A new fermentative hydrogen-producing strain FS2011 was isolated from an effluent of bio-hydrogen production reactor, and identified as Bacillus amyloliquefaciens on the basis of 16S rDNA gene sequence. The strain could utilize various carbon and nitrogen sources to produce hydrogen in a broad range of initial pH (5.29-7.38). Phosphate buffer concentration and fermentation temperature significantly affected hydrogen production and cell growth. The maximum hydrogen yield of 2.26 mol/mol was observed at glucose concentration of 10 g/l, beef extract concentration of 2 g/l initial pH 6.98, phosphate buffer of 20 mmol/l, and 35 degrees C, indicating FS2011 was a high-efficiency hydrogen-producing bacterium. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.