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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 new natural compound produced by Streptomyces, which has strong anti-rheumatoid arthritis activity in vivo and may develop into a new drug with independent intellectual property rights in China. In order to determine the content of ebosin in the fermentation broth, this study established a method for the detection of ebosin based on its inhibitory activity on ICE enzyme (IL-1β-converting enzyme). Methods: Fluorescent labeled ICE enzyme substrate Ac-Trp-Glu-AlaAsp-AMC was reacted with ebensin-containing fermentation broth. Samples without ebosin and recombinant ICE enzyme were used as controls. The plate detector measures the fluorescence intensity of the product and determines the content of ebosin in the fermentation broth. Results: The fermentation broths cultured in shake flask and fermentor were determined by the above method. The results showed that the method could effectively determine the content of ebosin in the fermentation broth. Conclusion: This method is an effective method to detect the level of ebosin fermentation.