Paola Di Donato Associate Researcher

1. Agro-industry vegetable residues valorisation : a waste biorefinery Paola Di Donato Associate Researcher (paola.didonato@icb.cnr.it) Institute of Biomolecular Chemistry Via Campi Flegrei 34, (80078) Napoli - ITALY November 20th, 2012 Città della Scienza, Naples, Italy

2. 31 institutes  Padova Institute of Biomolecular Chemistry Pozzuoli, Napoli 45 institutes Rome  Naples  Sassari  31 institutes  Catania November 20th, 2012 Città della Scienza, Naples, Italy

3. Born in 1967 as “Istituto per la Chimica e Fisica delle Molecole di Interesse Biologico” (ICMIB), “Institute for the Chemistry and Physics of Biologically Interesting Molecules”. • In 2002 it moved to the new building in Pozzuoli (NA) and took the name of “Istituto di Chimica Biomolecolare (ICB-CNR)”. Institute of Biomolecular Chemistry that today includes also the Roma, Catania, Sassari and Padova branches. • Main research activities in ICB are in the fields of organic chemistry, biochemistry and microbiology with main topics including environmental, pharmaceutical, food and biotechnological applications. November 20th, 2012 Città della Scienza, Naples, Italy

4. Agro-industry residues: a waste biomass According the Directive 2001/77/EC of the European Community biomass is “the biodegradable fraction of products, waste and residues from agriculture (including vegetal and animal substances), forestry and related industries, as well as the biodegradable fraction of industrial and municipal waste” According the Directive 2001/77/EC of the European Community biomass is “the biodegradable fraction of products, waste and residues from agriculture (including vegetal and animal substances), forestry and related industries, as well as the biodegradable fraction of industrial and municipal waste” Source: http://www.ecomena.org/biomass-resources-in-middle-east/ November 20th, 2012 Città della Scienza, Naples, Italy

5. Residual biomass: yearly production in Italy (source: C.R.P.A., Animal Production Research Centre, Italy)   November 20th, 2012 Città della Scienza, Naples, Italy

6. Agro-industry residual biomass in Italy: yearly amounts (source: Itabia )   November 20th, 2012 Città della Scienza, Naples, Italy

7. drugs nutraceuticals cosmetics biopolymers biofuels Agro-industry residual biomass: a resource that can be valorised Environmental problem Economical issue …. but a significant fraction of biomolecules is lost after industrial processing, being discarded into the residual matter….. proteins, fibres, polysaccharides, sugars, polyphenols, carotenoids • by-products that can represent the starting material for other processes • raw materials not in competition with food chain November 20th, 2012 Città della Scienza, Naples, Italy

8. Food industry wastes (vegetables processing residues) • Agriculture and crop residues BIOMASS • Bioprocesses (bacteria, enzymes) • Chemical Processes • Physical Processes • Biofuels • Antioxidants • Biopolymers Energy and chemicals Agro-industry residual biomass: a resource that can be valorised The biorefinery approach Transformation technologies Adapted from Kamm, B.; Kamm, M. (2004). Principles of Biorefineries. Appl. Microbiol. Biotechnol. (AMB), 64: 137-145. November 20th, 2012 Città della Scienza, Naples, Italy

9. Agro-industry wastes biorefinery: ongoing research at the Institute of Biomolecular Chemistry dry waste gross composition Carrot Tomato Lemon Fennel Data from INRAN (Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione) November 20th, 2012 Città della Scienza, Naples, Italy

10. II generation bioethanol Agro-industry wastes biorefinery: ongoing research at the Institute of Biomolecular Chemistry Polysaccharides recovery Antioxidants recovery November 20th, 2012 Città della Scienza, Naples, Italy

11. Alkali treatment at r.t. Polysaccharide Isolation of tomato’s wastes polysaccharides Tomato waste peels Chemical and physical characterization of isolated polysaccharides MW (Da)>1.000.000; High viscosity; High thermal resistance; [a]D25-0,189.0 Glc/Xil/Gal/GalN/GlcN/Fuc (1:0.9:0.5:0.4:0.2:tr) • Collaboration with Dr. Mario Malinconico and Dr. Barbara Immirzi (ICTP – CNR) Container and object construction Innovative material for containers or for green house applications November 20th, 2012 Città della Scienza, Naples, Italy

12. biotechnological applications Alkali treatment at r.t. Biodegradable film for solarization or mulching applications • Collaboration with Dr. Mario Malinconico and Dr. Barbara Immirzi (ICTP – CNR) Polysaccharide • biological properties Inhibition of NF-kB/DNA binding activity Incorrect regulation of NF-κB has been linked to cancer, inflammatory and autoimmune diseases, et cetera Isolation of tomato’s wastes polysaccharides Tomato waste peels November 20th, 2012 Città della Scienza, Naples, Italy

13. vegetable wastes polysaccharides extraction: ongoing research fennel residues lemon wastes (pomace) carrot residues • Maceration • Ultrasound assisted • Microwaves assisted alkali/acid extraction • Structural analysis (MW; monomer composition; spectroscopic analysis) • Thermogravimetrical analysis • Biotechnological applications • Biological properties POLYSACCHARIDES November 20th, 2012 Città della Scienza, Naples, Italy

14. II generation bioethanol Agro-industry wastes biorefinery: ongoing research at the Institute of Biomolecular Chemistry Polysaccharides recovery Antioxidants recovery November 20th, 2012 Città della Scienza, Naples, Italy

15. vegetable wastes polyphenols extraction: ongoing research tomato residues fennel residues lemon wastes (pomace) carrot residues • Maceration • Ultrasound assisted • Microwaves assisted MeOH; EtOH; MeOH/H2O; EtOH/H2O • Structural analysis • Antioxidant and radical scavenging activities • Biological activity assay • Biotechnological applications • Biological properties POLYPHENOLS November 20th, 2012 Città della Scienza, Naples, Italy

16. Vegetable wastes polyphenols extraction work in progress….. Project PON01_01966 Integrated agro-industrial chains with high energy efficiency for the development of eco-compatible processes of energy and biochemicals production from renewable sources and for the land valorisation (EnerbioChem) Naples University “Federico II” November 20th, 2012 Città della Scienza, Naples, Italy

17. II generation bioethanol Agro-industry wastes biorefinery: ongoing research at the Institute of Biomolecular Chemistry Polysaccharides recovery Antioxidants recovery November 20th, 2012 Città della Scienza, Naples, Italy

18. Agro-industry wastes: production of enzymes for second generation bioethanol from extremophiles Bilateral Project Italy-Turkey “Extremophiles for Next-Generation Biofuels ” Dr. Barbara Nicolaus Prof. Ebru Toksoy Oner • Vegetable wastes under investigation: potato peels, carrot, tomato, sugar beet pulp, starch and sugar beet molasses November 20th, 2012 Città della Scienza, Naples, Italy

19. II generation bioethanol Waste Biomass* Simultaneous Saccharification and Fermentation Starch degradation and sugar Fermentation by engineered yeasts: expression ofraw starch degrading enzymesin Saccharomyces spp glucose CH3CH2OH * Not in competition with food chain Agro-industry wastes: production of second generation bioethanol I generation bioethanol Starch (energy crops) liquefaction by thermostable amylases (starch cooking) T = 90-100°C dextrins saccharification by thermostable amylases T = 65°C glucose Yeasts Fermentation (Saccharomyces spp) T = 45°C CH3CH2OH November 20th, 2012 Città della Scienza, Naples, Italy

20. Agro-industry wastes as culture media for extremophiles’ fermentation Geobacillus thermoleovorans (strain PIZZOT) synthetic media tomato lemon Agro-industry wastes: production of enzymes for second generation bioethanol from extremophiles Amylolytic thermophile raw starch degrading amylase (starch digestion to glucose without cooking pre-treatment) wastes as culture media for amylase production Di Donato P., G. Fiorentino G., Anzelmo G., Tommonaro G., Nicolaus B., Poli A. 2011 “Waste Biomass Valorization” 2: 103-111 Optical microscopy observation of raw starch before (a) and after (b) digestion by G. thermoleovorans amylase November 20th, 2012 Città della Scienza, Naples, Italy

21. In conclusion….. by means of an integrated biorefinery approach waste biomass from agro-industry can be valorised to give • Fibers and polysaccharides for biodegradable polymers production • Biomolecules for several applications: antioxidants and bioactive molecules for pharmaceutical, cosmetic or food applications • Microorganisms and enzymes for second generation biofuels production November 20th, 2012 Città della Scienza, Naples, Italy

22. Thanks to…. Dott. Barbara Nicolaus Dott. Annarita Poli Dott. Licia Lama Dott. Giuseppina Tommonaro Prof. Ebru Toksoy Oner “If you want to go fast, go alone.  If you want to go far, go together” African proverb November 20th, 2012 Città della Scienza, Naples, Italy

23. Thanks for your attention! Dott. Paola Di Donato paola.didonato@icb.cnr.it paola.didonato@uniparthenope.it Grönlund-Nisunen - Wave of Matter 2012