For the resolution of L-carnitine from DL-carnitine, resting cells of Enterobacter sp. NH-104, which had a higher capacity of D-carnitine decomposition, were harvested at maximal specific activity of D-carnitine decomposition of 47.05 unit/mg cell. The cells were frozen at $-80^{circ}C$ to assess functions as enzyme sources. Optimal concentration of cells and DL-carnitine were 17 g/$ell ; and ; 20 g/ell$, respectively, and reaction buffer was best at 75 mM of Tris. HCl. Optimal temperature and pH were $36^{circ}C$ and 8.2, respectively. When the reaction at optimal conditions was carried out for 14 h, the optical purity was 98.21 %, and the quantity and yield of remaining L-carnitine were 4.432 g/<REX>$ell$</TEX> and 44.32%, respectively.