Lignocellulosic biorefinery pathways to biobased chemicals and materials 1 st Int. Forest Biorefinery Conference, Thunder Bay, Canada May 9-11, 2017 Richard Gosselink, Carmen Boeriu, Paulien Harmsen, Jeroen Hugenholtz
Contents Wageningen Food & Biobased Research Biorefinery value chains Lignin valorization to materials and chemicals Microbial production of biochemicals
2 Partners Wageningen Research 2,410 FTE of faculty and staff 11,000 students Revenue in 2015: 635 million
Wageningen Food & Biobased Research In-depth knowledge of the entire agri-food chain Connecting agri-food with chemistry, materials and energy production Market oriented R&D approach Multi-disciplinary applied R&D project teams Up-scaling: from lab to pilot > 200 employees with a yearly turnover of 30M Sustainable Food Chains Biobased Products Healthy & Delicious Foods
Research Topics BU Biobased Products Biorefinery & Sustainable value chains Biomass sourcing & logistics Biomass resources & characterization Biomass pretreatment (mechanical/chemical) Composite materials Sustainable building materials Sustainable Chemistry Bioplastics, coatings and packaging Fine chemicals (plasticizers, polymer building blocks) Polymeric foams Fire retardants Bioconversion Fermentation (H 2, ethanol, lactate, ABE, fatty acids) Bio-catalysis
Chemicals and materials driven biorefineries Developing biobased chemicals will increase the profitability of second generation biofuels production Biobased chemicals and materials driven biorefineries can also be created alongside traditional vegetable oil, starch, sugar and paper producers Agri-food industries are diversifying their product slate; increasingly engaging in non-food products
Composition of lignocellulosic feedstocks (wt% dm) Origin Species Carbohydrates Lignin C6 sugars C5 sugars Hardwoods Mixed (stem) 60-75 40-50 16-20 18-25 Softwoods Grasses Agricultural residues Mixed (stem) Sugar cane bagasse 60-67 40-50 15-18 27-33 60-70 33-36 20-25 19-24 Corn cobs 75 40 30-34 15 Wheat straw 55-60 30-35 20-23 16-21 Rice husks 50-55 30-35 20-22 20-22
Pretreatment of lignocellulose Fibres: pulp, paper, building materials Dissolving cellulose: textile Sugars: molecules for conversion Make the polysaccharides accessible to catalysts Using low/high ph, high temperatures, oxidative agents, mechanical forces (explosion) Catalysts for polysaccharide hydrolysis: enzymes or acid Micro-organisms can use the monosaccharides in fermentation processes
Pretreatment technologies Chemi-mechanical Green biorefineries Alkaline hydrolysis Autohydrolysis (only water) Organosolv pretreatment using acetic acid Thermal pretreatment Mild pretreatment (assisted by enzymes)
Enzymatic pretreatment (see poster)
CIMV organosolv fractionation process Acetic/formic acid RAW MATERIAL (wood, straw, bagasse ) Since 2006 @ 150 kg/h Mechanical conditioning Organic refining SUGARS & LIGNINS Organic acid solutions Organic Acid recycling RAW PULP Delignification / Peracids Lignin / sugars separation Washing / Neutralisation Bleaching / Washing LIGNINS C 5 SUGARS Cellulose C 6 PULP M. Delmas, Chem. Eng. Technol. 2008,31,No.5,792-797 12
Intermediate products Cellulose (glucose) Hemicelluloses (xylose and arabinose) Lignins 13
Further value chain assessment PU elastomer coatings Rigid PU foams Bio-ethanol for fuel or PVC plasticizer Biobased phenol-formaldehyde resins Itaconic acid for paint
Drivers for lignin valorization Additional revenues beyond energy value Development of sustainable processes and products Biobased and circular economy Unique functionality Aromatic structure Polymer properties Crosslinking ability (softwood lignin) UV stability Flame retardance Anti-microbial Hydrophobicity Bulk versus niche markets
Aromatic chemicals & materials derived from lignin Polymerisation Depolymerisation Binders/resins Fractionation Oligomeric fragments Chemical/ Enzymatic upgrading (Bio-)catalysis Monomeric chemicals Composites Coatings Surfactants Bitumen (asphalt, roofing) Marine fuels Polymer building blocks Confidential 16
Examples for lignin applications Application Scale of operation TRL Main challenges Bio-asphalt Demonstration 6-7 Costs reduction, industrial handling Thermoset resins Pilot / demo 5-6 Reactivity Polyurethanes Pilot 5-6 Viscosity, reactivity Carbon fibres Pilot 5-7 Strength performance Marine fuels Pilot 5-6 Sulphur-free, viscosity Aromatic chemicals R&D Commercial for vanillin 2-4 9 Desulphurisation, coke formation
3.666 1.000 0.708 1.945 1.840 1.071 2.074 Lignin fractionation Aim: produce homogeneous and purified lignin fractions with tailored molar mass and properties Solvent fractionation Solvents with increasing Hansen solubility parameters Internal std Guaiacyl-OH COOH Aliphatic OH Syringyl-OH p-hydroxyl-oh Condensed OH 150 148 146 144 142 140 138 136 134 ppm Lignin fractions ranges from Mw 1000 to 7000 g/mol Lower molar mass fractions increases in purity and functionality Gosselink, R.J.A., J.C. van der Putten, D.S. van Es, Fractionation of technical lignin, WO2015/ 178771
Laccase-mediated oxidation of lignin Explore the effect of lignin composition on enzymatic oxidative polymerisation Lignin substrates: LMW fractions of 5 technical lignins Laccase Acetone/water 50:50
Laccase-mediated oxidation of lignin Enzymatic modification of lignins in water/acetone Lignin oxidative polymerization Kinetics Products DP = f(s, G, H) Fitigau et al, 2013, ABP
Bio-asphalt Bulk / low value application Substitution of fossil bitumen (200 500 / ton) Manufacturing at lower temperature (lower CO 2 ) Infra project Zeeland (NL): 50% substitution of bitumen by lignin from lab to demonstration 2 public demonstration roads in The Netherlands Challenges: costs lignin raw material, industrial handling
Hydrothermal lignin depolymerization Biomass Focus: Biorefinery lignins Aromatics and phenolics O H Turn lignin into high value aromatics (BTX) and building blocks (phenol) Approach: Selective catalytic hydrothermal depolymerisation without external hydrogen; Prevent re-condensation Van Es, D.S., Van der Klis, F., Van Haveren, J., Gosselink, R.J.A., Method for the 22 depolymerization of lignin, WO2014/168473
Relative Abundance Hydrothermal lignin conversion RT: 4.00-23.00 100 95 90 85 80 75 70 65 10.84 NL: 7.59E6 TIC MS Lignin1603 11 60 55 50 45 40 35 30 25 20 12.02 14.68 20.42 15 10 5 0 13.04 15.29 8.54 8.62 16.09 20.37 14.03 11.17 16.27 22.32 12.59 19.10 8.84 17.80 4.04 19.19 7.22 13.29 9.90 20.47 7.08 5.81 7.27 4.38 21.00 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Time (min) Lignin depolymerization to monomers is 20% over Pd/C in water Oligomeric lignin fragments 60-80% Char formation <10% Limited product distribution 1 compound in 50% (Guaiacol) Monomers further converted into BTX, cyclohexanone
Fermentation challenges Maximize gas transfer & cooling capacity Prevent substrate and product inhibition Minimize costs for product recovery Minimize substrate costs Maximize performance of micro-organism
Microbial production of MC-Fatty Acids Cheap waste streams as substrate for MCFA production Various microbial platforms for MCFA production Cryptococcus curvatus Yarrowia lipolytica Pseudomonas putida (hydroxy-fa s/pha) Various Microalgae Metabolic Engineering Tailor-made chain length Tailor-made saturation level
X Fatty acid degradation through the β-oxidation pathway. X Unsaturated fatty acids 26
Pseudomonas for hydroxy-fa production Oleic acid C18:1 Linoleic acid C18:2 Linolenic acid C18:3 Palmitic acid C16:0 Tetradecanoic acid C14:0 Partial degradation Pseudomonas putida KT 2442 X PHA X TesB 3OH-C6 3OH-C8 3OH-C10 3OH-C14 27
Anaerobic fermentation to alcohols ABE (acetone/butanol/ethanol) and IBE (isopropanol/butanol/ethanol) production by Clostridia CRISPR/CAS introduction in C. beijerinckii Conversion of cellulose and hemicellulose Chemical and enzymatic conversion of alcohols to highvalue alcohols/aldehydes/esters/etc. (Anaerobic) Production of flavours, surfactants, fatty acids Conversion of C1-gasses/syngas to various alcohols by mixed anaerobic cultures
Waste streams to ABE
C1 (CO/CO 2 ) to alcohols
Production of organic acids at low ph Approach: Select potential host based on required process condition, High product and substrate tolerance Ability to grow on envisaged substrate Suitable metabolism Rapid growth/substrate conversion rate Simple medium requirements Genetic accessibility Engineer production pathway in selected host
Example: Lactic acid production Lactic acid: food & feed conservation, monomer for PLA Production at neutral ph -> salt (gypsum) as by-product Strains isolated with high tolerance to lactic acid and sugar (glucose + xylose) at low ph on mineral medium Best strain: Monascus ruber Engineered from rapid lactic acid consumer into rapid lactic acid producer
Biobased Antifungals/Antimicrobials Protective Cultures Direct application in food, on crops Protective cultures for ingredient production Protective cultures for Ferment production Antifungals from abundant biomass Various (modified) chitosans from chitin Aromatics (ferulic acid, cinnamic acid, etc) Plant components (terpenes/polyphenols)
Protective cultures for Strawberry Juice
Conclusions Biorefinery industries exits and there is more focus on chemicals and materials Development of sustainable processes is key Combination of enzymatic and chemical pretreatments gives benefits Mixture of products is possible and key Lignin has large potential to be used in materials and for chemicals Need for larger volume lignin production at reasonable costs Focus both on bulk and niche (high value) applications Fermentation processes can be tuned to produce biobased chemicals (fatty acids, organic acids, alcohols) from waste/side streams
Thank you! For collaboration or more information please contact: Richard.gosselink@wur.nl Jeroen.hugenholtz@wur.nl