粉的抗癌活性有了大幅度的提高，具有较高的药用价值。 关键词： 五谷虫；脂肪酸；酶法水解；抗肿瘤 Preparation of oriental latrine fly larvina fatty acids and studies of anti-tumor active component 1 2 3 MAI Bo , QIAN Junqing , ZHOU Wenwu (College of pharmaceutical science, Zhejiang University of technology , Hangzhou 310014 , China) Abstract ：Many researches of traditional Chinese medicine and naturally occurring drugs shows that the lipids and fatty acids can inhibit or kill various kinds of cancer and tumor cells. It was generally believed that the carboxy group had the anti-tumor activity through the studies between the QSAR and the anti-tumor effects of fatty acids. Oriental latrine fly larvina contained many free fatty acids and lipids which can inhibit the cancer and tumor cells, so it was necessary to get high purity of fatty acids which had higher anti-tumor activity by the enzymolysis of lipids. The petroleum ether extract of oriental latrine fly larvina powder was used in two-phase system to get free fatty acids. First we obtained a better enzymolysis conditions as the use of the enzyme,