吡咯伯克霍尔德氏菌JK_SH007抗菌蛋白的分离纯化_任嘉红.pdfVIP

微生物学通报 JUN 20, 2010, 37(6): 872−880 Microbiology China © 2010 by Institute of Microbiology, CAS tongbao@ 简 报 吡咯伯克霍尔德氏菌JK-SH007 抗菌蛋白的分离纯化 1,2 1 1 1* 任嘉红 吴小芹 刘辉 叶建仁 (1. 210037) (2. 046011) 摘 要: 对拮抗细菌吡咯伯克霍尔德氏菌(Burkholderia pyrrocinia) JK-SH007 菌株抗菌蛋白进行了 初步研究。研究发现, JK-SH007 菌株的抗菌粗蛋白对热和蛋白酶K 不敏感, 碱性环境不利于抑菌 蛋白活性的发挥。JK-SH007 菌株的无菌发酵滤液经硫酸铵沉淀、透析(3.5 kD) 、冷丙酮沉淀 (−20°C) 、Sephadex G-50 葡聚糖凝胶分子筛层析及DEAE-Sephadex A-25 离子交换层析分离纯化, 得到了蛋白组分A- Ⅱ-2, 经SDS 聚丙烯酰胺凝胶电泳检测不是单一组分, 该复合组分具有明显抑 制 3 种杨树溃疡病病原真菌金黄壳囊孢(Cytospora chrysosperma) 、拟茎点霉(Phomopsis macro- spore) 、七叶树壳梭孢(Fusicoccum aesculi)生长的作用。 关键词: 杨树溃疡病, 吡咯伯克霍尔德氏菌, 抗菌蛋白, 分离纯化, 拮抗细菌 Isolation and Purification of Antifungal Protein from Burkholderia pyrrocinia JK-SH007 REN Jia-Hong1,2 WU Xiao-Qin1 LIU Hui1 YE Jian-Ren1* (1. College of Forest Resources and Environment, Nanjing Forestry University, Nanjing, Jiangsu 210037, China) (2. Department of Biochemistry, Changzhi College, Changzhi, Shanxi 046011, China) Abstract: A preliminary study on the antagonistic protein produced by Burkholderia pyrrocinia JK-SH007 was carried out. The results showed it was stable to heat, not sensitived to proteinase K and partially sensitive to alkali. Cell culture was obtained after centrifuging and filtering (0.22 μm) of fer- m