Isolation and characterization of an alkaline protease from the marine shipworm bacterium
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Bacterial isolates from the gland of Deshayes of the marine shipworm (Psiloteredo healdi) produced extracellular protease activity when cultured with 1% cellulose. A protease with a relative molecular mass of 36,000 daltons as determined by SDS-PAGE and a pI of 8.6 was isolated from the medium and purified to electrophoretic homogeneity. No carbohydrate appeared to be associated with the protein. The enzyme was activated and stabilized by relatively high salt concentrations (>0.2M). Below 0.1M salt, significant protein aggregation occurred, as well as autohydrolysis of the protease, both of which resulted in the loss of activity. The specific activity of the enzyme was 65,840 proteolytic units/mg with azocasein substrate of optimal temperature (42°C), pH (9.0), and salt concentration (0.20M NaCl). The activity was stable up to 40°C, from pH 3.0 to pH 11.9, and from 0.1M to 3.5M NaCl. These stabilities, as well as the protease's stability in the presence of chelators, oxidizing agents, and heavy metals, suggest the enzyme has potential for use in relatively low temperature (40°C) industrial applications.
KeywordsEnzyme Cellulose Heavy Metal Molecular Mass Salt Concentration
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