SusChEM: Collaborative Research: Efficient biological activation and conversion of short-chain hydrocarbons

SusChEM：合作研究：短链碳氢化合物的高效生物活化和转化

• 批准号: 1938893
• 负责人: Clifford Henderson
• 金额: $ 15.79万
• 依托单位: University of South Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-31 至 2021-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1938893&HistoricalAwards=false
• 关键词: SusChEM; Collaborative; Research; Efficient; biological

Advances in the recovery of oil and gas reserves, through the use of fracking and horizontal well drilling, for example, have dramatically increased the amount of natural gas that can be economically harvested in the US. These reserves also contain compounds, know as natural gas liquids (NGLs), that are expensive to convert to useful products using current refinery technology. Fortunately, many microorganisms are capable of consuming NGLs for biomass and energy production using pathways that are much more efficient and less expensive. The biological mechanisms of these conversions will be studied in order to design organisms that exhibit even greater yield of product. By eliminating the generation of waste products and enhancing the effectiveness of these microbes, the resulting technology will be more widely deployed at fracking sites, increasing the energy security of the US while decreasing the negative environmental impact of natural gas extraction. This project will also provide STEM research opportunities to traditionally underrepresented groups, including to high school students through outreach to high schools in Houston, through the Science Academy of South Texas, and through the McNair program at Penn State.By cloning and repurposing poorly understood alkane-metabolizing enzymes, this project aims to develop metabolically engineered microorganisms capable of converting short-chain alkanes to target products such as butanol. Harnessing alkylsuccinate synthases for such biocatalytic applications requires functionally expressing their partner "activase" enzymes, that require iron-sulfur cluster incorporation, in a microbial host amenable to metabolic engineering. Screens for functional expression of known or putative enzyme pairs and for activation by non-cognate activases will be executed. Enzyme directed evolution strategies to achieve alkylsuccinate synthase activation will also be employed. Functional, heterologous partners will be thoroughly characterized, providing new insights into activase-alkylsuccinate synthase interaction, and new opportunities for biotechnological applications of these and related enzymes. The overall research plan combines protein and metabolic engineering to establish a novel engineering platform for biocatalytic processes with enhanced carbon and energy efficiencies. If successful, this project could transform the commercial processing of NGL away from large, capital- and energy-intensive refinery processing to smaller-scale processes with high mass and energy efficiency. This would in turn eliminate a major source of waste involved in natural gas processing.

例如，通过使用水力压裂和水平井钻探，石油和天然气储量的开采取得了进展，极大地增加了美国可以经济地开采的天然气量。这些储量还含有被称为液化天然气 (NGL) 的化合物，使用当前的炼油技术将其转化为有用的产品的成本很高。幸运的是，许多微生物能够通过更高效、更便宜的途径消耗 NGL 来生产生物质和能源。我们将研究这些转化的生物学机制，以便设计出具有更高产量的生物体。通过消除废物的产生并提高这些微生物的有效性，由此产生的技术将在水力压裂现场得到更广泛的部署，从而提高美国的能源安全，同时减少天然气开采对环境的负面影响。该项目还将为传统上代表性不足的群体提供 STEM 研究机会，包括通过向休斯顿的高中、南德克萨斯科学院和宾夕法尼亚州立大学的麦克奈尔项目进行推广，向高中生提供研究机会。 -代谢酶，该项目旨在开发能够将短链烷烃转化为丁醇等目标产物的代谢工程微生物。利用烷基琥珀酸合酶进行此类生物催化应用需要在适合代谢工程的微生物宿主中功能性地表达其伙伴“激活酶”，这需要铁硫簇掺入。将进行已知或推定酶对的功能表达和非同源激活酶激活的筛选。还将采用酶定向进化策略来实现烷基琥珀酸合酶激活。功能性异源伙伴将被彻底表征，为激活酶-烷基琥珀酸合酶相互作用提供新的见解，并为这些酶和相关酶的生物技术应用提供新的机会。总体研究计划将蛋白质和代谢工程结合起来，为生物催化过程建立一个新颖的工程平台，提高碳和能源效率。如果成功，该项目可以将液化天然气的商业加工从大型、资本和能源密集型炼油厂加工转变为具有高质量和能源效率的小规模加工。这反过来又会消除天然气加工过程中产生的主要废物来源。