cellulase and acetylcellulose

This study was conducted to realize the potential application of cellulose acetate to enzymatic processing, followed by microbial ethanol fermentation. To eliminate the effect of steric hindrance of acetyl groups on the action of cellulase, cellulose acetate was subjected to deacetylation in the presence of 1N sodium hydroxide and a mixture of methanol/acetone, yielding 88.8-98.6% at 5-20% substrate loadings during a 48h saccharification at 50°C. Ethanol fermentation using Saccharomyces cerevisiae attained a high yield of 92.3% from the initial glucose concentration of 44.2g/L; however, a low saccharification yield was obtained at 35°C, decreasing efficiency during simultaneous saccharification and fermentation (SSF). Presaccharification at 50°C prior to SSF without increasing the total process time attained the ethanol titers of 19.8g/L (5% substrate), 38.0g/L (10% substrate), 55.9g/L (15% substrate), and 70.9g/L (20% substrate), which show a 12.0-16.2% improvement in ethanol yield.

Topics: Acetylation; Biotechnology; Carbohydrate Metabolism; Cellulase; Cellulose; Ethanol; Fermentation; Glucose; Saccharomyces cerevisiae; Temperature; Time Factors

An enzyme hydrolyzing beta-1,4 bonds in cellulose acetate was purified 10.5-fold to electrophoretic homogeneity from a culture supernatant of Neisseria sicca SB, which assimilate cellulose acetate as the sole carbon and energy source. The enzyme was an endo-1,4-beta-glucanase, to judge from the substrate specificity and hydrolysis products of cellooligosaccharides, we named it endo-1,4-beta-glucanase I (EG I). Its molecular mass was 50 kDa, 9 kDa larger than EG II from this strain, and its isoelectric point was 5.0. Results of N-terminal and inner-peptide sequences of both enzymes, and a similarity search, suggested that EG I contained a carbohydrate-binding module at the N-terminus and that EG II lacked this module. The pH and temperature optima of EG I were 5.0-6.0 and 45 degrees C. It hydrolyzed water-soluble cellulose acetate (degree of substitution, 0.88) and carboxymethyl cellulose. The Km and Vmax for these compounds were 0.296% and 1.29 micromol min(-1) mg(-1), and 0.448% and 13.6 micromol min(-1) mg(-1), respectively. Both glucanases and cellulose acetate esterase from this strain degraded water-insoluble cellulose acetate synergistically.

Topics: Adsorption; Amino Acid Sequence; Biodegradation, Environmental; Carboxylic Ester Hydrolases; Cellulase; Cellulose; Chromatography, DEAE-Cellulose; Electrophoresis, Polyacrylamide Gel; Glucans; Hydrolysis; Kinetics; Molecular Sequence Data; Neisseria; Sequence Alignment; Sequence Homology, Amino Acid; Substrate Specificity

An enzyme catalyzing hydrolysis of beta-1,4 bonds in cellulose acetate was purified 18.3-fold to electrophoretic homogeneity from a culture supernatant of Neisseria sicca SB, which can assimilate cellulose acetate as the sole carbon and energy source. The molecular mass of the enzyme was 41 kDa and the isoelectric point was 4.8. The pH and temperature optima of the enzyme were 6.0-7.0 and 60 degrees C. The enzyme catalyzed hydrolysis of water-soluble cellulose acetate (degree of substitution, 0.88) and carboxymethyl cellulose. The Km and Vmax for water-soluble cellulose acetate and carboxymethyl cellulose were 0.242% and 2.24 micromol/min/mg, and 2.28% and 12.8 micromol/min/mg, respectively. It is estimated that the enzyme is a kind of endo-1,4-beta-glucanase (EC 3.2.1.4) from the substrate specificity and hydrolysis products of cellooligosaccharides. The enzyme and cellulose acetate esterase from Neisseria sicca SB degraded water-insoluble cellulose acetate by synergistic action.

Topics: Cellulase; Cellulose; Chromatography, DEAE-Cellulose; Chromatography, Ion Exchange; Culture Media; Electrophoresis, Polyacrylamide Gel; Esterases; Hydrogen-Ion Concentration; Hydrolysis; Kinetics; Neisseria; Oligosaccharides; Soil Microbiology; Substrate Specificity; Temperature