National Technical University of Athens NaOH ALKALINE School of Chemical Engineering Unit of Environmental Science & Technology PRETREATMENT FOR IMPROVEMENT OF ENZYMATIC DIGESTIBILITY OF WHEAT STRAW AS AL TERNATIVE SUGAR J. Novakovic, N. Kontogianni, E.M. Barampouti, S. Mai, D. SOURCE Malamis, M. Loizidou 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Agricultural Waste Biorefjnery Forster-Carneiro, T., Berni, M. D., Dorileo, I. L., & Rostagno, M. A. (2013). Biorefjnery study of availability of agriculture residues and wastes for integrated biorefjneries in brazil. Resources, Conservation and Recycling, 77 , 78-88. 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
-Lignocellulosic material The major barrier to the enzymatic hydrolysis of carbohydrates towards the production of fermentable sugars https://www.e- education.psu.edu/egee439/node/606 corn stover on a fjeld credit: USDOE-NRE https://www.greenoptimistic.com 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Pretreatments Removal of lignin is favorable to reducing the recalcitrance of lignocellulose for enzymatic attack. 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
tested A/A Reagent T Solid :Liquid Time (w/w) i H 2 O 2 (5%) 50°C 1:20 1h NaOH (pH=11,5) ii H 2 O 2 (10%) 50°C 1:20 1h NaOH (pH=11,5) iii NaOH 0.5M 50°C 1:10 96 h iv NaOH 0.5M 121°C 1:10 1h v CH 5 N 25 %w/w 50°C 1:10 96 h vi CH 5 N 25 %w/w 121°C 1:10 1h vii Na 2 CO 3 0.5M 50°C 1:10 96 h viii Na 2 CO 3 0.5M 121°C 1:10 1h ix NH 3 25 %w/w 50°C 1:10 96 h x NH 3 25 %w/w 121°C 1:10 1h 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Saccharifjcation of alkaline pretreated straw %cellulose 100 300 degradation 90 250 80 m 70 200 g 60 % g 50 150 l u c 40 o 100 s 30 e / 20 g 50 10 s t r 0 0 a 1 2 3 4 5 6 7 8 9 10 11 w Pretreatment methods (i) alkaline peroxide 5%, (ii) alkaline peroxide 10%, (iii) NaOH 0.5M , (iv) NaOH 0.5M autoclaving, (v) methylamine 25 %w/w, (vi) methylamine 25 %w/w autoclaving, (vii) Na 2 CO 3 0.5M, (viii) Na 2 CO 3 0.5M autoclaving, (ix) ammonia 25 %w/w, (x) ammonia 25 %w/w autoclaving. 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
AIM Optimisation of the NaOH pretreatment process along with the enzymatic hydrolysis in order to obtain more fermentable sugars and glucose recovery 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Raw material - Wheat straw Origin: From Aspropyrgos province, Greece Parameter Value (% w/w) Cellulose 33.8 Hemicellulose 45.1 Lignin 16.4 Klason lignin 15.4 Acid-soluble lignin 1.0 Ash 4.7 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Optimisation Parameters Particle Size Enzyme Loading Recycling of alkaline solution Pretreatment Kinetics 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
PARTICLE SIZE 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Particle Size- Experimental procedure Comminution NaOH Enzymatic pretretment Hydrolysis • FRITSCH Cutting mill Pulverisette • NaOH 0.5M • 10% w/w dry 15 solid • S/L: 1/10 w/w • Particle size • 15μL g -1 dry • Autoclaved at 1mm solid 121°C for 1 h • Particle size 10- (CellicCT ec2) 20mm • 50°C and 300 rpm for 96 h 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Particle Size 100 1mm 90 80 70 60 50 40 30 20 % 10 0 Conclusion 600 The particle size in the mg/g pretreated straw 1mm 500 range of 1 to 20mm 400 didn’t afgect signifjcantly 300 the performance of the 200 pretreatment scheme 100 0 TOC Glucose Efgect of particle size on enzymatic hydrolysis of pretreated straw 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
ENZYME LOADING 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Enzyme Loading- Experimental procedure Comminution NaOH Enzymatic pretretment Hydrolysis • Particle size 10- 20mm • NaOH 0.5M • 10% w/w dry • S/L: 1/10 w/w solid • 50°C and 300 • Autoclaved at rpm for 96 h 121°C for 1 h • Cellic CTec2 5, 10, 15 and 20 μL/g pretreated straw 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Enzyme Loading Conclusion 15 μL/g pretreated straw was selected as the optimum cellulase dosage % c e llu lo s e d e g r a d a t io n 0 5 10 15 20 % cellulose degradation TOC (mg/g pretreated straw) Efgect of enzyme loading on the straw solubilization and Glucose (mg/g pretreated straw) cellulose degradation during enzymatic hydrolysis 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
ALKALINE SOLUTION RECYCLING 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Alkaline solution Recycling- Experimental procedure Comminution NaOH Enzymatic pretretment Hydrolysis • Particle size 10- 20mm • NaOH solution • 10% w/w dry recycled solid • S/L: 1/10 w/w • 50°C and 300 • Autoclaved at rpm for 96 h • Cellic 15 μL/g 121°C for 1 h pretreated straw 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Alkaline solution Recycling %TS %cellulose %AIL %ASL %hemicellulose Conclusion Cycle hydrolysis degradation degradation degradation degradation No The alkaline solution 36.6 ± 0.1 2.1 99.5 51.7 could be recirculated 1 2.74 ± 0.26 86.36 ± ± 2.84 ± 2.59 1 2 6 4 1 at least 4 times 30.1 ± 0.2 3.5 99.4 26.3 2 4.12 ± 1.47 79.51 ± ± 2.93 ± 2.31 before being 6 1 8 8 4 “saturated” 26.8 ± 0.3 5.2 99.4 15.6 3 7.34 ± 1.61 72.21 ± ± 3.65 ± 3.26 although after each 0 2 3 1 7 30.0 ± 0.7 2.4 99.4 44.0 experiment it 4 8.50 ± 2.44 76.47 ± ± 5.62 ± 6.19 8 8 5 1 2 needed to be 14.0 ± 0.1 1.3 99.3 23.6 supplemented with 5 5.05 ± 2.63 65.96 ± ± 3.18 ± 4.36 3 3 6 8 5 some fresh solution. Solid fractions’ degradation after the difgerent cycles of pretreatment of wheat straw. 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
PRETREATMENT KINETICS 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Pretreatment Kinetics - Experimental procedure Comminution NaOH Enzymatic pretretment Hydrolysis • Particle size 10- 20mm • NaOH 0.5M • 10% w/w dry solid • S/L: 1/10 w/w • 50°C and 300 • 50°C for 0-96 h rpm for 96 h • Cellic 15 μL/g pretreated straw 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Pretreatment Kinetics Conclusion In the time interval of 3-5h of pretreatment, the delignifjcation effjciency reached its maximum and remained statistically constant the rest 90h. Timeline of total organic carbon (g/L) and delignifjcation effjciency (%) during NaOH 0.5M pretreatment at 50°C 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
SCALE-UP 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Scaling-up - Experimental procedure 5g/batch 3000g/batch 200g/batch V reactor = 250mL V reactor = 200L V reactor = 4L 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
Scaling-up NaOH pretreatment step Saccharifjcation step 300 120 %Delignifjcation VFA (mg/gstraw) Glucose (mg/g straw) 250 100 Phenols (mg/gstraw) 200 80 150 60 100 40 50 20 0 0 5 200 3000 5 200 3000 Capacity (g straw/batch) Capacity (g/batch) 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
CONCLUSIONS NaOH Enzymatic Hydrolysis Straw 1kg pretreatment 50 o C Hydrolysate 50 o C, 5h CelliCT ec 10.5μL/g straw 7kg mg/L TSS 23000 NaOH 0.5M 6.25kg VSS 21390 TOC 30507 VFA 6387 Phenols 127 mg/L Glucose 37444 TSS 3000 Exhausted VSS 2700 Alkaline Solution TOC 29256 1,25kg VFA 976 Phenols 125 7 th International Conference on Sustainable Solid Waste Management, 26-29 June 2019, Heraklion, Crete Island, Greece
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