Biotechnology |
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Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29 |
Jing LI, Qian YANG, Li-hua ZHAO, Shu-mei ZHANG, Yu-xia WANG, Xiao-yu ZHAO |
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China; Institute of Microbiology of Heilongjiang Academy of Sciences, Harbin 150010, China; Heilongjiang University Press, Harbin 150040, China |
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Abstract An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl (DEAE)-52 cellulose and gel filtration chromatography on Bio-Gel® P-100. The protein was absorbed on DEAE-cellulose and Bio-Gel® P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), pI value 5.69 by isoelectric focusing (IEF)-PAGE, and 97.81% purity by high performance liquid chromatography (HPLC). B29I exhibited inhibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia sclerotiorum. The 50% inhibitory concentrations (IC50) of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B29I also demonstrated an inhibitory effect on conidial spore germination of Fusarium oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germinated spores.
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Received: 26 October 2008
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