The unicellular green alga Chlamydomonas reinhardtii can acclimate to a wide range of CO(2) concentrations through the regulation of a CO(2)-concentrating mechanism (CCM). By proteomic analysis, here we identified the proteins which were specifically accumulated under high-CO(2) conditions in a cell wall-less strain of C. reinhardtii which release their extracellular matrix into the medium. When the CO(2) concentration was elevated from the ambient air level to 3% during culture, the algal growth rate increased 1.5-fold and the composition of extracellular proteins, but not intracellular soluble and insoluble proteins, clearly changed. Proteomic analysis data showed that the levels of 22 of 129 extracellular proteins increased for 1 and 3 d and such multiple high-CO(2)-inducible proteins include gametogenesis-related proteins and hydroxyproline-rich glycoproteins. However, we could not prove the induction of gametogenesis under high-CO(2) conditions, suggesting that the inductive signal might be incomplete, not strong enough or that only high-CO(2) conditions might be not sufficient for the cell stage to proceed to the formation of sexually active gametes. However, these gametogenesis-related proteins and/or hydroxyproline-rich glycoproteins may have novel roles outside the cell under high-CO(2) conditions.
The effect of monochromatic light on growth, photosynthesis, and hydrocarbon production was tested in Botryococcus braunii Bot-144 (race B), which produces triterpenoid hydrocarbons. The growth was higher in order of red, blue, and green light. The color of red light-grown cells became more orange-yellow and their shape dominantly changed to grape-like with long branches. Photosynthetic carbon fixation activity was higher in order of blue, red, and green light-grown cells, but photosystem activities showed no difference. In the pulse-chase experiments with 14 CO 2 , no major difference was observed in the production of lipids, hydrocarbons, polysaccharides, or proteins among the three kinds of cells, although hydrocarbon production was slightly lower in green light-grown cells. These results indicate that blue and red light were more effective for growth, photosynthetic CO 2 fixation, and hydrocarbon production than green light, and that red light is the most efficient light source when calculated based on photoenergy supplied.
Optimization of the light conditions for biofuel production by the microalga Botryococcus braunii BOT-22 (race B) was performed using monochromatic red light. The lipid and sugar contents were approximately 40% and 20-30% of the cell dry weight, respectively, and about half of the lipids were liquid hydrocarbons. The half-saturation intensities for the production rate of lipids, hydrocarbons, and sugars were 63, 49, and 44μmolm(-2)s(-1), respectively. Fluorescence microscopic images of Nile Red-stained cells showed an increased number of intracellular neutral lipid granules due to increased light intensity. After 16days of incubation in the dark, lipid and sugar, but not hydrocarbon content decreased. Growth, metabolite production, and photosynthesis were saturated at 100, 200 and 1000μmolm(-2)s(-1), respectively. These results indicate that photosynthetically captured energy is not used efficiently for metabolite production; thus, improvements in metabolic regulation may increase hydrocarbon production.
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