A hydrogen-producing strain of bacteria suitable for immobilization was isolated from anaerobic sludge obtained from a methane fermentation plant. The isolated strain, CFPA-20 was identified as a novel species of the genus Clostridium by phylogenetic analysis of the 16S rRNA sequence. The changes in free energy of interaction of adhesion to polymer resin and self-aggregation were both negative. This indicated that CFPA-20 was thermodynamically favored for immobilization. CFPA-20 grew at a temperature range of 25-37 degrees C and at a pH range of 4.5-9.0. Immobilization of CFPA-20 on block copolymer polyethylene glycol-b-polypropylene glycol gave a radically improved hydrogen production yield (2.91 mol/mol-glucose) and a maximum hydrogen production rate (568 mL/L-culture/h) compared to the non-immobilized isolate. In addition, the biofilm of CFPA-20 acquired tolerance for volatile fatty acids. Further investigation into this mechanism may ultimately improve the hydrogen production capacity of CFPA-20. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.