The pretreated sugarcane bagasse (SCB) was detoxified by laccase, NaBH4 and washing before anaerobic degradation. The detoxified SCB was used for biohydrogen fermentation by MJ1. The biohydrogen productions from the detoxified samples by laccase and NaBH4 were low (below 7 mM). As for detoxification by washing, the biohydrogen production reached 44 mM, nearly 10 times higher than the non-detoxified sample. Co-culture strategy (DSM 1313 and MJ1) was further adopted to enhance the biohydrogen production. The biohydrogen production of 119.7 mM was reached by optimizing the fermentation conditions, which was 2.7 times higher than monoculture and 25 times higher than non detoxified sample. The biohydrogen productions from detoxified sample indicated that the detoxifications of acid pretreated SCB by laccase and NaBH4 were not suitable for biohydrogen fermentation by MJ1 and washing was an effective detoxification method. Co-culture of DSM 1313 and MJ1 was an efficient fermentation strategy for biohydrogen production from pretreated SCB. The present study provides a new insight into the detoxification research on biohydrogen production. (C) 2018 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.