Dept of Nano-Bioscience and Chemical Engineering, Ulsan National Institute of Science and Technology, Ulsan, South Korea.
Bioresour Technol. 2013 Jan;127:429-34. doi: 10.1016/j.biortech.2012.09.086. Epub 2012 Sep 29.
A bioluminescent Escherichia coli bioreporter strain to detect hydrolysate related phenolics was developed by cloning the aaeXAB promoter from E. coli upstream of the luxCDABE genes. E. coli str. DH5α carrying this plasmid (pDMA3) was responsive to sub-inhibitory concentrations of plant hydrolysate-related phenolics, such as ferulic and vanillic acids, responding to these compounds at concentrations as low as 9.8 and 4.9 mg/L, respectively. Experiments with a mixture of the compounds showed similar responses as with single compound tests, with a minimum detectable concentration of 19.5mg/L. Finally, tests using rice straw hydrolysates were conducted, with E. coli str. DH5α/pDMA3 showing a maximum induction of 33-fold and a minimum detectable phenolic concentration of 9.3mg/L, based upon Folin-Ciocalteu's reagent. These results demonstrate that this bioreporter maintains its sensitivity even with hydrolysate samples and that it can be potentially applied within biofuel industries to detect phenolics present within plant hydrolysates.
通过将大肠杆菌的 aaeXAB 启动子克隆到 luxCDABE 基因的上游,开发了一种用于检测水解物相关酚类的生物发光大肠杆菌生物报告菌株。携带该质粒(pDMA3)的大肠杆菌 str. DH5α 对植物水解物相关酚类物质(如阿魏酸和香草酸)的亚抑制浓度有反应,对这些化合物的反应浓度低至 9.8 和 4.9mg/L 。用化合物混合物进行的实验与单化合物测试具有相似的反应,最低检测浓度为 19.5mg/L 。最后,对稻草水解物进行了测试,大肠杆菌 str. DH5α/pDMA3 表现出最大诱导 33 倍,基于福林-希卡尔特试剂的最低可检测酚浓度为 9.3mg/L 。这些结果表明,即使在水解物样品中,该生物报告器仍保持其灵敏度,并且可以在生物燃料行业中潜在地应用于检测植物水解物中存在的酚类物质。