一种简便快速检测细菌黑芥子酶对应蛋白带的方法

Abdulhadi Albaser, فيجترا لونغ ان, نمرة نسار, نورالهدى عبدالقادر, جون روسيتر
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引用次数: 1

摘要

黑芥子酶具有重要的科学和医学意义。不幸的是,从微生物中检测和纯化黑芥子酶需要使用高成本的底物(硫代葡萄糖苷),如sinigrin和昂贵的仪器,如快速蛋白质液相色谱和/或离子交换色谱。在这项工作中,我们只使用了20 mL的细菌培养物,补充了紫荆素(10 mM),获得了部分纯化的黑芥子酶。将粗蛋白提取液装于天然聚丙烯酰胺凝胶上,鉴定并洗脱推定的黑芥子酶带。这一步骤成功地减少了细菌粗提取物中待进一步分析的蛋白质条带的数量。目前的方法描述了一种简单、快速和经济有效的分离和检测活性细菌黑芥子酶的方法。此外,我们的方法可以作为一个纯化步骤。
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A simple & Rapid Method For Detecting Bacterial Myrosinase Corresponding Protein Band
Myrosinases have significant scientific and medical implications. Unfortunately, detection and purification of myrosinase from microbes requires the use of highly cost substrates (glucosinolates) such as sinigrin and expensive instruments such as Fast Protein Liquid Chromatography and or ion exchange chromatography. In this work, we used only 20 mL of bacterial culture supplemented with sinigrin (10 mM) to obtain partially purified myrosinase. The crude protein extract was loaded onto native polyacrylamide gel and putative myrosinase band was identified and eluted. This step successfully minimised the numbers of protein bands of bacterial crude extracts to be further analysed. The current method describes a simple, rapid and cost effective protocol for isolation and detection of active bacterial myrosinases. Furthermore, our method can be used as a purification step.
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