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

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.