Department of Biochemistry and Molecular Biology, Michigan State University, MI 48824, USA.
Department of Chemistry, Michigan State University, East Lansing, MI 48824, USA.
Bioresour Technol. 2013 Oct;146:310-316. doi: 10.1016/j.biortech.2013.07.088. Epub 2013 Jul 25.
Triacylglycerols (TAGs) from microalgae can serve as feedstock for the production of biofuels. To gain a comprehensive understanding of TAG metabolism in algae through genetic and molecular approaches, and to improve algal biofuel production, efficient and quantitative phenotyping methods focusing on TAGs are required. Towards this end, a facile ultrahigh performance liquid chromatography-mass spectrometry protocol was developed for TAG profiling, achieving identification and quantification of intact TAG molecular species in two algae. TAG profiling was performed in Chlamydomonas reinhardtii and Nannochloropsis oceanica grown in nitrogen (N)-replete or N-depleted medium. For the quantification of algal TAGs and fatty acids, two sets of internal standards were developed by taking advantage of the presence of pheophytin and specific fatty acids in algal samples. Comparison of algal TAG levels was simplified by using these internal standards for TAG analysis, paving the way for high-throughput mutant screening.
微藻中的三酰基甘油（TAG）可用作生物燃料生产的原料。为了通过遗传和分子方法全面了解藻类中的 TAG 代谢，并提高藻类生物燃料的产量，需要开发针对 TAG 的高效、定量表型分析方法。为此，开发了一种简单的超高效液相色谱-质谱联用协议，用于 TAG 分析，可在两种藻类中鉴定和定量完整的 TAG 分子种类。在氮（N）充足或耗尽的培养基中培养莱茵衣藻和海洋盐藻，进行 TAG 分析。为了定量分析藻类的 TAG 和脂肪酸，利用藻类样品中叶绿素和特定脂肪酸的存在，开发了两组内标。通过使用这些内标进行 TAG 分析，简化了藻类 TAG 水平的比较，为高通量突变体筛选铺平了道路。
