EAPSI: The enzymatic conversion of methane to methanol

EAPSI：甲烷酶促转化为甲醇

• 批准号: 1415005
• 负责人: Ann Kowalski
• 金额: $ 0.51万
• 依托单位: Kowalski Ann
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1415005&HistoricalAwards=false
• 关键词: EAPSI; enzymatic; conversion; methane; methanol

Advanced drilling techniques have increased access to natural gas, which is often emphasized as a global energy source. However, high costs for steam reforming and pipeline infrastructure can make natural gas production impractical at remote locations. Some bacteria use methane, the primary component of natural gas, as an energy source by converting methane to methanol with the enzyme methane mono-oxygenase (MMO). In collaboration with Dr. Kini, an expert in mini-enzyme design at the National University of Singapore, this project will investigate the potential for improvement to an existing MMO enzyme. The project will require research in improvements to modeling enzymes like MMO, and investigation of methods to immobilize MMO on a solid surface. The economical conversion of methane to methanol at industrial scale provides two significant benefits: reducing potent greenhouse gas emissions from methane flaring and generating methanol as an efficient fuel source and commodity chemical.Starting with an active fragment of the enzyme, computationally redesigned for solubility, the project will investigate further improvement of the construct via directed evolution - purposeful mutations to the structure of the protein that alter and may enhance desirable qualities like stability and activity. First, the expressed fragments must be loaded with Cu+2 at the active sites. Next, propylene epoxidation activity can be tested by HPLC and compared to that of the wild type. The design space can then be probed by directed evolution. Enzyme fragments with considerable activity will be further stabilized for industrial use by immobilization to a host scaffold. The result will be a solid-state biocatalyst with retained activity and enhanced qualities for industrial application: stability after multiple recycles, performance under harsh conditions, and immobilization for ease of separation. This NSF EAPSI award is funded in collaboration with the National Research Foundation of Singapore.

先进的钻井技术增加了天然气的获取，天然气经常被强调为全球能源。然而，蒸汽重整和管道基础设施的高成本可能使偏远地区的天然气生产变得不切实际。一些细菌使用甲烷（天然气的主要成分）作为能源，通过用甲烷单加氧酶（MMO）将甲烷转化为甲醇。与新加坡国立大学的微型酶设计专家Kini博士合作，该项目将研究改进现有MMO酶的潜力。该项目将需要研究改进MMO等模拟酶的方法，并研究将MMO固定在固体表面上的方法。以工业规模将甲烷经济地转化为甲醇提供了两个显著的益处：减少甲烷燃烧产生的温室气体排放，并将甲醇作为一种有效的燃料来源和商品化学品。从酶的活性片段开始，通过计算重新设计溶解度，该项目将研究通过定向进化进一步改进该结构-对蛋白质结构进行有目的的突变，改变并可能增强所需的质量，如稳定性和活性。首先，表达的片段必须在活性位点负载Cu+2。接下来，可以通过HPLC测试丙烯环氧化活性，并与野生型进行比较。设计空间，然后可以探索定向进化。具有相当大活性的酶片段将通过固定到宿主支架上而进一步稳定以用于工业用途。其结果将是一种固态生物催化剂，具有保留的活性和增强的工业应用质量：多次循环后的稳定性，在恶劣条件下的性能，以及易于分离的固定化。这个NSF EAPSI奖是与新加坡国家研究基金会合作资助的。