EAGER: Enabling Direct Microbial Biotransformation of Methane and Derived Methanol to Valuable Biochemicals and Biofuels by Yarrowia Lipolytica

甲烷资源丰富，来自天然气、煤炭、石油和牲畜沼气，也是主要的温室气体之一。甲烷主要用作产生热量和电力的燃料，并通过合成气体路线生产甲醇，这一路线的处理成本很高，因为它是非常能源密集型的。作为化学过程中的副产品或牲畜的沼气产生的甲烷通常在释放到环境之前被释放或燃烧。因此，有效地捕获和利用甲烷可以为生产有价值的产品和缓解全球变暖提供丰富的原料。这项急切的提议寻求探索尚未开发的微生物生物转化路线，以利用甲烷和衍生甲醇合成有价值的生物基产品。拟议的研究将具有变革性，在室温和环境压力下实现丰富的甲烷和衍生甲醇的直接微生物转化为有价值的生物基产品，所需能量比传统化学转化少得多，并同时捕获甲烷以减缓全球变暖。该项目还将把研究纳入对田纳西大学不同的本科生和K-12学生群体产生重大影响的教育推广活动中。这项急切的提议的目标是阐明和激活产油酵母(如解脂亚罗维菌)在生物化学和生物燃料合成中利用甲烷和甲醇的独特途径。在这项提案中，PI团队将使用系统生物学工具来解决以下基本问题：1)解脂耶尔森菌利用甲醇的途径涉及哪些基因？2)解脂耶尔森菌能否利用其不同的CYP450单加氧酶中的任何一种将甲烷转化为甲醇并在甲烷上生长？此外，PI团队将应用代谢工程和合成生物学工具来利用解脂耶尔森菌独特的新陈代谢，从甲烷和甲烷衍生分子中合成有价值的生物基产品。如果成功，该项目将揭示真核生物(如解脂酵母)中鲜为人知的独特的甲烷和甲醇利用途径，并为发现同时具有产油和甲烷营养特性的其他非传统酵母铺平道路。

1360867Trinh, Cong T. Methane is abundant and comes from natural gas, coal, petroleum, and biogas of livestock; it is also one of the major and potent greenhouse gases. Methane is primarily used as a fuel for generating heat and electricity, and to produce methanol via the synthesis gas route, which is expensive to process as it is very energy intensive. Methane generated as a byproduct in a chemical process or as biogas of livestock is often either released or flared before being released to the environment. Therefore, effectively capturing and using methane can provide an abundant feedstock to produce valuable products and alleviate global warming. This EAGER proposal seeks to explore the untapped microbial biotransformation route to utilize methane and derived methanol for the synthesis of valuable biobased products. The proposed research will be transformative by achieving the direct microbial transformation of abundant methane and derived methanol to valuable biobased products at room temperature and ambient pressure with much less energy than conventional chemical transformation, and by simultaneously capturing methane to abate global warming. This project will also integrate the research into educational outreach activities that have significant impact on a diverse group of undergraduate and K-12 students at The University of Tennessee. The goal of this EAGER proposal is to elucidate and activate the unique methane- and methanol-utilizing pathways of oleaginous yeasts (such as Yarrowia lipolytica) for biochemicals and biofuels synthesis. In this proposal, the PI team will use systems biology tools to address the following fundamental questions: 1) What genes are involved in the methanol-utilizing pathway of Y. lipolytica? and 2) Can Y. lipolytica utilize any of its diverse set of CYP450 monooxygenases to convert methane to methanol and enable growth on methane? Furthermore, the PI team will apply metabolic engineering and synthetic biology tools to harness the unique metabolism of Y. lipolytica to synthesize valuable biobased products from methane and methane-derived molecules. If successful, this project will shed light on unique methane- and methanol-utilizing pathways that are poorly understood in eukaryotes (e.g., Y. lipolytica) and pave the way for discovering other non-conventional yeasts that have both oleaginous and methanotrophic characteristics.