Adjusted AFEX Pret reatment Conditions and enzymatic Hydrolysis for Poplar.


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Adjusted AFEX Pret reatment Conditions and enzymatic Hydrolysis for Poplar Balan Venkatesh, Shishir Chundawat, Derek Marshall and Bruce E. Dale Branch of Concoction Designing and Material Science BCRL, Michigan State College (www.everythingbiomass.org) 16 th November, 2006
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Adjusted AFEX Pret reatment Conditions 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) 16 th November, 2006

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Ammonia Fiber Expansion (AFEX) Pretreatment Process Ammonia Recovery Gaseous Recycle Ammonia Treated Biomass Heat Biomass Expansion Reactor Conditions we watchful : Temperature (70-180 o C) Moisture substance of biomass (40-80%) Ammonia to biomass ratio(1:1-3:1) Residence time (5-30 min.) More subtle elements in Understanding AFEX presentation Friday, Nov. seventeenth 12. 50 p.m. (672b), HSF, mainland 7

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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 o C, 1:1 (BM:Ammonia), 5 min. habitation time Enzyme Loading : 15 FPU cellulase, 64 pNPGU of b-glucosidase Reaction condition : 50 o C, 90 rpm, 168 h HLP-High Lignin Poplar

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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 Ash 1.07 0.8 Extractives 3.56 3.4 Conversions for Two clumps of Poplar HLP-High Lignin Poplar LLP-Low Lignin Poplar AFEX Conditions : 100 o C, 1:1 (BM:Ammonia), 10 min. habitation time

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Strategy : Low temperature pretreatment was not viable, subsequently went for higher temperature Modified conditions like splashing and autoclaving before AFEX Tried diverse compound blends and surfactant impact Understand the debasement item framed amid the pretreatment by LC-MS and LC-UV systems. High throughput AFEX (lesser warmth exchange issue) for extensive variety of screening conditions

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Reaction conditions and Enzyme Loading Substrate : HLP-High lignin poplar, LLP-low lignin poplar AFEX Condition : 100 o C/180 o C, 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, An Additive (Tween 80)

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To summarize…… 60FPU of cellulase stacking

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3D Hydrogen holding in cellulose 2D

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Synergy Experiments on High Lignin Poplar (HLP) 160 Enzyme Combinations 31.3 mg Cellulase (C) + 64 pNPGU b G (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

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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 o C, 70 % dampness, 10 min. 1:1 (alkali : Biomass) Sample Autoclaved before 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

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Representative Chromatogram AFEX treated HLP 1, formic corrosive; 2, malonic corrosive; 3, lactic corrosive; 4, acidic corrosive; 5, maleic; 6, succinic; 7, fumaric; 8, t-aconitic corrosive; 9, levulinic Acid; 10, glutaric corrosive; 11, itaconic; 12, 2-hydroxy-2-methylbutyric corrosive; 13, gallic corrosive; 14, 5HMF; 15, 2-furoic corrosive; 16, furfural; 17, adipic corrosive; 18, 3,4-dihydroxylbenzoic corrosive; 19, 3,4-dihydroxybenzaldehyde; 20, 4-hydroxylbenzoic corrosive; 21, 2,5-dihydroxylbenzoic corrosive; 22, 4-hydroxybenzaldehyde; 23, vanillic corrosive; 24, homovanilic corrosive; 25, caffeic corrosive; 26, syringic corrosive; 27, 4-hydroxyacetophenone; 28, vanillin; 29, 4-hydroxycoumaric corrosive; 30, syringaldehyde; 31, benzoic corrosive; 32, ferulic corrosive; 33, sinapic corrosive; 34, 3-hydroxy-4-methoxycinnamic corrosive; 35, salicyclic corrosive; 36, 4-hydroxycoumarin; 37, o-toluic corrosive and 38, p-toluic corrosive. Untreated treated HLP The examples were investigated by LC-MS for subjective purposes just. Be that as it may, the fixations decided utilizing UV identification may show a high predisposition because of unidentified co-eluting parts.

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Comparing Degradation Products: * Unknown mixes co-elute alongside lactic corrosive Collaborative work with Dr. Kevin at Baylor Institute of Technology Ref .:Dilute corrosive hydrolysate information from Chen et al., 2006

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High Throughput AFEX Unit

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Untreated and AFEX treated Poplar tests at diverse conditions Untreated 140 o C, 1:1 140 o C, 1:1.5 140 o C, 1:2 140 o C, 1:1 140 o C, 1:1.5 140 o C, 1:2 160 o C, 1:1 160 o C, 1:1.5 160 o C, 1:2 160 o C, 1:1 160 o C, 1:1.5 160 o C, 1:2 180 o C, 1:1 180 o C, 1:1.5 180 o C, 1:2 180 o C, 1:1 180 o C, 1:1.5 180 o C, 1:2 Water un-drenched preceding AFEX Water doused before AFEX *All examinations finished with 70% dampness and 30 minutes occupant time

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Some preparatory results……… 72h Hydrolysis Enzymatic hydrolysis : 50 o C, 90 rpm US-un-splashed High Lignin Poplar

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Conclusion : Combination of autoclaving took after by AFEX gives better Glucan yield both for high and low lignin poplar Collective sugar yield for high and low lignin poplar was compressed for the best AFEX treated poplar At lifted temperature and in the vicinity of smelling salts, some sugar debasement items like natural corrosive is framed, taking into account LC-MS/LC-UV thinks about Introduction to High through put AFEX (new device to screen the distinctive food stock)

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Acknowledgment CAFI group for their important proposals Genencor for supplying their compounds Dr. Kevin and Dr. Walsum for doing LC-MS and LC-UV investigation DOE for financing

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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

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