Mercury (Hg) and arsenic (As) are toxic and harmful heavy metals, with exceedance rates of 1.6% and 2.7%, respectively, in soils across China. Compared to soils contaminated with Hg or As alone, co-contaminated soils pose complex environmental risks and are difficult to remediate. Biochar is widely used as a soil amendment to adsorb and immobilize pollutants such as heavy metals. However, only a few studies have explored the efficiency of biochars produced from different crop straws to reduce the bioavailability of heavy metals in co-contaminated soils, and the effects on soil biological properties are often overlooked. The aim of this study was to investigate changes to the physicochemical properties, enzyme activities, and heavy metal bioavailability of an industrial soil co-contaminated with Hg and As upon the addition of different biochars from reed, cassava, and rice straws (REB, CAB, and RIB, respectively). The soil was amended with 1% biochar and planted with spinach in pots for 30 days. RIB was more effective than REB and CAB in increasing the soil pH, organic matter content, and cation exchange capacity. RIB and CAB exhibited similar positive effects on the soil dehydrogenase, catalase, invertase, and urease activities, which were higher than those of REB. The exchangeable fraction of both metals decreased upon biochar addition, and the residual fraction showed the opposite trend. All biochar amendments reduced the bioconcentration factors of heavy metals (especially Hg) in plants and decreased the metal bioavailability in soil. RIB is the optimal amendment for the stabilization of soil co-contaminated with Hg and As.