Enzymatic Engineering and Biotechnology

Course Leader: Dr Zied Zaray

Home Institution: Higher Institute of Biotechnology, University of Sfax, Tunisia

Course pre-requisites: None.

Course Overview
The lecture gives an in-depth insight into methodologies and approaches regarding enzymatic engineering and the use of enzymes in industrial processes. Enzymes are presently used in many industrial sectors and several of these are discussed in the course. The course will in particular detail consider the Screening and production of industrial-relevant enzymes (more particularly lipolytic enzymes) from a different source and processes within the sectors of the food industry, small molecule synthesis, bioenergy production, environmental biotechnology, biomedical....,

In particular, a good part of the course will be devoted to the purification, biochemical and molecular characterization of its enzymes with the aim of orienting them towards biotechnological applications.

Learning Outcomes
By the end of this course students should be able to:
1) Describe and explain how enzymes are used in a broad spectrum of industrial processes and describe how different enzymes can be modified for optimal performance in these processes.
2) Describe and explain the fundamental chemical, technical and physical properties that govern an efficient use of lipolytic enzymes in industrial processes.
3) Plan and initiate an industrial process in which the use of an enzyme is a central part.
4) Present the planning, analysis and evaluation of an industrial process using enzyme technology in the form of a technical report.

Course Content
Unit I- Introduction to Enzymes
General introduction and historic background- General Terminology, Nomenclature and Classification of Enzymes. Criteria of purity of enzymes- Specific activity. Enzyme units-Katal and IU. Enzyme activity- chemical nature of enzymes.
Hydrolases: glycosidases, peptidases, lipases
Oxidoreductases: oxidases, peroxidases
Transferases: glycosyltransferases
Isomerases: carbohydrate epimerases
Unit II Enzyme Kinetics and Regulation
Factors affecting the enzyme activity- Concentration, pH and temperature. Kinetics of a single-substrate enzyme catalysed reaction, Michealis-Menten Equation, Km, Vmax, L.B Plot, Kcat. Kinetics of Enzyme Inhibition.
Feedback Regulation, Organisation of enzymes in the cell. Enzymes in the cell, localization, compartmentation of metabolic pathways, enzymes in membranes, concentrations. Mechanisms of enzyme degradation.
Unit III- Lipolytic enzymes
Lipases/ Specificity of lipases/Mechanism of action of lipases. The digestive lipases/The classic pancreatic lipases/The gastric lipase. Characterization of the lipase-colipase system
Phospholipases /Phospholipases A2 (PLA2)/The cytosolic PLA2: cPLA2/The secret PLA2: sPLA2/SPLA2 of mammals/The pancreatic PLA2: sPLA2-IB.
Importance and physiological role/Physiological roles of sPLA2-IB/Physiological roles of lipases.
Unit IV Industrial uses of Enzymes (Applied Enzymology)
(The most detailed example: lipases and phospholipases).
Industrial Enzymes- Thermophilic enzymes, amylases, lipases, proteolytic enzymes in meat and leather industry, enzymes used in various fermentation processes, Lipases in oleochemistry , Synthesis of esters, Protein Engineering to Alter Fatty Acid Selectivity of Lipase…
Unit V Clinical uses of Enzymes (Applied Enzymology)
Lipases and treatment of pancreatic insufficiency, Anti-inflamatory agents, Isoenzymes like LDH, Cholinesterases, Phosphatases. Immobilization of enzymes, ELISA. Enzyme Engineering and site directed mutagenesis, Designer enzymes…
Unit VI Enzyme Structure activity Relationship and Drug Discovery- Properties of Enzymes.
Case study of LIPASES and PHOSPHOLIPASES.

Instructional Method
Teaching is organized in lectures, seminars, videos, quizzes, and project work. A large part of the course is pursued as individual projects and in project groups of 2-3 students (homework using Journal Resources)

Required Course Materials
Data show and internet

Assessment
During the teaching, the evaluations will be in the form of classroom quizzes and exams, homework (project groups of 2-3 students and individual Evaluation Exercise) and formative evaluation.
At the end of the course the evaluations will be in the form of final written exam.