Aims: Most of the researches investigating immobilized fungi in chromate [Cr(VI)] bioremediation have used dead cells to adsorb Cr(VI). Therefore, the aim was to identify a Cr(VI)-reducing fungus with the ability of reducing the toxic Cr(VI) into the much less toxic Cr(III) and to apply the immobilized living fungus in continual reduction of Cr(VI). Methods and Results: Cr(VI) reduction occurred using both free fungi and immobilized living Rhizopus sp. LG04. The Cr(VI) bioreduction by the free fungi was achieved mainly by bioreduction coupled with a small amount of biosorption on the cell surfaces. LG04 spores immobilized with 3% polyvinyl alcohol and 3% sodium alginate produced the most stable and efficient biobeads. When the LG04 biobeads were washed and transferred into fresh medium containing 42 mg l(-1) of Cr(VI), the biobeads could be reused to reduce Cr(VI) for more than 30 cycles during an 82-day operation period. Interestingly, as the cycles increased, the time required for complete reduction stabilized at approximately 2 5 days, which was faster than that obtained using the free fungi (4 5 days). The pH value of the solution decreased from 6 60 +/- 0 10 to 3 85 +/- 0 15 after each reduction cycle, which may be because the metabolic products of the fungus changed the environmental pH or because there was an accumulation of the organo-Cr(III) complex. Conclusions: The results indicate that using the immobilized living fungus for the removal of Cr(VI) has the advantages in being stable, long-term treatment, easy to re-use and less biomass leakage. Significance and Impact of the Study: To our knowledge, this study reports the first successful use of immobilized living Rhizopus for the repeated reduction of Cr(VI).