Source: ENCHI CORPORATION submitted to NRP
PROOF-OF-CONCEPT FOR LIGNOCELLULOSE PROCESSING TO FUELS USING ENGINEERED THERMOPHILIC BACTERIA AND COTREATMENT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
SMALL BUSINESS GRANT
Reporting Frequency
Annual
Accession No.
1022823
Grant No.
2020-33610-32002
Cumulative Award Amt.
$99,830.00
Proposal No.
2020-00528
Multistate No.
(N/A)
Project Start Date
Sep 1, 2020
Project End Date
May 31, 2021
Grant Year
2020
Program Code
[8.8]- Biofuels and Biobased Products
Recipient Organization
ENCHI CORPORATION
610 LINCOLN ST STE 1
WALTHAM,MA 024512189
Performing Department
(N/A)
Non Technical Summary
Enchi Corporation proposes an SBIR project aimed at an innovative approach for low-cost processing of lignocellulose to fuels and chemicals based on engineered thermophilic bacteria combined with milling during fermentation (cotreatment). This approach, termed C-CBP, avoids the two process steps responsible for the high cost of current technology: thermochemical pretreatment and added enzymes. We target the application of C-CBP with the shortest path to commercialization: corn stover to ethanol.The potential of C-CBP for disruptive reductions in the cost of lignocellulose processing is indicated by peer-reviewed technoeconomic analysis carried out in collaboration with the National Renewable Energy Laboratory (NREL), in which an advanced scenario featuring C-CBP is compared to an NREL-defined base-case featuring dilute acid pretreatment and added fungal cellulase with coproduced electricity. Compared to the base case, the advanced scenario has an 8-fold shorter payback period and economic feasibility at 10-fold smaller scale. Extensions of this analysis show project payback periods for performance parameters in Phase I and Phase II.Phase I aims to provide an initial proof of concept that the various components of the C-CBP approach can be functionally integrated, and will enable Phase II activities aimed at realizing commercially-viable performance. Tasks include: 1. Corn stover fermentation using thermophilic cocultures and cotreatment; 2. Technoeconomic analysis; and 3. Preparation of the Phase II proposal. A large-scale ethanol producing company will provide corn stover and participate in Task 2. A consultant will also be included to provide expertise on biomass milling for cotreatment and estimating costs for the equipment to be used at large scale.
Animal Health Component
50%
Research Effort Categories
Basic
(N/A)
Applied
50%
Developmental
50%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
51140991100100%
Knowledge Area
511 - New and Improved Non-Food Products and Processes;

Subject Of Investigation
4099 - Microorganisms, general/other;

Field Of Science
1100 - Bacteriology;
Goals / Objectives
Enchi Corporation proposes an SBIR project aimed at an innovative approach for low-cost processing of lignocellulose to fuels and chemicals based on engineered thermophilic bacteria combined with milling during fermentation (cotreatment). This approach, termed C-CBP, avoids the two process steps responsible for the high cost of current technology: thermochemical pretreatment and added enzymes. We target the application of C-CBP with the shortest path to commercialization: corn stover to ethanol. Phase I aims to provide an initial proof of concept that the various components of the C-CBP approach can be functionally integrated, and will enable Phase II activities aimed at realizing commercially-viable performance.Task 1. Demonstrate performance. Realize performance parameter values or similar values with equivalent payback period using a co-culture of engineered C. thermocellum and T. thermosaccharolyticum.Task 2. Technoeconomic analysis. Analyze process economics based on performance parameters achieved in Phase I.Task 3. Prepare the Phase II proposal.
Project Methods
Culture of C. thermocellum is described in many publications, but a notable reference is Ghosh S, Holwerda EK, Worthen RS, Lynd LR, Epps BP. 2018. Biotechnol Biofuels. Rheological properties of corn stover slurries during fermentation by Clostridium thermocellum. 11:246. doi: 10.1186/s13068-018-1248-z.Cotreatment, one of the key methods used herein, is described in Balch ML, et al. 2017. "Lignocellulose fermentation and residual solids characterization for senescent switchgrass fermentation by Clostridium thermocellum in the presence and absence of continuous in situ ball-milling." Energy Environ. Sci., 10, 1252.Strain and process optimization of a related thermophile is described in Herring CD et al. "Strain and bioprocess improvement of a thermophilic anaerobe for the production of ethanol from wood." Biotechnol Biofuels. 2016. 9:125. doi: 10.1186/s13068-016-0536-8.

Progress 09/01/20 to 05/31/21

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 a technician who 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. 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 with 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 reviewers of the Phase II SBIR proposal that continues the work of this Phase I project. What do you plan to do during the next reporting period to accomplish the goals?This project will be continued as a Phase II SBIR under award 2021-33610-35655 "Development of C-CBP to enable cash-positive conversion of corn stover to biofuels co-located at a corn ethanol mill."

Impacts
What was accomplished under these goals? The technical objective of Phase I, to show proof-of-concept for C-CBP by demonstrating performance with corn stover, has been achieved. Multiple performance values previously shown separately, such as solubilization, yield and titer, have now been demonstrated in a single integrated fermentation with realistic substrate. We combined advances in strain engineering and bioprocess development. The highest ethanol titer produced by thermophilic bacteria from corn stover was increased more than 60% with advances in strain engineering and bioprocess development. Cotreatment has been successfully integrated into the bioprocess but further experimentation is warranted. Our results give strong support for the technical feasibility of C-CBP. In the Phase I application, it was stated that three components were needed to reach the Phase I objectives: 1) a compatible co-culture of engineered thermophiles; 2) cotreatment integrated with fermentation; 3) a bioreactor configuration compatible with 1 & 2 and able to ferment >105 g/L corn stover. All three of these components were produced and integrated in the Phase I project. Previous experience with strain and bioprocess improvement suggests that continued improvements will follow with continued effort and funding.

Publications