cellulase and hydroxyethylcellulose

The production of endo-beta-1,4-glucanase by a Bacillus strain isolated from a hot spring in Zimbabwe was studied in batch culture, chemostat culture, and carbon dioxide-regulated auxostat (CO2-auxostat). The bacteria produced the enzyme in the presence of excess glucose or sucrose, but not under carbon-limited conditions in a chemostat using mineral medium. There was a specific growth rate dependent linear increase in enzyme production in glucose excess, nitrogen-limited chemostat cultures. A high specific growth rate of 2.2 h-1 and a high rate of enzyme production of 362 nkat (mg dry mass.h)-1 were attained under nutrient rich conditions in the CO2-auxostat. The bacteria had the highest specific growth rate and endo-beta-1,4-glucanase enzyme production at 50 degrees C. The maximum specific growth rate and the rate of enzyme production increased when yeast extract and tryptone were added in increasing amounts to the mineral medium used for cultivation in separate experiments. Increasing the glucose concentration in the CO2-auxostat cultures increased the rate of enzyme production but did not affect the specific growth rate.

Topics: Bacillus; Bacterial Proteins; Carbon Dioxide; Cell Division; Cellulase; Cellulose; Culture Media; Environment; Glucose; Peptones; Sucrose; Temperature; Zimbabwe

New soluble chromogenic substrates were prepared for specific and rapid assays of endo-1,4-beta-xylanases and endo-1,4-beta-glucanases. A soluble beechwood 4-O-methyl-D-glucurono-D-xylan was dyed with Remazol brilliant blue R, and hydroxyethylcellulose was coupled to Ostazin brilliant red H-3B. The assays are based on photometric measurements of the enzyme-released dyed fragments soluble in the presence of organic solvents which precipitate the original substrates and their high-molecular-weight fractions. The assays are advantageous for rapid analyses of large amount of samples and also permit evaluation of the activities of both enzymes in the presence of exo-beta-glycanases and beta-glycosidases, at a high level of reducing compounds and viable cells, on the cell surface and on cell membranes and organelles.

Topics: Aspergillus niger; Cellulase; Cellulose; Chromogenic Compounds; Coloring Agents; Endo-1,4-beta Xylanases; Glycoside Hydrolases; Solubility; Substrate Specificity; Trichoderma; Xylans

A simple, highly sensitive zymogram technique for detection of endo-1,4-beta-glucanases and endo-1,4-beta-xylanases in polyacrylamide gels after electrophoresis or isoelectric focusing was developed. The detection employs transparent agar replicas containing soluble covalently dyed polysaccharides, hydroxyethylcellulose dyed with Ostazin brilliant red H-3B and beechwood 4-O-methyl-D-glucurono-D-xylan dyed with Remazol brilliant blue R, as the respective substrates. The high sensitivity of the detection is achieved by selective removal of depolymerized dyed substrates from the agar replicas by solvents which neither solubilize nor precipitate the original nondegraded dyed polysaccharides present in the agar gel.

Topics: Aspergillus niger; Cellulase; Cellulose; Chemical Phenomena; Chemistry; Coloring Agents; Electrophoresis, Polyacrylamide Gel; Endo-1,4-beta Xylanases; Glycoside Hydrolases; Hydrogen-Ion Concentration; Isoelectric Focusing; Substrate Specificity; Trichoderma; Xylans