A novel strategy for improving volatile fatty acids (VFAs) bioproduction from waste activated sludge (WAS) was developed by coupling pretreatments with conditioning (CPC). Agaricus bisporus substrate (ABS) was used as external carbohydrate additive source of conditioning step. Pretreatment was studied in three ways: alkaline, alkaline-thermal and ultrasonic. WAS hydrolysis and protein degradation was distinctly improved by CPC treatments, resulting in a considerable enhancement of VFAs yield. The maximal VFAs yield was 614 +/- 71,712 +/- 49 and 598 +/- 19 mg COD/gVSS at pre-optimized alkaline, alkaline-thermal and ultrasonic CPC treatments, respectively, with an increase of 35%, 50% and 38% compared to the yields of pretreated WAS fermentation. Fourier transformed infrared spectroscopy and three-dimensional excitation-emission matrix fluorescence spectroscopy indicated that a synergistic effect occurred in co-digesting WAS and ABS. The conditioning of carbohydrate with feasible pretreatment provided large room for the digestibility improvement and the operation cost reduction in the whole WAS treatment system. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.