INGLESE

English

Metabolic engineering of microbial cells. Design of experiments (DoE) for screening and optimization of results. Hints to biotechnological processes: fermentation development, upstream and downstream processes.

1-“Biotechnology”- De Gruyter
2- Laboratory protocols
3- Slides of lessons are available in pdf format in dedicated teams folder

LEARNING OBJECTIVES. The course aims to provide knowledge of Metabolic Engineering approaches, with a focus on model-driven ones, approaches that allow the study and understanding of biological systems more easily and the identification of metabolic targets. It will also focus on the use of a “design of experiment” approach, to understand how to reduce the number of experiments when optimizing a biotechnological process and finally on the scale up of processes and the transition to controlled conditions. The course integrates principles of metabolic engineering, strain engineering, upstream and downstream processing, experimental design, and bioreactor operations with extensive laboratory activities, in accordance with the Dublin Descriptors outlined below.

KNOWLEDGE AND UNDERSTANDING.
Students will acquire a comprehensive understanding of the principles of metabolic engineering and microbial strain engineering, the design and optimization of upstream and downstream processes, the application of Design of Experiments (DoE) for process optimization, and the operation of bioreactors and different fermentation strategies. They will also gain knowledge of the main downstream processing techniques, including solid-liquid separation, filtration, and solvent precipitation for product recovery.

APPLYING KNOWLEDGE AND UNDERSTANDING.
Students will be able to apply theoretical knowledge to engineer a microbial strain; design, implement and optimize microbial growth and metabolite production in laboratory-scale cultures; perform upstream and downstream processing operations. They will also acquire practical experience in analyzing process performance and product recovery.
MAKING JUDGEMENTS.
Students will develop the ability to critically analyze experimental results obtained during strain engineering, process optimization, fermentation, and downstream processing. They will be able to identify critical process parameters, evaluate the effectiveness of different bioprocess strategies, interpret experimental data, and select appropriate approaches to improve productivity, scalability, and product recovery.

COMMUNICATION SKILLS.
Students will be able to accurately use scientific terminology related to metabolic engineering, bioprocess technology, fermentation systems, and downstream processing. They will be capable of effectively communicating experimental procedures, analyzing and presenting laboratory results, and discussing bioprocess development strategies using appropriate scientific language.

LEARNING SKILLS.
Students will develop independent learning skills that will enable them to critically evaluate scientific literature in the fields of metabolic engineering, industrial biotechnology, and bioprocess engineering. They will acquire the ability to continuously update their knowledge of emerging technologies and methodologies for the development, optimization, and scale-up of microbial bioprocesses.

Knowledge provided by courses on Microbiology, Molecular Biology, Industrial Biochemistry and Biotechnology (possibly Chemical Biology of Fermentations).

Theoretical lessons and laboratory experiences in 6 lessons for a total of 32 hours.
Frontal and practical lessons will be split up:
Frontal lesson in the classroom: 8 total hours
Practical lessons: 24 total hours in 4 times “Laboratorio Centro Grandi Apparecchiature” (via de Crecchio n 7, 80138, Naples)

Achievement of the course learning objectives is certified through passing an exam, graded on a scale of thirty (30).
The exam consists of an oral examination, to be taken on scheduled dates. The exam is graded on a scale of thirty and is considered passed with a minimum score of 18/30.
The oral examination has an average duration of 30 minutes and consists of an interview structured as follows: i) general questions, to assess the student's command of fundamental concepts and the breadth of the knowledge acquired; ii) in-depth thematic questions, to verify the student's ability for critical analysis and methodological rigor; iii) interdisciplinary connections, to test the student's ability to link the different topics of the course into an overall view.
The final grade will be expressed on a scale of thirty.
Grading criteria:
<18 Fail: the student does not demonstrate results consistent with the descriptors of knowledge and understanding, application, judgement, communication, and learning skills.
18–21 Sufficient level: the student meets the basic descriptors relating to knowledge and understanding of the fundamental content and shows an initial ability to apply it in simple contexts.
22–24 Fully sufficient level: the student meets the descriptors of applied knowledge and understanding, showing the ability to correctly apply key concepts and beginning to critically analyze situations relevant to the course topics.
25–26 Good level: the student demonstrates independent judgement in evaluating and comparing relevant scenarios, applies knowledge effectively, and communicates results clearly, reflecting more consolidated competences.
27–29 Very good level: the student meets the descriptors of all five areas at an advanced level, with mastery of the content, the ability to critically evaluate complex cases, and strong communication skills.
30 Excellent level: the student excels in all the Dublin descriptors, demonstrating the ability to learn continuously and creatively, comprehensive knowledge and understanding, confident and critical application of knowledge, and effective communication skills, while achieving an outstanding standard in both the written and oral examinations.
Honours (cum laude) may be awarded when, in addition to the above, the student demonstrates exceptional originality, depth of analysis in the oral discussion, and performance that exceeds expectations.

Attendance is mandatory, with a maximum of 25% of allowed absences
Supporting material:
- slides of lectures are available online in pdf format and scientific publications on the topics
Office hours are held on the days indicated in the course syllabus; additional appointments may be scheduled by email upon request.

Theoretical lessons:
Metabolic engineering principles, methodologies and applications in microbial cells and model driven strain engineering. Processing of complex renewable materials. Design of experiments: principles of DoE and set up of the protocol for process optimization. Bioreactor and fermentation strategies. Scale up of the process from bottles to controlled conditions in bioreactors. Solid liquid separation, filtration, solvent precipitation.
Practical lessons in the lab
-Strain engineering/characterization
-Application of the DoE protocol identified during the course in small scale flask experiments to identify optimal conditions for strain growth and production of the metabolite of interest. Scale up of the process in bioreactor. Recovery of biomass/ added value metabolite.