Progress 09/01/09 to 02/29/12

Outputs
OUTPUTS: The main goal of this project was to develop an integrated process for producing polyitaconic acid through the fermentation of a biomass-based sugar feedstock. Currently, there is no commercial source of itaconic acid in the United States. Itaconix Corporation, the prime for this award, is a company which requires itaconic acid as the feedstock for the production of their suite of bio-based products. Tasks for this project were addressed by a multidisciplinary team comprised of scientists from two universities, a small business, and supplemented by experts contracted specifically for this project. By the close of this project, the research team successfully developed a stream-integrated approach for the cost effective production of a specialty water soluble polymer polyitaconic acid (PIA). Specifically, work focused on producing "minimally treated carbohydrate" from wood biomass for microbial conversion to itaconic acid (IA). Aspergillus terreus is known to produce itaconic acid from either C6 or C5 sugars. This project proposed the development of a process that used northeastern hardwoods as the source of C5 sugars. This process included; hemicellulose extraction, sugar hydrolysis and detoxification of the hydrolyzate, fermentation, and the recovery of itaconic acid from the fermentation broth followed by polymerization. The project also included an economic analysis for a commercial scale plant and a cradle to gate Life Cycle Analysis (LCA) of the impact on resource use. The major project achievements include the following. The extraction of hemicellulose from hardwood biomass was achieved and a partial detoxification of the crude extract was performed. The effects of xylan extract pre-treatment conditions and potential inhibitors on growth and IA production by A. terreus was quantified and the analysis of the final extract composition and the fermentation of minimally pretreated xylan extract were performed. The optimization of a laboratory scale fermentation for the conversion of xylose to itaconix acid was achieved. The optimization of large scale sporulation and the completion of the analysis of interfering impurities in the fermentation broth were achieved. Bench-scale recovery trials with model and experimental broths were achieved and the transfer kinetics for the extraction and stripping of experimental broths were determined. The potential impurity effects on the polymerization of IA using spiked solutions were assessed. A prototype synthesis and polymerization of esters of IA was achieved. A process design and economic analysis on the production of PIA from northeast wood biomass was performed, and an economic analysis of the integrated process was completed. The biodegradability testing of PIA was completed, and a Life Cycle Analysis was performed. From a more macroscale perspective this project was successful in developing new products made from lignocellulosic biomass. Yet it was determined that the economics of a stand alone green field PIA plant co-located with a biomass energy plant were not favorable. Alternative co-locations offering greater economic synergy continue to be explored. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The main goal of this project was to develop an integrated process for producing polyitaconic acid through the fermentation of a biomass-based sugar feedstock. While the various processes to perform this transformation existed separately or as a limited methodology, integrating these processes had never before been done with PIA. In addition, each stream associated with a processing step needed optimization in order to provide the maximum efficiency, productivity, and performance necessary to yield a total value chain that could be competitive with petrochemical processes. This project was performed by a multi-disciplinary team of researchers. The University of Maine, Orono provided expertise in the area of hemicellulose extraction with demonstrated capabilities in xylan production from hardwood within a kraft pulping plant. The University of Massachusetts, Lowell provided expertise in optimizing industrial fermentation processes. Itaconix brought the capabilities needed to optimize the extraction of organic acids from fermentation broth, and to purify the extract to a polymerizable grade. Itaconix also had the technology to polymerize itaconic acid into polyitaconic acid, a water soluble polymer with marketable potential as a dispersant, detergent, and superabsorbent. Itaconix also provided the expertise needed to perform the majority of the fermentation tasks required during the first 18 months of the project. Itaconix worked with the University of Maine to investigate the effects of extraction conditions on fermentation and overall process integration. Early on it was realized that certain substances in the wood extract interfered to varying degrees with itaconic acid production during fermentation. Therefore the dozen or more potential inhibitors present in the extract were identified and potential treatment options were examined. Itaconix also took the lead in investigating enzyme hydrolysis as an alternative or potentially complementary approach to obtaining a complete monomeric sugar preparation for fermentation. An outline of the major tasks performed during this project is provided below along with the accomplishments achieved for each activity. The results from this project have the potential to re-establish the production of itaconic acid (possibly from woody biomass) in the US. During this project the research team was successful in developing a stream-integrated approach to the cost effective production of a specialty water soluble polymer polyitaconic acid. Specifically, work focused on producing "minimally treated carbohydrate" from wood biomass for microbial conversion to itaconic acid. In addition, prototype samples of bio-latex and bio-adhesive have been made and partnerships for commercial development are being pursued. A list of patents and invited talks that are a direct result of this project is provided below.

Publications

• 5) Speaker: 2nd Annual Next Generation Bio-Based Chemicals Summit, San Diego,CA. Title: From wood to detergents and paints Date: February 16th 2011 Presenter: Yvon Durant.
• 6) Invited speaker: 94th Canadian Chemistry Conference, Montreal, Canada. Title: From wood to wood coatings Presenter: Yvon Durant, Date: June 6th 2011.
• 7) Invited speaker: TAPPI, Orono, Me. Title: Biobased Latex for Paper coatings. Presenter: Yvon Durant October 6th 2011.
• 1) Emulsion Polymerization of Esters of Itaconic Acid, United States Patent Application 20110144265 Inventors: Durant, Yvon (Lee, NH, US)
• 2) Vinyl Acid monomer recovery. Patent US application number: 13081187. Application Date: June 16th 2011.
• 3) Making a Wood Varnish out of Wood. University of New Hampshire Materials Science Seminar, Presenter: Yvon Durant, Date: October 16th 2010.
• 4) Seminar: University of New Hampshire. Chemistry Department Title: From wood to wood coatings Presenter: Yvon Durant, Date: February 8th 2011.

Progress 09/01/10 to 08/31/11

Outputs
OUTPUTS: Our collaborators, the University of Maine and University of Massachusetts are 95% complete on their tasks and will be done by August 31. Our most intensive tasks, Recovery of Itaconic Acid from A. Terreus and Engineering Integration are both around the 75-80% mark. We are making great strides and have quantified the inhibitors to the fungi, and have found the optimum methodology for esterification of the polymer for creating biolatexes by emulsion polymerization. We have been giving presentations on our work at a number of national and international Biomass Symposia as well as receiving a patent on the discoveries made during this project. Patent Title: Emulsion Polymerization Of Esters Of Itaconic Acid Document Type and Number: U.S. Patent Application 20110144265 Kind Code: A1 Inventors: Durant, Yvon (Lee, NH, US) Publication Date: 06/16/2011 Filing Date: 12/16/2010 Abstract: Esters of itaconic acid are polymerized in aqueous medium in the presence of seed particles. The seed particles may absorb the monomer and provide for polymerization to a desired particle size. The polymers that are produced may be employed in adhesives, paints, and paper formulations. Seminar : University of New Hampshire. Materials Science program Title: Making a Wood Varnish out of Wood Date: 10-06-2010 Presenter: Yvon Durant. Seminar : University of New Hampshire. Chemistry Department Title: From wood to wood coatings Date: 02-08-2011 Presenter: Yvon Durant. Speaker: 2nd Annual Next Generation Bio-Based Chemicals Summit, San Diego, CA. Title: From wood to detergents and paints Date: 02-16-2011 Presenter: Yvon Durant. Invited speaker: 94th Canadian Chemistry Conference, Montreal, Canada. Title: From wood to wood coatings Date: 06-06-2011 Presenter: Yvon Durant. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Seminar : University of New Hampshire. Materials Science program Title: Making a Wood Varnish out of Wood Date: 10-06-2010 Presenter: Yvon Durant. Seminar : University of New Hampshire. Chemistry Department Title: From wood to wood coatings Date: 02-08-2011 Presenter: Yvon Durant. Speaker: 2nd Annual Next Generation Bio-Based Chemicals Summit, San Diego, CA. Title: From wood to detergents and paints Date: 02-16-2011 Presenter: Yvon Durant. Invited speaker: 94th Canadian Chemistry Conference, Montreal, Canada. Title: From wood to wood coatings Date: 06-06-2011 Presenter: Yvon Durant. PROJECT MODIFICATIONS: The original objective of Task 1 was to extract northeast hardwood to maximize dissolution of hemicellulose polymers for hydrolysis to monomer sugars, predominantly xylose, while producing the minimum amount of impurities that inhibit fermentation of the sugars to itaconic acid. During the course of the research the initial objective was broadened to include mixed northeastern softwoods as well as corn gluten feed. Both the northeastern softwood and the corn gluten feed are high in 6-carbon sugars such as glucose and mannose which we found to be more readily converted to Itaconic acid during fermentation by Aspergillus terreus than the 5-carbon sugars such as xylose and arabinose.

Impacts
Goal/Objective The University of Maine was responsible for producing extracts from biomass that have a high concentration of sugars (100 grams per liter) suitable for the conversion to itaconic acid by fermentation using the fungus (mold) Aspergillus terreus. The process investigated consists of the extraction of carbohydrates or complex sugar polymers from the biomass source by using water or dilute acid. The polymeric sugars in the raw extract solution are first concentrated, and then the molecular weight of the sugar polymers is reduced by acid hydrolysis or cleavage of the carbohydrate macromolecules into component sugars. Finally, the extract is purified by removal of substances that are toxic to the Aspergillus terreus fungus used in fermentation. Substances known to be toxic to Aspergillus terreus are lignin and organic acids such as acetic acid as well as furfural. The original objective of Task 1 was to extract northeast hardwood to maximize dissolution of hemicellulose polymers for hydrolysis to monomer sugars, predominantly xylose, while producing the minimum amount of impurities that inhibit fermentation of the sugars to itaconic acid. During the course of the research the initial objective was broadened to include mixed northeastern softwoods as well as corn gluten feed. Both the northeastern softwood and the corn gluten feed are high in 6-carbon sugars such as glucose and mannose which we found to be more readily converted to Itaconic acid during fermentation by Aspergillus terreus than the 5-carbon sugars such as xylose and arabinose. Figure 1. Laboratory and Pilot Scale Extraction Reactors (not shown) Findings/Conclusions Sugars are readily removed by extracting biomass with either water or dilute acid. Depending upon the final pH of the extract, the sugars are removed in the polymeric (or macromolecular form) and as simple sugars. An attractive biomass material evaluated as a possible carbohydrate source was never-dried corn gluten feed which could be readily extracted with dilute acid. These conditions result in raw extracts with high concentrations of 6-carbon sugars, predominately glucose and mannose. During the extraction and hydrolysis processes, substances are formed in the extract such as soluble lignin, acetic acid, and furfural that are toxic to Aspergillus terreus. Thus, to use the extract as a source of sugars these substances must be removed by purification of the extract.

Publications

• No publications reported this period

Progress 09/01/09 to 08/31/10

Outputs
OUTPUTS: Consortium: Itaconix, LLC, (Principal); University of Massachusetts, Lowell; University of Maine, Orono WORKPLAN- Collaborator Tasks Accomplished Over the First Year of the Biomass Grant U. Maine: High Xylose Feedstock Devel. for Microbial Conversion to IA Hemicellulose Extraction from Hardwood Biomass (80% complete) Experiments Designed to Upgrade the Quality of the Crude Extract (75% complete) Analysis of Final Extract Composition (79% complete) Preliminary Process Design and Economic Analysis of Extract Production (25% complete) U.Mass: Strain Development and Fermentation Optimize pellet formation of Aspergillus terreus (19% complete) Large Scale Sporulation (10% complete) Fermented Material (IA) for Purification and polymerization experiments (initial experiments started) Potential inhibitors of xylan extract (5% complete) Nutrient Optimization of IA production (15% complete) Strain Optimization and genetic engineering (initial experiments started) Pilot Runs Itaconix: Recovery Feedstock/metabolite impurity analysis (90% complete) Liquid Liquid Extraction data acquisition and FTIR (70% complete) Bench scale trials with model and real broths (40% complete) Transfer kinetics of HFLM Ext/stripping with real broths (80% complete) Scale Up/Skid Development/Testing at UNH (80% complete) Mobile recovery unit integration (50% complete) Itaconix: Polymerization Commercial Stock Potential Impurity Assessment (80% complete) Synthesis and Polymerization of Esters of Itaconic Acid (70% complete) Recovery Skid Operation and Polymerization integration (40% complete) Life Cycle Analysis (5% complete) Economic Analysis of Integrated Process (30% complete) Biodegradability testing of Poly Itatonic Acid (60% complete) PARTICIPANTS: Itaconix Senior/Key Person PI (Yvon Durant)Polymerization, Co- PI (Jennifer Durant)Recovery, Other Personnel: Research Mgr (Theresa Ridgeway), R A (Elif Alyamac), R A (Yangbin Chen), R A (Brian Luthi), R A (Andrew Bryce) University of Maine, Orono - Dr. Joe Genco University of Massachusetts, Lowell - Dr. Carl Lawton TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Through this grant, we are working to develop a stream-integrated approach to the production of a large industrial commodity, polyitaconic acid. Studies are being carried out at the University of Maine with the objective of providing the University of Massachusetts and Itaconix with minimally treated carbohydrate from wood biomass for use in microbial conversion to Itaconic Acid. Analysis on the samples from the water extracted Northeast hardwood chips have been completed by the group at University of Maine, Orono. The data show that the amount of lignin in the liquor increases as the concentration level increases. The data indicate that if we concentrate to 30% solids, we can achieve total sugar concentrations of 200 grams/liter; and xylose concentrations of 160 grams per liter. The Recovery Unit group headed by Dr. Jennifer Durant is tasked with the recovery of itaconic acid from fermentation broth. Their focus has been on the use of Hollow Fiber technology to accomplish this goal. Based on softwood extract composition data from U. Maine, Dr. Durants group has been examining the effect of the carbon source on the relative itaconic acid titer. The parent A. terreus strain responds similarly to xylose, mannose and mixtures thereof. We have also developed a method for large scale sporulation. The Recovery Unit has determined three potential solvent systems. The extraction systems are composed of an amine, modifier, and diluent. The modifier plays a critical role in IA extraction. To confirm the observed equilibrium loading data, we have also investigated the acid:amine complex within the solvent system. Inspection of the carbonyl and carboxylate bands suggest that the interaction involves a 1:1 rather than 1:2 complex. In developing the recovery method, a thorough understanding of the impurities present is necessary. Based on our working list we are determining which substances are problematic and developing methods to remove these substances at the appropriate stage. We have moved forward with the viable solvent systems to test the Hollow Fiber modules. Model solutions have been used to determine the rates of extraction and stripping. Real broths have also been tested at this point. The effect of flow rates, concentrations, and impurities on the overall performance of the product recovery is being investigated. The Integration and Polymerization of Itaconic Acid group headed by Dr. Yvon Durant, has been examining the effect of the carbon source (glucose, xylan, xylan and glucose, mannose, and xylan with mannose) on the relative itaconic acid titer. Polymerization experiments on esters of itaconic acid have been done to assess the potential to synthesize polymers based on the alkyl esters of itaconic acid. We have been able to demonstrate a 37% solid content and a 99% conversion rate using the IA derived monomer, Edi-n-butylitaconate. Next Steps: Softwood versus Hardwood Extraction studies, Optimization of pellet formation of Aspergillus terreus, Large scale sporulation, Fermented material for purification and polymerization experiments, Nutrient optimization of IA production, Strain optimization,Pilot runs

Publications

• No publications reported this period