A fast growing strain of Chlorella saccharophila was isolated from the marine water of New Zealand and grown in heterotrophic conditions using glucose or glycerol as a carbon source. Biomass production was found to be higher in culture fed with glucose (2.14 +/- 0.08 g L-1) as compared to glycerol (0378 +/- 0.04 g L-1). Lipid accumulation was similar for both carbon sources, at approximately 22% of dry cell weight. However, carotenoid yield was higher with glycerol (0.406 +/- 0.0125 mg g(-1)) than with glucose (0.21 +/- 0.034 mg g(-1)). Further optimization of the growth of the isolate gave maximal carotenoid production of 16.39 +/- 1.19 mg g(-1), total carotenoid, containing 11.32 +/- 0.64 mg g(-1), zeaxanthin and 5.07 +/- 0.55 mg g(-1) beta-carotene. Comparison of various chemical and physical carotenoid extraction methods showed that ultrasonication was required for maximum extraction yields. The new strain has potential for biofuel, with carotenoid co-production. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.