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Maximum Total Time for Talk = 25 minutes

Maximum Total Time for Talk = 25 minutes. Comparative Sugar Recovery Data from Application of Leading Pretreatment Technologies to Corn Stover and Poplar. Charles E. Wyman, Dartmouth College/University of California Bruce E. Dale, Michigan State University

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Maximum Total Time for Talk = 25 minutes

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  1. Maximum Total Time for Talk = 25 minutes

  2. Comparative Sugar Recovery Data from Application of Leading Pretreatment Technologies to Corn Stover and Poplar Charles E. Wyman, Dartmouth College/University of California Bruce E. Dale, Michigan State University Richard T. Elander, National Renewable Energy Laboratory Mark T. Holtzapple, Texas A&M University Michael R. Ladisch, Purdue University Y. Y. Lee, Auburn University Mohammed Moniruzzaman, Genencor International John N. Saddler, University of British Columbia 28th Symposium on Biotechnology for Fuels and Chemicals Nashville, Tennessee May 1, 2006 Biomass Refining CAFI

  3. CAFI Background • Biomass Refining Consortium for Applied Fundamentals and Innovation organized in late 1999 and early 2000 • Included top researchers in biomass hydrolysis from Auburn, Dartmouth, Michigan State, Purdue, NREL, Texas A&M, U. British Columbia, U. Sherbrooke • Mission: • Develop information and a fundamental understanding of biomass hydrolysis that will facilitate commercialization, • Accelerate the development of next generation technologies that dramatically reduce the cost of sugars from cellulosic biomass • Train future engineers, scientists, and managers. Biomass Refining CAFI

  4. CAFI Approach • Developing data on leading pretreatments using: • Common feedstocks • Shared enzymes • Identical analytical methods • The same material and energy balance methods • The same costing methods • Goal is to provide information that helps industry select technologies for their applications • Also seek to understand mechanisms that influence performance and differentiate pretreatments • Provide technology base to facilitate commercial use • Identify promising paths to advance pretreatment technologies Biomass Refining CAFI

  5. USDA IFAFS Project Overview: CAFI 1 • Multi-institutional effort funded by USDA Initiative for Future Agriculture and Food Systems Program for $1.2 million to develop comparative information on cellulosic biomass pretreatment by leading pretreatment options with common source of cellulosic biomass (corn stover) and identical analytical methods • Aqueous ammonia recycle pretreatment - YY Lee, Auburn University • Water only and dilute acid hydrolysis by co-current and flowthrough systems - Charles Wyman, Dartmouth College • Ammonia fiber explosion (AFEX) - Bruce Dale, Michigan State University • Controlled pH pretreatment - Mike Ladisch, Purdue University • Lime pretreatment - Mark Holtzapple, Texas A&M University • Logistical support and economic analysis - Rick Elander/Tim Eggeman, NREL through DOE Biomass Program funding • Completed in 2004 Biomass Refining CAFI

  6. Hydrolysis Stages Cellulase enzyme Stage 1 Pretreatment Stage 2 Enzymatic hydrolysis Residual solids: cellulose, hemicellulose, lignin Biomass Solids: cellulose, hemicellulose, lignin Chemicals Dissolved sugars, oligomers Dissolved sugars, oligomers, lignin Biomass Refining CAFI

  7. Key Features of CAFI Pretreatments

  8. CAFI 1 Feedstock: Corn Stover • NREL supplied corn stover to all project participants (source: BioMass AgriProducts, Harlan IA) • Stover washed and dried in small commercial operation, knife milled to pass ¼ inch round screen Biomass Refining CAFI

  9. Consistent Mass Balance Approach as Applied to AFEX Enzyme (15 FPU/g of Glucan) Ammonia Hydrolyzate 99.0 lb AFEX Stover Treated Liquid Hydrolysis Wash 38.5 lb glucose Stover System 100 lb (Ave. of 4 runs) 18.9 lb xylose 101.0 lb Residual (dry basis) Solids Solids washed out 36.1 lb glucan 2 lb 39.2 lb 21.4 lb xylan Very few solubles from pretreatment—about 2% of inlet stover 95.9% glucan conversion to glucose, 77.6% xylan conversion to xylose 99% mass balance closure includes: (solids + glucose + xylose + arabinose ) Biomass Refining CAFI

  10. Calculation of Sugar Yields • Comparing the amount of each sugar monomer or oligomer released to the maximum potential amount for that sugar would give yield of each • However, most cellulosic biomass is richer in glucose than xylose • Consequently, glucose yields have a greater impact than for xylose • Sugar yields in this project were defined by dividing the amount of xylose or glucose or the sum of the two recovered in each stage by the maximum potential amount of both sugars • The maximum xylose yield is 24.3/64.4 or 37.7% • The maximum glucose yield is 40.1/64.4 or 62.3% • The maximum amount of total xylose and glucose is 100%. Biomass Refining CAFI

  11. Increasing pH Overall Sugar Yields from Corn Stover at 60 FPU/g Glucan *Cumulative soluble sugars as total/monomers. Single number = just monomers. Biomass Refining CAFI

  12. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  13. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  14. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  15. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  16. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  17. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  18. Sugar Yields from Corn Stover at 15 FPU/g Glucan Maximum possible Dilute acid Controlled pH Flowthrough ARP Lime AFEX Biomass Refining CAFI

  19. Sugar Yields from Corn Stover at 15 FPU/g Glucan Biomass Refining CAFI

  20. Pretreatment Model Aspen Plus Bioethanol Plant Model 2001 NREL Design Case 2000 Metric Tons Stover (dry)/Day Stover Cost: $35/ton Enzyme Cost: ~$0.15/gal ethanol CAFI Economic Estimates Thermodynamics Process Analogies Chemistry Design Methods CAFI Researcher Updated Model Basis and Feedstock Basis in “CAFI 2” Project Biomass Refining CAFI

  21. Enzymes CO Water 2 Hydrolysis Feed Stover EtOH Pretreatment Recovery + Handling Fermentation Syrup + Solids Chemicals Water Boiler Steam + Gene rator Power General Process Flow Diagram Poplar Biomass Refining CAFI

  22. Capital Cost Estimates Basis: 2000 metric tons (dry basis) corn stover/day, assumes only monomers fermented Biomass Refining CAFI

  23. Minimum Ethanol Selling Price (MESP) Assumptions: 2.5 years construction, 0.5 years start up, 20 year plant life, zero net present value when cash flows are discounted at 10% real after tax rate Biomass Refining CAFI

  24. Effect of Oligomer Conversion Biomass Refining CAFI

  25. Observations for Corn Stover • All pretreatments were effective in making cellulose accessible to enzymes • Lime, ARP, and flowthrough remove substantial amounts of lignin and achieved somewhat higher glucose yields from enzymes than dilute acid or controlled pH • However, AFEX achieved slightly higher yields from enzymes even though no lignin was removed • Cellulase was effective in releasing residual xylose from all pretreated solids during enzymatic hydrolysis in Stage 2 • Xylose release by cellulase was particularly important for the high-pH pretreatments by AFEX, ARP, and lime, with about half being solubilized by enzymes for ARP, two thirds for lime, and essentially all for AFEX • The projected costs were similar due to the high yields and similar capital costs for the overall processes Biomass Refining CAFI

  26. Publication of Results from CAFI 1 • Bruce Dale of the CAFI Team arranged for and edited a special December 2005 issue of Bioresource Technology entitled “Coordinated Development of Leading Biomass Pretreatment Technologies” to document these results: • Wyman CE, Dale BE, Elander RT, Holtzapple M, Ladisch MR, Lee YY. 2005. “Coordinated Development of Leading Biomass Pretreatment Technologies,” Bioresource Technology 96(18): 1959-1966, invited. • Lloyd TA, Wyman CE. 2005. “Total Sugar Yields for Pretreatment by Hemicellulose Hydrolysis Coupled with Enzymatic Hydrolysis of the Remaining Solids,”Bioresource Technology96(18): 1967-1977, invited. • Liu C, Wyman CE. 2005. "Partial Flow of Compressed-Hot Water Through Corn Stover to Enhance Hemicellulose Sugar Recovery and Enzymatic Digestibility of Cellulose,”Bioresource Technology96(18): 1978-1985, invited. • Mosier N, Hendrickson R, Ho N, Sedlak M, Ladisch MR. 2005. “Optimization of pH Controlled Liquid Hot Water Pretreatment of Corn Stover,” Bioresource Technology96(18): 1986-1993, invited. • Kim S, Holtzapple MT. 2005. “Lime Pretreatment and Enzymatic Hydrolysis of CornStover,”Bioresource Technology96(18): 1994-2006, invited. • Kim TH, Lee YY. 2005. “Pretreatment and Fractionation of Corn Stover by Ammonia Recycle Percolation Process,”Bioresource Technology96(18): 2007-2013, invited. • Teymouri F, Laureano-Perez L, Alizadeh H, Dale BE. 2005. “Optimization of the Ammonia Fiber Explosion (AFEX) Treatment Parameters for Enzymatic Hydrolysis of Corn Stover,” Bioresource Technology96(18): 2014-2018, invited. • Eggeman T, Elander RT. 2005. “Process and Economic Analysis of Pretreatment Technologies,” Bioresource Technology96(18): 2019-2025, invited. • Wyman CE, Dale BE, Elander RT, Holtzapple M, Ladisch MR, Lee YY. 2005. “Comparative Sugar Recovery Data from Laboratory Scale Application of Leading Pretreatment Technologies to Corn Stover,”Bioresource Technology96(18): 2026-2032, invited. Biomass Refining CAFI

  27. DOE OBP Project: April 2004 Start • Funded by DOE Office of the Biomass Program for $1.88 million through a joint competitive solicitation with USDA • Using identical analytical methods and feedstock sources to develop comparative data for corn stover and poplar • Determining more depth information on • Enzymatic hydrolysis of cellulose and hemicellulose in solids • Conditioning and fermentation of pretreatment hydrolyzate liquids • Predictive models • Added University of British Columbia to team through funding from Natural Resources Canada to • Capitalize on their expertise with xylanases for better hemicellulose utilization • Evaluate sulfur dioxide pretreatment along with those previously examined: dilute acid, controlled pH, AFEX, ARP, lime • Augmented by Genencor to supply commercial and advanced enzymes Biomass Refining CAFI

  28. Tasks for the DOE OBP Project • Pretreat corn stover and poplar by leading technologies to improve cellulose accessibility to enzymes • Enzymatically hydrolyze cellulose and hemicellulose in pretreated biomass, as appropriate, and develop models to understand the relationship between pretreated biomass features, advanced enzyme characteristics, and enzymatic digestion results • Develop conditioning methods as needed to maximize fermentation yields by a recombinant yeast, determine the cause of inhibition, and model fermentations • Estimate capital and operating costs for each integrated pretreatment, hydrolysis, and fermentation system and use to guide research Biomass Refining CAFI

  29. CAFI 2 Corn Stover • 2nd pass harvested corn stover from Kramer farm (Wray, CO) • Collected using high rake setting to avoid soil pick-up • No washing • Milled to pass ¼ inch round screen Biomass Refining CAFI

  30. CAFI 2 Standard Poplar • Feedstock: USDA-supplied hybrid poplar (Alexandria, MN) • Debarked, chipped, and milled to pass ¼ inch round screen Biomass Refining CAFI

  31. Hydrolysis Stages Cellulase enzyme Stage 1 Pretreatment Stage 2 Enzymatic hydrolysis Residual solids: cellulose, hemicellulose, lignin Biomass Solids: cellulose, hemicellulose, lignin Chemicals Dissolved sugars, oligomers Dissolved sugars, oligomers, lignin Stage 3 Sugar fermentation Biomass Refining CAFI

  32. CAFI 2 Pretreated Substrate Schedule Biomass Refining CAFI

  33. Increasing pH Overall Yields for Corn Stover at 15 FPU/g Glucan *Cumulative soluble sugars as total/monomers. Single number = just monomers. Biomass Refining CAFI

  34. Increasing severity Effect of Pretreatment Severity on Enzymatic Hydrolysis of Dilute Acid Pretreated Poplar CBU:FPU = 2.0 Digestion time =72hr 2% glucan concentration 50 FPU/ gm original glucan Biomass Refining CAFI

  35. Effect of Protein Loadings on Cellulose Hydrolysis of Poplar Solids Biomass Refining CAFI Digestion time =72hr

  36. Effect of Protein Loadings on Cellulose Hydrolysis of Poplar Solids Biomass Refining CAFI Digestion time =72hr

  37. Effect of Protein Loadings on Cellulose Hydrolysis of Poplar Solids Biomass Refining CAFI Digestion time =72hr

  38. CAFI 2 Initial Poplar • Feedstock: USDA-supplied hybrid poplar (Arlington, WI) • Debarked, chipped, and milled to pass ¼ inch round screen • Not enough to meet needs Biomass Refining CAFI

  39. CAFI 2 Initial Poplar • Feedstock: USDA-supplied hybrid poplar (Arlington, WI) • Debarked, chipped, and milled to pass ¼ inch round screen • Not enough to meet needs Biomass Refining CAFI

  40. C - Cellulase (31.3 mg/g glucan) X - Xylanase (3.1 mg/g glucan) A - Additive (0.35g/g glucan) UT - Untreated AFEX condition 24 h water soaked 1:1 (Poplar:NH3) 10 min. res. time AFEX Optimization for High/Low Lignin Poplar

  41. Differences Among Poplar Species* * Based on information provided by Adam Wiese, USDA Rheinlander, WI Biomass Refining CAFI

  42. Fermentation of Dilute Acid Treated Corn Stover Ca(OH)2 S. cerevisiae 424A(LNH-ST) H2SO4 B corn stover Fermentation 30C Ethanol 175C Liquid A pH 1.2 pH 6.0 Solids Cells pH 1.2 A B Stream g/L g/L 80% of theoretical Inhibitor consumed consumed

  43. Fermentation of Hot Water Treated Corn Stover Ca(OH)2 S. cerevisiae 424A(LNH-ST) Water Enzyme B corn stover Fermentation 30C Ethanol 190C 50C A pH 4.5 pH 6.0 Cells + Solids A B Stream g/L g/L 95% of theoretical No Xylanase below threshold consumed consumed

  44. Fermentation of SO2 Treated Corn Stover SO2 Ca(OH)2 Enzyme Ca(OH)2 S. cerevisiae 424A(LNH-ST) B corn stover Fermentation 30C 180C 50C Ethanol A ~pH 1 pH 4.8 pH 6.0 Cells Solids A B Stream g/L g/L 96% of theoretical No Xylanase below threshold consumed consumed

  45. Observations • The yields can be further increased for some pretreatments with enzymes a potential key • Mixed sugar streams will be better used in some processes than others • Oligomers may require special considerations, depending on process configuration and choice of fermentative organism • Initial data on conditioning and fermentation shows mostly good yields • All pretreatments gave similar results for corn stover • Initial hydrolysis results for poplar are not as good, with one variety more recalcitrant than other Biomass Refining CAFI

  46. Planned Work • Maximize yields with standard poplar for each pretreatment • Evaluate differences with initial poplar at optimal conditions for standard poplar • Develop fermentation data with hydrolyzate for each material • Upgrade technoeconomic model with corn stover and poplar • Identify key features that distinguish performance of different pretreatments Biomass Refining CAFI

  47. Acknowledgments • US Department of Agriculture Initiative for Future Agricultural and Food Systems Program, Contract 00-52104-9663 • US Department of Energy Office of the Biomass Program, Contract DE-FG36-04GO14017 • Natural Resources Canada • All of the CAFI Team members, students, and others who have been so cooperative Biomass Refining CAFI

  48. Quang Nguyen, Abengoa Bioenergy Mat Peabody, Applied CarboChemicals Gary Welch, Aventinerei Greg Luli, BC International Paris Tsobanakis, Cargill Robert Wooley, Cargill Dow James Hettenhaus, CEA Steve Thomas, CERES Lyman Young, ChevronTexaco Kevin Gray, Diversa Paul Roessler, Dow Julie Friend, DuPont Jack Huttner, Genencor Don Johnson, GPC (Retired) Dale Monceaux, Katzen Engineers Kendall Pye, Lignol Farzaneh Teymouri, MBI Richard Glass, National Corn Growers Association Bill Cruickshank, Natural Resources Canada Robert Goldberg, NIST Joel Cherry, Novozymes Ron Reinsfelder, Shell Andrew Richard, Sunopta Carl Miller, Syngenta Carmela Bailey, USDA Don Riemenschneider, USDA CAFI DOE Project A&I Advisory Board:Meetings Every 6 Months

  49. Questions?? Louisiana Rice Hulls Pile

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