Ateneo - UPO

Module I, Part "Food Biotechnology":
- Food biotechnology: introduction and significance
- Characteristics of microorganisms useful in biotechnology; the cell as a "molecular factory"
- Fermentation processes of interest to the food industry
- Beneficial microorganisms and the concept of "microbial starters"
- Prebiotics, probiotics, synbiotics, and postbiotics
- Fermented foods
- Industrial fermentation: classification and characteristics
- Fermentation plants: bioreactors and downstream processing; fermentation control parameters
- Fermentation reaction kinetics
- Enzyme and cell immobilization
- Biomass and Single-Cell Proteins (SCP)
- Examples of primary and secondary metabolites of food interest obtained via fermentation or recombinant technology: alcohols, sweeteners, neutral polysaccharides, organic acids, amino acids, flavorings, pigments
- Examples of bioconversion and bioremediation in the food sector (overview)
- Enzymes: significance, production, and applications
- Second-generation food biotechnology: recombinant DNA; MOGMs (Microorganisms Obtained via Genetic Modification) and GMOs
- Relevant national and international regulations (overview)
- Overview of analytical biotechnology
- Useful web resources in the field of biotechnology

Module I, Part "Quality Control (with laboratory)":
- Analytical methods for determining parameters listed on food product nutritional labels
- Official analytical methods for regulated food matrices such as: drinking water, wine, milk, olive oil, milled grain products, and pasta
- Instrumental analysis applied to food products, such as electrophoresis, GC-MS, HPLC-MS, and isotope ratio analysis - Laboratory exercises involving the analysis of wine, honey, fermented milk, and oil.

Slides used in the course, provided by the Teacher.
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.

Quality Control with laboratory:
ANALISI DEI PRODOTTI ALIMENTARI, Cabras, Tuberoso, Ed. Piccin, Padova, 2014.
For laboratory activities, handouts will be provided by the teacher.

This module must be considered an integral part of Course FA0443 – Quality and Safety in the Pharmaceutical and Food Biotechnology Sector.
The section on food biotechnology aims to provide students of the CTF (Chemistry and Pharmaceutical Technology) program with fundamental knowledge in this field (covering molecular biology basics; beneficial microorganisms and microbial starters; fermentation processes; bioproduction and recovery of bioproducts of interest to the food, nutraceutical, and pharmaceutical sectors; biotechnological production of flavors and additives; enzymes and their food applications; recombinant biotechnology, MOGMs, and GMOs; and an overview of relevant regulations). This knowledge will enable students to manage quality and safety assessments for biotechnological processes and products, while also providing the technical-scientific foundation for the biotechnological production of high-value-added molecules, ingredients, and materials for the food and pharmaceutical industries. The quality control section, which includes laboratory work, allows students to acquire basic knowledge of food analysis, enabling them to understand the significance of these analyses and their scope of application. Laboratory exercises aim to guide students and assess their ability to apply their knowledge to the analysis of simple and complex food matrices, including in the context of food fraud detection. Students must demonstrate the ability to correctly determine and interpret analytical data and to communicate the results using appropriate methods.

We recommend the acquisition of knowledge about the fields of Organic Chemistry, Biochemistry and Food chemistry, fundamental for the understanding of the lessons of this course. The basic knowledge in Quantitative Drug Analysis laboratory (Analisi dei Farmaci) is welcomed for laboratory experiences.

The teaching method involves in-person lectures complemented by the discussion of specific case studies. Specialized seminars may be organized in collaboration with faculty members and industry experts. For the quality control module, which includes a laboratory component, lectures are supplemented by laboratory activities. Students are required to perform analyses by following and interpreting the provided protocols, documenting the steps taken in a laboratory notebook, performing the necessary calculations to express results in the correct units of measurement, and completing analysis reports. Instructions will also be provided on how to draft a comprehensive report on the complex food product analyzed. Module materials will be published on the DIR platform and made available to students.

The module, comprising two parts, is designed as a unified program aimed at rounding out the food-related knowledge required by a Pharmaceutical Chemist to handle issues in the food and nutraceutical sectors—specifically regarding quality and safety control (analysis) and the biotechnological production of bioactive ingredients and food products. Laboratory activities involving the analysis of food matrices will take place in January. Students with disabilities, Specific Learning Disorders (SLD), or Special Educational Needs (SEN) may request specific services and tools by contacting the Student Career Development and Coordination Staff and consulting the dedicated page on the University website: https://uniupo.it/it/servizi/servizi-studenti-disabili-e-dsa. Once they have contacted the University staff, students with disabilities, SLD, or SEN may also contact the course instructor to discuss the specific arrangements for exams and other educational aspects.

Student assessment is based on an oral exam covering all course topics, aimed at evaluating students' knowledge and critical skills—with particular reference to the role of the production process, variability management, and the technological strategies employed in the development and industrial production of biopharmaceuticals and "bioactive" ingredients for foods and dietary supplements.
Students must prepare a short PowerPoint presentation (maximum 4-5 slides) introducing an original in-depth topic of their choice—one not covered during the course but linked to one or more course themes through an interdisciplinary approach—and present it to the examination board. The discussion with the instructors will start from the presented topic and extend to all disciplines involved, integrating scientific information from the fields of pharmaceutical technology, pharmaceutical-food science, and applied microbiology.
Please note that for the second part of Module I (Quality Control with Laboratory), assessment includes a mandatory mid-term test—a prerequisite for laboratory access—covering all topics addressed in the preparatory lectures (including safety regulations). This test is structured as a written exam consisting of 7 multiple-choice questions, 2 calculation exercises, and 6 open-ended questions; notably, two of the open-ended questions require students to select the most appropriate analytical methods from those studied to solve a specific analytical problem. The mid-term test grade contributes to the final module assessment; if the result is unsatisfactory, it will be supplemented by oral questions during the laboratory period or at the end of the course via a specific oral exam session. The oral component typically involves three questions: a description of instrumental analysis methods, a description of an analysis performed in the laboratory (with particular emphasis on safety regulations), and a question regarding the selection of analytical methods. Evaluation will focus on both knowledge of the topics covered and the ability to critically discuss the course's core themes from an interdisciplinary perspective. Laboratory activities will also be assessed based on submitted analysis reports, the accuracy of calculations, and the completeness of the laboratory notebook. Finally, the ability to present the analytical data obtained from the complex matrix in a written report will be evaluated.
The overall grade calculation will take into account the varying university credit values of the individual modules. A minimum level of knowledge (a passing grade) for each module is required to pass the exam and earn the associated credits.

Part I (Food Biotechnology, Prof. Arlorio): History and significance of food biotechnology. Molecules/macromolecules of biotechnological interest. Characteristics of microorganisms/prokaryotic and eukaryotic organisms. Transcription, translation, and gene regulation (overview). The cell as a "molecular factory." Food fermentations: alcoholic, homo- and hetero-lactic, malolactic, propionic, acetic, butyric and isobutyric, citric. Beneficial microorganisms and starter cultures in food production. Fungi of food interest (filamentous fungi, yeasts, cultivated and wild edible fungi). Lactic acid bacteria, bifidobacteria. Prebiotics, probiotics, and synbiotics. Bioactive milk peptides; prebiotic oligosaccharides. Industrial production of starter cultures and probiotics. Bacteriocins: characteristics and classification. Cyanobacteria and microalgae; production of biomass and pigments. Fermented foods: microbiological, biochemical, fermentative, and biotechnological aspects (milk and dairy products; alcoholic beverages; fermented vegetables; leavened baked goods; cocoa, coffee, and tea; salami; other fermented foods). Industrial fermentation: classification (continuous and batch). Fermentation plant design; fermenters and applications. Fermentation control parameters. Fermentation kinetics and yields (overview). Enzyme/cell immobilization and applications. Unit operations (recovery, separation, purification): downstream processing. Biomass, Single Cell Protein (SCP); red yeast rice, Pruteen, Quorn. Primary and secondary metabolites of interest to the food industry (via fermentation or recombinant methods): alcohols, amino acids, organic acids, neutral polysaccharides, other metabolites. Production of sweeteners (aspartame, glucose/maltose/fructose syrups, isomalt). Examples of bioconversions and bioremediation in the food sector (overview). Aromas: receptors and flavor perception. "Biotechnological" aromas: lactones, terpenes, vanillin. Production of cyclodextrins. Natural pigments; biotechnologically produced carotenoids. Enzymes: significance and production; enzyme classification. Enzymes as process markers. Examples of enzymatic applications in the food sector (glucose syrups, milk delactosation, meat tenderizing, extraction technologies, chitosan production, clarification and stabilization of juices and alcoholic beverages, accelerated cheese ripening, acrylamide reduction, other applications). Enzymes from extremophilic microorganisms. Second-generation food biotechnology: recombinant DNA. Genetically modified microorganisms (GMMs): r-chymosin and r-BGH. Plant biotechnology: manipulation and transformation techniques. Food GMOs: Flavr-Savr™ tomato, Bt™ maize, Roundup Ready™ soybean, Golden Rice, other GMOs. Biotechnology in the animal sector (cloning, salmonids, mammals, other applications). Relevant national and international regulations (overview). Biotechnology in food analysis (overview). Relevant web resources.
"Quality control and laboratory" section (Prof. Bordiga): significance and competencies. Water: heating methods, Marcusson method, and Karl Fischer method. Proteins: total nitrogen (Kjeldahl)—principle, equipment, safety regulations, correction factors. Protein extraction; SDS-PAGE electrophoresis; mono- and two-dimensional urea-PAGE; isoelectric focusing. Lipids: Soxhlet extraction; Gerber method for lipids in milk and dairy products. Characterization methods: refractive index, saponification value, iodine value. Quality assessment methods: acidity, peroxide value, and p-anisidine value. Olive oils: regulations, panel test, spectrophotometric analysis, esterified fatty acids (GC), and unsaponifiable matter analysis. Milk: specific gravity, cryoscopic index, dry matter, acidity, peroxidase and phosphatase activity, soluble whey proteins. Cheese analysis. Sugars: reducing sugars (Fehling's method), sucrose (polarimetry/Clerget method), refractometry, and Brix degrees. Fiber: composition; crude fiber, detergent fiber, and dietary fiber (total, soluble, and insoluble). Cereals and derivatives (flours, bread, and pasta): moisture, ash, gluten, acidity; bleaching agents and ascorbic acid. Wine: alcoholic strength, dry extract, total/fixed/volatile acidity, proline, polyphenols; NMR and mass spectrometry methods for assessing sugar origin. Drinking water: regulations for drinking water and mineral water. Determination of ammonia (Nessler and phenol-hypochlorite methods), nitrites (Griess method), and nitrates. Oxidizability value, BOD, COD. Hardness, chlorides, chlorine, sulfides, iron, chromium. Analysis of additives: determination of benzoic acid in sugary drinks. Enzymatic analysis: significance, sample preparation, methods, and operational protocols. Tests to evaluate the antioxidant activity of foods. Instrumental analysis: applications of HPLC techniques with SPE extraction and purification; applications of GC techniques, aroma extraction via SPME. Atomic absorption: principles and applications.
LABORATORY: Basic analysis of wine, fermented milk, honey, and olive oil. Enzymatic kits for the determination of lactose, sucrose, fructose, and glucose. Food analysis: moisture, pH, ash, protein, and acidity. Lipid fraction: acidity and peroxides. Polarimetry for honey. Protein electrophoresis: extraction, SDS-PAGE, and image analysis. Polyphenols: Folin and DMAC methods.
Regarding the gender dimension, teaching approaches will be employed to ensure equal benefits for women and men across all actions and activities and to eliminate gender inequalities, including through the use of practical examples and selected case studies.

"Food Biotechnology" section: students must demonstrate the acquisition of basic, specific, and advanced knowledge regarding the application of first- and second-generation biotechnologies to food, nutraceutical, and pharmaceutical production models. They must show an understanding of key biotechnology topics within the nutraceutical-food sector and the resulting products, while demonstrating a high level of technical-scientific communication skills and appropriate terminology. Students must demonstrate knowledge of biotechnology-related production techniques and the associated regulatory framework, as well as the ability to select the most suitable technique for managing applied work scenarios (at both laboratory and industrial scales—the latter as part of an interdisciplinary technical-scientific team). Specific expertise will be gained regarding genetically modified microorganisms/organisms and their derived products—suitable for use in both the food and pharmaceutical sectors—as well as the underlying European regulatory landscape. Finally, students must demonstrate the ability to plan a biotechnology project applied to the food and nutraceutical sectors.

"Quality Control (with laboratory)" section: acquisition of basic and specific knowledge regarding analytical methods applicable to food analysis; understanding of the theoretical principles of instrumental analytical techniques and their specific applications in the food sector (including fraud detection); and acquisition of appropriate scientific terminology in this context. Regarding the practical application of acquired knowledge, students must demonstrate the ability to identify the analytical methods best suited to solving a specific problem and to understand the information that a given analytical method can provide. As part of the laboratory activities, students must be able to correctly understand and apply the provided analytical protocols, obtain the relevant analytical data, perform the necessary calculations to express the data in the correct units of measurement, and interpret the results in light of any regulations specific to the matrix under examination, exercising independent judgment in evaluating the experimental data. They must also demonstrate the ability to present the analytical results in a written report that is understandable even to non-specialists.