Kuss S, Tanner E E L, Ordovas-Montanes M, Compton R G
Department of Chemistry , Physical and Theoretical Chemistry Laboratory , University of Oxford , South Parks Road , Oxford , OX1 3QZ , UK . Email:
Department of Zoology , University of Oxford , Woodstock Road , Oxford , OX2 6GG , UK.
Chem Sci. 2017 Nov 1;8(11):7682-7688. doi: 10.1039/c7sc03498a. Epub 2017 Sep 20.
The colorimetric identification of pathogenic and non-pathogenic bacteria in cell culture is commonly performed using the redox mediator ,,','-tetramethyl--phenylene-diamine (TMPD) in the so-called oxidase test, which indicates the presence of bacterial cytochrome c oxidases. The presented study demonstrates the ability of electrochemistry to employ TMPD to detect bacteria and quantify the activity of bacterial cytochrome c oxidases. Cyclic voltammetry studies and chronoamperometry measurements performed on the model organism result in a turnover number, calculated for single bacteria. Furthermore, trace amounts of cytochrome c oxidases were revealed in aerobically cultured , which to our knowledge no other technique is currently able to quantify in molecular biology. The reported technique could be applied to a variety of pathogenic bacteria and has the potential to be employed in future biosensing technology.
在细胞培养中,通常使用氧化还原介质N,N,N',N'-四甲基对苯二胺(TMPD)在所谓的氧化酶试验中对致病和非致病细菌进行比色鉴定,该试验表明细菌细胞色素c氧化酶的存在。本研究展示了电化学利用TMPD检测细菌并量化细菌细胞色素c氧化酶活性的能力。对模式生物进行的循环伏安法研究和计时电流法测量得出了单个细菌的周转数。此外,在需氧培养的大肠杆菌中发现了痕量的细胞色素c氧化酶,据我们所知,目前分子生物学中的其他技术无法对其进行量化。所报道的技术可应用于多种致病细菌,并且有潜力用于未来的生物传感技术。