Biodiesels (fatty acid methyl esters) derived from oleaginous microbes (microalgae, yeast, and bacteria) are being actively pursued as potential renewable substitutes for petroleum diesel. Here, we report the engine performance characteristics of biodiesel produced from a microalgae (Chaetoceros gracilis), a yeast (Cryptococcus curvatus), and a bacteria (Rhodococcus opacus) in a two-cylinder diesel engine outfitted with an eddy current brake dynamometer, comparing the fuel performance to petroleum diesel (#2) and commercial biodiesel from soybeans. Key physical and chemical properties, including heating value, viscosity, density, and cetane index, for each of the microbial-derived biofuels were found to compare favorably to those of soybean biodiesel. Likewise, the horsepower, torque, and brake specific fuel consumption across a range of engine speeds also compared favorably to values determined for soybean biodiesel. Analysis of exhaust emissions (hydrocarbon, CO, CO₂, O₂, and NO_x) revealed that all biofuels produced significantly less CO and hydrocarbon than petroleum diesel. Surprisingly, microalgae biodiesel was found to have the lowest NO_x output, even lower than petroleum diesel. The results are discussed in the context of the fatty acid composition of the fuels and the technical viability of microbial biofuels as replacements for petroleum diesel.

Wax esters are produced in certain bacteria as a potential carbon and energy storage compound. The final enzyme in the biosynthetic pathway responsible for wax ester production is the bifunctional wax ester synthase/acyl-coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS/DGAT), which utilizes a range of fatty alcohols and fatty acyl-CoAs to synthesize the corresponding wax ester. We report here the isolation and substrate range characterization for five WS/DGAT enzymes from four different bacteria: Marinobacter aquaeolei VT8, Acinetobacter baylyi , Rhodococcus jostii RHA1, and Psychrobacter cryohalolentis K5. The results from kinetic studies of isolated enzymes reveal a differential activity based on the order of substrate addition and reveal subtle differences between the substrate selectivity of the different enzymes. These in vitro results are compared to the wax ester and triacylglyceride product profiles obtained from each organism grown under neutral lipid accumulating conditions, providing potential insights into the role that the WS/DGAT enzyme plays in determining the final wax ester products that are produced under conditions of nutrient stress in each of these bacteria. Further, the analysis revealed that one enzyme in particular from M. aquaeolei VT8 showed the greatest potential for future study based on rapid purification and significantly higher activity than was found for the other isolated WS/DGAT enzymes. The results provide a framework to test prospective differences between these enzymes for potential biotechnological applications such as high-value petrochemicals and biofuel production.