Synthesis of sucrose-6-acetate in non-aqueous medium from sucrose using lipase was studied.The transesterification conditions including reaction medium,acyl donor,enzyme,water content and temperature were investigated.Results showed that immobilized-lipase Lipozyme TLIM could effectively catalyzed sucrose and vinyl acetate to produce sucrose-6-acetate in the medium composing of DMSO and tert-butyl alcohol.A mole yield of 89.3% was reached when the transesterification was performed as follows;volume ratio of DMSO to tert-butyl alcohol 1:4,mole ratio of vinyl acetate to sucrose 10:1,concentration of sucrose 0.06 mmoL/mL,concentration of,enzyme 80 mg/mL,reaction temperature of 30 ℃ and reaction time of 9 h.The structure of the products was also characterized as sucrose-6-acetate by FTIR and NMR.