The bifunctional enzyme chitosanase-cellulase produced by the gram-negative microorganism Myxobacter sp. AL-1 is highly similar to Bacillus subtilis endoglucanases.
The gram-negative bacterium Myxobacter sp. AL-1 produces chitosanase-cellulase activity that is maximally excreted during the stationary phase of growth. Carboxymethylcellulase zymogram analysis revealed that the enzymatic activity was correlated with two bands of 32 and 35 kDa. Ion-exchange-chromatography-enriched preparations of the 32-kDa enzyme were capable of degrading the cellulose fluorescent derivatives 4-methylumbelliferyl-beta-D-cellobioside and 4-methylumbelliferyl-beta-D-cellotrioside. These enzymatic preparations also showed a greater capacity at 70 degrees C than at 42 degrees C to degrade chitosan oligomers of a minimum size of six units. Conversely, the beta-1,4 glucanolytic activity was more efficient at attacking carboxymethylcellulose and methylumbelliferyl-cellotrioside at 42 degrees C than at 70 degrees C. The 32-kDa enzyme was purified more than 800-fold to apparent homogeneity by a combination of ion-exchange and molecular-exclusion chromatography. Amino-terminal sequencing indicated that mature chitosanase-cellulase shares more than 70% identity with endocellulases produced by strains DLG, PAP115, and 168 of the gram-positive microorganism Bacillus subtilis.
Pedraza-Reyes
M
M
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato, Apdo. Postal 187, Guanajuato 36000, Guanajuato, Mexico. pedrama@quijote.ugto.mx
Gutiérrez-Corona
F
F
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000378:metabolism; D002266:Carboxymethylcellulose Sodium / Q000378:metabolism; D002475:Cellobiose / Q000031:analogs & derivatives / Q000378:metabolism; D002480:Cellulase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D002686:Chitin / Q000031:analogs & derivatives / Q000378:metabolism; D048271:Chitosan; D002852:Chromatography, Ion Exchange; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D008969:Molecular Sequence Data; D009231:Myxococcales / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014312:Trisaccharides / Q000378:metabolism
C109060:4-methylumbelliferyl-beta-D-cellotrioside; D014312:Trisaccharides; D002686:Chitin; D002475:Cellobiose; D006923:Hymecromone; C068475:4-methylumbelliferylcellobioside; D002482:Cellulose; D048271:Chitosan; D006026:Glycoside Hydrolases; C019868:chitosanase; D002480:Cellulase; D002266:Carboxymethylcellulose Sodium