Flocculation of Candida tropicalis HY200 was systemically investigated to elucidate its mechanism, and used for cell cycles in repeated-batch cultivations for the production of xylitol from xylose. Flocculation occurred only after the late exponential phase of growth in the culture media and buffer within the narrow pH range of 3.0-5.0. The flocculation was completely inhibited by treatments of cells with proteases and partially reduced by treatments with carbohydrate-hydrolyzing enzymes and by the presence of mannose and glucose. The addition of calcium ions significantly enhanced the flocculation during cultivation, which was completely abolished by the addition of EDTA. The flocculent yeast HY200 provided repeated-batch cultivations employing cell recycles by flocculation over 6 rounds of cultivation for the production of xylitol from xylose, resulting in a relatively high productivity of averaged 4.6 g xylitol/l h over six batches and maximal 6.3 g xylitol/l h in the final sixth batch. Cell recycle by flocculation was fast and convenient, which could be applicable for the industrial scale of xylitol production.