Li Jing, Yang Qian, Zhao Li-hua, Zhang Shu-mei, Wang Yu-xia, Zhao Xiao-yu
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China.
J Zhejiang Univ Sci B. 2009 Apr;10(4):264-72. doi: 10.1631/jzus.B0820341.
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 (IC(50)) of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 micromol/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.
从枯草芽孢杆菌B29菌株的培养物中分离出一种抗真菌蛋白。分离过程包括在二乙氨基乙基（DEAE）-52纤维素上进行离子交换色谱以及在Bio-Gel P-100上进行凝胶过滤色谱。该蛋白能吸附在DEAE-纤维素和Bio-Gel P-100上。纯化后的抗真菌组分被命名为B29I，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）测得其分子量为42.3 kDa，通过等电聚焦（IEF）-PAGE测得其pI值为5.69，通过高效液相色谱（HPLC）测得其纯度为97.81%。B29I对尖孢镰刀菌、立枯丝核菌、串珠镰刀菌和核盘菌的菌丝生长具有抑制活性。其对尖孢镰刀菌和立枯丝核菌的抗真菌活性的50%抑制浓度（IC50）分别为45和112 μmol/L。B29I还对尖孢镰刀菌的分生孢子萌发具有抑制作用，并能抑制芽管伸长，还会导致部分萌发孢子出现畸变、肿胀和破裂。
