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Journal of Bacteriology, November 1999, p. 6642-6649, Vol. 181, No. 21
Department of Life Science and Biotechnology,
Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, Shimane 690-8504, Japan
Received 7 June 1999/Accepted 18 August 1999
The extracellular chitosanase (34,000 Mr)
produced by a novel gram-negative bacterium Matsuebacter
chitosanotabidus 3001 was purified. The optimal pH of this
chitosanase was 4.0, and the optimal temperature was between 30 and
40°C. The purified chitosanase was most active on 90% deacetylated
colloidal chitosan and glycol chitosan, both of which were hydrolyzed
in an endosplitting manner, but this did not hydrolyze chitin,
cellulose, or their derivatives. Among potential inhibitors, the
purified chitosanase was only inhibited by Ag+. Internal
amino acid sequences of the purified chitosanase were obtained. A PCR
fragment corresponding to one of these amino acid sequences was then
used to screen a genomic library for the entire choA gene
encoding chitosanase. Sequencing of the choA gene revealed an open reading frame encoding a 391-amino-acid protein. The N-terminal amino acid sequence had an excretion signal, but the sequence did not
show any significant homology to other proteins, including known
chitosanases. The 80-amino-acid excretion signal of ChoA fused to green
fluorescent protein was functional in Escherichia coli.
Taken together, these results suggest that we have identified a novel,
previously unreported chitosanase.
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Purification, Characterization, and Gene Analysis of a
Chitosanase (ChoA) from Matsuebacter chitosanotabidus
3001
*
Corresponding author. Mailing address: Department of
Life Science and Biotechnology, Faculty of Life and Environmental
Science, Shimane University, 1060 Nishikawatsu, Matsue, Shimane
690-8504, Japan. Phone and fax: 81-852-32-6587. E-mail:
kawamuka{at}life.shimane-u.ac.jp.
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