Purification and characterization of chitinase from Alcaligenes xylosoxydans.
Extracellular chitinase from Alcaligenes xylosoxydans was purified to electrophoretic homogeneity using affinity and gel filtration chromatography. The molecular mass of chitinase was estimated to be 45 kDa and 44 kDa by SDS-PAGE and gel-filtration, respectively. The enzyme was optimally active at 50 degrees C (over 30 min) and pH 5. Activity staining after PAGE showed a single band. The Km for chitin was 3 g l-1. Cu2+ and Na+ at 5 mM inhibited chitinase activity to 25% while Ca2+, Mg2+ and Ba2+ had no effect at the same concentration. The purified enzyme degraded mycelia of Aspergillus niger.
Vaidya
Rajiv
R
Department of Microbiology and Biotechnology Centre, Faculty of Science, M.S. University of Baroda, Vadodara-390 002, Gujarat, India.
Roy
Satarupa
S
Macmil
Simone
S
Gandhi
Sapan
S
Vyas
Pranav
P
Chhatpar
H S
HS
D000421:Alcaligenes / Q000737:chemistry* / Q000145:classification / Q000201:enzymology; D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002688:Chitinases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000737:chemistry; D008970:Molecular Weight; D025282:Mycelium / Q000187:drug effects; D013045:Species Specificity; D013696:Temperature
D004791:Enzyme Inhibitors; D008670:Metals; D002688:Chitinases