When the mycelium of Streptomyces griseus HUT 6037 was suspended in 0.5% NaCl solution containing 14C-glucosamine, peptidoglycan precursors accumulated in the cells. While UDP-7Vacetylglucosamine accumulated in the largest amount among the precursors, extracellularly added and intracellularly accumulated UDP-N-acetylglucosamine were not used to synthesize streptomycin and were probably used for peptidoglycan formation. On the other hand, correlation was recognized between accumulation of glucosamine-6- phosphate (GlcN-6P) and treptomycin formation. Addition of an inhibitor of peptidoglycan synthesis such as enduracidin, vancomycin or cycloserine to a mycelium-suspended culture changed the ratio of accumulated peptidoglycan precursors. When streptomycin formation was stimulated by addition of enduracidin or vancomycin, intracellular GlcN-6P remarkably increased and then decreased rapidly. On the contrary, when cycloserine was added to the culture, no increase ofGlcN- 6P was observed and streptomycin formation was not stimulated. These results suggest that an increase in the intracellular concentration of GlcN-6P is required for activation or induction of the system for utilizing GlcN-6P for streptomycin formation.