Source: Terragia Biofuel Incorporated submitted to NRP
DEVELOPMENT OF C-CBP TO ENABLE CASH-POSITIVE CONVERSION OF CORN STOVER TO BIOFUELS CO-LOCATED AT A CORN ETHANOL MILL
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1030895
Grant No.
2023-33610-40261
Cumulative Award Amt.
$193,242.94
Proposal No.
2023-03880
Multistate No.
(N/A)
Project Start Date
May 15, 2023
Project End Date
Nov 14, 2023
Grant Year
2023
Program Code
[8.8]- Biofuels and Biobased Products
Recipient Organization
Terragia Biofuel Incorporated
15 Thayer Drive
Hanover,NH 03755
Performing Department
Terragia Biofuel Incorporated
Non Technical Summary
Terragia Biofuel Inc.proposes a Phase II SBIR project aimed at an innovative approach for low-cost processing of lignocelluloseto fuels and chemicals based on engineered thermophilic bacteria combined with milling during fermentation (cotreatment). Thisapproach, termed C-CBP, avoids the two process steps responsible for the high cost of current technology: thermochemicalpretreatment and added enzymes. We target the opportunity for C-CBP with the shortest path to commercialization: corn stoverto ethanol. Phase I efforts nearly doubled ethanol titers from corn stover, successfully demonstrated simultaneous conversion ofC6 and C5 sugars, and showed that components of the C-CBP approach can be functionally integrated.Peer-reviewed technoeconomic analysis carried out in collaboration with the National Renewable Energy Laboratory (NREL)has shown that an advanced cellulosic ethanol plant employing C-CBP has an 8-fold shorter payback period and economicfeasibility at 10-fold smaller scale than conventional scenarios. Given the cost savings and scale-insensitivity of C-CBP, wehypothesize that it is possible to build a demonstration facility for which revenues exceed operating expenses, co-located at anexisting corn ethanol plant. Toward this end, objectives of the Phase II proposal are:1. Improve the technical performance of ethanol production from corn stover via C-CBP through a combination of bioprocessand strain improvement;2. Develop a low-cost growth medium for C-CBP making use of streams available within a corn ethanol mill;3. Develop a detailed design for a demonstration plant co-located at a POET corn ethanol mill that will cost less than $5 millionto build and have revenues that exceed operating expenses.4. Demonstrate key features of C-CBP at 60 gallon scale at POET's Research Center in Scotland, South Dakota.5. Perform a technoeconomic and market analysis of catalytic conversion of ethanol to hydrocarbon blendstocks, co-located at acorn ethanol plant.POET LLC will provide materials needed for work on objectives 1 and 2, and will participate in activities pursuant to theremaining objectives. Vertimass LLC will work with Enchi and POET on objective 5. An experienced industry consultant and IPfirm will provide Technical and Business Assistance.The anticipated results of this project include the development of a small business and commercialization of a highly innovativetechnology. This Phase II project represents a critical step toward enabling low-cost conversion of corn-stover to ethanol andwould lead to increased revenue and job creation for both corn farmers and fuel producers. Commercialization of C-CBP will bean important step toward realizing the long-anticipated benefits of expanded use of cellulosic feedstocks with respect to ruraleconomic development and climate stabilization.
Animal Health Component
80%
Research Effort Categories
Basic
10%
Applied
80%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
40240101100100%
Knowledge Area
402 - Engineering Systems and Equipment;

Subject Of Investigation
4010 - Bacteria;

Field Of Science
1100 - Bacteriology;
Goals / Objectives
1. Improve the technical performance of ethanol production from corn stover via C-CBP through a combination of bioprocessand strain improvement;2. Develop a low-cost growth medium for C-CBP making use of streams available within a corn ethanol mill;3. Incorporating results from 1 & 2, develop a detailed design for a demonstration plant co-located at a POET corn ethanolmill that will cost less than $5 million to build and have revenues that exceed operating expenses.4. Demonstrate key features of C-CBP at 60 gallon scale at POET's Research Center in Scotland, South Dakota.5. Perform a technoeconomic and market analysis of catalytic conversion of ethanol to hydrocarbon blendstocks co-locatedat a corn ethanol plant.
Project Methods
We will perform strain and bioprocess improvement using methods described in Herring et al., 2016 (Biotechnol Biofuels. 2016.9:125.). Technoeconomic analysis will be performed using methods similar to those described in Lynd et al., 2017 (Curr OpinBiotechnol. 45:202-211.)Corn stover will be supplied by POET LLC. Fermentations will be conducted at 0.5 - 2.0 L scale in Sartorius Biostat A pHcontrolledreactors or in small scale 20-50 ml serum bottle cultures. Genetic engineering methods will be used as previouslydescribed (Shaw et al. 2012, Herring et al. 2016, Hon et al. 2020). The sugar composition of corn stover and solid fermentationresidues will be determined by acid hydrolysis and subsequent HPLC (Sluiter et al. 2008). Total solubilized sugars infermentation broths (both mono- and polymeric) will be determined by a similar method of acid hydrolysis and HPLC calledLiquid QS (Beri et al. 2020).

Progress 05/15/23 to 11/14/23

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 continued work, 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? Nothing Reported

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. 6. Different stover samples were obtained with varying carbohydrate content and performance wastested with our bacteria. 7. Work was performed to study the performance decrease observed at high solids loadings of corn stover. An analytical system was established to fractionate stover hydrolysateto identify what components are inhibitory at the levels of cell growth and cellulase activity. 8. Work was performed tosupport a patent application showinghigh ethanol production frommixes of cellulosic and starch compoents of agricultual materials, such as mixes of corn stover with corn kernels.Technoeconomic analysis was conducted to study the commercial feasability of such a bioprocess.

Publications