Purification and some properties of a novel chitosanase from Bacillus subtilis KH1.

This is the authors' abstract. We don't add key points for this paper.

One of at least two chitosanases secreted in the culture filtrate of Bacillus subtilis KH1 was purified by two sequential DEAE Sepharose CL-6B chromatographies, followed by Sephacryl S-100 HR gel chromatography. The purified enzyme was homogenous as judged by SDS-PAGE. It showed an estimated molecular weight and pI of 28,000 and 8.3, respectively. The enzyme drastically reduced the viscosity of highly deacetylated chitosan substrates, with the subsequent formation of chitooligosaccharides [(GlcN)(n), n=2-6]. No activity toward carboxymethylcellulose (CMC), chitobiose (GlcN)(2), or chitotriose (GlcN)(3) was detected. Separation and quantification of products of hydrolysis of 10% (w/v) solutions of chitooligosaccharides, (GlcN)(n), n=2-6, by HPLC showed the splitting of (GlcN) (n), n=4-6, in an endo-splitting manner. Oligomers comprising higher units than the starting substrate were also detected, indicating transglycosylation activity. The amino terminal sequence of this enzyme (A-G-L-N-K-D-Q-K-R-R) is identical to that of the chitosanase derived from Bacillus pumilus BN262 and to the deduced amino terminal sequences of Bacillus subtilis 168 and Bacillus amyloliquefaciens UTK chitosanases.