In Vitro Suppression of Mycelial Growth of Fusarium oxysporum by Extracellular Chitosanase of Sphingobacterium multivorum and Cloning of the Chitosanase Gene csnSM1 |
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Authors: | Yoshinori MATSUDA Yuichiro IIDA Takeshi SHINOGI Koji KAKUTANI Teruo NONOMURA Hideyoshi TOYODA |
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Institution: | (1) Laboratory of Plant Pathology and Biotechnology, Faculty of Agriculture, Kinki University, Nara 631–8505, Japan, JP;(2) Pharmaceutical Research and Technology Institute, Kinki University, Higashi-Osaka 577–8502, Japan, JP |
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Abstract: | A chitosan-degrading bacterium, isolated from field soil that had been amended with chitin, was identified as Sphingobacterium multivorum KST-009 on the basis of its bacteriological characteristics. The extracellular chitosanase (SM1) secreted by KST-009 was a
34-kDa protein and could be purified through ammonium sulfate precipitation, gel permeation column chromatography and SDS
polyacrylamide gel electrophoresis. A chitosanase gene (csnSM1) was isolated from genomic DNA of the bacteria, and the entire nucleotide sequence of the gene and the partial N-terminal
amino acid sequence of the purified SM1 were determined. The csnSM1 gene was found to encode 383 amino acids, 72 N-terminal amino acid residues were processed to produce the mature enzyme
during the secretion process. Germinated microconidia of four formae speciales (lycopersici, radicis-lycopersici, melonis, and fragariae ) of Fusarium oxysporum were treated with SM1. Chitosanase treatment caused morphological changes, such as swelling of hyphal cells or indistinctness
of hyphal cell tips and cessation or reduction of mycelial elongation.
Received 2 May 2001/ Accepted in revised form 21 June 2001 |
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Keywords: | : Sphingobacterium multivorum chitosanase SM1 chitosanase gene csnSM1 Fusarium oxysporum |
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