Plant Science Division, Research School of Biology, The Australian National University, Canberra, ACT, Australia.
Bioresour Technol. 2011 Feb;102(3):3343-51. doi: 10.1016/j.biortech.2010.11.051. Epub 2010 Nov 16.
The Chlamydomonas reinhardtii starch-less mutant, BAF-J5, was found to store lipids up to 65% of dry cell weight when grown photoheterotrophically and subjected to nitrogen starvation. Fourier transform infrared spectroscopy was used as a high-throughput method for semi-quantitative measurements of protein, carbohydrate and lipid content. The fatty acids of wild-type and starch mutants were identified and quantified by gas chromatography mass spectrometry. C. reinhardtii starch mutants, BAF-J5 and I7, produce significantly elevated levels of 16:0, 18:1(Δ9), 18:2(Δ9,12) and 18:3(Δ9,12,15) fatty acids. Long-chain saturated, mono- and polyunsaturated fatty acids were found under nitrogen starvation. Oleosin-like and caleosin-like genes were identified in the C. reinhardtii genome. However, proteomic analysis of isolated lipid bodies only identified a key lipid droplet associated protein. This study shows it is possible to manipulate algal biosynthetic pathways to produce high levels of lipid that may be suitable for conversion to liquid fuels.
当培养为光异养型并经历氮饥饿时,莱茵衣藻无淀粉突变体 BAF-J5 的细胞内脂质积累量可达干重的 65%。傅里叶变换红外光谱(FTIR)被用作一种高通量的方法,用于对蛋白质、碳水化合物和脂质含量进行半定量测量。野生型和淀粉突变体的脂肪酸通过气相色谱质谱联用(GC-MS)进行鉴定和定量。淀粉突变体 BAF-J5 和 I7 产生显著升高水平的 16:0、18:1(Δ9)、18:2(Δ9,12)和 18:3(Δ9,12,15)脂肪酸。在氮饥饿条件下发现了长链饱和、单不饱和和多不饱和脂肪酸。在莱茵衣藻基因组中鉴定到了油蛋白样和钙蛋白样基因。然而,对分离的脂滴的蛋白质组学分析仅鉴定到一个关键的脂滴相关蛋白。本研究表明,操纵藻类生物合成途径以生产可能适合转化为液体燃料的高含量脂质是可行的。