Applied molecular biotechnologies

A.Y. 2020/2021
11
Max ECTS
112
Overall hours
SSD
BIO/10
Language
Italian
Learning objectives
The aim of the course is:
-to teach the operational and critical skills necessary for the correct analytical and / or preparative methodology in the field of molecular biotechnology (UD1).
- to teach the fundamentals about the chemical and biochemical composition of foods, including their modifications during processing and storing (UD2).
Expected learning outcomes
The student will be able to apply techniques and methodologies related to the most frequent experimental problems, such as: separation of cellular organelles, extraction and analysis of proteins from plant material, extraction and analysis of nucleic acids, enzymes purification, design and execution of DNA amplification, gene cloning. In addition, the student will be able to critically front the issues related to the use of molecular biotechnologies for the production and analysis of foods.
Course syllabus and organization

Single session

Responsible
Lesson period
Second semester
Didactic Unit Molecular Methodologies:
The program and the reference material will not change.
Learning verification procedures and assessment criteria
The exam will be in written form using EXAM.NET + SEB platform, in webconference mode as indicated in UNIMI guidelines or, where the regulation will allows it, in attendance, always written.

Didactic Unit Molecular food biotechnologies:
Teaching methods: The lessons will be held on the Microsoft Teams platform and can be followed both synchronously on the basis of the second semester timetable and asynchronously because they will be recorded and left available to students on the same platform. The schedule of lessons and all the details of the activities will be published through notices on the ARIEL board before the start of the lessons. Any updates will also be communicated through notices on the ARIEL board.
The methods and criteria for participating to any possible face-to-face lessons and practical labs as established by the University, will be published in due time on the ARIEL notice board of the course.
Program and reference material: The program and reference material will not change.
Learning verification procedures and assessment criteria: the exam will take place orally using the Microsoft Teams platform, according to the indications provided by the University.
Course syllabus
Didactic Unit Molecular Methodologies:
The program includes the following topics:
- How to work in a laboratory of molecular biotechnology (safety, behavior, equipment, etc.).
- Preparation of solutions (buffer systems, biological buffers, etc.).
- Solubility of proteins and nucleic acids (extraction and precipitation)
- Spectrophotometric techniques (quantification and spectroscopic analysis of nucleic acids and proteins, fluorimetry).
- Electrophoretic techniques: one- and two-dimensional electrophoresis of proteins, electrophoresis of nucleic acids (genomic DNA, restriction fragments, RNA), blotting.
- Immunochemical techniques (Western and dot blotting, ELISA).
- Chromatographic techniques: traditional and MiniSpin (gelfiltration, ion exchange, hydrophobic interaction, affinity).
- Essays of enzyme activity.
- The enzymes in molecular biology (nucleases, ligases, polymerases, modifying enzymes).
- Restriction enzyme analysis
- PCR: end-point and real-time (design of oligonucleotides, optimum temperature, titration of MgCl2).
- Ligation of a restriction fragment in a plasmid cloning. Transformation and blue-white screen.
- Bioinformatics: bioinformatics in gene cloning and PCR (databases, search of sequences for alignment, position in the genome, etc.).

Didactic Unit Molecular food biotechnologies:
1. Elements of food chemistry and biochemistry: Description of the molecular components that make up food and raw materials. The intrinsic properties of the main classes of molecules (proteins, polysaccharides, lipids) which constitute some primary foods (milk, meat, egg, cereals, legumes) and their behavior in tissue systems (eg muscle, seeds, etc.) will be described. Food matrices. Biotechnological foods and products with high biotechnological content derived from food. Elements of molar nutraceutical.
2. Biological and biotechnological modifications of food: Molecular description of the role of the main chemical reactions and structural modifications to which the major macromolecular components of food are exposed during food processing and storage (physical, chemical and biological processes), also in relation to the ability of macromolecules to interact with each other, with the solvent, and with small molecules of nutritional relevance.
3. Biotechnological methods for food analysis: Food analysis based on the characterization of informational macromolecules (DNA and proteins). General principles of food analysis and fields of application of biotechnological approaches. Extraction and isolation of macromolecules (DNA, RNA and proteins) from the food material for analytical and diagnostic purposes. Resolution of the problems related to the interferences of the substances present in the food matrix. Methods and technologies used for the analysis of nucleic acids from food matrices. Methods and technologies for the analysis of proteins present in food. Use of tests for the determination of enzymatic activities and use of immunochemical techniques.
Prerequisites for admission
The course requires consolidated knowledge of the structural properties of proteins and nucleic acids which can be acquired in the "Biochemistry" and "Molecular Biology" courses.
Teaching methods
Didactic Unit Molecular Methodologies:
Frontal lessons: 24 hours
Single seat laboratory exercises: 48 hours
Laboratory exercises are compulsory

Didactic Unit Molecular food biotechnologies:
Frontal lessons 40 hours (5 CFU)
Teaching Resources
Didactic Unit Molecular Methodologies:
Course slides uploaded to the ARIEL platform.
Supplementary material distributed on the ARIEL platform

Didactic Unit Molecular food biotechnologies:
Books: Gigliotti-Verna. Biotecnologie alimentari, Piccin Editore.
Simpson-Nollet. Food Biochemistry and Food Processing. Blackwell Publisher.
Course slides uploaded to the ARIEL platform.
Supplementary material distributed on the ARIEL platform
Assessment methods and Criteria
Didactic Unit Molecular Methodologies:
It is mandatory to write laboratory reports before the exam. To verify learning achievements, students will go through a final written exam, in which six questions are proposed. The achievement of the objectives established by the course will be evaluated considering the candidate's ability to accurately define the topics, to offer an overall systematic view of the topics, and familiarity with the specific language.
It is mandatory to sign up at the exam within the deadline indicated on the SIFA.
The evaluation is expressed in thirtieths.
Results are communicated to the students by an automatically-generated e-mail.

Didactic Unit Molecular food biotechnologies:
Oral exam based on the presentation and discussion of the topics covered during the course. The exam can be taken, by appointment with the teacher, to be requested by e-mail, after registering for an appeal on the SIFA platform.
The learning verification will be evaluated according to the following criteria:
1. Proof of understanding of the topics covered during lectures and laboratory exercises
2. Ability to expose, in a critical and integrated way, the concepts at the molecular level of the role of the main chemical reactions and structural modifications that the major macromolecular components of food undergo during the transformation and conservation of food (physical, chemical processes, and biological), also in relation to the ability of macromolecules to interact with each other and with the solvent.
3. Knowledge of laboratory methodologies for the realization and understanding of biotechnological interventions on foods
4. Correct terminology
5. Completeness of the answers
Molecular food biotechnologies
BIO/10 - BIOCHEMISTRY - University credits: 5
Lessons: 40 hours
Professor: Scarafoni Alessio
Molecular methodologies
BIO/10 - BIOCHEMISTRY - University credits: 6
Single bench laboratory practical: 48 hours
Lessons: 24 hours
Professor: Magni Chiara
Professor(s)
Reception:
Contact the professor
DeFens, sez. Scienze Chimiche e Biomolecolari
Reception:
Thursday afternoon only by appointment (by e-mail)
Building 21040, first floor