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Bioseparation, enzyme kinetic and gene regulation

Presentation

Programme (detailed contents):

 

Fractional precipitation: definition, classification, salting-in, salting-out, solvent or polymer precipitation. Centrifugation and ultracentrifugation: definition, classification, sedimentation theory, equipments. Membrane procedure: microfiltration, ultra and nano filtration, reverse osmosis, dialysis. Electrophoresis (PAGE, isoelectric focusing, 2-D and capillary electrophoresis)

Enzyme classification, thermodynamic for enzymology, reaction rate, kinetic equation, action mechanism (Michaelis-Menten), inhibition (competitive, non competitive, partial…), activation, allostery (sequential and concerted models), steady-state kinetics of multireactant enzymes, effect of pH and temperature.

 

Examples of gene regulations in prokaryotes and eukaryotes

 

Organisation:

 

Lectures and tutorials in parallel and lab work. Some lectures are given by the students.

 

Main difficulties for students:

 

Mathematical description of biological phenomena.

 

New concepts and weak background

Objectives

At the end of this module, the student will have understood and be able to explain (main concepts):

 

- the mechanisms of separation techniques usually used in biochemistry-biotechnology, more particularly in the case of biocatalysts: techniques of precipitation (proteins, nucleic acids), techniques of centrifugation and ultracentrifugation, membrane techniques (MF, UF, NF) and electrophoretic techniques;

- the bases of the measure of the reaction rate of an enzyme, the various models allowing the description of the behaviour of an enzyme, from the simplest to the most complex, the effect of physico-chemical parameters such as the pH or the temperature

- the main pathways of gene expression regulation

 

The student will be able to:

 

- choose the most suited method of bioseparation to a context by knowing correctly its functioning                 

- to use from a practical point of view the equipment and the techniques adapted in the purification of proteins and to the control of their purification (low and medium pressure chromatography, electrophoresis)

- to establish complex enzymatic equations of reaction rate by means of models.

- to determine experimentally the various kinetic parameters of an enzyme as well as its optimal conditions of functioning

- explain and analyse data concerning gene expression regulation in prokaryotic and eukaryotic organisms

Form of assessment

The evaluation of outcome prior learning is made as a continuous training during the semester. According ot the teaching, the assessment will be different: as a written exam, an oral exam, a record, a written report, peers review...

Additional information

Organic chemistry (I2BECH11), structural biochemistry (I2BEBC11), transport phenomena and reaction in fluids (I2BETF11). Analytical method (I2BEAN11). Molecular biology.

 

Separation, purification, enzyme kinetic, gene regulation