Isolation and preliminary characterization of an extracellular protease of Cytophaga sp.

Abstract

The extracellular protease(s) of Cytophaga sp. is present naturally as a complex with acidic polysaccharides of the slime layer. The protein was separated from the slime by selective precipitation of the polysaccharide with a cationic detergent and the enzyme was purified about 100-fold by precipitation with alcohol and by chromatography on DEAE-cellulose. Inhibitor studies indicated that this alkaline protease was not of the trypsin/chymotrypsin type. Ca2+ had a strong stabilizing effect on the enzyme. The complex of basic protein (enzyme) and acidic slime polysaccharides has led to the conclusion that the enzyme is perhaps not truly extracellular but rather is surface bound. The slime layer would appear to have a role in attaching the bacterial cells to insoluble substrates as well as ensuring that degradative enzymes are kept in close contact with the substrate.