Microbial biodiesel production by direct methanolysis of oleaginous biomass.

Biodiesel is usually produced by the transesterification of vegetable oils and animal fats with methanol, catalyzed by strong acids or bases. This study introduces a novel biodiesel production method that features direct base-catalyzed methanolysis of the cellular biomass of oleaginous yeast Rhodosporidium toruloides Y4. NaOH was used as catalyst for transesterification reactions and the variables affecting the esterification level including catalyst concentration, reaction temperature, reaction time, solvent loading (methanol) and moisture content were investigated using the oleaginous yeast biomass. The most suitable pretreatment condition was found to be 4gL(-1) NaOH and 1:20 (w/v) dried biomass to methanol ratio for 10h at 50°C and under ambient pressure. Under these conditions, the fatty acid methyl ester (FAME) yield was 97.7%. Therefore, the novel method of direct base-catalyzed methanolysis of R. toruloides is a much simpler, less tedious and time-consuming, process than the conventional processes with higher FAME (biodiesel) conversion yield.

Microbial oil produced from biodiesel by-products could enhance overall production.

Glycerol and rapeseed meal, two major by-products of biodiesel production, have been tested for possible use as low-cost raw materials for the production of microbial bio-oil using the oleaginous yeast Rhodosporidium toruloides. Using fed-batch fermentation with crude glycerol and a novel nitrogen rich nutrient source derived from rapeseed meal as feed, it was shown that 13 g/L lipids could be produced, compared with 9.4 g/L when crude glycerol was used with yeast extract. When 100 g/L pure glycerol was used, the final lipid concentration was 19.7 g/L with the novel biomedium compared to 16.2 g/L for yeast extract. The novel biomedium also resulted in higher lipid yields (0.19 g lipid/g glycerol consumed compared to 0.12 g/L) suggesting it provides a better carbon to nitrogen balance for accumulating lipids. FAMEs produced from the microbial lipids indicated a high degree of unsaturation confirming that the fatty acids produced from the novel biomedium have potential for biodiesel production.

Enhancing the value of nitrogen from rapeseed meal for microbial oil production.

Rapeseed meal, a major byproduct of biodiesel production, has been used as a low-cost raw material for the production of a generic microbial feedstock through a consolidated bioconversion process. Various strategies were tested for the production of a novel fermentation medium, rich in free amino nitrogen (FAN): commercial enzymes (CEs) (2.7 mg g⁻¹ dry meal), liquid state fungal pre-treatment (LSF) using Aspergillus oryzae (4.6 mg g⁻¹), liquid state fungal pre-treatment followed by fungal autolysis (LSFA) (9.13 mg g⁻¹), liquid state pre-treatment using fungal enzymatic broth (EB) (2.1 mg g⁻¹), but the best strategy was a solid state fungal pre-treatment followed by fungal autolysis (34.5 mg g⁻¹). The bioavailability of the nitrogen sources in the novel medium was confirmed in fed-batch bioreactor studies, in which 82.3g dry cell L⁻¹ of the oleaginous yeast Rhodosporidium toruloides Y4 was obtained with a lipid content of 48%. The dry cell weight obtained was higher than that obtained using conventional yeast extract, due to a higher total nitrogen content in the novel biomedium. The fatty acids obtained from the microbial oil were similar to those derived from rapeseed oil.