Anthropogenic activities have transformed the global geochemical cycling of heavy metals (HMs). Many physical, chemical and biological methods are used to reduce the toxicity of HMs to humans, plants and environment. This study aimed to investigate the immobilization and phytotoxicity reduction of HMs after application of bamboo biochar (BB) in mine-polluted soil in Feng county (FC) and Tongguan (TG). The results showed that BB application to contaminated soil immobilized HMs (Zn, Pb, Cd and Cu). The soil pH and EC increased and the bioavailability of HMs decreased in FC and TG, whereas Pb and Cu increased in TG soil. The addition of BB reduced HMs uptake in the shoot/root of Brassica juncea. Physiological responses showed that BB application improved the shoot/root growth, dry biomass, and enhanced the chlorophyll (a and b) and carotenoid concentrations in Brassica. The incorporation of BB improved the soil health and accelerated enzymatic activities (β-glucosidase, alkaline phosphatase and urease) in HMs polluted soils. Antioxidant activities (POD, PPO, CAT and SOD) were also used as biomarkers to determine the negative effects of HMs on the growth of Brassica. Overall, the immobilization potential and phytotoxicity reduction of HMs were confirmed by BCF, TF and MEA for both soils.