SBIR Phase I: Engineering an Industrial Synthetic Microorganism for Lower-Cost Chemical Production

这项小企业创新研究（SBIR）项目的更广泛影响和商业潜力将是一项基于发酵的技术，能够将美国丰富的天然气供应转化为有价值的基本化学品，这将促进经济和安全利益，同时鼓励制造业创新。这个项目的重点是研究能够通过发酵将天然气转化为绿色化学品的工程微生物。天然气是化工生产中最便宜的原料。使用这种廉价的原材料将为绿色制造方法开辟数十亿美元的燃料和化学市场，而这些方法以前被认为无利可图。这些项目的商业化将增加清洁技术、制造业和建筑业的创新、投资和就业增长。如果成功，天然气发酵技术将减少碳污染；对目标化学品生产的生命周期分析表明，与目前的石油和煤炭工艺相比，二氧化碳排放量减少了六倍。最后，这个项目的目标是发现新的酶，可以在处理环境中的碳氢化合物泄漏中找到应用。该SBIR一期项目建议将乙烷消耗途径设计成工业微生物，以降低制造化学品的成本。该解决方案的核心是从环境DNA样本中识别乙烷氧化酶。绝大多数细菌从未被培养过，但最近的技术进步现在允许从不同的环境中提取数百万以前未知的DNA片段并进行测试。筛选环境DNA样本的新功能已经确定了生物基化学物质的新途径。该提案概述了一种基于选择的策略，以丰富可以氧化乙烷的酶的大型文库。我们第一阶段研究的结果将是世界上第一个在乙烷上生长的工业菌株。该菌株将作为一个平台：(1)利用定向进化加速乙烷的生长速度；(2)生产工业产品，包括一种有价值的四碳构建块化学物质；(3)开发一种消耗甲烷的途径。这种强大的选择方案可能会导致发现全新种类的酶，这些酶可能通过新的催化机制起作用。在经过充分研究的菌株中成功地、功能性地表达这种酶，将加速这些迷人而复杂的酶的生化表征。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be a fermentation-based technology capable of converting abundant American natural gas supplies into valuable building-block chemicals that will advance economic and security interests while encouraging innovation in manufacturing. This project focuses on engineering microorganisms that can convert natural gas into green chemicals via fermentation. Natural gas is the least expensive raw material available for chemical production. The use of this inexpensive raw material will open up multi-billion-dollar fuel and chemical markets for green manufacturing methods that have been previously disregarded as unprofitable. Commercialization of these projects will increase innovation, investment, and job growth in the clean technology, manufacturing, and construction industries. If successful, natural gas-based fermentation technology will reduce carbon pollution; a lifecycle analysis for production of a target chemical shows a six-fold reduction of carbon dioxide emissions compared to the current petroleum and coal process. Finally, this project aims to discover new enzymes that may find applications in treating hydrocarbon spills in the environment. This SBIR Phase I project proposes to engineer an ethane-consuming pathway into an industrial microorganism in order to reduce the cost of manufacturing chemicals. The solution centers on identifying ethane-oxidizing enzymes from environmental DNA samples. The vast majority of bacteria have never been cultured, but recent technological advances now allow millions of previously unknown DNA fragments to be extracted from diverse environments and tested. Screening environmental DNA samples for novel functionality has identified new pathways to bio-based chemicals. This proposal outlines a selection-based strategy to enrich a large library for enzymes that can oxidize ethane. The outcome of our Phase I research will be the world's first industrial strain engineered to grow on ethane. This strain will serve as a platform for: (1) accelerating the growth rate on ethane, using directed evolution, (2) producing industrial products, including a valuable four-carbon building block chemical, and (3) developing a pathway that consumes methane. This powerful selection scheme could lead to the discovery of entirely new classes of enzymes, which may function via novel catalytic mechanisms. Successful, functional expression of such an enzyme in a well-studied strain would accelerate biochemical characterization of these fascinating and complicated enzymes.