The hydrogen production utilizing photosynthetic and anaerobic bacteria in two-stage approach has many drawbacks, such as shortage of raw materials and complexity of operations. Accordingly, we aimed to develop a simple one-stage H-2 production protocol using the depolymerization of maize straw cellulose as a cheap carbon source. R. sphaeroides HY01 and its mutant (Hup(-)) were studied regarding their H-2 production under different culture conditions. Further study using two model sugars, their combination, and straw hydrolysate as carbon sources was conducted to determine the effects of substrate on H-2 production. When using the straw hydrolysate as carbon source, the pH remained in a range of 7.1-7.6, whereas it dropped to 5.4-7.4 when using the model sugars, and the former biomass value was greater. The H-2 production performance of the mutant was significantly better than that of HY01. One-step photo-fermentative H-2 production was superior when using straw hydrolysate as opposed to the simple model sugars, and its yield was up to 4.62 mol H-2.mol(-1) reducing sugar. (C) 2018 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.