高速逆流色谱法从哈茨木霉发酵液中分离纯化抑菌活性成分  

Isolation and Purification of Antifungal Components from Trichoderma harzianum Ferment Broth by High-Speed Counter-Current Chromatography

作　　者：沙莎[1] 刘雷[2] 潘顺[1] 王为民[1] 

机构地区：[1]浙江省生物计量及检验检疫技术重点实验室/中国计量学院生命科学学院,杭州310018 [2]澳大利亚南十字星大学植物科学研究中心

出　　处：《中国生物防治学报》2013年第1期83-88,共6页Chinese Journal of Biological Control

基　　金：浙江省科技厅重大科技专项(2008C02007-2)

摘　　要：本文采用高速逆流色谱法(HSCCC),以正己烷:乙酸乙酯:甲醇:水=7:3:5:5(v/v/v/v)作为两相溶剂体系,直接从哈茨木霉菌Trichoderma harzianum发酵液的乙酸乙酯提取液中分离出对立枯丝核菌Rhizoctonia solani有抑制作用的主要活性成分,经FTICR质谱及核磁共振鉴定为木霉菌素trichodermin(每324.3 mg的乙酸乙酯提取物中分离得到4 mg),对水稻纹枯病菌Thanatephorus cucmeris菌丝生长毒力EC50及EC90分别为0.69μg.mL 1、3.02μg.mL 1,此方法可用于扩大制备木霉菌素trichodermin。此外高速逆流色谱法还得到了另一对立枯丝核菌Rhizoctonia solani有明显抑制效果的活性混合组分。We developed a high-speed counter-current chromatography （HSCCC） method and identified the major antifungal components isolated from Trichoderma harzianum broth after extraction with ethyl acetate. The HSCCC conditions were optimized and a two-phase solvent system containing n-hexane：ethyl acetate：methanol：water =7：3：5：5 （v/v/v/v） was selected for the investigation. One pure and active compound was elucidated as trichodermin using NMR and FTICR mass spectroscopy. Trichodermin exerted a potent activity in vitro against mycelial growth of Thanatephorus cucmeris with EC50=0.69 μg·mL^-1 and EC90=3.02 μg·mL^-l. This optimized HSCCC method can be scaled up to produce a larger amount of purified trichodermin.

关 键 词：高速逆流色谱 哈茨木霉 木霉菌素 立枯丝核菌 水稻纹枯病 

分 类 号：S482.2[农业科学—农药学] S435.111.42[农业科学—植物保护]