Chlamydomonas reinhardtii possesses many potential advantages to be exploited as a biocatalyst in microbial fuel cells (MFCs) for electricity generation. In the present study, we performed computational studies based on flux balance analysis (FBA) to probe the maximum potential of C. reinhardtii for current output and identify the metabolic mechanisms supporting a high current generation in three different cultivation conditions, i.e., heterotrophic, photoautotrophic and mixotrophic growth. The results showed that flux balance limitations allow the highest current output for C. reinhardtii in the mixotrophic growth mode (2.368 A/gDW), followed by heterotrophic growth (1.141 AlgDW) and photoautotrophic growth the lowest (0.7035 A/gDW). The significantly higher mediated electron transfer (MET) rate in the mixotrophic mode is in complete contrast to previous findings for a photosynthetic cyanobacterium, and was attributed to the fact that for C. reinhardtii the photophosphorylation improved the efficiency of converting the acetate into biomass and NADH production. Overall, the cytosolic NADH-dependent current production was mainly associated with five reactions in both mixotrophic and photoautotrophic nutritional modes, whereas four reactions participated in the heterotrophic mode. The mixotrophic and photoautotrophic metabolisms were alike and shared the same set of reactions for maximizing current production, whereas in the heterotrophic mode, the current production was additionally contributed by the metabolic activities in the two organelles: glyoxysome and chloroplast. In conclusion, C. reinhardtii has a potential to be exploited in MFCs of MET mode to produce a high current output. (C) 2014, The Society for Biotechnology, Japan. All rights reserved.