The aim of this study was to explore the utilization of an intractable waster of Eichhornia crassipes in laccase production by Pycnoporus sanguineus SYBC-L1. E. crassipes as the sole carbon and nitrogen source was confirmed to produce laccase (7.26 U/g dry substrate). The fermentation medium for the maximum enzyme production was optimized and the laccase was then purified and characterized. The optimized culture medium was 25.1% E. crassipes, 13.9% sawdust, 1.5 mM CuSO4, 40 μM gallic acid, 65% moisture content and initial pH 6.0. A maximum laccase activity of 32.02 U/g dry substrate was detected at 9th day, which was 4.5-fold compared to the initial medium. The molecular mass of the purified Lac-S was 58.4 kDa. The optimum activity of Lac-S for DMP was at pH 3.0 and 70°C. Lac-S showed not only high catalytic activities at low temperature, but also good stabilities toward pH and temperature. The residual catalytic activities of Lac-S were 30%, 40% and 50% at 0°C, 10°C and 20°C, and the half-lives at 50°C, 60°C and 70°C were 21.7, 9.7 and 1.5 h, respectively. The results provide a significant basis for E. crassipes further utilization and Lac-S specific application in harsh industry.