建立一种有效的薄层层析生物自显影方法，用于检测磷酸烯醇丙酮酸对结核分枝杆菌H37Ra磷酸葡萄糖异构酶的抑制作用。
Establishment of an effective TLC bioautographic method for the detection of Mycobacterium tuberculosis H37Ra phosphoglucose isomerase inhibition by phosphoenolpyruvate.
作者信息
Paradowska Katarzyna, Polak Beata, Chomicki Adam, Ginalska Grażyna
机构信息
a Chair and Department of Biochemistry and Biotechnology, Medical University of Lublin , Lublin , Poland and.
b Department of Physical Chemistry , Medical University of Lublin , Lublin , Poland.
出版信息
J Enzyme Inhib Med Chem. 2016 Dec;31(6):1712-7. doi: 10.3109/14756366.2016.1151012. Epub 2016 Mar 17.
A bioautographic assay based on thin layer chromatography was developed for phosphoenolpyruvate (PEP) detecting as a known but rarely studied inhibitor of phosphoglucose isomerase (PGI). The protocol with NADP(+)/NBT/PMS (β-nicotinamide adenine dinucleotide phosphate/nitrotetrazolium blue chloride/phenazine methosulfate) staining was capable of detecting Mycobacterium tuberculosis H37Ra PGI inhibition using PEP. According to this method, visibly brighter spots (zones) against purple background are observed in the area of inhibition of the above-mentioned enzyme activity. The detection limit for PEP as an inhibitor of Mycobacterium tuberculosis H37Ra PGI was 226 μg per spot/zone. Noteworthy is that we are the first authors to have successfully used a bioautographic assay to detect Mycobacterium tuberculosis H37Ra PGI inhibition by PEP.
基于薄层色谱法开发了一种生物自显影测定法，用于检测磷酸烯醇丙酮酸（PEP），它是一种已知但很少被研究的磷酸葡萄糖异构酶（PGI）抑制剂。采用NADP(+)/NBT/PMS（β-烟酰胺腺嘌呤二核苷酸磷酸/氯化硝基四氮唑蓝/吩嗪硫酸甲酯）染色的实验方案能够检测出PEP对结核分枝杆菌H37Ra PGI的抑制作用。根据该方法，在上述酶活性抑制区域可观察到相对于紫色背景明显更亮的斑点（区域）。作为结核分枝杆菌H37Ra PGI抑制剂的PEP的检测限为每斑点/区域226μg。值得注意的是，我们是首次成功使用生物自显影测定法检测PEP对结核分枝杆菌H37Ra PGI抑制作用的作者。
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