algae to biofuels:
from strain isolation
to strain engineering

Algae to biofuel conversion research at LEAP currently concentrates on the following areas:

1. Isolation of new microalgae strains and screening for use in biofuels research.
2. Improving the productivity of mass cultures.
3. Metabolism of hydrocarbons: Dunaliella.

Isolation of new microalgae strains by LEAP will help development towards the production of oil and the production of hydrocarbons. The labor-intensive procedure to isolate as many unicellular strains as possible begins with a dual spatial and temporal approach, sampling from a variety of habitats (e.g., freshwater, brackish, marine, hypersaline) in a wide range of locations including New York, Salton Sea, the Great Salt Lake, and the southwestern states of the United States at different times throughout the year. Aliquots from enrichment samples are directly plated before being transferred in series to index plates. Unialgal strains are then isolated on individual plates.

Samples from various locations are transferred in the lab to plates. Samples are further transferred to index plates and then unialgal strains are separated onto individual plates.

In addition to the traditional isolation method, a cell sorter is used for high-throughput isolation of strains.

Shown is the BD FACSAria System II cell sorter operated by Dr. Duc Tran and Mr. Sixto Portilla.

To check for lipid content, the screening process uses fluorescent dyes. Nile red fluorescence is used to detect triacylglyceride.

TAG detection: Chlorophycea, Dunaliella salina

TAG detection: Chlorophycea, Haematococcus pluvialis

TAG detection: Chlorophyceae, Haematococcus pluvialis

Using a plate reader, a TAG screening is performed to obtain analytical data.

acknowledgements

Funding for this research provided by the Air Force Office of Scientific Research (AFOSR) and the United States Department of Energy (DOE).

Some water samples have been provided as part of a collaboration with the Boy Scouts of America and cadets in the Sea-Legs sailing program.