From Nature towards Nature: molecules, mechanisms and processes for sustainability, energy, food quality and health Istituto di Chimica Biomolecolare UOS di CATANIA - Via Paolo Gaifami, 18 Phenomics, the concepts behind: monitoring the expression of selected secondary metabolites for chemotaxonomy, traceability, quality and safety studies1 Sustainable processes for biodiesel production (PCT/IB2014/058783) 1L. Siracusa, G.Ruberto. Plant polyphenol profiles as a tool for traceability and valuable support to biodiversity. Chapter II (15-33) In Polyphenols in Plants: Isolation, Purification and Extract Preparation , 1st Edition (April 2014) . Elsevier Books, ISBN 9780123979346. Biodiesel is a non-toxic and biodegradable alternative fuel obtained from renewable sources. There are several key challenges arising with biodiesel optimization processes which must still be addressed efficiently. The main problem is the inevitable production of glycerol as the co-product (about 10% of produced biodiesel), which must be removed and whose formation impacts on the expense of the entire process. The use of glycerol-derivatives that may be exploited as biodiesel additives is being explored. saffron (C. sativus) L.Siracusa, F.Gresta, G.Avola, E.Albertini, L.Raggi, G.Marconi, G.Ruberto. Agronomic, chemical and genetic variability of saffron of different origin by LC-UV-VIS-DAD and AFLP analyses. Genetic Resources Crop Evolution 2013, 60, 711-721. Soybean oil: 1.0 g TBME: 3.2 mL (Oil/TBME 1:10) Catalyst: 1.0% w/w One-pot microwave assisted catalytic transformation of vegetable oil into glycerol-free Biodiesel R O “long storage” tomato (L. esculentum) MW: power/time ))) O O O R + CH OH 3 O O O O OH O R O O OH O O O OH O O acid-catalyst O O 70:30 O O O MW: power/time oregano (O.vulgare) ))) O MW: power/time O ))) + + CH3OH 3 acid-catalyst T.Tuttolomondo,S. La Bella,M.Licata,G.Virga,C.Leto,A.Saija,D.Trombetta, M.Cristani, A.Speciale, E.M. Napoli, L.Siracusa, A.Pasquale, G.Curcuruto, G.Ruberto. Biomolecular Characterization of Wild Sicilian Oregano –Phytochemical Screening of Essential oils and Extracts And Evaluation of Their Antioxidant Activities. Chemistry & Biodiversity 2013,10,411433. Laura Siracusa, Cristina Patanè, Giovanni Avola, Giuseppe Ruberto. Polyphenols as chemotaxonomic markers in Italian “long storage” tomato genotypes. J. Agric. Food Chem. 2012, 60, 309-314. Isolation and characterization of new natural products and phytocomplexes endowed with peculiar and interesting biological activities A pool of polyphenols, namely dihydrostilbens and flavonoids, have been isolated from the leaves of Sicilian liquorice (Glycyrrhiza glabra L.). The lipid extract containing the aforesaid components showed antioxidant and anti-inflammatory properties, which have been associated to the polyphenol compounds. In particular, both the extracts and the pure compounds showed a selective anti-inflammatory activity against COX-2 rather than COX-1, indicating these molecules as potential new anti-inflammatory components with limited side effects. The figure shows the docking studies of one of the most active dihydrostilbenes isolated from the leaves of liquorice with COX-1 (A) and COX-2 (B) O O R O The microwave assisted catalytic decomposition of TBME gives methanol and isobutene which participate to the transesterification of triglycerides and the etherification of the obtained glycerol. The three independent reactions occur simultaneously and efficiently in presence of the same catalyst and under the same reaction conditions. Complete conversion into the corresponding methyl esters and a mixture of mono- and di- tert-butyl glycerol ether in a ratio 30:70. As desired, negligible traces of glycerol were detected. References: 1. M. L. Testa, V. La Parola, L. F. Liotta, A. M. Venezia; J. Mol. Catal. A-Chem., 2013, 367, 69 2. C. Drago, L. F. Liotta, V. La Parola, M. L. Testa and G. Nicolosi Fuel, 2013, 113, 707-711 Citrus genetic improvement One of the most used procedures is the production of new varieties by crossing the best extant species. In collaboration with the CNR Institute of Bioscience and Bioresources (Palermo) and CRA-ACM of Acireale we have analysed the secondary metabolic profiles (polyphenols from the juices and the peel essential oils) of the new fruits and those of the parents. HPLC profiles of the flavonoid content of a series of new hybrids References: Biondi, D.M.; Rocco, C.; Ruberto, G. New dihydrostilbene derivatives from the leaves of Glycyrrhiza glabra and evaluation of their antioxidant activity. J. Nat. Prod., 2003, 66, 477-480. Biondi, D.M.; Rocco, C.; Ruberto, G. Dihydrostilbene derivatives from Glycyrrhiza glabra L. leaves. J. Nat. Prod. 2005, 68, 1099-1102. Siracusa, L.; Saija, A.; Cristani, M.; Cimino, F.; D’Arrigo, M.; Trombetta, D.; Rao, F.; Ruberto, G. Phytocomplexes from liquorice (Glycyrrhiza glabra L.) leaves: chemical characterization and evaluation of their antioxidant, anti-genotoxic and anti-inflammatory activity. Fitoterapia 2011, 82, 546-556. References: Biocatalytic procedures to retrieve high added value molecules from fish wastes 1. Ruberto, G.; Rapisarda, P. Essential oils of new pigmented Citrus hybrids: Citrus sinensis L. Osbeck x C. clementina Hort ex Tanaka. J. Food Sci. 2002, 67, 2778-2780. 2. Tusa, N.; Abbate, L.; Renda, A.; Ruberto, G. Polyphenols distribution in juices from Citrus allotetraploid somatic hybrids and their sexual hybrids J. Agric. Food Chem. 2007, 55, 9089-9094. 3. Fabroni, S.; Ruberto, G.; Rapisarda, P. Essential oil profiles of new Citrus hybrids, a tool for genetic citrus improvement. J. Essent. Oil Res. 2012, 24(2), 159-169. 4. Abbate, L.; Tusa, N.; Fatta Del Bosco, S.; Strano, T.; Renda, A.; Ruberto, G. Genetic improvement of Citrus fruits: New somatic hybrids from Citrus sinensis (L.) Osb. and Citrus limon (L.) Burm. F. Food Res. Int. 2012, 48, 284-290. 5. Fatta Del Bosco, S.; Abbate, L.; Tusa, N.; Strano, T. ; Renda, A.; Ruberto, G. Genetic improvement of Citrus fruits: The essential oil profiles in a Citrus limon backcross progeny derived from somatic hybridization. Food Res. Int. 2013, 50, 344-350. COOEt Mechanism of antioxidant activity Omega-3 ALA Sequential Proton Loss Electron Transfer CH3 EPA H 3C CH3 COOEt COOEt DHA We discovered that oxidant radicals (e.g. NO2, CO3•-, dpph, etc.) react with phenols in ionizing solvents through electron-transfer from the phenoxide anions. This mechanism was baptized, “Sequential Proton Loss Electron Transfer“ (SPLET). Quenching of radicals via SPLET can be very active in those cellular compartments at higher pHs. The discovery of this mechanism was achieved by studying the reactivity of cinnamic acids (ACs) with dpph in methanol. We observed that the bimolecular rate constants decreased as the concentration of ACs was increased (see Fig. 2. This is because ionization of the carboxylic acid group represses the ionization of the phenolic OH and reduces, therefore, the concentration of phenoxide anions in solution (see Fig. 3). High added value molecules from Olive Mill Waste Water Green routes SPLET Fig. 1 Fig. 2 Fig. 3 References 1. Litwinienko, G.; Ingold, K. U. J. Org. Chem. 2003, 68, 3433 – 3438 Abnormal Solvent Effects on Hydrogen Atom Abstractions. 1. The Reactions of Phenols with 2,2-Diphenyl-1-picrylhydrazyl (dpph•) in Alcohols 2. Foti, M. C.; Daquino, C.; Geraci, C. J. Org. Chem. 2004, 69, 2309 – 2314 Electron-Transfer Reaction of Cinnamic Acids and Their Methyl Esters with the DPPH Radical in Alcoholic Solutions 3. Litwinienko, G.; Ingold, K. U. J. Org. Chem. 2004, 69, 5888 – 5896 Abnormal Solvent Effects on Hydrogen Atom Abstraction. Resolution of the Curcumin Antioxidant Controversy. The Role of Sequential Proton Loss Electron Transfer. References: PO FESR 2007/2013 “Valorizzazione di Biomasse Residuali per Applicazioni nel Settore Farmaceutico e Cosmetico” PON02 “ Valorizzazione Biomolecolare ed Energetica di biomasse residuali del settore Agroindustriale ed Ittico “ (BIO4BIO) Essential oils as therapeutic option in oncology (Patent N° MI2014A000075) Functional food: fruit and vegetable juice blends for health Green Drink Orange Drink 1 Orange Drink 2 celery 10% strawberry 40% golden apple 20% pomegranate 30% kiwi 70% golden apple 30% pomegranate 40% white grape 40% strawberry 50% carrot 10% pomegranate 30% celery 10% golden apple 20% pomegranate 30% white grape 40% black grape 40% carrot 15% pomegranate 30% kiwi 15% golden apple 30% Red Drink 1 Red Drink 2 Red Drink 3 For each «drink» and for single components of fruit and vegetable, tests were carried out to define the antioxidant profile (ABTS, DPPH, FRAP, RNS, and total phenols), the hypoglycemic activity, and the content of biophenols belonging to classes of phenolic acids, ellagitannins, flavonoids, and anthocyanins. Among the 28 juice blends, six have higher values of antioxidant activity compared to that obtained by adding the contribution of individual components in the mix (synergistic effect). This is in agreement with the sensorial analysis. Part of the benchtop prototype used for making mixed “drinks”. This prototype has the peculiarity of not providing any preliminary processing of natural matrix used (such as peeling or removing large seeds), which makes it suitable for use in “Horeca” (Hotel-Restaurant-Café) facilities. References: POR Regione Sicilia Obiettivo Operativo 4.1.1 Linea di Intervento 220.127.116.11 “Realizzazione di un prototipo di macchina automatica da banco di nuova concezione per la produzione istantanea di miscele di centrifugati di frutta e ortaggi freschi ad elevato potere salutistico” MTT Lung cancer Salvia officinalis GC-FID-MS Several essential oils are under study for their potential use as anticancer agents1 alone or in combination with conventional drugs. Moreover their antibiofilm2 activity is under evaluation. References: 1. Privitera G, Napoli E, Luca T, Ruberto G and Castorina S. In vitro Anti-Proliferative Effect of Salvia officinalis Essential Oil and its Three Main Components on Human Lung Cancer Cells . American Journal of Phytomedicine and Clinical Therapeutics , 2014 in press. 2. Schillaci D, Napoli, Cusimano M, Vitale M, and Ruberto G. Origanum vulgare subsp. Hirtum Essential Oil Prevented Biofilm Formation and Showed Antibacterial Activity against Planktonic and Sessile Bacterial Cells. J. Food Protection, 76 (10), 1747-1752, 2013.