Elucidation of Alkene Metabolism in Two Sulfate-Reducing Isolates Via Metabolite Profiling and Transcriptomics

通过代谢物分析和转录组学阐明两种硫酸盐还原分离株中的烯烃代谢

• 批准号: 1329890
• 负责人: Amy Callaghan
• 金额: $ 83.19万
• 依托单位: University of Oklahoma Norman Campus
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1329890&HistoricalAwards=false
• 关键词: Elucidation; Alkene; Metabolism; Two; Sulfate

Intellectual MeritAnaerobic bacteria and archaea play key roles in biogeochemical cycling, the biotransformation of pollutants, and in both subsurface production and oxidation of methane, an important biofuel. The past 30 years of research have yielded novel mechanisms by which anaerobes activate hydrocarbons in the absence of oxygen. However, there is still a great deal to be learned regarding the enzymes catalyzing reactions in the requisite biochemical pathways. Alkenes are ubiquitous hydrocarbons derived from both natural and anthropogenic sources. Although more reactive than aromatic hydrocarbons and alkanes, little is known regarding the biochemistry of anaerobic alkene biotransformation. This project aims to elucidate the fundamental mechanisms of microbial activation and degradation of alkenes in two model strains of sulfate-reducers, Desulfatibacillum alkenivorans AK-01 and Desulfococcus oleovorans Hxd3, by coupling transcriptomics with metabolite profiling. The bacteria are not only model organisms for anaerobic hydrocarbon degradation, they are also two of only three sulfate-reducing, hydrocarbon-utilizing microbes (the other is the archaeon Archaeoglobus fulgidus) for which complete (i.e. closed) genome sequences are available. The project will shed light on the novel biochemistry of anaerobic alkene activation and degradation by a class of microorganisms that is both environmentally and economically important.Broader ImpactsAlkenes are widely present in the environment, and are generated from both natural and industrial processes. These chemicals are economically and environmentally important. This project aims to increase understanding of the activation and biotransformation of alkene substrates and environmental contaminants with similar structures, as well as the biochemistry and physiology of the relevant organisms involved in biogeochemical cycling, bioremediation, and biofuel production. As part of this project, research activities will be integrated into the education of high-school, undergraduate, and graduate students by providing laboratory research experience and mentoring. Students will be trained in aspects of molecular biology, anaerobic cultivation and monitoring of microorganisms, metabolite profiling, microbial ecology, and "omics". Research will be incorporated into existing department courses, and future efforts are aimed toward developing a "Nucleotide Sequence Analysis" class that integrates the project's omics datasets into classroom learning, with the long-term goal of implementing a Biotechnology program at the University of Oklahoma. Educational outreach to the community will be coordinated through the K-20 Center for Education and Community Renewal, which partners faculty and graduate students with K-12 teachers and students and collaborates with over 800 Oklahoma schools and industrial partners to enhance learning across all educational levels. Goals to integrate diversity into NSF programs, projects and activities will be achieved through the McNair Scholars program, which prepares undergraduate students from underrepresented ethnicities for doctoral studies through research opportunities and activities. Efforts to disseminate project results and enhance scientific and technological understanding will be achieved via participation in the OU Annual Student Research and Performance Day, Science Café of Norman, and BioBlitz! Oklahoma.

厌氧细菌和古生菌在生物地球化学循环、污染物的生物转化以及甲烷的地下生产和氧化过程中发挥关键作用，甲烷是一种重要的生物燃料。过去30年的研究已经产生了新的机制，通过这种机制，厌氧菌在缺乏氧气的情况下激活碳氢化合物。然而，关于在必要的生化途径中催化反应的酶，仍然有大量的知识需要了解。烯烃是一种普遍存在的碳氢化合物，来源于自然和人为来源。虽然比芳香烃和烷烃更具活性，但对厌氧烯烃生物转化的生物化学知之甚少。本项目旨在通过将转录组分与代谢产物谱相结合的方法，阐明两株硫酸盐还原菌--嗜碱脱硫杆菌AK-01和油链球菌Hxd3中烯烃的微生物活化和降解的基本机制。这些细菌不仅是厌氧降解碳氢化合物的模式生物，也是仅有的三种硫酸盐还原利用碳氢化合物微生物(另一种是古古细菌)中的两种，可以获得完整的(即封闭的)基因组序列。该项目将揭示一类对环境和经济都具有重要意义的微生物对厌氧烯的活化和降解的新生物化学。这些化学品在经济上和环境上都很重要。该项目旨在增进对具有相似结构的烯烃底物和环境污染物的活化和生物转化，以及参与生物地球化学循环、生物修复和生物燃料生产的相关生物的生物化学和生理学的了解。作为该项目的一部分，研究活动将通过提供实验室研究经验和指导，整合到高中、本科生和研究生的教育中。学生将接受分子生物学、微生物厌氧培养和监测、代谢物分析、微生物生态学和组学方面的培训。研究将被纳入现有的系级课程，未来的努力旨在开发一门将该项目的组学数据集整合到课堂学习中的“核苷酸序列分析”课程，长期目标是在俄克拉荷马大学实施一个生物技术项目。面向社区的教育推广将通过K-20教育和社区更新中心进行协调，该中心将教职员工和研究生与K-12教师和学生合作，并与800多所俄克拉荷马州学校和行业合作伙伴合作，加强所有教育级别的学习。将多样性纳入NSF计划、项目和活动的目标将通过麦克奈尔学者计划实现，该计划通过研究机会和活动为来自未被充分代表的民族的本科生准备博士学习。通过参加俄亥俄州立大学一年一度的学生研究和表演日、诺曼科学咖啡馆和生物闪电战，将努力传播项目成果和增进对科学技术的理解！俄克拉荷马州。