In this study, a microalga Tetraselmis subcordiformis (synonym: Platymonas subcordiformis)based photoanode was prepared by a novel method developed in our lab. The optimal photocurrent density of microalgae photoanode, 37 mu A/cm(2), was achieved under illumination of 145 mu mol s(-1) m(-2) at anode potential of 0.5 V vs Ag vertical bar AgCl vertical bar sat. KCl, immobilized cell density of 2.08 x 10(6)/cm(2) and BQ concentration of 300 mu mol/L. The results of measurements showed that oxygen evolution peak, hydrogen evolution peak and photocurrent response were all synchronous to light impulse in a three-electrode system. It revealed that there occurred a process of photo-bioelectrochemical water-splitting. Hydrogen can be produced by the method. The investigation for whole photo-bioelectrochemical process also indicated that the electrons for hydrogen evolution had two sources, microalgal metabolic process in dark condition and photosynthetic water oxidation. The photo-hydrogen evolution was twice more than hydrogen evolution in dark condition. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.