Objective:Synthesis of L-2,6-dimethylphenylalanine and L-2,4,6-trimethylphenylalanine through enzymatic resolution. Methods:Racemic 2,6-dimethylphenylalanine and 2,4,6-trimethylphenylalanine were synthesized through reaction of diethyl acetamidomalonate with 2,6-dimethylbenzyl bromide or 2,4,6-trimethylbenzyl bromide,respectively. The DL-amino acids were then enzymatically resoluted by L-amino acylase. Results:Racemic 2,6-dimethylphenylalanine and 2,4,6-trimethylphenylalanine were synthesized through reaction of diethyl acetamidomalonate with 2,6-dimethylbenzyl bromide and 2,4,6-trimethylbenzyl bromide in yield of 87.9% and 86.0% respectively;DL-2,6-dimethylphenylalanine was successfully resoluted by L-amino acylase,but it was failed in the resolution of 2,4,6-trimethylphenylalanine. Conclusion:L-amino acylase can be used in the resolution of DL-2,6-dimethylphenylalanine,but it is ineffective in the resolution of 2,4,6-trimethylphenylalanine.