β-Glucosidase biosynthesis inThermomonospora curvata

Abstract

β-Glucosidase biosynthesis was induced inThermomonospora curvata during early exponential growth on cellobiose, cellulose and protein-extracted lucerne fibre in mineral salts minimal medium at 53°C. Only about 3% of the total culture β-glucosidase was found in cell-free fluid when the actinomycete was grown on cellobiose or purified cellulose. A variety of non-lytic agents including detergents, salts, proteinase and electroporation failed to release the cellbound enzyme. However, cells grown on the fibre released more than 50% of their β-glucosidase. The maximal amount of extracellular accumulation was dependent on the initial concentration of fibre in the medium. Thermal instability at 53°C was the major cause for low exracellular β-glucosidase activity in cellobiose- and cellulose-grown cultures. In cultures grown on the fibre, the extracellular enzyme was stabilized against thermal denaturation by the composition of the cell-free fluid, but was degraded by transient proteinase activity. Proteolysis decreased the average beta-glucosidase specific activity from about 460mU/mg extracellular protein to about 80mU/mg within 1 day after the appearance of the proteinase.