【摘要】 为了确定破囊壶菌Aurantiochytrium sp.的最佳破壁方法,分别使用反复冻融法、超声法、高速匀浆法、碱热法和研磨法对破囊壶菌进行破壁处理。为了解各方法的破壁效果,使用显微镜观察破壁前后的细胞个数变化,并利用SEM(Scanning Electron Microscope)观察细胞的形态变化。为探究不同破壁方法对油脂提取的影响,首先使用尼罗红荧光染色法对油脂含量进行了定性检测,随后采用正己烷提取油脂。为探究不同破壁方法对油脂组分的影响,使用气相色谱法分析了油脂的组成情况。结果显示:(1)碱热的破壁效果最好,破壁率达到95.51%;(2)碱热破壁后得到的油脂产率最高,达到61.33%,远高于其他4种方法 ;(3)碱热破壁后得到的油脂中DHA含量最高、达到41.33%,相应的DHA产量也远高于其他方法、达到2.192 g/L。从制备高纯度DHA的角度出发,碱热法是最佳的破壁方法。更多还原

【Abstract】 In order to select the best method for cell disruption of Aurantiochytrium sp.,five cell disruption methods,including repeated freezing and melting,ultrasonication,high-speed homogenization,alkaline heating and trituration,were used to break up the cell wall of Aurantiochytrium sp.,then cell disruption efficiency of different methods and their influences on oil extraction and oil composition were analyzed and compared. In order to characterize the cell disruption efficiency of different methods,cell number was estimated by microscope observation,and cell morphology was observed using SEM. In order to explore the influences of different cell disruption methods on oil extraction,oil distribution was qualitatively analyzed using Nile red staining,then crude oil was extracted by n-hexane. In order to explore the influences of different cell disruption methods on oil composition,the specific composition of oils was analyzed using GC(Gas Chromatography). Results showed that :(1)Alkaline heating had the best effect on cell disruption for its highest efficiency(95.51%)and biggest cell morphology changes.(2)The extracted crude oil after alkaline heating was the most and up to 61.33% of the dry biomass weight.(3)Highest DHA content in oil was 41.33% after alkaline heating,of which DHA production was much higher than the other methods and up to 2.192 g/L. From the perspective of high purity DHA production,alkaline heating is the most promising method for cell disruption of Aurantiochytrium sp.更多还原