82576 - PROTEIN ENGINEERING

Learning outcomes

At the end of the course, the Student will be able to analyze the kinetics of enzyme-catalyzed reactions, and will be confident with the methodologies of enzyme engineering. In addition, the Student will be able to evaluate the feasibility of industrial processes relying on the use of natural or artificial enzymes.

Course contents

Review of kinetics. Enzymatic catalysis, Michaelis-Menten and Briggs-Haldane equations. Enzyme inhibitors: competitive, non-competitive, uncompetitive. Mechanism of action of serine proteases and DNA polymerases. Linus Pauling's paradigm and catalytic antibodies. Rational protein engineering: construction of thermostable enzyme variants and grafting of heterologous catalytic activities. Directed evolution of enzymes. Error-prone PCR and construction of random mutant libraries of enzyme-encoding genes. DNA shuffling. Evolution of enzymes and isolation of variants resistant to organic solvents, thermostable mutants, or mutants exhibiting altered specificity. Cold-active enzymes and their evolution. Protein chimeras and the enzymatic assay of blood glucose. In vivo evolution of enzymes and production of variants with the desired phenotype.

Readings/Bibliography

Scientific articles.

Teaching methods

PowerPoint presentations. The slides will be uploaded in "Virtuale" before the lectures will be given. 

Assessment methods

The written exam (duration 120 minutes) includes: i) evaluation of the laboratory report, worth a maximum of 6 points; ii) 18 multiple-choice questions (3 answers per question), worth a total of 18 points; ii) 2 open-ended questions, worth a total of 6 points. These questions are designed to test: i) the student's ability to critically analyze a topic; ii) the ability to connect different themes; iii) the student's ability to identify future prospects for existing technologies. The use of artificial intelligence tools is not permitted during the exam.

Students with learning disorders and/or temporary or permanent disabilities: please contact the office responsible (https://site.unibo.it/studenti-con-disabilita-e-dsa/en/for-students ) as soon as possible, so they can propose acceptable adjustments.
The request for adaptation must be submitted to the lecturer at least 15 days before the exam date. The lecturer will assess the appropriateness of the adjustments, taking into account the teaching objectives.

Office hours

See the website of Alejandro Hochkoeppler