邵慧敏^1,2, 王 聪³, 汪 辉¹, 刘天中¹
1.中国科学院 青岛生物能源与过程研究所 藻类生物技术中心;2.中国科学院大学;3.中国科学院 青岛生物能源与过程研究所 公共实验室

摘要:
利用代谢物组学技术有助于更为深入地了解不分枝丝状微藻黄丝藻细胞的生物学特性。本研究比较了3种黄丝藻细胞破碎方法(冻融法、超声破碎法、液氮研磨法)以及基于气相色谱-质谱联用技术的3种黄丝藻代谢物提取系统(冷甲醇、热乙醇、甲醇/氯仿法), 以期为黄丝藻细胞的代谢组学分析奠定基础。结果显示: 液氮研磨法处理下的黄丝藻细胞破碎效果最佳; 根据提取到的代谢物的种类角度出发, 冷甲醇法的提取效果要明显优于其他2种提取方法。在黄丝藻细胞中共检测到40种代谢物, 包括9种氨基酸、17种有机酸、12种糖类和2种醇类物质。
关键词:黄丝藻代谢物组学细胞破壁代谢物提取
DOI：10.11759/hykx20170828002
分类号:
基金项目:青岛市应用基础研究计划项目(16-5-1-68-jch); 青岛市海洋经济创新发展产业链项目资助

Comparison of metabolome extraction strategies from Tribonema minus
SHAO Hui-min,WANG Cong,WANG Hui,LIU Tian-zhong
Abstract:
Metabolomics Technology contributes to the study on the biological characteristics of filamentous microalgae Tribonema minus. In this study, in order to find a reliable cell disruption and extraction methodology and lay foundation for metabolomics of T. minus, three ways of cell disruption method (freeze-thawing, ultrasonic and liquid nitrogen grinding), and three different extraction method based on freezing-thawing in methanol extraction, boiling ethanol extraction, methanol/chloroform extraction were compared for their extraction efficiency of metabolites using gas chromatography-mass spectrometry (GC-MS) as the evaluation standard. The results showed that ultrasonic and liquid nitrogen grinding was helpful on the fragmentation of filaments microalgae T. minus, while there is no obvious effect on the cell disruption after freezing-thawing. Moreover, compared with ultrasonic, the majority filaments of T. minus were broken into pieces after liquid nitrogen grinding, and the intracellular components were released effectively. Therefore, liquid nitrogen grinding is the best method for breaking cells. For the extraction system, the total metabolite number of cold methanol extraction is much more than those of boiling ethanol extraction and methanol/chloroform extraction. In particular, the extraction efficiency of cold methanol extraction is higher than those of boiling ethanol extraction and methanol/chloroform extraction in amino acids, organic acids and sugar. Using cold methanol extraction method, 40 kinds of metabolites from T. minus were detected and identified, including 9 kinds of amino acids, 16 kinds of organic acids, 11kinds of sugar and 2 kinds of alcohols. Accordingly, cold methanol extraction is an efficient extraction method for further analysis of metabolites from T. minus. In conclusion, our study provided evidence that the optimal conditions for T. minus sample preparation are cell disruption with liquid nitrogen grinding, and extraction of metabolites by cold methanol method.
Key words:Tribonema sp.metabolomicscell lysisMetabolite extraction





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