Purification and characterization of an extracellular endoglucanase from the marine shipworm bacterium.

Bacterial cultures isolated from the gland of Deshayes of marine shipworm (Psiloteredo healdi) produced extracellular endoglucanase activity when cultured with 1% cellulose. An endoglucanase of subunit relative molecular mass 58,000, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was purified to homogeneity from cell-free culture medium. Similarly, the relative molecular mass of the native enzyme was 60,100 as determined by gel permeation chromatography. No carbohydrate appeared to be associated with the purified protein. The action of the purified enzyme on various cellodextrins was also studied. Only interior glucosyl linkages of cellodextrin chains larger than cellotriose were cleaved by the enzyme and the centermost bond of cellohexaose was preferentially cleaved. The Km values of the purified endoglucanase were 0.12 mM for cellotetraose, 0.05 mM for cellopentaose, and 0.11 mM for cellohexaose. Glucose, cellobiose, and cellotriose did not inhibit enzymatic activity.