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. (C) 2010 Elsevier Ltd. All rights reserved.