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Microalgae are being investigated as potential candidates for biodiesel production since they can be grown without competition with food production, have an inherently fast growth rate, and can have a high lipid content under different nutrient limiting conditions. However, large scale production will best be carried out with indigenous strains, well adapted to local conditions. This study reports on the characterization of the novel microalga Chlorella sp. PCH90, isolated in Quebec. Its molecular phylogeny was established and lipid production studies as a function of the initial concentrations of nitrate, phosphate, and sodium chloride were carried out using response surface methodology. Under the appropriate conditions this microalga could produce up to 36% lipid and grew well in both synthetic medium and secondary effluent from a wastewater treatment plant at both 22 and 10 °C. Thus, this strain is promising for further development as a potential biofuels producer under local climatic conditions.

Hydrogen production from glucose via single-stage photofermentation was examined with the photosynthetic bacterium Rhodobacter capsulatus JP91 (hup-). Response surface methodology with Box–Behnken design was used to optimize the independent experimental variables of glucose concentration, glutamate concentration and light intensity, as well as examining their interactive effects for maximization of molar hydrogen yield. Under optimal condition with a light intensity of 175 W/m 2 , 35 mM glucose, and 4.5 mM glutamate, a maximum hydrogen yield of 5.5 (±0.15) mol H 2 /mol glucose, and a maximum nitrogenase activity of 246 (±3.5) nmol C 2 H 4 /ml/min were obtained. Densitometric analysis of nitrogenase Fe-protein expression under different conditions showed significant variation in Fe-protein expression with a maximum at the optimized central point. Even under optimum conditions for hydrogen production, a significant fraction of the Fe-protein was found in the ADP-ribosylated state, suggesting that further improvement in yields might be possible.

Biological hydrogen production presents a possible avenue for the large scale sustainable generation of hydrogen needed to fuel a future hydrogen economy. Amongst the possible approaches that are under active investigation and that will be briefly discussed; biophotolysis, photofermentation, microbial electrolysis, and dark fermentation, dark fermentation has the additional advantages of largely relying on already developed bioprocess technology and of potentially using various waste streams as feedstock. However, the major roadblock to developing a practical process has been the low yields, typically around 25%, well below those achievable for the production of other biofuels from the same feedstocks. Moreover, low yields also lead to the generation of side products whose large scale production would generate a waste disposal problem. Here recent attempts to overcome these barriers are reviewed and recent progress in efforts to increase hydrogen yields through physiological manipulation, metabolic engineering and the use of two-stage systems are described.

A future hydrogen economy requires the sustainable production of renewable hydrogen. One method to produce hydrogen from stored renewable energy could be through reformation of bioethanol. However, chemically catalyzed reformation processes, although well studied, still present a number of significant technical challenges. Here, bioreformation of ethanol to hydrogen by photofermentation with the photosynthetic bacterium Rhodopseudomonas palustris is described. Cultures were shown to tolerate up to 2% ethanol. An RSM (response surface methodology) was carried out in which three key factors, light intensity, and ethanol and glutamate concentrations were varied. The results showed that nearly 2mol of H2 could be obtained from one mole of ethanol, 33% of that theoretically possible.

Photofermentation by the photosynthetic bacterium Rhodopseudomonas palustris has been used to convert the crude glycerol fraction from biodiesel production to hydrogen as a means of converting this large resource to useful energy. In the present study response surface methodology was applied to investigate the interactive effects among several important process parameters; light intensity, and the concentrations of crude glycerol and glutamate, on the stoichiometric conversion of crude glycerol to hydrogen. Under optimal conditions, a light intensity of 175 W/m 2 , 30 mM glycerol, and 4.5 mM glutamate, 6.69 mol hydrogen/mole of crude glycerol were obtained, a yield 96% of theoretical. Determination of nitrogenase activity and expression levels showed that there was relatively little variation in levels of nitrogenase protein with changes in process variables whereas nitrogenase activity varied considerably, with maximal nitrogenase activity (228 nmol of C 2 H 4 /ml/min) at the optimal central point.