Department of Biology, U.S. Air Force Academy, 2355 Faculty Drive, Colorado Springs, CO 80840, USA.
Mil Med. 2021 Jan 25;186(Suppl 1):801-807. doi: 10.1093/milmed/usaa367.
Per- and polyfluoroalkyl substances (PFAS) are a class of synthetic compounds used industrially for a wide variety of applications. These PFAS compounds are very stable and persist in the environment. The PFAS contamination is a growing health issue as these compounds have been reported to impact human health and have been detected in both domestic and global water sources. Contaminated water found on military bases poses a potentially serious health concern for active duty military, their families, and the surrounding communities. Previous detection methods for PFAS in contaminated water samples require expensive and time-consuming testing protocols that limit the ability to detect this important global pollutant. The main objective of this work was to develop a novel detection system that utilizes a biological reporter and engineered bacteria as a way to rapidly and efficiently detect PFAS contamination.
The United States Air Force Academy International Genetically Engineered Machine team is genetically engineering Rhodococcus jostii strain RHA1 to contain novel DNA sequences composed of a propane 2-monooxygenase alpha (prmA) promoter and monomeric red fluorescent protein (mRFP). The prmA promoter is activated in the presence of PFAS and transcribes the mRFP reporter.
The recombinant R. jostii containing the prmA promoter and mRFP reporter respond to exposure of PFAS by activating gene expression of the mRFP. At 100 µM of perfluorooctanoic acid, the mRFP expression was increased 3-fold (qRT-PCR). Rhodococcus jostii without exposure to PFAS compounds had no mRFP expression.
This novel detection system represents a synthetic biology approach to more efficiently detect PFAS in contaminated samples. With further refinement and modifications, a similar system could be readily deployed in the field around the world to detect this critical pollutant.
全氟和多氟烷基物质（PFAS）是一类用于各种应用的合成化合物。这些 PFAS 化合物非常稳定，在环境中持久存在。由于这些化合物已被报道会影响人类健康，并已在国内和全球水源中检测到，因此 PFAS 污染是一个日益严重的健康问题。军事基地发现的受污染的水对现役军人、他们的家人和周围社区构成了潜在的严重健康威胁。以前对受污染水样中的 PFAS 进行检测的方法需要昂贵且耗时的测试方案，这限制了检测这种重要全球污染物的能力。这项工作的主要目的是开发一种新的检测系统，该系统利用生物报告基因和工程细菌作为快速、高效检测 PFAS 污染的方法。
美国空军学院国际基因工程机器团队正在对 Rhodococcus jostii 菌株 RHA1 进行基因工程改造，使其包含由丙烷 2-单加氧酶 alpha（prmA）启动子和单体红色荧光蛋白（mRFP）组成的新 DNA 序列。在 PFAS 存在下，prmA 启动子被激活，并转录 mRFP 报告基因。
含有 prmA 启动子和 mRFP 报告基因的重组 Rhodococcus jostii 对 PFAS 的暴露作出反应，通过激活 mRFP 报告基因的表达来激活基因表达。在 100 µM 的全氟辛酸存在下，mRFP 的表达增加了 3 倍（qRT-PCR）。未接触 PFAS 化合物的 Rhodococcus jostii 没有 mRFP 表达。
这种新的检测系统代表了一种合成生物学方法，可更有效地检测受污染样品中的 PFAS。通过进一步的改进和修改，类似的系统可以很容易地在世界各地的现场部署，以检测这种关键污染物。
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