Modified AFEX Pret reatment Conditions and enzymatic Hydrolysis for Poplar - PowerPoint PPT Presentation

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Modified AFEX Pret reatment Conditions and enzymatic Hydrolysis for Poplar
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Modified AFEX Pret reatment Conditions and enzymatic Hydrolysis for Poplar

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  1. Modified AFEX PretreatmentConditions and enzymatic Hydrolysis for Poplar Balan Venkatesh, Shishir Chundawat, Derek Marshall and Bruce E. Dale Department of Chemical Engineering and Material Science BCRL, Michigan State University (www.everythingbiomass.org) 16th November, 2006

  2. Ammonia Fiber Expansion (AFEX) Pretreatment Process Ammonia Recovery Gaseous Recycle Ammonia Ammonia Treated Biomass Heat Biomass Expansion Reactor • Conditions we wary: • Temperature (70-180 oC) • Moisture content of biomass (40-80%) • Ammonia to biomass ratio(1:1-3:1) • Residence time (5-30 min.) More details in Understanding AFEX presentation Friday, Nov. 17th 12. 50 p.m. (672b), HSF, continental 7

  3. Component CS Poplar Glucan 34.4 43.8 Xylan 22.8 14.5 Arabinan 4.2 0.61 Mannan 0.6 3.94 Galactan 1.4 1.02 Lignin 11.0 29.12 Protein 2.3 nd Ash 6.1 1.07 Extractives 8.5 3.56 Conversions on Different Feed stock Corn Stover (CS) Poplar (HLP) AFEX Conditions: 90 oC, 1:1 (BM:Ammonia), 5 min. residence time Enzyme Loading: 15 FPU cellulase, 64 pNPGU of b-glucosidase Reaction condition: 50oC, 90 rpm, 168 h HLP- High Lignin Poplar

  4. Component HLP LLP Glucan 43.8 45.1 Xylan 14.5 17.8 Arabinan 0.61 0.6 Mannan 3.94 1.7 Galactan 1.02 1.5 Lignin 29.12 21.4 Protein nd nd Ash 1.07 0.8 Extractives 3.56 3.4 Conversions for Two batches of Poplar HLP- High Lignin Poplar LLP- Low Lignin Poplar AFEX Conditions: 100 oC, 1:1 (BM:Ammonia), 10 min. residence time

  5. Strategy: • Low temperature pretreatment was not effective, hence went for higher temperature • Modified conditions like soaking and autoclaving prior to AFEX • Tried different enzyme combinations and surfactant effect • Understand the degradation product formed during the pretreatment by LC-MS and LC-UV techniques. • High throughput AFEX (lesser heat transfer problem) for wide range of screening conditions

  6. Reaction conditions and Enzyme Loading Substrate : HLP-High lignin poplar, LLP-low lignin poplar AFEX Condition : 100 oC/180 oC, 1:1 (BM:Ammonia) Enzyme Loading: mg of protein/g of glucan Abbreviation : C-Spezyme CP Cellulase, X-Multifect Xylanase, P-Pectinase, F- Depol 720 Feruloyl esterase, A-Additive (Tween 80)

  7. To summarize…… 60FPU of cellulase loading

  8. 3D Hydrogen bonding in cellulose 2D

  9. Synergy Experiments on High Lignin Poplar (HLP) 160 Enzyme Combinations 31.3 mg Cellulase (C) + 64 pNPGU bG (bG)/g of Glucan and/or Xylanase (X 10, 25, 50, 100) (3.1, 7.75, 15.5, 31.0 mg)/g of Glucan Pectinase (P10, 24, 50, 100) (5.6, 14, 28, 56 mg)/g of Glucan Feruloyl esterase (F 10, 25, 50, 100) (1, 2.5, 5, 10 mg)/g of Glucan

  10. Overall Sugar Yield for HLP and LLP 25.7 28.7 18.1 14.9 13.7 7.9 71.3 42.8 71.3 74.3 37.2 66.9 AFEX Condition: 180 oC, 70 % moisture, 10 min. 1:1 (ammonia : Biomass) Sample Autoclaved prior to AFEX 168 h hydrolysis LLP – Low Lignin Poplar HLP – High Lignin Poplar MSY – Maximum Sugar Yield 15 - 15 FPU of cellulase 60 – 60 FPU of cellulase

  11. Representative Chromatogram AFEX treated HLP 1, formic acid; 2, malonic acid; 3, lactic acid; 4, acetic acid; 5, maleic; 6, succinic; 7, fumaric; 8, t-aconitic acid; 9, levulinic Acid; 10, glutaric acid; 11, itaconic; 12, 2-hydroxy-2-methylbutyric acid; 13, gallic acid; 14, 5HMF; 15, 2-furoic acid; 16, furfural; 17, adipic acid; 18, 3,4-dihydroxylbenzoic acid; 19, 3,4-dihydroxybenzaldehyde; 20, 4-hydroxylbenzoic acid; 21, 2,5-dihydroxylbenzoic acid; 22, 4-hydroxybenzaldehyde; 23, vanillic acid; 24, homovanilic acid; 25, caffeic acid; 26, syringic acid; 27, 4-hydroxyacetophenone; 28, vanillin; 29, 4-hydroxycoumaric acid; 30, syringaldehyde; 31, benzoic acid; 32, ferulic acid; 33, sinapic acid; 34, 3-hydroxy-4-methoxycinnamic acid; 35, salicyclic acid; 36, 4-hydroxycoumarin; 37, o-toluic acid and 38, p-toluic acid. Untreated treated HLP The samples were analyzed by LC-MS for qualitative purposes only. However, the concentrations determined using UV detection may exhibit a high bias due to unidentified co-eluting components.

  12. Comparing Degradation Products: * * Unknown compounds co-elute along with lactic acid Collaborative work with Dr. Kevin at Baylor Institute of Technology Ref.:Dilute acid hydrolysate data from Chen et al., 2006

  13. High Throughput AFEX Unit

  14. Untreated and AFEX treated Poplar samples at different conditions Untreated 140 oC, 1:1 140 oC, 1:1.5 140 oC, 1:2 140 oC, 1:1 140 oC, 1:1.5 140 oC, 1:2 160 oC, 1:1 160 oC, 1:1.5 160 oC, 1:2 160 oC, 1:1 160 oC, 1:1.5 160 oC, 1:2 180 oC, 1:1 180 oC, 1:1.5 180 oC, 1:2 180 oC, 1:1 180 oC, 1:1.5 180 oC, 1:2 Water un-soaked prior to AFEX Water soaked prior to AFEX *All experiments done with 70% moisture and 30 minutes resident time

  15. Some preliminary results……… 72h Hydrolysis Enzymatic hydrolysis: 50 oC, 90 rpm US- un-soaked High Lignin Poplar

  16. Conclusion: • Combination of autoclaving followed by AFEX gives better Glucan yield both for high and low lignin poplar • Collective sugar yield for high and low lignin poplar was summarized for the best AFEX treated poplar • At elevated temperature and in the presence of ammonia, some sugar degradation products like organic acid is formed, based on LC-MS/LC-UV studies • Introduction to High through put AFEX (new tool to screen the different feed stock)

  17. Acknowledgement • CAFI team for their valuable suggestions • Genencor for supplying their enzymes • Dr. Kevin and Dr. Walsum for doing LC-MS and LC-UV analysis • DOE for funding

  18. Biomass Conversion Research Lab at Michigan State University Left to Right (Back): Shishir, Derek, Bryan, Ming Left to Right (Front): Elizabeth, Dr. Dale, Dr. Ven, Holly, Ben