The production of polyhydroxyalkanoates (PHAs) by Bacillus tequilensis biocatalyst using spent wash effluents as substrate was evaluated to increase the versatility of the existing PHA production process and reduce production cost. In this study, spent wash was used as a substrate for biohydrogen (H-2) production and the resulting acidogenic effluents were subsequently employed as substrate for PHA production. Maximum H-2 production of 39.8 L and maximum PHA accumulation of 40% dry cell weight was attained. Good substrate removal associated with decrement in acidification (53% to 15%) indicates that the VFA generated were effectively utilized for PHA production. The PHA composition showed presence of copolymer [P (3HB-co-3HV)] with varying contents of hydroxybutyrate and hydroxyvalerate. The results obtained suggest that the use of spent wash effluents as substrate can considerably reduce the production cost of PHA with simultaneous waste valorization. PHA synthesis with B. tequilensis and spent wash effluents is reported for the first time. (C) 2014 Elsevier Ltd. All rights reserved.