JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2009 Vol.10 No.4 P.264-272

http://doi.org/10.1631/jzus.B0820341

Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29

Author(s): Jing LI, Qian YANG, Li-hua ZHAO, Shu-mei ZHANG, Yu-xia WANG, Xiao-yu ZHAO
Affiliation(s): Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China; more
Corresponding email(s): yangq@hit.edu.cn
Key Words: Bacillus subtilis, Antifungal protein, Purification

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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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