Production of bio-based chemicals and polymers from industrial waste and byproduct streams
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1 Section II. Developments in biorefinery: feedstocks, processes and products (I) Production of bio-based chemicals and polymers from industrial waste and byproduct streams Apostolis Koutinas Group of Food Bioprocesses and Biorefineries Department of Food Science and Human Nutrition Agricultural University of Athens, Greece
2 Main research interests Agricultural University of Athens Department of Food Science and Human Nutrition Group of Food Bioprocesses and Biorefineries Biorefinery development using agriindustrial waste and by-product streams Separation of value-added co-products Bioprocess development using entirely renewable resources for the production of platform chemicals, biopolymers and microbial lipids Biorefinery and bioprocess design including techno-economic evaluation and life cycle assessment 214, 43,
3 Approach on industrial waste/by-product biorefinery development The case of food waste Extraction of valueadded co-products Food Wastes Upstream Processing Fermentation Downstream processing Bio-based products PHB Filtration L-L Extraction Distillation Crystallisation Drying Enzymatic Hydrolysis Microbial oil Carotenoids Solid State Fermentati on Fungi Bacteria l cellulos e Platform chemicals
4 Biorefinery/bioprocess design and techno-economic evaluation t 1 Fermentor Total Installed Capacity (m3) Bioreactor total installed capacity (m3) Fixed Capital Investment per L3 FCI per L of installed fermentor capacity($/l or k$/m ) installed bioreactor capacity($/l) t3 2 kt y 1 1 Molasses M 2 $ t 1 8 t5 t6 t7 1 t9 Sugarcane S t1 4 $ t 1 t11 1 kt y 4 t12 5 kt y 1 2 Glycerol G 2 $ t Fermentor Installed Capacity (L) Bioreactor totaltotal installed capacity (L) SA t4 t nb t2 t13 t14 t15 LA BDO PA PHB PDO 3HP SCO t16 t17 BA t18 t19 EtOH t2 Dheskali et al Bioresource Technology 224: t21 Bonatsos et al Biochemical Engineering Journal 116: t22 Koutinas et al Bioresource Technology 24:55-64 Dimou et al Biochemical Engineering Journal 116: t23 t24 t25 Koutinas et al Fuel 116:
5 Biorefinery development based on the valorisation of biodiesel industry byproducts for the production of antioxidant-rich fraction, protein isolate and poly(3-hydroxybutyrate)
6 Current biodiesel production process from oilseeds Hull Biodiesel Transesterification Crude glycerol Oil Crush Screw press and solvent extraction Oil 28, t/y (35-5% glycerol) Purification to more than 85% Sunflower seeds 7, t/y Oilseeds Biodiesel 28,14 t/y Seed residues Animal feed Sunflower meal (by-product 2) 42, t/y Glycerol (by-product 1) 2,912 t/y Animal feed or other low value applications
7 Base-case processing scenario Sunflower seed Sunflower oil Mechanical pressing and hexane extraction Transesterification Biodiesel Aspergillus oryzae Crude glycerol Carbon source Sunflower meal Solid state fermentation Enzymatic hydrolysis Microbial fermentation Nutrient-rich supplement Poly(3-hydroxybutyrate)
8 PHB production from whole sunflower meal hydrolysate (SFM) and crude glycerol PHB production stage FAN, IP (mg/l) Glycerol, TDW, PHB (g/l) Microbial growth stage 25 FAN ( ) In. Phosphorus ( ) Glycerol ( ) Total Dry Weight ( ) PHB ( ) Fermentation time (h) Crude glycerol Sunflower meal hydrolysate Microbial fermentation Polyhydroxybutyrate Kachrimanidou et al Bioresourse Technol. 172:121 13
9 Purification of PHAs using commercial enzymes Aspergillus oryzae Process I Sunflower meal Sunflower seed Solid state fermentation Remaining lignocellulose-rich solids Enzymatic hydrolysis Levulinic acid production Nutrient-rich supplement Biodiesel Oil Crude glycerol Microbial Bioconversion Commercial Pancreatin Heat treatment to deactivate enzymes non-pha cell lysis with commercial enzymes PHB or P(3HB-co-3HV) Removal of cell debris, colour and water
10 Advanced sunflower-based biorefinery Sunflower seeds Biodies el Partial Dehulling Oil Transesterification Mechanical pressing and hexane extraction Crude glycerol Microbial fermentation PHB Production Protei n isolate Nutrient supplemen t Partly dehulled or undehulled sunflower meal Protein-rich fraction Residual streams Enzymatic hydrolysis Remainin g stream Crude enzymes Antioxidan ts Aqueous extraction Liquid fraction Lignocelluloserich fraction Solid state fermentati on Remaining solids
11 PHB fermentation using the hydrolysate produced when all residual streams are employed Microbial growth stage 8 7 PHB production stage FAN and IP (mg/l) Glycerol, TDW and PHB (g/l) 6 FAN ( ) IP ( ) Glycerol ( ) Total Dry Weight 2 ( ) 1 PHB ( ) 15 Fermentation Time (h) This medium is efficient for enhanced PHB production Kachrimanidou et al Ind. Crops Prod. 71:16 113
12 Integration of multipurpose usage of crude enzymes produced on-site Protein-rich fraction Process II Sunflower meal Aqueous extraction Sunflower seed Antioxidants and Protein isolate or hydrolysate Remaining solids Fibre-rich fraction Solid state fermentation Aqueous stream Enzymatic hydrolysis Remaining lignocelluloserich solids Nutrient-rich supplement Biodiesel Oil Crude glycerol Levulinic acid production Microbial Bioconversion Heat treatment to deactivate enzymes PHB or P(3HB-co-3HV) non-pha cell lysis with crude on-site enzymes Removal of cell debris, colour and water
13 Evaluation of hydrolyzed solution of bacterial cells as fermentation substrate for PHB production The lysate from enzymatic nonphb cell lysis could be used as fermentation feedstock supplemented with phosphate salts Cupriavidus necator DSM 7237 FAN: 329 mg/l Glycerol: 2 g/l TDW: 5.75 g/l PHB content : 2%
14 Profitability assessment of different processing scenarios Process type Current practice - utilisation of SFM and crude glycerol as animal feed Process I - Net Present Value 1 year (million $) 31.5 PHB production from crude glycerol and part of SFM Lower than SFM utilisation as animal feed Process II (low range of market prices for biorefinery products) - fractionation of SFM to produce protein isolate and antioxidants Lower than 31.5 PHB production from crude glycerol and remaining SFM fractions Process III (high range of market prices for biorefinery products) - - fractionation of SFM to produce protein isolate and antioxidants - PHB production from crude glycerol and remaining SFM fractions Higher than 31.5
15 NPV range estimated based on low and high market prices for the protein isolate (1,25 2,5 $/t) and antioxidants (6 15 $/kg) Current Practice NPV = 31.5 million $ Low NPV value estimated when the protein isolate and antioxidants market prices are 1,25 $/t and 6 $/kg, respectively Lower than 31.5 million $ High NPV value estimated when the protein isolate and antioxidants market prices are 2,5 $/t and 15 $/kg, respectively Higher than 31.5 million $ 25 $/t A n tio x Protein id a n ts isolate: f r a c tio125 n ( $ /t) Protein isolate: 125 $/t Antioxidants: 6 15 $/t P r o tein I s o la te ( $ /t) Antioxidants: 6 $/t N P V 1 y ea r ( m illio n $ )
16 Biorefinery development based on the valorisation of spent sulphite liquor for the production of lignosulphonates, succinic acid and antioxidant-rich fraction
17 Spent sulphite liquor (SSL) production in the sulphite wood pulping process SO2 RECOVERY WOOD CHIPS COOKING CHEMICALS SO2/MeHSO3 (Me: Ca, Mg, Na, NH4) SSL Characterisation Value ph BLOW TANK WATER WASHING DIGESTER THIN LIQUOR CELLULOSE FIBRES SSL MULTIPLE EFFECT EVAPORATION PROCESS St Dev 2.7 Density (g/ml) Viscosity (cp) Dry Matter (g-dm/l) Lignosulphonates (g/l) Ash % (g/g-dm) Phenolics % (g/g-dm) Carbohydrates (g/l) Xylose (g/l) Galactose (g/l) Glucose (g/l) Mannose (g/l) Arabinose (g/l) Acetic Acid (g/l) THICK LIQUOR
18 Integrated biorefinery based on current pulp and paper mills Lignosulphonates Phenolic Extract Fermentation SSL Nanofiltration Fireproof biopolymers Phenolic extraction with ethyl acetate Bioprocess optimisation Techno-economic evaluation Life Cycle Analysis Succinic acid production Polybutylene succinate Succinic acid separation and purification
19 2 Fermentation with LS addition Fermentation with SSL 15 Actinobacillus succinogenes 1 Inhibition begins LS Concentration (g/l) Basfia succiniciproducens Succinic Acid Concentration (g/l) Succinic Acid Concentration (g/l) Effect of SSL and extracted lignosulphonate (LS) concentration on succinic acid production 2 Fermentation with LS addition Fermentation with SSL 15 1 Inhibition begins LS Concentration (g/l)
20 Pretreatment of spent sulphite liquor (SSL) SSL Nanofiltration Fermentation Phenolic extraction with ethyl acetate
21 Fed-batch fermentations in bench-top bioreactor Pure mixed sugars Nanofiltrated SSL Total sugars (g/l) Succinic acid (g/l) 35 Total sugars, succinic acid (g/l) Total sugars, succinic acid (g/l) Total sugars (g/l) Succinic acid (g/l) Time (h) 3 By-products (g/l) Time (h) Pure mixed sugars 2 Time (h) Formic acid (g/l) Acetic acid (g/l) Total by-products (g/l) Lactic acid (g/l) 3 Lactic acid (g/l) Formic acid (g/l) Acetic acid (g/l) Total by-products (g/l) 25 By-products (g/l) B. succiniciproducens 4 Final SA concentration : 34.2 g/l Yield:.65 g/g Productivity:.59 g/l/h 2 4 Time (h) Nanofiltrated SSL with.5 kda MWCO Final SA concentration : 33.8 g/l Yield:.58 g/g Productivity:.48 g/l/h
22 ontinuous cultures for the production of succinic acid from SS Lactic acid Formic acid Ace tic acid,19 SSL,14 2,9 15,4 1 Dilution Rate (1/h) 25 Concentrations (g/l) 3 Succinic acid,24 35 Total Sugars -, ,6 Time (h) Observations Succinic acid concertation varied between 2-25 g/l when synthetic xylose was consumed. Xylose accumulation was observed when dilution rate was increased The optimum dilution rate observed was.4 h-1 when pretreated SSL was used The maximum succinic acid concentration achieved was 2.7 g/l at dilution rate.4 h-1.
23 Direct crystallization scheme for succinic acid purification Fermentation Broth Centrifugation Activated carbon Ion-Exchange Resins Acidification Process 1 Evaporation Process 2 Crystallization Sample Yield (%) Purity (%) Drying Synthetic fermentation broth Actual fermentation broth up to 8 up to 99.7 Succinic acid crystals
24 Cost evaluation of bio-based succinic acid production Two different fermentation strategies Continuous Succinic acid production via fermentation using spent sulphite liquor as substrate Three production capacities 5 (t/y) 3 (t/y) Fed-batch 1 (t/y)
25 Succinic acid production flow diagram Spent Sulphite liquor Nutrients Upstream Nano filtration Sterilization Inoculum Fermentation Fermentation Direct Crystallization Downstream Succinic acid crystals
26 Cost evaluation of bio-based succinic acid production 6 Succinic acid total production cost Continuous procces 4,19 3,56 4,8 3,37 3 1,5 Cost ($/kg) 4,5 4,34 3,92 Fed-batch process 1 % contribution of individual costs on Total Production Cost Continuous procces 8 Fed-batch process 67, ,57 39,7 4 2 Contribution (%) Capacities (t/y) 22,56 9,77 9,38 )+C(la C(utility a te ria C(raw m FCI
27 Thank you for your attention
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