Biofuel cells (BFCs) use enzymes and microbial cells to produce energy from bioavailable substrates and treat various wastewaters, and cathodic oxygen reduction is a key factor governing the efficiency of BFCs. In this study, we demonstrated that a green alga, Chlamydomonas reinhardtii, could directly mediate oxygen reduction. Cyclic voltammogram analysis revealed that the C. reinhardtii biofilm formed on a solid electrode was responsible for oxygen reduction without dosing of electron mediator. Furthermore, 4-electron oxygen reduction pathway was found in this self-sustained, light-responded BFC. The results of this study could expand our understanding and viewpoints of biocathode catalysis, which is essential for novel catalyst design and development for BFCs. Biotechnol. Bioeng. 2013; 110: 173–179. © 2012 Wiley Periodicals, Inc.