Mubeen Umarah, Giraldi Lais Albuquerque, Jüppner Jessica, Giavalisco Patrick
Max Planck Institute of Molecular Plant Physiology;
Max Planck Institute of Molecular Plant Physiology.
J Vis Exp. 2019 Aug 8(150). doi: 10.3791/59547.
Microalgae have been the focus of research for their applications in the production of high value compounds, food and fuel. Moreover, they are valuable photosynthetic models facilitating the understanding of the basic cellular processes. System wide studies enable comprehensive and in-depth understanding of molecular functions of the organisms. However, multiple independent samples and protocols are required for proteomics, lipidomics and metabolomics studies introducing higher error and variability. A robust high throughput extraction method for the simultaneous extraction of chlorophyll, lipids, metabolites, proteins and starch from a single sample of the green alga Chlamydomonas reinhardtii is presented here. The illustrated experimental setup is for Chlamydomonas cultures synchronized using 12 h/12 h light/dark conditions. Samples were collected over a 24 h cell cycle to demonstrate that the metabolites, lipids and starch data obtained using various analytical platforms are well conformed. Furthermore, protein samples collected using the same extraction protocol were used to conduct detailed proteomics analysis to evaluate their quality and reproducibility. Based on the data, it can be inferred that the illustrated method provides a robust and reproducible approach to advance understanding of various biochemical pathways and their functions with greater confidence for both basic and applied research.
微藻因其在高价值化合物、食品和燃料生产中的应用而成为研究热点。此外，它们还是有助于理解基本细胞过程的重要光合模型。全系统研究能够全面深入地了解生物体的分子功能。然而，蛋白质组学、脂质组学和代谢组学研究需要多个独立样本和实验方案，这会带来更高的误差和变异性。本文介绍了一种强大的高通量提取方法，可从莱茵衣藻的单个样本中同时提取叶绿素、脂质、代谢物、蛋白质和淀粉。所示的实验装置适用于在12小时光照/12小时黑暗条件下同步培养的衣藻。在24小时的细胞周期内收集样本，以证明使用各种分析平台获得的代谢物、脂质和淀粉数据高度一致。此外，使用相同提取方案收集的蛋白质样本用于进行详细的蛋白质组学分析，以评估其质量和可重复性。基于这些数据，可以推断所示方法提供了一种强大且可重复的方法，能够更有信心地推进对各种生化途径及其功能的理解，无论是基础研究还是应用研究。
