Two addnl. strains (KT 251 and KT 253) of soil bacteria capable of resolving N-acetyl-DL-tryptophan (I) by their acylase were isolated from soil by using a synthetic medium containing I as the sole source of C.Both strains could be cultivated in various synthetic media with DL-tryptophan, AcOH, or PhCH2CO2H as the sole source of C.The asymmetric hydrolysis of I was carried out as described above for N-benzoyl-DL-threonine.KT 251 yielded 39% L-tryptophan (II) and 57% N-acetyl-D-tryptophan (III), whereas KT 253 yielded 29%, II and 61% III.