Cellulolytic activity of Thermomonospora curvata: optimal assay conditions, partial purification, and product of the cellulase.
Thermomonospora curvata produces cellulases active against both cotton fibers (designated C(1) activity) and carboxymethylcellulose (C(x) activity). In reaction systems employing optimal substrate concentration, pH, and temperature, hydrolysis rates (measured by the release of soluble reducing sugars) were initially linear and decreased on prolonged incubation, although only a small amount of substrate (1 to 2%) had been hydrolyzed. Persistence of this lower rate, even after addition of fresh enzyme (in the C(1) assay system), indicated alteration of cellulose susceptibility to hydrolysis rather than enzyme inactivation. Partial purification by (NH(4))(2)SO(4) precipitation and exclusion chromatography resolved cellulase activity into two fractions. The sole product of purified cellulase activity on ground cotton fibers appears to be cellobiose.
D000192:Actinomycetales / Q000201:enzymology / Q000378:metabolism*; D000645:Ammonium Sulfate; D002474:Cell-Free System; D002482:Cellulose / Q000378:metabolism*; D011232:Chemical Precipitation; D002845:Chromatography; D002854:Chromatography, Paper; D002855:Chromatography, Thin Layer; D004187:Disaccharides / Q000032:analysis / Q000096:biosynthesis*; D006026:Glycoside Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D003368:Gossypium / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008747:Methylcellulose / Q000378:metabolism; D013696:Temperature
D004187:Disaccharides; D002482:Cellulose; D008747:Methylcellulose; D006026:Glycoside Hydrolases; D000645:Ammonium Sulfate