高效液相色谱-质谱联用法高通量筛选及检测哈茨木霉菌发酵液中peptaibols类抗菌肽  

High Throughput Screening and Detection of Peptaibol Antibiotics from the Fermentation Broth of Trichoderma harzianum by LC-MS/MS

作　　者：潘顺[1] 刘雷[2] 付贤树[1] 郑文瑾[1] 王为民[1] 

机构地区：[1]中国计量学院生命科学学院,浙江省生物计量及检验检疫技术重点实验室,杭州300018 [2]澳大利亚南十字星大学植物科学研究中心,新南威尔士州2480

出　　处：《中国药学杂志》2012年第22期1849-1855,共7页Chinese Pharmaceutical Journal

基　　金：浙江省科技厅重大科技专项资助项目(2008C02007-2)

摘　　要：目的高通量筛选及检测哈茨木霉菌(Trichoderma harzianum)CGMCC No.1780发酵液中peptaibols类抗菌肽。方法以4,4'-(对二甲氨基)二苯基甲烷(TDM)特异性显色法做快速定性鉴定,以对立枯丝核菌(Rhizoctonia solani)菌丝生长的抑制作用做活性追踪;采用大孔吸附树脂、葡聚糖凝胶及高效薄层层析硅胶板分离纯化哈茨木霉菌发酵液中peptaibols类抗菌肽;利用LC-MS/MS分析peptaibols类抗菌肽序列结构。结果从哈茨木霉菌发酵液中分离出5组peptaibols类抗菌肽,经LC-MS/MS鉴定为20种peptaibols,其中包括5种新的peptaibols。结论 Peptaibols类抗菌肽是哈茨木霉菌主要抗生活性次生代谢产物之一,且种类较多,利用LC-MS/MS可快速有效鉴定哈茨木霉菌发酵液中pep-taibols类抗菌肽的序列结构。OBJECTIVE To identify peptaibol antibiotics from the fermentation broth of Trichoderma harzianum CGMCC No. 1780 （China General Microbiological Culture Collection）. METHODS Peptaibol antibiotics were isolated using a bioassay-guided fractionation against Rhizoctonia solani. To purify the peptaibols, the fermentation broth was fractionated by polystyrene resin, sephadex and high performance thin-layer chromatography （HTLC）. A specific chromogenic reaction with 4,4＇-methylenebis （N,N-dimethylani- line） was used for the preliminary identification of peptaibols. The sequences of the peptaibol antibiotics were determined by LC-MS/MS analysis. RESULTS Twenty peptaibol antibiotics were identified from the fermentation broth of T. harzianum and five of the peptai- bols were novel. CONCLUSION The fermentation broth of T. harzianum is a rich source of peptaibol antibiotics warranting further study and the LC-MS/MS method is rapid and effective to analyze the peptaibols.

关 键 词：哈茨木霉菌 Peptaibols抗菌肽 高效液相色谱-质谱联用法 高通量筛选 

分 类 号：Q814[生物学—生物工程]