Purification and characteristics of an autolytic chitinase of Piromyces communis OTS1 from culture medium.
An autolysis chitinase was purified from the cultural medium of the anaerobic fungus Piromyces communis OTS1 by ammonium sulfate precipitation, affinity chromatography with regenerated chitin, chromato-focusing, gel filtration, and chromato-focusing again. The optimal pH and temperature were 6.0 and 50 degrees C, respectively, for a 20-min assay. The chitinase was stable from pH 6.0 to 8.0, but was unstable at 70 degrees C for 20 min. The molecular mass of chitinase was estimated by SDS-PAGE to be 44.9 kDa, and its pI was 4.4. The enzyme activity, which was of the 'endo' type, was inhibited by Hg2+ and allosamidin. The chitinase hydrolyzes chitin powder and fungal cell walls at a higher rate than an artificial chitin substrate. It can be concluded that extracellular chitinase is similar to cytosolic chitinase, but they are not the same protein.
Sakurada
M
M
Laboratory of Animal Nutrition and Biochemistry, Animal Science Division, Faculty of Agriculture, Miyazaki University, Nishi 1-1, Gakuen Kibana-dai, Miyazaki 889-21, Japan.
Morgavi
D P
DP
Komatani
K
K
Tomita
Y
Y
Onodera
R
R
D002688:Chitinases / Q000302:isolation & purification*; D003470:Culture Media; D005658:Fungi / Q000201:enzymology*; D008970:Molecular Weight; D013379:Substrate Specificity
D003470:Culture Media; D002688:Chitinases