James, GabrielHocart, CharlesHillier, WarwickChen, HancaiKordbacheh, FarzanehPrice, Graeme (Dean)Djordjevic, Michael2015-12-080960-8524http://hdl.handle.net/1885/28793The 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, 18:1D9 , 18:2D9,12 and 18:3D9,12,15 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.Keywords: Biosynthetic pathway; Caleosin; Chlamydomonas reinhardtii; Dry cells; Elevated level; Gas chromatography-mass spectrometry; GC/MS; High-throughput method; Lipid bodies; Lipid content; Lipid droplets; Neutral lipid; Nitrogen starvation; Polyunsaturated fat Biofuel; Chlamydomonas reinhardtii; Fatty acid; GC/MS; Neutral lipid201110.1016/j.biortech.2010.11.0512016-02-24