Improving energetics of triacylglyceride extraction from wet oleaginous microbes.

Oleaginous microbes can upgrade carbon to lipids, which can be used as a feedstock to produce renewable replacements for petroleum-based compounds. Efficient extraction of lipids from oleaginous microbes typically involves dewatering and drying of the biomass. Problematically, drying often requires an amount of energy approaching that available from the cells. Here, we report an approach for the high efficiency extraction of triacylglycerides (TAG) from wet oleaginous microbes, bypassing the drying process. Solvent candidates for extraction of wet oleaginous biomass were identified using ASPEN's databases to determine an activity based selectivity coefficient. Optimal extraction conditions were determined which resulted in >91% extraction of TAG from yeast, bacteria, and microalgae. Experimental data was integrated into system models to evaluate the energetics of the processes compared to traditional extraction methods. The net energy ratio (NER) of a traditional dry solvent extraction is 0.84, whereas the approach presented here has a NER of 0.34 for yeast.

Characterization of a fatty acyl-CoA reductase from Marinobacter aquaeolei VT8: a bacterial enzyme catalyzing the reduction of fatty acyl-CoA to fatty alcohol.

Fatty alcohols are of interest as a renewable feedstock to replace petroleum compounds used as fuels, in cosmetics, and in pharmaceuticals. One biological approach to the production of fatty alcohols involves the sequential action of two bacterial enzymes: (i) reduction of a fatty acyl-CoA to the corresponding fatty aldehyde catalyzed by a fatty acyl-CoA reductase, followed by (ii) reduction of the fatty aldehyde to the corresponding fatty alcohol catalyzed by a fatty aldehyde reductase. Here, we identify, purify, and characterize a novel bacterial enzyme from Marinobacter aquaeolei VT8 that catalyzes the reduction of fatty acyl-CoA by four electrons to the corresponding fatty alcohol, eliminating the need for a separate fatty aldehyde reductase. The enzyme is shown to reduce fatty acyl-CoAs ranging from C8:0 to C20:4 to the corresponding fatty alcohols, with the highest rate found for palmitoyl-CoA (C16:0). The dependence of the rate of reduction of palmitoyl-CoA on substrate concentration was cooperative, with an apparent K(m) ~ 4 µM, V(max) ~ 200 nmol NADP(+) min(-1) (mg protein)(-1), and n ~ 3. The enzyme also reduced a range of fatty aldehydes with decanal having the highest activity. The substrate cis-11-hexadecenal was reduced in a cooperative manner with an apparent K(m) of ~50 µM, V(max) of ~8 µmol NADP(+) min(-1) (mg protein)(-1), and n ~ 2.

Biodiesel production by simultaneous extraction and conversion of total lipids from microalgae, cyanobacteria, and wild mixed-cultures.

Microalgae have been identified as a potential biodiesel feedstock due to their high lipid productivity and potential for cultivation on marginal land. One of the challenges in utilizing microalgae to make biodiesel is the complexities of extracting the lipids using organic solvents followed by transesterification of the extracts to biodiesel. In the present work, reaction conditions were optimized that allow a single step extraction and conversion to biodiesel in high yield from microalgae. From the optimized conditions, it is demonstrated that quantitative conversion of triglycerides from several different microalgae and cyanobacteria could be achieved, including from mixed microbial biomass collected from a municipal wastewater lagoon. Evidence is presented that for some samples, significantly more biodiesel can be produced than would be expected from available triglycerides, indicating conversion of fatty acids contained in other molecules (e.g., phospholipids) using this approach. The effectiveness of the approach on wet algae is also reported.