The influence of variable concentrations of the nutritive components of Gauze 1 medium on the β-lactamase synthesis by Streptomyces galbus (F) subsp. achromogenes—p154–59 has been studied. Twelve carbon, ten nitrogen, and four phosphate sources have been examined and the data has been used for carrying out Complete Factor Experiment (CFE 23). As a result of our experiments a complete optimisation of the fermentation medium has been obtained which enhances ten-fold β-lactamase synthesis by Streptomyces galbus.