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The role of protease produced by a heterotrophic bacterium during growth was investigated withAeromonas salmonicida , the pathogen of fish furunculosis, strain A-7301 and its protease-deficient mutant NTG-1 induced by mutagenesis. Strain A-7301 produced extracellular protease in a mixed amino acid medium (composed of Gly, Ala, Val, Ile, Leu, Thr, Ser, Cys, Met, Phe, Tyr, Lys, Arg, Pro, His, Try, Asp, Asn, Glu, and Gln at equal concentrations of 0.1 g/liter). Its multiplication rate was limited by the amounts of amino acids present, whereas strain NTG-1 showed no protease production despite considerable growth similar to that of A-7301. There was no difference between A-7301 and NTG-1 in amino acid requirements for growth, and seven amino acids (Gly, Ala, Val, Thr, Cys, Met, and His) were found to be indispensable. A defined level of the mixed amino acids (0.4 to 0.5 g/liter) was needed for A-7301 to initiate a large production of protease. Neither of the strains grew well in a casein medium, to which no amino acids were added. However, when a protease fraction obtained from extracellular products of A-7301 by DEAE-cellulose column chromatography was added, NTG-1 successfully reproduced in the casein medium. These results indicate that the extracellular protease plays an important role in supplyingA. salmonicida cells with available amino acids as nutrients and that higher growth is closely associated with protease production which stimulates further reproduction.

Significance of Extracellular Protease for Growth of a Heterotrophic Bacterium, Aeromonas salmonicida