Purification and characterization of an endo-1,4-beta-glucanase from Neisseria sicca SB that hydrolyzes beta-1,4 linkages in cellulose acetate.
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.
Moriyoshi
Kunihiko
K
Department of Biochemistry, Osaka Municipal Technical Research Institute, Japan.
Ohmoto
Takashi
T
Ohe
Tatsuhiko
T
Sakai
Kiyofumi
K
D002480:Cellulase / Q000737:chemistry* / Q000302:isolation & purification; D002482:Cellulose / Q000031:analogs & derivatives* / Q000378:metabolism*; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D009343:Neisseria / Q000201:enzymology*; D009844:Oligosaccharides / Q000737:chemistry; D012988:Soil Microbiology; D013379:Substrate Specificity; D013696:Temperature
D003470:Culture Media; D009844:Oligosaccharides; C005062:acetylcellulose; D002482:Cellulose; D004950:Esterases; D002480:Cellulase