Enzymatic Detection of Petroleum in Seafood

描述（由申请人提供）：石油泄漏会造成巨大的环境破坏，对公众健康构成严重威胁。石油含有许多有毒化合物，如多芳烃（PAHs）。一些多环芳烃化合物具有高度致癌性，并导致人类DNA突变。当石油泄漏到海洋中时，多环芳烃化合物迅速在海洋环境中扩散，最终在海洋生物中积累，威胁到我们的海鲜供应。美国食品和药物管理局规定，受石油污染的海产品对人类健康构成威胁，不应捕捞或出售。因此，在漏油事件发生后，需要仔细监测以评估受污染区域的程度，确保海鲜供应不含多环芳烃化合物，并监测受影响区域恢复正常状态，以便重新开放封闭区域。不幸的是，尽管有这种迫切需要，但目前使用的最常用的检测方法已被证明在检测海产品中多环芳烃化合物的潜在危险水平方面有些不准确和不可靠。需要新的替代测试方法。我们提出的研究将创造改进的测试方法来检测海产品中的石油成分。我们将开发新的基于酶的检测方法，利用微生物中的双加氧酶检测海产品中的多环芳烃化合物。当样品中存在多环芳烃化合物时，这些酶将用于产生颜色变化。我们简单的酶测试可以直接用于微孔板，这是一种广泛接受的，廉价的，高通量的食品测试应用格式。为了方便过渡到我们的新酶的格式，我们的分析将被设计为工作很好与目前接受的海鲜加工方法。我们还将研究缩短样品处理步骤的方法，以提高测试的吞吐能力。我们的新测试套件将为海产品生产商和政府机构（如FDA和NOAA）提供急需的具有成本效益，可靠和敏感的工具，以在石油泄漏后检测海产品中的石油。

DESCRIPTION (provided by applicant): Oil spills can cause tremendous environmental damage and are a serious threat to public health. Petroleum contains many toxic compounds such as polyaromatic hydrocarbons (PAHs). Some PAH compounds are highly carcinogenic and cause DNA mutations in humans. When petroleum is spilled into the sea, PAH compounds rapidly spread through the marine environment and eventually accumulate in marine life and threaten our seafood supply. The FDA has mandated that petroleum-contaminated seafood is a human health threat and should not be harvested or sold. Therefore, after an oil spill, careful monitoring is needed to assess the extent of the contaminated areas, ensure that seafood supplies are free of PAH compounds and monitor the return of affected areas to a normal state to allow re-opening of closed areas. Unfortunately, despite this critical need, the most currently used testing method has been shown to be somewhat inaccurate and unreliable for the detection of potentially hazardous levels of PAH compounds in seafood. New alternative testing methods are needed. Our proposed research will create improved tests to detect petroleum components in seafood. We will develop new enzyme-based tests to detect PAH compounds in seafood using dioxygenases enzymes from microbes. These enzymes will be used to generate a color change when PAH compounds are present in the sample. Our simple enzyme tests can be directly used in microplates, a widely-accepted, inexpensive, high-throughput format for food testing applications. To ease the transition to our new enzymatic format, our assays will be designed to work well with currently accepted seafood processing methods. We will also investigate ways to shorten the sample processing steps to increase the throughput-capacity of the test. Our new test kits will provide seafood producers and government agencies such as the FDA and NOAA with much-needed cost-effective, reliable and sensitive tools to detect petroleum in seafood after oil spills. PUBLIC HEALTH RELEVANCE: Oil spills are major threats to public health and the environment. Petroleum contains many toxic and carcinogenic substances; these substances can get into seafood near oil spill sites. It is important to test for toxic oil substances in seafood after an oil spill, but current methods are not adequate for thorough testing. Our proposed research will produce novel enzyme-based tests for oil contamination in seafood. The improved reliable properties of our new method will greatly assist the FDA to ensure seafood safety.