Progress 09/01/21 to 02/15/23

Outputs
Target Audience:The intended audience for the information generated under this project includes potential investors who would provide future financing for R&D and commercialization, or potential commercial partners, such as biofuel producers and technology providers in the biofuel space. Grant program managers and peer reviewers, along with the wider scientific community are also part of the intended audience. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This SBIR funding supported two technicians, one of whom was a new college graduate at the outset, and who received training in lignocellulose bioprocess research, particularly biomass processing and analysis, cultivation of anaerobic thermophilic bacteria, bioprocess optimization and troubleshooting, fermentation analysis and data reporting. A junior scientist was hired, who had previously worked on a similar subject in an academic setting. Under this project, he is receiving training in R&D focused on technology commercialization. The Project Director continued his professional development under this award by further advancing his ability to lead and coordinate grant funded research while also furthering business-oriented commercialization activities. How have the results been disseminated to communities of interest?Results of this project have been communicated as confidential information to the CSO and SVP of Research at POET as well as members of the Enchi Corporation Board of Directors, leadership of the DOE Center for BioEnergy Innovation, members of Lee Lynd's academic lab at Dartmouth College and our TABA consultant, bioengineering consultant Jeff Lievense. What do you plan to do during the next reporting period to accomplish the goals?Focus Areas for Continuation of the Project under Terragia Biofuel Inc. Identify underlying reasons for reduced performance at high solids and develop strategies to overcome them Continue to develop a low-cost growth medium for C-CBP making use of streams available within a corn ethanol mill Demonstrate key features of C-CBP at 60 gallon scale at POET's Research Center in Scotland, South Dakota.

Impacts
What was accomplished under these goals? 1. Improved strains were generated for T. thermosaccharolyticum and C. thermocellum. Ethanol titers from corn stover fermentations were increased beyond values from Phase I. Limiting factors in high solids fermentation were idenfied and overcome with bioprocess improvements. An assay was developed to continue to troubleshoot high solids fermentations. Cofermentation of corn stover and corn kernels was tested and ethanol titers approaching the target values were obtained. 2. Expensive nutrients (yeast extract) were replaced with less expensive alternatives, and vitamin requirements were identified. Process materials from a commercial corn ethanol facility were tested and found to be compatible with our process. 3. A design for a low-capital demonstration facility coloated at a commercial corn ethanol plant was modeled, and various scales were analyzed on the basis of the total fermentation capacity, revenues vs fixed operating costs and labor costs. 4. Two project leaders visited POET LLC and discussed plans for a demonstration of our process at POET's facility. 5. Technoeconomic analysis was conducted to compare scenarios with and without catalytic conversion of ethanol to hydrocarbons, and current performance levels and for projected future performance.

Publications

Progress 09/01/21 to 08/31/22

Outputs
Target Audience:The intended audience for the information generated under this project includes potential investors who would provide future financing for R&D and commercialization, or potential commercial partners, such as biofuel producers and technology providers in the biofuel space. Grant program managers and peer reviewers, along with the wider scientific community are also part of the intended audience. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This SBIR funding supported two technicians, one of whom was a new college graduate at the outset, and who received training in lignocellulose bioprocess research, particularly biomass processing and analysis, cultivation of anaerobic thermophilic bacteria, bioprocess optimization and troubleshooting, fermentation analysis and data reporting. A junior scientist was hired, who had previously worked on a similar subject in an academic setting. Under this project, he is receiving training in R&D focused on technology commercialization. The Project Director continued his professional development under this award by further advancing his ability to lead and coordinate grant funded research while also furthering business-oriented commercialization activities. How have the results been disseminated to communities of interest?Results of this project have been communicated as confidential information to the CSO and SVP of Research at POET as well as members of the Enchi Corporation Board of Directors, leadership of the DOE Center for BioEnergy Innovation, members of Lee Lynd's academic lab at Dartmouth College and our TABA consultant, bioengineering consultant Jeff Lievense. What do you plan to do during the next reporting period to accomplish the goals? We will continue to troubleshoot declining conversion of corn stover at increasing solids concentrations, pursuing both understanding of the limitations, as well as testing of strategies to overcome them. We will also continue to use adaptive evolution to generate improved strains of C. thermocellum and T. thermosaccharolyticum. We will also work to document and analyze process performance in the laboratory with materials and conditions that are expected in an industrial environment (facility co-located at an existing corn ethanol plant). A low-cost media formulation will be developed and the impact of the new formulation will be evaluated using technoeconomic models. Revisions to the formulation will be made to reduce the operating costs as much as possible. We will complete efforts to design a demonstration plant co-located at a POET corn ethanol mill, drawing conclusions about the efficacy of such a plant to reduce risks associated with scale up and demonstrate our operational capabilities at the lowest possible capital cost. Demonstration of key features of C-CBP at 60 gallon scale is planned at POET's Research Center in Scotland, South Dakota. We will complete the technoeconomic analysis of catalytic conversion of ethanol to hydrocarbon blendstocks co-located at a corn ethanol plant, with the aim of determining whether this approach could increase the profitability or marketability of corn stover conversion.

Impacts
What was accomplished under these goals? Progress was made on Objectives 1-5 as follows: 1. For economical conversion of corn stover to ethanol, we need to be able to show good fermentation performance at high concentrations of corn stover (i.e. high solids levels). Using the new fermentation technique developed in Phase I, we have increased ethanol concentrations approximately 20% above levels previously achieved. Also, we discovered a process parameter that has a major impact on process performance at high solids levels. However, we still observe a decline in stover conversion corresponding to the level of solids. Troubleshooting this problem is an ongoing subject of our research and development. Improvements in our strains of bacteria were made in order to increase the conversion performance and thereby reduce costs. Genetic engineering was used to improve the yield of ethanol for C. thermocellum, the bacterium responsible for deconstruction of cellulose. We have also used adaptive laboratory evolution to increase the performance of T. thermosaccharolyticum, the bacterium responsible for conversion of hemicellulose to ethanol. 2. The process of developing low-cost nutrient formulations for the bacteria was initiated. We discovered that small amounts of vitamins greatly increase the ability of one of our bacteria to use low-cost inorganic sources of nitrogen. We will use this information to develop a low cost formulation in the second half of the project. 3. A detailed process was designed for a plant that converts stover to ethanol, collocated at an existing corn ethanol plant. Technoeconomic analysis was performed, incorporating realistic cost factors for capital equipment and operating expenses. The analysis has been shared with our industrial partner POET and discussion is ongoing about the scale of such a facility that would generate revenue equal to operating costs. 4. Discussions are planned with our industrial partner POET to demonstrate our process in a realistic operating environment in the second half of the project. 5. Technoeconomic modeling was performed to study the feasibility of fermenting corn stover to produce ethanol as an intermediate, then catalytically convert the ethanol to hydrocarbons that could be used in Sustainable Aviation Fuel (SAF). The analysis was performed in partnership with Vertimass, a company seeking to commercialize the catalytic technology, and discussions of the results with POET are in progress.

Publications