Ateneo - UPO

Part I:
- Food biotechnology: introduction
- Characteristics of microorganisms useful in biotechnology: the cell as “ molecular factory”
- Fermentations in food area
- Virtuous microorganisms and “microbial starters”
- Prebiotic, probiotics and synbiotics
- Fermented foods
- Industrial microbiology: classificationd of fermentations and characteristics
- Bioreactors and downstream processes
- Fermentation’s kinetics
- Enzymes and cells immobilization
- Biomasses, Single Cell Proteins (SCP)
- Examples of primary and secondary metabolites: alcohols, sweteners, gums, organic acids, aminoacids, flavorings pigments.
- Examples of bio-conversions and bio-remediations (overview)
- Enzymes: classification, production and applications.
- Recombinant technology in food area: MGMOs e GMOs
- Regulatory framework (overview)
- Analytical biotechnology: overview
- Web resources

Part II
- Methods of analysis useful for the determinations of parameters present on nutrition labels (proteins, saturated and unsaturated lipids, simple and complex carbohydrates, fiber, minerals).
- Official methods of analysis for some foods such as: drinking water, wine, milk, olive oil, flour, pasta.
- Instrumental analyses applied to foods, such as electrophoresis and capillary electrophoresis, GCxGC-MS, HPLC-MS, MALDI-TOF, atomic spectroscopy, PCR and Real-time PCR.
- Laboratory activities focused on the analysis of drinking water, olive oil, wine, milk candies and chocolate.
- Laboratory activities relate to a complex food (cheese), defining its nutritional label, its characterization and evaluating the ripening.

Slides used in the course, provided by the Teachers
Food Biotechnology
“BIOTECNOLOGIE ALIMENTARI”, Gigliotti-Verga; Piccin Editore
Industrial microbiology and fermentations
CHIMICA DELLE FERMENTAZIONI E MICROBIOLOGIA INDUSTRIALE, Marzona, II Edizione. PICCIN Editore.
Applied Biotechnologies:
BIOTECNOLOGIE DI BASE, Colin Ratledge, Bjørn Kristiansen; Zanichelli Editore
Deep insights in biotechnology:
BIOTECNOLOGIA MOLECOLARE, Glick, Pasternak; Zanichelli Editore.
Food Analysis:
ANALISI DEI PRODOTTI ALIMENTARI, Cabras, Tuberoso, Ed. Piccin, Padova, 2014
Lab. activities:
Notes and protocols from the teacher

The knowledge gained from the course will be useful to the student in Chemistry and Pharmaceutical Technology to complete the knowledge in “food area” .
The first part, related to food biotechnologies, provides the students with the basic knowledge in the field of biotechnology in the food industry (molecular biology bases; virtuous microorganisms and microbial starter, fermentation, bio-production and recovery of bio-products used in food, nutraceutical and pharmaceutical industries; production of flavourings and additives through biotechnology, food enzymes and applications; recombinant biotechnology, GMO and GMO; notes on the regulatory field).
In the part related to the Food Analyses the student will acquire the basic knowledge in food analysis, in order to understand the meaning of each analysis and its scope. The laboratory exercises aim to prove students in food analysis and to verify their ability to apply the knowledge related to analysis of simple and complex food matrices. The student must demonstrate the ability to correctly obtain and discuss the analytical data and to use the appropriate method of communication of the results obtained

We recommend the acquisition of knowledge about the courses of Organic Chemistry, Biochemistry and Food chemistry, technology and contaminants, fundamental for the understanding of the lessons of this course. The basic knowledge in Quantitative Drug Analysis laboratory (Analisi dei Farmaci I and II) is welcomed for laboratory experiences. The compulsory prerequisites for the exams anr the corses of Applied Biochemisty and Food chemistry, technology and contaminants.

The teaching method used provides lessons in presence, completed by the discussion of particular case studies. Specific seminars may be organized in collaboration with teachers and industry experts.
For the Analysis module, the face-to-face lessons are accompanied by calculation exercises, related to laboratory analysis analysis methods. As for laboratory activities, students must carry out the analyzes following and interpreting the protocols provided, reporting the steps carried out in the laboratory notebook, carrying out the appropriate calculation operations for the expression of the result in the correct unit of measurement and filling in analysis reports. The method of drafting a complete report on the complex food analyzed will also be indicated.
The material used will be published on DIR and made available to students.

The course, even if structured in two parts, must to be considered a whole course finalized to complete the knowledge and the skills in "food area" of the chemist expert in pharmaceutical sciences, both considering the analysis of foods (quality and safety assessment) as well as the production of new bioactive ingredients/food by mean of biotechnology methodologies.
The lab activities are scheduled in two parts, the first including the analyses of simple foods (wine matrices, milk, oil, water, candy) will take place in January, the second related to the characterization of a complex food (cheese) will take place in March.

For the Food Biotechnology module, the assessment of learning is based on an oral exam that focuses on the topics of the course. The exam focuses on a question that concerns the first part of the course (introduction, fermentations, starter and virtuous microorganisms, probiotics), one on the second part of the course (industrial fermentations and bioproducts) and one on the third part of the course (GMOs and biotechnological products ).
For the Food Analysis part the assessment of the learning starts as an evaluation in itinere as written test at the end of theory lessons, mandatory for the laboratory admission, related to all topics described in the preparatory lessons (including lab safety rules). This evaluation consists in a written test with 7 multiple choice questions, 2 calculation exercises and 6 open questions, between these two questions in particular require that the student selects the most suitable analyses in order to solve a specific analytical problem. The evaluation of the in itinere test is part of the final evaluation of the course and, in case of unsatisfactory results, will be integrated with oral questions during the period in the laboratory or at the end of the course as a specific exam. Oral questions are typically 3: one to provide a description of an instrumental analytical method, one to describe an analysis performed in the laboratory with specific focus on safety rules and a question on the selection of methods of analysis. Related to the activities carried out in the laboratory, the evaluation includes the preparation of analysis reports, the correctness of the calculations and completeness of the laboratory notebook. Finally, it will evaluate the presentation skills of analytical data obtained on the complex food in a written report form.

Part I (Food Biotechnology, Prof. Arlorio)
History and meaning of food biotechnology. Molecules/macromolecules of biotechnological interest. Features of microorganisms (prokaryotes and eukaryotes), characteristics. Transcription, translation, gene expression and regulation. Cell as "molecular factory." Food fermentations: alcoholic, homo- and hetero-lactic malo-lactic, propionic, acetic, butyric, and isobutyric, citric fermentation. Virtuous and starter microorganisms in food productions. Fungi of food interest (filamentous fungi, yeasts, edible mushrooms). The lactic acid bacteria. Bifidobacteria. Prebiotics, probiotics and synbiotics. Bioactive peptides of milk; prebiotic oligosaccharides.
Industrial production of starters cultures and probiotic microorganisms. Bacteriocins: features and classification. Cyanobacteria and microalgae, biomass and pigments production. Fermented foods: microbiological, biochemical, biotechnological and fermentation features (milk/dairy products, alcoholics, fermented vegetables; leavened bakery products, cocoa, coffee and tea, sausages, other fermented foods).
Industrial microbiology: classification of fermentations (continuous, discontinuous). Bio-reactors. Fermentation parameters. Kinetics, yields of fermentation. Immobilized enzymes/cells. Application of immobilized systems. Operations Units for recovery, separation and purification of fermented products (downstream processing). Biomass, Single Cell Protein (SCP). Red yeast rice, Pruteen, Quorn. Primary and secondary metabolites obtained by fermentation: alcohols, amino acids, organic acids, neutral polysaccharides, other bioproducts. Production of sweeteners (aspartame, glucose syrup/ maltose/fructose and isomalt). Examples of bio-conversion and bio-remediation in food.
Flavourings. Receptors and taste/aroma perception. Examples of flavourings from biotechnology: lactones, terpenes, vanillin. Cyclodextrins production. Pigments: carotenoids productions.
Enzymes: meaning and production, classification. Enzymes as markers of food processes. Enzyme applications in food technology (production of glucose syrups; milk delactosation, meat tenderizing, extraction technologies, production of chitosan, clarification and stabilization of juices and alcoholic beverages, accelerated ripening of cheese, enzymatic reduction of acrylamide, other applications). Enzymes from extremophiles microorganisms.
Food Biotechnology of the second generation: recombinant DNA. GMMO: r-Chym and r-BGH. Plant biotechnology: GMO foods: Flavr-SavrTM tomato, Bt corn, Soja Round-up Ready, Golden Rice, other GMOs. Food biotechnology in animal area (cloning, salmonids, mammals, other experimental modifications). National and international regulatory framework. Outline on Analytical Biotechnology in food area.
Web resources useful in food biotechnology.

Part II (Food Analysis and Quality Control, Prof. Coisson)
The food analysis and quality control: importance and principal fields.
Moisture: oven drying, Marcusson and Karl Fischer methods.
Proteins: total nitrogen (Kjeldahl), principles, instruments, safety procedures, correction factors. Extraction, SDS-PAGE and native-PAGE electrophoresis, mono and two-dimensional; isoelectrofocusing.
Lipids: extraction in Soxhlet apparatus, Gerber methods for dairy products. Methods for characterization: refractive index, saponification number, iodine number. Quality evaluation: acidity, peroxide number, p-anisidine number.
Olive oils: European regulations. Panel test. Spectrophotometric analysis. FAME (Fatty Acid Methyl Esters) and unsaponifiable fraction analyses (sterols, superior alcohols, erythrodiol/uvaol).
Milk: density, cryoscopic index, dry matter, acidity, peroxidase and phosphatase tests, soluble not-denatured whey proteins. Cheese analysis.
Sugars: reducing sugars (Fehling), sucrose with polarimetric method (Clerget). Refractive index and Brix degrees.
Cereals and derived products (flours, bread and pasta): moisture, ashes, gluten, acidity, bleaching contaminants, ascorbic acid. Methods for identification of common wheat and eggs in pasta.
Wine: ethanol percentage, dry matter, total, fixed and volatile acidity, proline, polyphenols, NMR and mass spectrometry methods for evaluation of sugars origin.
Water: legislation for drinking water and mineral water. Ammoniac (Nessler and phenol-ipochlorite), nitrites (Griess), nitrates, oxidative Kubel number, BOD, COD., hardness, chlorides, chlorine, sulfides, iron and chrome
Additives: quantification of benzoic acid in non-alcoholic beverages.
Enzymatic analysis: aims, sample preparation, examples of methods and protocols.
Total antioxidant activity methods.
DNA analysis: PCR and PCR-related techniques (AFLP, RFLP, RAPD, Real time). HPLC applications and previous extraction and purification by SPE; GC-techniques applications, aroma extraction by SPME.
Metal analysis: Atomic absorption spectrometry, theory and applications.
Practical experiences
Analyses of drinking water, wine, milk, olive oil. Benzoic acid quantification in non-alcoholic beverages, sucrose in chocolate and candies.
Characterization of a complex food (cheese): moisture, ashes, proteins, lipids (Soxhlet and Van Gulik); titrimetric acidity and lactic acids (enzymatic). Fat fraction: acidity, peroxide number, oxidation index, FAME composition by GC. Proteins electrophoresis: extraction, SDS-PAGE and urea-PAGE, image analysis. Quantification of biogenic amines by HPLC. DNA analysis: extraction, clean-up and amplification

Analysis part: Acquisition of basic and specific knowledge on useful methods in food analysis, knowledge of the theoretical principles of analytical instrumental methods and specific applications in the food sector; acquisition of an appropriate scientific language on the matter. In relation to the ability to apply the acquired knowledge, the student will demonstrate that he / she knows how to identify the methods of analysis more suitable to the resolution of a specific problem and the information that the method of analysis is able to provide.
As part of the laboratory activities, the student must be able to understand and correctly apply the provided analysis protocols, obtain the analytical data, solve the calculations necessary in order to obtain the data in the correct measurement unit, interpret the data obtained following any specific legal rules, conducting with independent judgment the evaluation of the experimental data obtained.
Ability to expose the analytical data obtained in a written report, suitable for the understanding also of non-professionals.
Food Biotechnology module: the student must demonstrate the acquisition of basic (as well as specific and advanced) knowledge on the application of “first” and “second” generation biotechnologies to food, nutraceutical and pharmaceutical products; knowledge about the main themes of biotechnology applied in nutraceutical-food field, showing to have understood a technical language and good technical-scientific exposure capacity. The student will demonstrate learning knowledge about the production techniques related to biotechnology, the related regulatory framework and to master the possible choice of the most useful technique for managing an applied work situation (at laboratory and industry level, in this last case as a part of an interdisciplinary scientific technical team).
Particular expertise will be acquired in the field of genetically modified microorganisms / organisms, as well as in the field of the products derived from them which can be used both in the food and pharmaceutical areas, as well as the knowledge about the European regulatory status correlated to GMO's. Finally, the student must demonstrate the ability to plan a biotechnological project applied to the food and nutraceutical sector.