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一种快速显色定量检测微生物样品中棒酸的方法

, PP. 246-251

Keywords: 棒酸,棒状链霉菌,头孢硝噻吩,显色法,&beta,-内酰胺酶

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Abstract:

为满足从棒状链霉菌突变库中筛选棒酸高产株的需求,基于显色法原理设计了一种快速检测棒酸含量的方法.该方法的反应机制是β-内酰胺酶水解其黄色底物头孢硝噻吩(λmax=390nm),从而生成红色产物(λmax=486nm).由于棒酸是β-内酰胺酶不可逆抑制剂,在上述反应中棒酸含量的高低将影响底物和产物的量,因此可以通过测定底物和产物的比值(A390/A486)最终测定棒酸的含量.本实验证实,该方法对棒酸的灵敏度为50μg/L,最佳检测窗口为0.05~10mg/L,且该方法的检测结果与HPLC法相关性很好.通过筛选一个小规模棒状链霉菌突变株库,证实该显色法可以快速识别和评价棒酸高产突变株.因此该方法若用于工业实践中的大规模突变株筛选,将节省大量的时间与人力.另外,还利用该显色反应开发了平板检测法,直接根据颜色差异定性判断平板菌落棒酸产量高低.该显色法是一种快速、方便、可靠的棒酸产量检测方法,适用于高通量筛选棒酸高产株.

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