Chair of Chemistry of Biogenic Resources TU München - R&D activities - Chair of Chemistry of Biogenic Resources Schulgasse 18, 94315 Straubing, Germany Sieber@tum.de www.rohstoffwandel.de
Locations Freising Niederbayern Straubing Main site >1000 m 2 Laboratory > 300 m 2 Pilot plant hall Foto: Stefan Müller-Naumann Department of Chemistry, Garching Center for catalysis Center for industrial biotechnology
Situation of modern chemistry Vision: Biomass substitutes mineral oil as raw material for industriall chemistry Wood, straw, vegetable oil Polymeres, dyes, surfactants, nutrients, solvents, pharmaceuticals, biofuels
Raw materials of industrial chemistry Crude oil is the most important raw material, but it is limited Image Source: Maurer, Henkel
il C2 C3 C4 C5 C6 Mineral oil refinery Distillation, Cracking, Reforming Distillation, Hydrotreating, Reforming, Cracking, Alkylation
Biorefinery 3 rd Generation LCB Enzyms 1 Treatment 1 Enzyme 2 Treatment 2 E3 T3 E4 T4 C5 sugars C6 sugars Proteins Lignin Low quantity substances Residual substrates Polyols & Derivatives Acetone Xylene Amino acids & Derivatives Phenols Dyes, sterins, etc. Energy recovery (combustion or fermentative utilization)
R&D goals 1. Development of new products based on plant biomass 2. Development of catalysts as well as processes for the conversion (Chemo- and Biotransformation) of plant biomass 3. Development of methods for HT- catalyst engineering 2 1 Plant biomass 3 3 Robotics 3 Intermediate/ Monomer 2 Biopolymer/ Material 1 2
Processes Catalyst development Microorganisms Metabolic Engineering Homogeneous Catalysts Synthesis of Metal complexes Enzymes Proteindesign and -evolution Heterogeneous Catalysats Surface modification Process design Yesterday Today Tomorrow A B E F G C D Fermentation A B E F C D A B E F Chemo-enzymatic Cascade reactions
Bioconversion of lignocellulosic biomass
Bioconversion of lignocellulosic biomass Lignocellulosic biomass (LCB) Enzymatic saccharification
Bioconversion of lignocellulosic biomass ~ 25 % ~ 20 % ~ 40 % Hemicellulose Lignin Cellulose Physico-chemical methods Pretreatment Saccharolytic / lignolytic enzymes Hydrolysates ligo- /Monsaccharides
Microbial polysaccharides
Microbial polysaccharides Strain Selection Research areas Metabolic engineering Applications Filamentous fungi Fermentation & Downstream Polyacrylat Xanthan No agent 0,02% 0,02% Bacteria Food-Pharma- Cosmetics
Microbial polysaccharides xanthan Polysaccharides as flocculation agent Function: - Improvement of sedimentation (e.g. in sewage sludge) Sedimentation of clay suspension Current product: - Standard is polyacrylate => not biodegradable, petro derived Essential properties: - High molecular weight - Charged Polyacrylate 0,02 % Xanthan 0,02 % No agent
Microbial polysaccharides - xanthan Polysaccharide produced from plant pathogen bacteria Xanthomonas campestris pv. campestris - Leaf plotch Blattfleckenkrankheit, Ölfleckenkrankheit, black rot Schwarzfäule H Cellulose chain as backbone, Mannose and glucuronic acid side H H H chains, H Mw of >10 6 Da - H Shear thinning, thixotropic H H β-d-mannose β-1,4 D-Glucose - H H H H H H H H H β-1,2 D-Mannose β-1-4-d-glucuronic acid H H at rest in movement disordered viscous ordered free flowing
Microbial polysaccharides low acetyl xanthan Removal of acetate Improved viscosity Increased thermostability Improved interactions with other polymers => Coproduction of acetyl esterase during xanthan production
Depolymerisation of lignin
Depolymerisation of Lignin => Combination of chemical and enzymatic methods Fraunhofer Project Group BioCat RuCl 2 (PPh 3 ) 3 ADH/GST
Enzymatic cleavage of Lignin s beta-aryl-ether Enzymes focused on: Lig D, Lig F and Lig G* Reference substance: Guaiacol glycerol guaiacol ether (GGE)* Tasks Expression and purification of the LigD, F, G enzyme system Determination and optimization of enzyme activities Adoption to complex lignin substrates *Sato et al., Appl Environ Microbiol 75 (16), 2009
Enzymatic cleavage of Lignin s beta-aryl ether LigD-, LigF- and LigG- cell extracts Synthesis of codon-optimized gene sequences (Sphingomonas spec.) by Geneart GmbH Protein overexpression in E.coli BL21 (DE3) Enzyme-assays with cell extracts and GGE cleavage of 0.151 mm GGE in 150 min at 30 C, ph 8 (assay II) HPLC detection of 2 compounds GGE- concentration [mm] 0,3 0,25 0,2 0,15 0,1 0,05 0 I: 50 µl CE, II: 100 µl CE; 0.3 mm GGE 0 150 reaction time [min] without enzyme Assay I Assay II Guaiacol glycerol guaiacyl ether ß-hydroxypropiovanillone guaiacol
Thank you for your attention!