D-Arabitol was ﬁrst prepared from n-glucose using Candida famnta R28. The reaction gave 5.0% D-arabitol from 10.0% r>-glucose. D-Arabitol was then almost completely converted to D-xylulose using Acetobacter aceti IFO 3281. Finally, D-lyxose was prepared from D-xylulose enzymatically using L-ribose isomerase from toluene-treated cells of Acinetobucter sp. strain DL-28. The isomerization reaction progressed steadily and the concentration of D-xylulose increased from 1.0 to 10.0%. About 70% of D-xylnlose was converted to D-lyxose in all cases. Separation of residual D-xylulose from the reaction mixture is very difficult to achieve by column chromatography, but n-xylulose could be selectively degraded easily using Saccharomyces cerevisiae IFO 0841.The product was crystallized and was conﬁrmed to be D-lyxose by HPLC, C-NMR spectra, IR spectra analysis, and optical rotation measurement.