RAPID: ACQUISITION OF A PROTEOMICS ANALYZER TO ELUCIDATE PATHWAYS OF PETROLEUM HYDROCARBON BIOREMEDIATION IN THE GULF OF MEXICO

项目负责人：Kartik chandran提案编号：1057414机构：哥伦比亚大学标题：获得蛋白质组学分析仪以阐明墨西哥湾石油烃生物修复途径NSF MRI RAPID项目将支持获得最先进的仪器，以阐明甲烷和复杂石油烃生物降解的微生物机制定量蛋白质组学水平。由此获得的机理信息将用于制定生物修复策略，以减轻墨西哥湾深水地平线石油泄漏造成的环境损害。“深水地平线”漏油事件严重损害了墨西哥湾的水生生物和环境健康。尽管如此，污染物流的特定化学成分为同时对主要有机物（石油碳氢化合物）和甲烷（据称是导致泄漏的爆炸原因）进行生物修复提供了一个新的机会。甲烷通常是甲烷氧化细菌（MOB）的主要能量底物。然而，由于参与甲烷氧化为甲醛的第一种酶（甲烷单加氧酶）具有广泛的底物特异性，MOB不仅可以氧化甲烷，还可以氧化其他有机底物，如长链脂肪族和芳香族化合物。然而，MOB不能从这种转化中获得能量，这种转化被称为“协同代谢”。因此，基于协同代谢的生物修复策略的特点是有限的转化能力，受到主要底物（甲烷）可用性的限制。在这个项目中，他们将在实验室规模的生物反应器中阐明MOB氧化甲烷（通过初级能量代谢）和石油碳氢化合物混合物（通过共代谢）的蛋白质组学规模机制。NSF核磁共振快速项目将支持获得定量蛋白质组学分析仪（Waters公司），该分析仪最初将在地球与环境工程（PI的家庭部门，Kartik Chandran博士）中安置六个月，然后部署在海湾现场。当用于与甲烷（目前刚刚燃烧或释放）一起被隔离或撇去的污染物的异地生物修复时，本项目开发的生物工艺技术可以同时专门用于处理这两种污染物。描述甲烷氧化和石油碳氢化合物协同代谢的数学模型将使用蛋白质组学数据结合化学剖面和微生物测量来构建和参数化。最后，将制定和测试作业策略、现场获得的甲烷和石油碳氢化合物混合物的非原位转化和矿化。该项目的成功应用将加速墨西哥湾广泛存在的石油烃和甲烷污染的生物处理。此外，开发的策略将有助于加速处理并最大限度地减少这种特殊污染物混合物的广泛传播，这是未来海上钻井作业的典型特征。这一工具将及时用于与海湾溢油有关的研究，与RAPID的供资要求相一致。

PI: Kartik ChandranProposal Number: 1057414Institution: Columbia UniversityTitle: OIA: Acquisition of a Proteomics Analyzer to Elucidate Pathways of Petroleum Hydrocarbon Bioremediation in the Gulf of MexicoThis NSF MRI RAPID project will support the acquisition of a state-of-the art instrument to elucidate the microbial mechanisms of methane and complex petroleum hydrocarbon biodegradation at the quantitative proteomics level. The mechanistic information thus obtained will be used to develop bioremediation strategies to alleviate the environmental insults that have resulted from the Deepwater Horizon oil spill in the Gulf of Mexico. The Deepwater Horizon oil spill has resulted in drastically impaired aquatic life and environmental health of the gulf. Nevertheless, the specific chemical composition of the contaminant stream presents a novel opportunity for simultaneous bioremediation of both the primary organics (the petroleum hydrocarbons) and the methane (the purported cause of the explosion that led to the spill). Methane is typically the primary energy substrate for methane oxidizing bacteria (MOB). However, owing to the broad substrate specificity of the first enzyme involved in methane oxidation to formaldehyde (methane monooxygenase), MOB can oxidize not only methane but also alternate organic substrates such as longer chain aliphatic and aromatic compounds. However, MOB cannot derive energy from such transformations, which are termed ?co-metabolic?. Co-metabolism based bioremediation strategies are therefore characterized by a finite transformation capacity, limited by availability of the primary substrate (in casu, methane). In this project, they will elucidate the proteomics scale mechanisms of MOB to oxidize methane (via primary energy metabolism) and the mix of petroleum hydrocarbons (via co-metabolism) in lab-scale bioreactors. This NSF MRI Rapid project will support the acquisition of a quantitative proteomics analyzer (Waters Corp.), which will be housed initially for six-months in Earth and Environmental Engineering (the home department of the PI, Dr. Kartik Chandran) and then deployed onsite in the Gulf. When used for ex-situ bioremediation of the sequestered or skimmed pollutants along with the methane (which is currently just being flared or released), the bio-process technology developed in this project could be specifically applied to address both contaminants simultaneously. Mathematical models that describe methane oxidation and co-metabolism of petroleum hydrocarbons will be constructed and parameterized using the proteomics data combined with chemical profiles and microbial measurements. Finally, operating strategies, ex-situ transformation and mineralization of methane and petroleum hydrocarbons mixes obtained on site will be developed and tested. Successful application of this project will enable the accelerated biological treatment of the widespread petroleum hydrocarbon and methane pollution in the Gulf of Mexico. Additionally, the developed strategies will help to accelerate treatment and minimize such widespread dissemination of this particular contaminant mix, which is typical of offshore drilling operations in the future.This instrument will be used for Gulf oil spill related research in a timely fashion consistent with RAPID funding requirements.