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Exploiting Omics Sciences for Food Safety, Quality and Nutrition” “Exploiting omics sciences for food safety, quality and nutrition” Gianfranco Mamone DiSBA annual conference - «POST EXPO: INSEGNAMENTI DELL’ESPOSIZIONE UNIVERSALE E SFIDE FUTURE SUL TEMA DELLA RICERCA NEL SETTORE BIO-AGROALIMENTARE» MILANO 24-25 Novembre 2015 The Institute of Food Sciences (ISA) research activities are organized in three main science programs: A) Food; B) Health; and C) Omics. multidisciplinary scientific approach to Food Biotechnology. relationship between nutrition, health and diseases. food quality and safety and relationship between food composition and human health. What are Omics Technologies and what types of information do they generate? Trascriptomics/ Genomics/genome Proteomics/proteome Transcriptome Metabolomics/metabolome (gene) (protein) (mRNA) (metabolite) Interactomics Lipidomics Peptidomics Nutrigenomics (molecular (lipids) (peptides) (gene and health ) interactio) Food chain production Transformation Final product Living source Raw material end-product FOOD + OMICS = FOODOMICS The term FoodOmics is an array of integrated analytical platforms that studies the Food and Nutrition domains (Cifuentes et al., 2009) omics sciences for food safety, quality and nutrition EXPO HOT TOPICS Due to globalization, food scenario is rapidly changing: There is an urgent demand for advance analytical techniques addressing food safety quality and nutrition : • Food safety: development of rapid and fast devices to ensure food safety in all aspects of the food production chain, dealing with the farm-to-fork continuum • Food quality Characterization of macro and micronutrients to provide a deeper knowledge of food composition and quality (texture, flavour, odour, and shelf life) • Food nutrition: Improvement of our understanding of the link between food and health. OMICS for FOOD ANALYSIS Discovery driven Hypothesis driven Proteomic/metabolomic Biosensoristic analysis by devices Mass spectrometry (target approach) (untargeted approaches) Bioinformatic tools Research activities at ISA-CNR Food quality and authenticity Identification of species – specific markers of fish by direct MALDI TOF MS * Dentex dentex 11519.86 100 Dentex fish (Mediterranean) * 11567.25 5761.00 5787.47 i.s. 6181.00i.s. i.s. 12361.10 100 6181.16 0 Pagrus pagrus 3000 5400 7800 10200 12600 15000 Mass (m/z) Pangasius (catfish) 5791.01 * Mekong river in Vietnam 11581.50 i.s. 5745.22 * 11488.02 0 3000 5400 7800 10200 12600 15000 Mass (m/z) Fraud detection in fresh and processed fish products Omic sciences for food quality and nutrition Novel Food and Celiac Disease Celiac Disease Autoimmune Pathogenesis Genetic predisposition gluten celiac disease + = Innovative strategies to detoxify wheat gluten Consiglio Nazionale delle Ricerche Istituto di Scienze dell’Alimentazione PROTEOMICS contributes to the development of novel gluten free foods Structural analysis of gliadin peptides Structural analysis of modified gliadin peptides Definition of the primary structure of Definition of structural modification induced by tissue immunodominnat epitopes in gliadins transglutaminase on immunodominant gliadin peptides PROTEOMICS Novel analytical approach aimed at studying complex mixture of proteins Methods Tandem mass spectrometric analyses of gliadin peptides Analysis of detoxified flour Structural analysis of modified gliadins extracted from enzymatic detoxified flour in order to assess the efficiency of the masking treatment A multidisciplinary study integrating immunobiology, proteomics, protein chemistry and food technology has been successfully performed with the aim to develop novel gluten free foods that will be soon on the market Gluten free flour Consiglio Nazionale delle Ricerche Istituto di Scienze dell’Alimentazione Results: Novel Food e Celiac disease Detoxified wheat flour baked into bread Gluten content less then 20 ppm detox control 5.8 ppm food composition and nutrition Determinatione of bioaccessibility food derived biomolecules In vitro model of intestinal uptake Gastric Pepsin phase Trypsin Duodenal Chymotrypsin Elastase MS and MSMS phase Carboxypeptidase BBM Intestinal Endo/exo- phase peptidases in vitro m/z digestion model Biomolecules identification food quality and nutrition BIOACTIVE PEPTIDES FROM MILK PROTEINS Defensive Antimicrobials Nervous systems Inmunomodulators Opioids Cardiovascular system Antihipertensives Antithrombotics GI system Casein-phosphopetides Opioids food safety As result of the rising globalization levels, food suppliers producers, manufacturers and regulatory agencies are all facing greater pressure to test more food products for more contaminants and their presence at lower level with great accuracy and in less time. The need for reliable methods allowing accurate structural identification and dosimetry of the offending food compounds has prompted researchers to develop new methods for the unambiguous characterization of contaminates Biosensor for food safety The development of an advanced optical biosensor for a simple and rapid determination of contaminants in foods to ensure the safety of raw materials and end-products. Biosensor for food safety Advantages of an optical biosensor in Food Safety • Fast response • Use of non-trained personnel for its use • Low cost and portable biosensor • High selectivity • High sensitivity • Extensive extraction and sample clean-up are not needed What is a biosensor? Biological Element Signal Transducer Aptamers Optical Enzyme Electro-Chemistry Protein Calorimetric Antibody Biosensors can be grouped according to their biological element or their transducer element. Conjugation PAT to protein carrier and antibody production EDC/NHS + BSA PAT Derivate BSA-PAT Injection of BSA-PAT Ab anti PAT IR Biosensor for food safety Patulin (PAT) is a strong mycotoxin produced by different fungal species, capabel of growing on a variety of food, Lowering of immune defences PAT Adverse effects in the stomach and intestine Neurotoxic Genotoxic in mammalian cells Induces mutations in bacteria and chromosomal aberrationsin yeast Damage to the DNA Usually apples and pears and baby food are contamined by patulin IR Biosensor for food safety Hormones and antibiotics in fed Penicillin G Estradiol Hormones and antibiotics are improperly used in livestock. Due to their high risk for human health, the EU has strictly forbidden the administration to food producing animals Less profit More profit Biosensor for food safety We developed fluorescence method to detect toxins, antibiotics and hormones in foods to ensure the quality and the safety of raw materials. These methods allow to determine an amount of patulin, penicillin G and estradiol, in real matrices, much lower than conventional techniques and much lower than the EU limit. EU MRL* Fluorescence (Maximum Residue Polarization Limits ) Limit Patulin 50 ug/L 0.06 ug/L Penicillin G 12 nM 0.4 pM Estradiol 20 pmol 0.5 pmol *MRLs established from the European Union (EU) Regulation 508/1999 Forthcoming scenarios Omic approaches are currently irreplaceable for deciphering the molecular code encrypted in a food, in order to tell us the history of foodstuff from the raw material to consumption. Omic sciences http://www.isa.cnr.it/web Thank you for your attention! ISA-CNR participation to EXPO Consiglio Nazionale delle Ricerche Istituto di Scienze dell’Alimentazione STRATEGY Bioactive Biological molecules activity Food Objective 1 Objective 2 Nutraceutics and Cosmeceutics (prototype) Marketing health and well-being of the population Objective 3 Objective 5 Objective 4 4. The search for epitopes In vitro model of the digestion/uptake of food epitopes mouth / stomach duodenum jejunum / ileum legume Legume protein receptors absorption Caco-2 cells maintain many morphological and functional traits of the mature enterocytes of the small intestine: polarization, microvillous structure, carrier- mediated transport systems, functional tight junctions and expression of BBM peptidases Peptidomic analysis transwell insert Apical and basolateral solutions analyzed by MALDI-TOF MS and µ- tight junction apical HPLC-ESI-Q-TOF cell monolayer MS/MS chamber porous membrane basolateral Activity assessment chamber Sustainability of food chain Omic sciences also contribute to respond a further urgent point emerged from EXPO: Sustainability of food chain For instance: recycles of agrifood waste Sustainability of the Food chain: intesa come rispetto dell’ambiente. Nel caso dell’alimentazione la riduzione degli sperchi riutilizzo degli scarti alimentari. Ottimizzazione delle risorse energetiche. Le scienze omiche possono dare valorizzazione deglòi scarti di produzione Further EXPO ‘HOT TOPICS’ in AGROFOOD: Sustainability of the food chain In the perspective of the valorization of biomolecules from agrofood waste At ISA CNR have been developed extraction strategies of bioactive peptides from waste cheese whey (million of tons remnant yearly). Hundreds of milk-derived peptides from Mozzarella di Bufala whey have been characterized by MS-based peptidomics Many of these peptides exhibit anti-hypertensive, anticariogenic, immunomodulating, antioxidant etc. properties Bioactive molecules have been extracted, isolated and characterized for their biological activities from many (both animal and plant origin) food waste sources (BE & SAVE Project, PON Smart Cities) Advantages of Protein as MREs in biosensors - Specificity - Reversibility - Wide range of affinity constants - Low-cost sensors - Water soluble - Wide range of analytes - Easy to mutate and express in hosts - Enzymes
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