A number of tryptophan-plus-tyrosine double auxotrophs have been isolated from a glutamate producing Arthrobacter globiformis excreting L-phenylalanine by two-step mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine. For the three potent mutants tested the medium of Alföldi was found to be the best. The optimum tryptophan, tyrosine and biotin concentrations for phenylalanine production of these mutants were 0.5 mmol/L, 0.1 mmol/L and 5 micrograms/L, respectively. At these levels strain TT-39 yielded 2.6 g phenylalanine per L of medium in flask culture with glucose (350 mmol/L) and NH4Cl (60 mmol/L).