小球藻细胞匀浆制备方法探究

Study on the preparation methods of homogenate chlorella cells

以普通小球藻为受试生物,研究了液氮匀浆法、研磨匀浆法、机械匀浆法、反复冻融法等4种匀浆方法的破壁效率,同时使用4种方法制备的匀浆液测定SOD活力和GSH含量,旨在筛选出破碎效率高、方便快捷且测定生理生化指标准确的方法。试验结果表明,4种方法的破碎率分别为81.43%、100.00%、92.41%、56.54%,SOD活力分别为47.06、48.58、0、25.05U/mgprot,其中研磨匀浆法与液氮破碎法的测定结果无显著差异(P>0.05);GSH含量分别为34.42、24.52、87.37、38.75mgGSH/gprot,研磨匀浆法、液氮破碎法、反复冻融法的测定结果无显著差异(P>0.05),与机械匀浆法差异显著(P<0.05)。结果说明,研磨匀浆法和液氮破碎法的破碎率高,生化指标测定准确。但由于研磨匀浆法耗费人工成本过高,且无法处理大量样本,相较而言,在上述方法中采用液氮破碎法是制备小球藻细胞匀浆液最好的方法。

With Chlorella. vulgaris as the test organism, four homogenizate methods,liquid nitrogen method, grinding method, mechanical method, and repeated freeze-thaw method were studied. The homogenation with the above methods was used to determined the SOD activity and GSH content, which aimed to select a method with high crushing efficiency, convenience, and accurate determination of physiological and biochemical indicators. The results showed that the crushing rates of the four methods were 81.43%, 100%, 92.41%, 56.54%, and the SOD activity were 47.06, 48.58, 0, 25.05 U/mgprot. There was no significant difference between liquid nitrogen method and grinding method(p>0.05);GSH content were 34.42, 24.52, 87.37, 38.75 mgGSH/gprot;There was no significant difference apart of mechanical method(p>0.05). The above results indicated that the grinding method and liquid nitrogen method have a high crushing rate, and accurate measurement of the biochemical indicators. However, grinding method consumed too much labor and cannot process a large number of samples, so the liquid nitrogen method was considered to be the best solution for preparing microalgal cell homogenation.