Production of higher alcohols liquid biofuel via acidogenic digestion and chemical upgrading of industrial biomass streams. DE-FG36-08GO18165 April 7, 2011 Algae Platform Peer Review G. Peter van Walsum Clayton Wheeler University of Maine This presentation does not contain any proprietary, confidential, or otherwise restricted information

Goal Statement Determine feasibility of converting pulp mill and sea weed processing streams to advanced fuels via carboxylate platform processing. Relevance to the Biomass Program: Low cost enzymatic hydrolysis Complete carbohydrate fermentation Integrated biochemical and thermochemical processing Improving economics of biomass processing industry: Kraft pulping, carrageenan extraction. 2

Quad Chart Overview Project start date: 10/’08 (03/’09) Project end date: 09/’11 (12/’11) Percent complete: 73% –Bt-F, G Enzyme cost + loading Autohydrolytic cultures –Bt-J Organism development Evolving microbial ecosystem –Bt-K Biological Process Integration Consolidated bioprocessing –Bt-L Biochemical/Thermo-chemical Integration Acidogenesis + TC upgrading Total project funding –DOE share: $713k –Contractor share: $181k Funding received in FY09: $118k Funding for FY10:$203k ARRA Funding: none Timeline Budget Barriers addressed Old Town Fuel + Fiber (OTFF) FMC Biopolymer Partners Stage A,B Exploratory

Alcohols Existing biomass processing industries: Hardwood pulp Algal carrageenan Acidogenic digestion Separation and conversion : L/L extraction Thermal conversion Esterification H 2 treatment Alkali / neutral pH treated carbohydrates Carboxylate salts Ketones Esters H2H2 Ketones, Esters, Organic acids Project Overview I: Ferment to carboxylic acids II:Upgrade to alcohols III:Process modeling

Overview, cont’d fermentation

1-Approach Existing industries: Infrastructure, adding value Alkali / neutral pH treated streams: Good digestibility Low inhibition likely Evolving mixed culture: Low cost Autohydrolytic Complete sugar utilization Optimized through evolution Conventional thermochemical conv.: Low technical risk Demonstrated, scaled technologies Still opportunities for innovation + optimization: integrations Higher value products Chemicals Higher alcohols preferable to ethanol as fuel Existing biomass processing industries: Hardwood pulp Algal carrageenan Acidogeni c digestion Separation and conversion : L/L extraction Thermal conversion Esterification H 2 treatment Alkali / neutral pH treated carbohydrates Carboxylate salts Ketones Esters H2 Ketones, Esters, Organic acids Alcohols

Project objectives: I) Convert pre-pulping hemicellulose extract and seaweed sludge (Algefiber TM )from carrageenan production into carboxylate salts via mixed culture evolving fermentation. II) Upgrade the carboxylate salts into chemicals and alcohols via one of three possible routes: CKA: carboxylates-ketones-alcohols, CEA: carboxylates-esters-alcohols CHEAcarboxylates-acids-esters-alcohols. III) Develop process models to evaluate the M + E balances and costs of three configurations. Assess process synergies with existing facilities. 7

8 1 –Project pathway Milestones: –Produce carboxylic acids at bench scale from both feedstocks. –Assemble and run larger ( 50 gal) fermentation. –Produce ketones, esters and alcohols from stock chemicals. –Produce ketones or esters, and alcohols from biomass. –Generate economic numbers with techno economic model. Go / no-Go decision points: –Task A.GN: Identify fermentation cond’s: Temperature, buffer type. –Tasks D. F. G. GN: Decide on continuation of CKA, CEA or CHEA past pure feedstock. –Tasks H. J. K.GN: Decide on continuation of CKA, CEA or CHEA to alcohol products.

9 2 - Technical Accomplishments/ Progress/Results Objective I: Fermentation to carboxylic acids. –Several feedstock samples characterized for carbohydrate, solids. –Fermentations completed on several samples of Algefiber TM and wood extracts at two temperatures, using calcium or ammonium buffers. –Carboxylic acids produced at up to 25 g/L in batch culture. –Verified autohydrolysis of oligomers. –50 gal fermentors built and in use.

Fermentation work: Algefiber ® + chicken Manure

Kelp results

Autohydrolysis in fermentor Xylan

2 - Technical Accomplishments/Progress/Results (cont’d) Objective II: Upgrade acids to mixed alcohols –Characterized ketonization of different cation carboxylates (Na +, Ca ++, Mg ++, NH 4 + ). –Converted stock carboxylate salts to ketones. –Converted fermentation salts to ketones. –Extracted acids from model systems and wood extracts. –Esterified carboxylic acids with ethanol and octanol. –Esterified ammonium acetate to octyl acetate. –Hydrogenated ketones to alcohols. –Developed kinetic model for hydrogenation. –Selected CKA (Ketone) upgrading pathway 13

Compounds investigated Ca(Ac) 2, Ca(Pr) 2 : –Dehydration: < 200 °C –Ketonization: 380 – 480 °C –Calcining: > 600 °C Mg(Ac) 2 (H 2 O) 4 : –Dehydration: < 200 °C –Ketonization + calcination 270 – 420 °C NaAc –Dehydration: < 200 °C –Ketonization: °C NH 4 Ac: –Decomposition to Acetamide < 200 °C

Ketonization of Fermentation (Algefiber TM ) salts

Hydrogenation kinetics: Arrhenius plots

Acid extractions: pH vs chain length

Objective III: Techno economic modeling –Developed ASPEN-Plus model of CKA, CEA, CHEA upgrading pathways –Compared yields, costs and energy balances of three pathways –Different strengths and weaknesses associated with each pathway Technical Accomplishments/Progress/Results (cont’d)

Production volumes CKACEACHEA Ethanol--10,208,06810,111,793 Propanol--976,292969,151 2-Propanol7,051, Pentanol731,446-- Total (gallons) 7,783,13011,184,36011,080,944 Annual production amount in gallons of alcohol produced for each pathway: 1000 tpd pulp mill.

Net energy production

And the winner is... Upgrading Scenario Capital Cost Yearly Total Alcohol Production (gallons) Cap. $/ annual gal $/Gal to achieve break-even point Cap. $/MJ CKA$31.4 MM7,783, CEA$39.5 MM11,184, CHEA$32.8 MM11,080,

Most important technical accomplishments: –Demonstrated autohydrolysis and fermentation in inhibitory wood extracts. –Demonstrated ketone production from biomass- derived carboxylates—including several longer C- chain molecules. –Identified catalytic action of water in ketone hydrogenation to 2-alcohols Technical Accomplishments/Progress/Results (cont’d)

Key Milestones and Status, Barriers addressed –Produce acids at bench scale from both feedstocks. –Assembled and running larger ( 50 gal) fermentation. –Narrowed conditions from 8 to 2. Working on increasing titer. –Barriers addressed: Bt-F,G (enzymes); Bt-J (organisms); Bt-K (consolidated bioprocessing) –Produce ketones, esters, alcohols from stock chemicals. Ketones and alcohols work well. Esters have had low yields –Produce ketones or esters, and alcohols from biomass. Ketones produced from Algefiber TM. Alcohols to come. –Barrier: Bt-L Biochemical/Thermo-chemical Integration –Economic numbers from techno economic model. Have estimates for 3 pathways. Mill integration to come Technical Accomplishments/Progress/Results (cont’d)

Ties to applications –Mixed culture CBP of undetoxified feedstocks: Process simplification: no hydrolysis step Operating cost savings: no acid, base, enzymes needed Robust operation: fewer shutdowns Cap cost savings: Plastic/concrete instead of SS –Demonstrated ketone production from biomass- derived carboxylates. Process works, easy separation of ketones from residues Lignin, other residues do not hinder ketonization –Identified catalytic action of water in ketone hydrogenation to alcohols. High yields, fast rates, good catalyst durability Technical Accomplishments/Progress/Results (cont’d)

Relevance Mission of the Biomass Program MYPP Renewable biomass resources: Wood hemicellulose + macro algae Cost-competitive: $3.30 – 3.40/gal (wood extracts, ~10 MMgal/yr) High-performance biofuels: Mixed alcohols. HCs possible Bioproducts and biopower: Chemical intermediates Program Goals: Reduce dependence on oil: blendable mixed alcohols 36 bgy by 2022 : High carbon retention in liquid products Applicable to bio wastes and energy crops.

26 3 – Relevance, Cont’d Contribution to goals of the Program Enable the production of biofuels nationwide Technology suitable to forested and coastal areas of the US. Reduce dependence on oil Producing higher alcohols--higher blend ratios than ethanol. Addressing critical issues Low enzyme cost: CBP has no enzyme costs. Improve catalyst performance: Ketonization catalyst (Ca++) works with biomass residues Hydrogenation works on relatively clean ketone condensate

27 3 – Relevance, Cont’d Technology development timeline 2012: $0.86/gal processing cost ethanol 2017: $1.56/gal gasoline, diesel blendstocks Cost estimates for processing wood extracts are: For a scale of a 1000 tpd pulp mill (Typical in North America) Mixed Alcohol production: MM gallons/year Capcost: $3 - $4/annual gallon mixed alcohols Processing cost: ~$1.60/gal mixed alcohols Downstream processing costs for Algefiber TM similar Feedstock for Algefiber TM lower cost. Fermentation and solids handling for Algefiber TM may be higher.

28 3 – Relevance, cont’d Applications of the expected outputs Working with OTFF on 50+ gal wood exact fermentation. Results on Algefiber TM too low in titer ( so far) Looking at raw kelp as feedstock (different project). Technology development transferable to other feedstocks

Critical Success Factors Technical Acid concentrations in fermentation, fermentation rates. Being improved through: continuous culture (adaptation) percolation processing (higher solids/liquid ratio) counter-current solids processing (reducing inhibition) Yield from ketonization reaction, products from other components. Yield is being assessed. Operating procedures improving (mixing, heating rates, condensation temperatures) Durability of catalyst in hydrogenation. So far so good, but not yet tested with biomass derived ketones.

Critical Success Factors, Cont’d Business Feedstock supply Industrial partners currently source their biomass. Capital costs, technology risk Project partners may enable co-location to diminish cost and risk Partners have expertise in biomass processing Partner motivations: FMC Biopolymer can achieve some avoided disposal costs. Oceans Approved (Separate project) interested in expanding market for kelp, using waste from processing. OTFF has $30M DOE biorefinery grant. Terrabon (IP holder) desires business-relevant research.

Critical Success Factors, Cont’d Market No fuel market for mixed 2-alcohols Others have interest in mixed alcohols: FT or C-4 alcohol fermentators—this should come eventually. Distill alcohols for fuel (ethanol, butanol) and chemical markets Conversion of mixed alcohols to gasoline range alkanes has been demonstrated by Terrabon (70 gal/ton biomass). Accessing chemical market FMC or Patriarch Partners (OTFF) may be able to use or market chemical products (Acids, Ketones, Esters) Distilled chemical products may be marketed as bio-derived

Critical Success Factors, Cont’d Regulatory Permitting Co-location could diminish permitting barriers FMC Biopolymer may benefit from waste reduction. No GMOs being used Legal Most IP on carboxylate platform held by Terrabon UMaine and Tarrabon negotiating a research license agreement.

Critical Success Factors, Cont’d Top 2-3 potential challenges Fermentation: Low titers, slow rates. Evolve cultures, counter current and sequenced processing Poor yield through ketonization (?) Optimize conditions, utilize all products Economies of scale too small. Demonstrate technology, apply at larger scale Target chemical markets

Critical Success Factors, Cont’d Advancing the state of technology and positively impacting the commercial viability –Cross-Cutting Analysis: Aspen modeling of processes. –Feedstock Supply R&D: Technology applicable to many feedstocks, mixed feedstocks. –Downstream Refining R&D: Demonstrating integrated process. –Environmental sustainability: – No GMOs. – Reducing wastes from existing industries. – May be viable at smaller scale than other technologies.

35 Future Work Increase acid titer from algaefiber through counter- current percolation processing. Continue 50-gal scale fermentations of wood extract to generate kg quantities of carboxylate salts. Operate in sequencing batch. Integrated production: Upgrade fermentation- derived salts to ketones and then to alcohols. Continue to refine ASPEN model and costing. Integrate pulp and carageenan mill interface into model. Decision points: Final economics will determine continued development.

36 Summary –Relevance: Demonstrating low hydrolysis and fermentation cost, integrated bio and thermochemical platform. –Approach: No breakthroughs needed. Focus on robust technology, cheap feedstock, existing industries. –Technical accomplishments: Demonstrated fermentation and downstream unit ops—need to demonstrate integrated process. –Success factors and challenges: Higher fermentation concentrations needed. –Technology transfer and future work: Working closely with OTFF. FMC option looks less promising (low titer). Terrabon relationship developing. Since 2009 review: 2009 we had just started project (1-2 months in). Have attained most milestones. Have down-selected to final demonstration pathway.

37 Additional Slides

Abbreviations used 38 Ac: Acetate CBP: Consolidated bioprocessing CKA: carboxylates-ketones-alcohols CEA: carboxylates-esters-alcohols CHEA: carboxylates-acids-esters-alcohols FT: Fischer Tropsch synthesis GMO: Genetically modified organism MYPP: DOE Multi year program plan OTFF: Old Town Fuel and Fiber—Kraft pulp mill

39 Responses to Previous Reviewers’ Comments Process complexity –The hydrolysis and fermentation are very simple compared to sterile cellulose fermentations –Downstream processing is complex, but unit operations are familiar and commercial (ex. hydrogenation) –Our down selection chose the simplest pathway (ketones  2- alcohols) Feedstock and culture variability –Feedstock is likely to be variable. Mixed culture is better than pure cultures for adapting to different inputs. –Operation in acidogenic phase is more stable than methanogenic stage, so should be more stable than regular AD.

40 Publications and Presentations Patent disclosure filed: Removal of lignin and other contaminants from recycled liquid-liquid extraction solvent by means of contacting with alkaline pulping liquors. Publications: Rakhi Baddam, M. Clayton Wheeler, G. Peter van Walsum. Anaerobic Fermentation of Hemicellulose Present in Pre- Pulping Extracts of Northern Hardwoods to Carboxylic Acids. Proceedings of the AIChE annual meeting, Nashville TN, November Aymn Abdulrahman, Adriaan van Heiningen, M. Clayton Wheeler, G. Peter van Walsum. Acetic Acid Removal from Pre-Pulping Wood Extract. Proceedings of the AIChE annual meeting, Nashville TN, November Mohit, Keith Hurley, G. Peter van Walsum, M. Clayton Wheeler. Thermal conversion of carboxylate salts derived from biomass fermentation to ketones. Proceedings of the AIChE annual meeting, Nashville TN, November Abigail Engelberth, M. Clayton Wheeler, G. Peter van Walsum. Exploring Three Pathways to Convert a Hemi-Cellulose Rich Pre-Pulping Extract Into Long-Chain Alcohols Via the MixAlco™ Process. Proceedings of the AIChE annual Meeting, Salt Lake City, UT, November Sampath Karunarathne, Clayton Wheeler, G. Peter van Walsum. Production of Carboxylic Acids From Acidogenic Fermentation of Algefiber™ (Sea Weed Sludge) Using a Mixed Culture of Marine Microorganisms. Proceedings of the AIChE annual Meeting, Salt Lake City, UT, November 2010.

41 Publications and Presentations cont’d Presentations: Rakhi Baddam, M. Clayton Wheeler, G. Peter van Walsum. Anaerobic Fermentation of Hemicellulose Present in Pre-Pulping Extracts of Northern Hardwoods to Carboxylic Acids. AIChE annual meeting, Nashville TN, November Aymn Abdulrahman, Adriaan van Heiningen, M. Clayton Wheeler, G. Peter van Walsum. Acetic Acid Removal from Pre- Pulping Wood Extract. AIChE annual meeting, Nashville TN, November Mohit, Keith Hurley, G. Peter van Walsum, M. Clayton Wheeler. Thermal conversion of carboxylate salts derived from biomass fermentation to ketones. AIChE annual meeting, Nashville TN, November Abigail Engelberth, M. Clayton Wheeler, G. Peter van Walsum. Alcohol Production from Pre Pulping Extracts: A modeling Assessment. 32 nd Annual Symposium on Biotechnology for Fuels and Chemicals, April Invited presentation. Sampath Karunarathne, Clayton Wheeler and Peter Van Walsum. Production of Carboxylic Salts via Mixed Culture Acidogenic Fermentation of Algefiber™ (Sea Weed Sludge) for Chemical Upgrading to Higher Alcohols Liquid Biofuel. 32 nd Annual Symposium on Biotechnology for Fuels and Chemicals, April Abigail. Engelberth, M. Clayton Wheeler, G. Peter van Walsum. Comparative economics and yields for mixed alcohols from pre-pulping hemicellulose extracts via three MixAlco TM pathways. 32 nd Annual Symposium on Biotechnology for Fuels and Chemicals, April Abigail Engelberth, M. Clayton Wheeler, G. Peter van Walsum. Exploring Three Pathways to Convert a Hemi-Cellulose Rich Pre-Pulping Extract Into Long-Chain Alcohols Via the MixAlco™ Process. AIChE annual Meeting, Salt Lake City, UT, November Sampath Karunarathne, Clayton Wheeler, G. Peter van Walsum. Production of Carboxylic Acids From Acidogenic Fermentation of Algefiber™ (Sea Weed Sludge) Using a Mixed Culture of Marine Microorganisms. AIChE annual Meeting, Salt Lake City, UT, November Mohit Bhatia, Adriaan van Heiningen,G. Peter van Walsum, Clayton Wheeler. Kinetics and Mechanism for Acetone Hydrogenation by Ru/Carbon. AIChE annual Meeting, Salt Lake City, UT, November 2010.