An overview of the main research activities of the Biomass and Bioenergy Lab

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1 An overview of the main research activities of the Biomass and Bioenergy Lab Speaker: Cristiano Varrone 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 / giulio.izzo@enea.it

2 IDROBIO Project: coupling dark and light fermentation 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 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 H 2 /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 m2 containing Rp. palustris AV33 isolated from Averno sediments. Max H2 production rate : 1,4 L / L / day

3 Continuous Flow Reactor using a mixed pools l H 2 / l day mol H 2 /mol gluc. max rate (l H 2 /l h) H 2 (%) Continuum 1300 h ,

4 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 The Approach: It can lead to the conversion of organic waste and feedstock into a host of valuable chemicals and energy One way to improve the efficiency of H 2 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 H 2 producing abilities from different substrates, and to characterize the microbial metabolism, in order to understand and optimize the whole process.

5 Synthetic Overview HYDROGEN PRODUCTION FROM: Isolation of hydrogen producing bacteria from vegetable waste for bioaugmentation: L/L/d Selection and acclimatation of microbial mixed pools for degradation of agri- and zootecnical waste: L/L/d Bioconversion of crude glycerol into H2 and ethanol from enriched activity sludge: 3 L/L/d

6 MSE-ENEA project: H 2 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 H 2 -producers and cellulolytic bacteria 1 V (leaf shaped vegetable waste); 2 VP (80% leaf shaped vegetable waste + 20% potato peels);. Test on cellulose and hydrolysis products DNA extraction glucose xylose cellobiose arabinose Identification: Sequencing of amplified 16S rdna PCR amplification 16S rdna gene

7 Cumulative H2 production (ml H2/gVS) Cumulative H2 production (ml H2/gVS) Improvement of the H 2 production from self-fermentation of Vegetal Waste performing Bioaugmentation V Fermentation time (h) Buttiauxella sp.4 Rahnella sp. 10 Raoultella sp. 47 Consortium Self-fermentation VP Effect of the artificial consortium on vegetal waste Fermentation time (h) Buttiauxella sp.4 Rahnella sp. 10 Raoultella sp. 47 Consortium Self-fermentation Marone et al., 2012 International Journal of Hydrogen; 37(7): Substrati Inoculo H 2 (%) L H 2 /l/d ml H 2 /g VS m 3 H 2 /t substrate Vegetal waste Self-fermentation Vegetal waste Bio-agumentation

8 Bioconversion of agri- and zootechnical waste Degradation of Organic Matter : 1 Phase Hydrolysis of macromolecules 2 Phase Digestion of hydrolisates 3 Phase Acidogenesis 4 Phase Methanogenesis Wood Vegetables BIOMASS Sugars Aminoacids Fatty acids H 2 /CO 2 BIOGAS CH 4 /CO 2 Manure Glycerol Carboxylic acids Alcohols Acetate Theoretic Yield from Glucose Energetic efficiency C 6 H 12 O 6 3CH 4+ 3CO % C 6 H 12 O H 2 O 2CH 3 COOH + 2CO 2 + 4H % 2CH 3 COOH 2CH 4 + 2CO 2 C 6 H 12 O H 2 O 2CO 2 + 2CH 4 + 4H %

9 Marea project: coupling H 2 and CH 4 production from agri- and zootecnical waste Substrati Inoculo H 2 (%) L H 2 /l/d Cheese whey Cow manure Glycerol consorzio IDROBIO consorzio IDROBIO consorzio IDROBIO ml H 2 /g VS m 3 H 2 /t substrate Serb. Alim. Liquam i Glicerolo Microorganism i idrolitici Alim entazione Q Al bruciatore Idrogeno,CO2 Brodo in ferm entazione Bioreattore 1 Q Al bruciatore Metano, CO2 Microorganism i m etanogeni Bioreattore 2 C.M = 66%, C.W. = 33% L/Ld (H 2 = 34-40%) Mixture Design Q Acqua Risc./Raff Pompa 1 Pompa 2 Al Bioreattore 2 Pompa 3 Digestato 3.86 L/Ld Enhanced production of methane by bioaugmentation of H 2 producing community

10 Statistical Optimization of Glycerol Fermentation In optimized conditions it was possible to obtain a max hydrogen production rate of more than L H 2 /L/day (yield > 0.94 mol H 2 / mol crude glycerol), while reaching a max EtOH concentration of almost 8g/L (yield ~ 1), without adding any vitamins, minerals, triptone or yeast extract. H 2 concentration in the biogas reached more than 50%, and the H 2 /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,

11 Scale-up Tests 3 L BioFlo 115 Benchtop Fermentor P max R max l R 2 Rate Yield (ml) (ml/h) (h) (ml/l/d) (mol/mol) Novaol Itabiol Pure Gly Modified Gomperzt equation Rate= Pmax/(l+Pmax/Rmax) N.S. Novaol

12 Future project: Pilot Plant Acqua Glicerolo Microorganism i Idrogeno Alla "torcia" We are now evaluating the possibility to set up a 1000L pilot plant, which might produce up to: Serb. Alim. Pompa 2 Bioreattore Etanolo (95 %) Acqua + ac. organici Kg EtOH/t of glycerol (with an estimated value of 350 /t glycerol) m 3 H 2 /t of glycerol (with an estimated value of 70 /t glycerol; based on national subsidies of 0,28 cents/kwh) Pompa 1 Acqua+etanolo+acidi organici Simulated industrial plant showed a production of 100 m 3 H 2 /d and L EtOH/d, correspoing to 80 GWh/anno from H 2 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: (ENEA patent nr. 735) ENTERPRISE EUROPE NETWORK: (Reference: 12 IT 56Z7 3PF3)

13 THANK YOU FOR YOUR ATTENTION