Chlamydomonas reinhardtii is a model alga for studying triacylglycerol (TAG) accumulation in the photosynthetic production of biofuel. Previous studies were conducted under photoheterotrophic growth conditions in medium supplemented with acetate and/or ammonium. We wanted to demonstrate TAG accumulation under truly photoautotrophic conditions without reduced elements. We first reidentified all lipid components and fatty acids by mass spectrometry, because the currently used identification knowledge relies on data obtained in the 1980s. Accordingly, various isomers of fatty acids, which are potentially useful in tracing the flow of fatty acids leading to the accumulation of TAG, were detected. In strain CC1010 grown under photoautotrophic conditions, TAG accumulated to about 57.5 mol% of total lipids on a mole fatty acid basis after the transfer to nitrogen-deficient conditions. The content of monogalactosyl diacylglycerol, sulfoquinovosyl diacylglycerol, and phosphatidylglycerol decreased drastically. The accumulated TAG contained 16:0 as the major acid and 16:4(4,7,10,13), 18:2(9,12), and 18: 3(9,12,15), which are typically found in chloroplast lipids. Additionally, 18:1(11) and 18:3(5,9,12), which are specific to extrachloroplast lipids, were also abundant in the accumulated TAG. Photosynthesis and respiration slowed markedly after the shift to nitrogen-deficient conditions. These results suggest that fatty acids for the production of TAG were supplied not only from chloroplast lipids but also from other membranes within the cells, although the possibility of de novo synthesis cannot be excluded. Under nitrogen-replete conditions, supplementation with a high concentration of CO 2 promoted TAG production in the cells grown photoautotrophically, opening up the possibility to the continuous production of TAG using CO 2 produced by industry.
Photosynthetic production of various forms of reduced carbon is the ultimate source of biological free energy, which enables biological, social, and industrial human activities. Agricultural production has been the major source of human nutrition, but biofuel is now attracting social interest as an alternative to fossil resources. Algal production of bioresources is a new perspective as a means to supply both biofuel and chemical materials, avoiding competition with nutritional resources (1). Among various algal products, triacylglycerol (TAG), hydrocarbons, and carbohydrates are the major targets of bioengineering. In many algae, TAG is known to accumulate under conditions of nutrient deficiency (2). A unicellular green alga, Chlamydomonas reinhardtii, is currently studied as a model organism for elucidating the metabolic pathway and signaling pathway of TAG accumulation. This alga has been used in studies of photosynthesis, reproduction, and motility, mainly because the methodology for genetic analysis is applicable (3, 4). In addition to classical crossing, molecular genetic manipulation (5, 6) is becoming useful, especially on the basis of the genomic information (7). Vari...