STTR Phase II: Development of a computational protein engineering platform and its application to methane activating enzymes

STTR第二阶段：计算蛋白质工程平台的开发及其在甲烷活化酶中的应用

• 批准号: 1534743
• 负责人: Barry Olafson
• 金额: $ 75万
• 依托单位: Protabit LLC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2018-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1534743&HistoricalAwards=false
• 关键词: STTR; Phase; II; Development; computational

The broader impact/commercial potential of this Small Business Innovation Research Phase II project will combine computational protein engineering (CPE) software, tools and methods into a "platform technology" that enables new products and technologies in a wide range of scientific areas, including industrial enzymes, pharmaceuticals, therapeutics, medical diagnostics and bioenergy. This project will validate the CPE platform in an application that has both commercial and environmental value: engineered enzymes for utilizing methane as a feedstock. The engineered methane activating enzymes developed will open up numerous new biological routes to chemicals and fuels from natural gas, capitalizing on abundant domestic shale gas reserves and improving U.S. energy independence. This CPE enzyme technology has the potential to significantly decrease the capital cost of small-scale Gas-to-liquid (GTL) plant, enabling the capture and monetization of otherwise economically-stranded natural gas. In North Dakota, more than a quarter of the total gas produced is flared or vented, wasting a valuable natural resource and needlessly emitting greenhouse gases. By facilitating the conversion of stranded or flared methane to fuels and high-value chemicals, this research can help reduce carbon footprint and spur domestic manufacturing, investment, and job creation. The objectives of this Phase II project are to develop and license a CPE Platform technology that will enable experimental biological scientists to harness the power of computational protein design and engineering; to validate the CPE Platform technology by engineering novel methane activating enzymes that have increased soluble expression and methane-oxidizing activity; and in collaboration with potential licensees of this technology, work to incorporate the engineered enzyme into their industrial host organisms. These organisms will be enabled to use methane as a feedstock for the bio-production of a wide range of end products, including liquid transportation fuels, commodity chemicals and high value fine chemicals. In Phase I, the Northwestern-Caltech-Protabit team succeeded in engineering the particulate MMO (pMMO) catalytic subunit (spmoB) to allow it to be solubly expressed in E. coli. This was the first demonstration of an active methane-oxidizing enzyme that can be solubly expressed and purified in significant quantities in a genetically-tractable recombinant host. Phase II will continue this work, with the objective of making the spmoB enzyme more effective by increasing its activity and solving certain other challenges for inserting it into an industrial host.

这个小企业创新研究第二阶段项目的更广泛的影响/商业潜力将把计算蛋白工程（CPE）软件、工具和方法结合到一个“平台技术”中，使新产品和技术在广泛的科学领域成为可能，包括工业酶、制药、治疗、医疗诊断和生物能源。该项目将在一个具有商业和环境价值的应用中验证CPE平台：利用甲烷作为原料的工程酶。开发的工程甲烷激活酶将开辟许多新的生物途径，从天然气中获得化学品和燃料，利用丰富的国内页岩气储量，提高美国的能源独立性。这种CPE酶技术有可能显著降低小型气制液（GTL）工厂的资本成本，从而实现经济上搁浅的天然气的捕获和货币化。在北达科他州，超过四分之一的天然气被燃烧或排放，浪费了宝贵的自然资源，并不必要地排放温室气体。通过促进将滞留或燃烧的甲烷转化为燃料和高价值化学品，这项研究有助于减少碳足迹，刺激国内制造业、投资和创造就业机会。第二阶段项目的目标是开发和许可CPE平台技术，使实验生物科学家能够利用计算蛋白质设计和工程的力量；通过设计提高可溶性表达和甲烷氧化活性的新型甲烷激活酶来验证CPE平台技术；并与这项技术的潜在授权方合作，努力将这种工程酶整合到它们的工业宿主生物中。这些生物将能够使用甲烷作为原料，生产各种最终产品，包括液体运输燃料、商品化学品和高价值精细化学品。在第一阶段，西北加州理工大学- protabit团队成功地设计了颗粒MMO （pMMO）催化亚基（spmoB），使其能够在大肠杆菌中可溶性表达。这是首次证明一种活性甲烷氧化酶可以在基因可处理的重组宿主中大量可溶性表达和纯化。第二阶段将继续这项工作，目标是通过增加spmoB酶的活性并解决将其插入工业宿主的某些其他挑战，使其更有效。