An overview of the main research activities of the “Biomass and Bioenergy Lab” Speaker: Cristiano Varrone Email: [email protected] Giulio Izzo, Giulia Massini, Antonella Signorini, Fabrizio De Poli, Antonella Marone, Floriana Fiocchetti, Andrea Aliboni, Chiara Patriarca, Silvia Rosa, Luciano Mentuccia, Elena De Luca, Cristiano Varrone. Laboratory of Biomass and Bioenergy Head of Laboratory: Dott. Giulio Izzo / [email protected] IDROBIO Project: coupling dark and light fermentation - 2.4 L bioreactor operated at 80 °C and inoculated withThermotoga neapolitana ; a yield of 3.9 mol H2/mol glucose was obtained, with a production rate of 51 mL/L/h - 3.8 L bioreactor with 1.8 L working volume (BFM medium; 10g/L glucose, 37°C; pH 6.8), inoculated with 1g of sediments from the Averno lake; a yield of 1.7 mol H2/mol glucose was obtained, with a production rate of 1.8 L H2/L/day and 54.6% of H2 content 50 L Tubular Photobioreactor tested under natural light conditions (in cooperation with UO CNR-ISE) , using Rp. palustris 42 OL grown on a synthetic medium . Mean H2 production rate = 17 mL/L/h. 50 L bioreactor for the culture of T. neapolitana , with 25 L working volume. 1.2 L planar Photobioreactor (in cooperation with UO ENEA) with a surface of 0.0730 m2 containing Rp. palustris AV33 isolated from Averno sediments. Max H2 production rate : 1,4 L / L / day Continuous Flow Reactor using a mixed pools l H2/ l day mol H2/mol gluc. max rate (l H2/l h) Continuum 1300 h 7-8 2-2,5 0.5- 0.6 H2 (%) 54-59 Dark Fermentation of waste products Hydrogen/biogas production by dark anaerobic fermentation of organic wastes is a promising strategy to obtain renewable and clean energy in a sustainable way It can lead to the conversion of organic waste and feedstock into a host of valuable chemicals and energy The Approach: One way to improve the efficiency of H2 production is to explore the potentials offered by the microbial biodiversity, both in natural and artificial environment, identify and to select bacterial strains with high H2 producing abilities from different substrates, and to characterize the microbial metabolism, in order to understand and optimize the whole process. Synthetic Overview HYDROGEN PRODUCTION FROM: Isolation of hydrogen producing bacteria from vegetable waste for bioaugmentation: 2.1-2.4 L/L/d Selection and acclimatation of microbial mixed pools for degradation of agri- and zootecnical waste: 2.8-3 L/L/d Bioconversion of crude glycerol into H2 and ethanol from enriched activity sludge: 3 L/L/d MSE-ENEA project: H2 production from vegetal waste isolation and characterization of meso-philic bacterial strains contained in the waste for bioaugmentation Vegetable waste Isolation of bacterial strains Selection of H2-producers and cellulolytic bacteria 1 – V (leaf shaped vegetable waste); 2 – VP (80% leaf shaped vegetable waste + 20% potato peels);. DNA extraction Test on cellulose and hydrolysis products glucose cellobiose Identification: Sequencing of amplified 16S rDNA arabinose xylose PCR amplification 16S rDNA gene Improvement of the H2 production from self-fermentation of Vegetal Waste performing Bioaugmentation 90 V 70 60 50 40 30 20 10 0 0 5 10 Buttiauxella sp.4 15 20 25 30 Fermentation time (h) Rahnella sp. 10 Raoultella sp. 47 35 40 Consortium 45 50 Self-fermentation 90 VP 80 Cumulative H2 production (ml H2 /gVS) Cumulative H2 production (ml H2 /gVS) 80 Effect of the artificial consortium on vegetal waste 70 60 50 40 30 20 10 0 0 5 Buttiauxella sp.4 10 15 20 25 30 Fermentation time (h) Rahnella sp. 10 Raoultella sp. 47 35 40 Consortium 45 50 Self-fermentation Marone et al., 2012 International Journal of Hydrogen; 37(7): 56125622. Substrati Inoculo H2 (%) L H2/l/d ml H2/g VS m3 H2/t substrate Vegetal waste Self-fermentation 10-12 0.557-0.748 18-22 1-1.3 Vegetal waste Bio-agumentation 26-28 2.1-2.4 67-86 4-4.8 Bioconversion of agri- and zootechnical waste Degradation of Organic Matter : 1°Phase Hydrolysis of macromolecules 2°Phase Digestion of hydrolisates 3°Phase 4°Phase Acidogenesis Methanogenesis Vegetables Wood BIOMASS H2/CO2 Sugars Aminoacids Fatty acids Manure Glycerol • • • • Theoretic Yield from Glucose C6H12O6 → 3CH 4+ 3CO2 C6H12O6 + 2 H2O → 2CH3COOH + 2CO2 + 4H2 2CH3COOH → 2CH 4 + 2CO2 C6H12O6 + 2 H2O → 2CO2 + 2CH 4 + 4H2 BIOGAS CH4/CO2 Carboxylic acids Alcohols Acetate Energetic efficiency 83.2% 33.5% 89.0% Marea project: coupling H2 and CH4 production from agri- and zootecnical waste Substrati Cheese whey Cow manure Glycerol Inoculo consorzio IDROBIO consorzio IDROBIO consorzio IDROBIO H2 (%) 44-46 L H2/l/d 2.8-3.0 10-12 0.220-0.230 ml H2/g VS m3 H2/t substrate 160-180 20-23 23-25 0.520-0.540 32-35 0.600-0.800 120-140 Al bruciatore Al bruciatore Idrogeno,CO2 Liquam i Glicerolo Serb. Alim. Alim entazione Enhanced production of methane by bioaugmentation of H2 producing community Bioreatt ore 2 Q Bioreatt ore 1 Pompa 2 Acqua Risc./Raff Pompa 1 Q 3.86 L/Ld Brodo in ferm entazione Q Mixture Design (H2 = 34-40%) Microorganism i m etanogeni Microorganism i idrolitici 30-33 C.M = 66%, C.W. = 33% 2.15-2.98 L/Ld Metano, CO2 Al B ioreattore 2 Pompa 3 Digestato Statistical Optimization of Glycerol Fermentation In optimized conditions it was possible to obtain a max hydrogen production rate of more than 2.150 L H2/L/day (yield > 0.94 mol H2 / mol crude glycerol), while reaching a max EtOH concentration of almost 8g/L (yield ~ 1), without adding any vitamins, minerals, triptone or yeast extract. H2 concentration in the biogas reached more than 50%, and the H2/medium ratio (mL/mL) was found to be around 4 (max value obtained was 4,5 at 18g/L of glycerol). Cristiano Varrone et al.,2012. International Journal of Hydrogen Energy (in press, http://dx.doi.org/10.1016/j.ijhydene.2012.02.106) Scale-up Tests 3 L BioFlo 115 Benchtop Fermentor Pmax Rmax l (mL) (mL/h) (h) Novaol 3720.9 154.1 5.64 0.998 2997.7 0.91 Itabiol 3428.1 146.7 4.20 0.984 2983.4 0.89 Pure Gly 3808.1 138.8 6.34 0.998 2706.5 0.91 N.S. Novaol 3451.3 174.4 6.48 0.996 3152.4 0.92 R2 Rate (mL/L/d) Yield (mol/mol) Modified Gomperzt equation Rate= Pmax/(l+Pmax/Rmax) Future project: Pilot Plant Alla "torcia" We are now evaluating the possibility to set up a 1000L pilot plant, which might produce up to: Idrogeno Acqua Glicerolo Microorganism i Etanolo (95 %) - 400-500Kg EtOH/t of glycerol (with an estimated value of 350€/t glycerol) Bioreattore - 200-250 m3 H2/t of glycerol (with an estimated value of 70€/t glycerol; based on national subsidies of 0,28 cents/kWh) Serb. Alim . Pom pa 2 Acqua + ac. organici Pom pa 1 Acqua+etanolo+acidi organici Simulated industrial plant showed a production of 100 m3 H2/d and 25000 L EtOH/d, correspoing to 80 GWh/anno from H2 and 540 GWh/anno from EtOH, with an energy efficiency of 39%. PATENT APPLICATION: Bioconversion of crude glycerol into hydrogen and ethanol (Number: RM2011A000480) Applying data: 13/09/2011. Inventor: Cristiano Varrone Link: http://brevetti.enea.it/tabella.php (ENEA patent nr. 735) ENTERPRISE EUROPE NETWORK: http://portal.enterprise-europe-network.ec.europa.eu/ (Reference: 12 IT 56Z7 3PF3) THANK YOU FOR YOUR ATTENTION