依博素检测方法的研究  

A quantitative assay for determining ebosin

作　　者：郭连宏[1] 张洋[1] 赵宏阳[2] 仲伟潭[2] 张雪霞[2] 李元[1] 

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所卫生部抗生素生物工程重点实验室,北京100050 [2]华北制药集团新药研究开发有限责任公司,石家庄050015

出　　处：《中国新药杂志》2015年第1期31-34,89,共5页Chinese Journal of New Drugs

基　　金：国家“重大新药创制”科技重大专项(2012ZX09301002-001-023-02)

摘　　要：目的:依博素是一种链霉菌产生的新天然化合物,体内具有较强抗类风湿关节炎活性,有可能发展成为具有我国自主知识产权的新药。为了确定发酵液中依博素的含量,基于其对ICE酶(IL-1β转化酶)具有抑制活性,本研究建立了一种检测依博素的方法。方法:以荧光标记的ICE酶底物Ac-Trp-Glu-AlaAsp-AMC与含依博素的发酵液进行反应,采用不加依博素及重组ICE酶的样品作为对照,用多标记微孔板检测仪测定产物荧光强度,确定发酵液中依博素的含量。结果:采用上述方法,分别对多批次摇瓶及发酵罐培养的发酵液进行了测定,结果显示,本方法可有效确定发酵液中依博素的含量。结论:本方法是检测依博素发酵水平的有效方法。Objective： Ebosin is a novel compound produced by Streptomyces with remarkable anti-rheumatic arthritis activity in vivo, which may be developed as a new drug treating rheumatic arthritis. Based on the inhibitive activity of ebosin on ICE （ interleukin-l-converting enzyme） , a quantitative assay was established for determining ebosin during fermentation. Methods： Ebosin in fermentation cultures was reacted with a substrate, fluorogenic Ac-Trp-Glu-Ala-Asp-AMC, and then the fluorescence intensity of the reaction sample was measured on Multilable Reader with the reaction sample without recombinant ICE and ebosin as the control. According to the results, the quantity of ebosin could be identified. Results： With this assay, a multiple samples cultured in both fermentation tanks and flasks were measured, which showed that this method was more effective. Conclusion： This is a useful method for quantitative analysis of ebosin in fermentation cultures.

关 键 词：依博素 ICE酶 检测方法 发酵 链霉菌 

分 类 号：R917[医药卫生—药物分析学]