Properties of chitosanase from Bacillus cereus S1.
Chitosanase from Bacillus cereus S1 was purified, and the enzymatic properties were investigated. The molecular weight was estimated to 45,000 on SDS-PAGE. Optimum pH was about 6, and stable pH in the incubation at 40 degrees C for 60 min was 6-11. This chitosanase was stable in alkaline side. Optimum temperature was around 60 degrees C, and enzyme activity was relatively stable below 60 degrees C. The degradations of colloidal chitosan and carboxymethyl cellulose (CMC) were about 30 and 20% relative to the value of soluble chitosan, respectively, but colloidal chitin and crystalline cellulose were not almost hydrolyzed. On the other hand, S1 chitosanase adsorbed on colloidal chitin completely and by about 50% also on crystalline cellulose, in contrast to colloidal chitosan, which it did not adsorb. S1 chitosanase finally hydrolyzed 100% N-deacetylated chitosan (soluble state) to chitobiose (27.2%), chitotriose (40.6%), and chitotetraose (32.2%). In the hydrolysis of various chitooligosaccharides, chitobiose and chitotriose were not hydrolyzed, and chitotetraose was hydrolyzed to chitobiose. Chitobiose and chitotriose were released from chitopentaose and chitohexaose. From this specificity, it was hypothesized that the active site of S1 chitosanase recognized more than two glucosamine residues posited in both sides against splitting point for glucosamine polymer.
Kurakake
M
M
Department of Food Science & Technology, Fukuyama University, Sanzou, Gakuenchou 1 banchi, Fukuyama, Hiroshima 729-0292, Japan.
Yo-u
S
S
Nakagawa
K
K
Sugihara
M
M
Komaki
T
T
D000327:Adsorption; D000595:Amino Acid Sequence; D001409:Bacillus cereus / Q000201:enzymology* / Q000254:growth & development; D002686:Chitin / Q000031:analogs & derivatives / Q000378:metabolism; D048271:Chitosan; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D013696:Temperature
D002686:Chitin; D048271:Chitosan; D006026:Glycoside Hydrolases; C019868:chitosanase