Enzymology and Experimental Biochemistry

Course content

The course comprises the theoretical and experimental work on enzymes, proteins and peptides in biological matrices. Focus is placed on experimental and theoretical understanding of the relations between structure and properties of the enzymes.

Topics included are:

  • Extraction techniques
  • Chromatographic systems
  • Solvent and buffer relations
  • Enzymatic assay design and development
  • Thermodynamics and kinetics in enzymology and biochemical analyses.
  • Binding between molecules in equilibrium reactions and thermodynamic stable complexes.
  • Receptor and effector sites, activators and inhibitors, and their effects on the active sites.
  • Native enzymes: holoenzymes, apoenzymes, co-factors, activation energy and kinetics



The combinations of such knowledge give the students basis for understanding and development of a relevant strategy of methods as used in analytical biochemistry.

Project

The students will perform an activity based purification of individual enzymes from a complex protein sample originating from Aspergillus strain. In groups they will design a purification strategy of the enzyme in question. The students will use ÄKTA FPLC-systems for the enzyme purifcation. 

Upon purification enzymatic properties of the enzyme must be determined.
These includes Vmax, Km and kcat, and interpretation of such data in relation to ideal catalytic effects. We will use the graphing program PRISM from GraphPad for the fitting and analysis of data.


 

Education

MSc Programme in Biotechnology

Learning outcome

The course provides a thorough and fundamental theoretical and practical basis for knowledge to and work with biomolecules, especially enzymes used in biotechnology. Focus is placed on the relations between structure and properties of biomolecules, strategies in use of actual new and traditional biochemical methods and understanding of the descriptions in original scientific papers.

After completing the course the student should be able to:

Knowledge

  • Understand physico-chemical-biochemical properties of biomolcules and especially enzymes.
  • Describe strategies to efficient isolation and purification of enzymes from complex biological systems
  • Describe analytical techniques used in biochemistry and for enzyme characterisation.
  • Understand the theoretical basis for changes of water as solvent, extraction and chromatographic systems in work with molecules of different polarity.


Skills

  • Evaluate the detection, quantification and linearity of experimental data in relation to published values (theoretical exercise).
  • Design experimental stategies in analytical biochemistry and enzymology based on structure and properties of analytes in complex matrix systems.
  • Apply theoretical principles of analytical biochemistry to carry out experimental isolation and purification of enzymes
  • Evaluate the performance of applied techniques in an enzyme related project carried out in a group (preferably 2-4 persons).



Competences

  • Cooperate with a fellow student on laboratory reports and a project report
  • Expand the knowledge to biochemistry by use of mathematics evaluations
  • Utilise the obtained knowledge in more comprehensive projects within research, development or industry.

Lectures and theoretical exercises: lectures will be used to present the theoretical background for the subjects considered with at least 3 hours per week.
Theoretical exercises are an integrated part of the lectures. Here students will work on answers of specific questions related to the theory and practical use of methods and technology. Laboratory work with knowledge to theory and practical use of the various instruments and biochemical methods are central parts of the course. The students will be in groups of 2-4 and spend about 9 hours per week with this part of the course. Four weeks will be devoted to the project focused on studies of a selected enzyme or inhibitor molecule. The combined theoretical and practical laboratory experiments with focus on strategies of methods in analytical biochemistry and enzymology will train the students in calculations on the produced data.

Practical Enzymology, 3rd edition by Hans Bisswanger (2019)

Practical enzymology will be used as basis for the course. In addition we work with selected papers during theoretical part of the course (to be found on Absalon). For your experimental project you are expeted to find relevant litterature yourself.

 

 

Competences within basic organic chemistry and basic biochemistry are recommended.

Academic qualifications equivalent to a BSc degree is recommended.

This course is identical to LKEK10081U Enzymology and Experimental Biochemistry.

Oral
Individual
Collective
Continuous feedback during the course of the semester
Feedback by final exam (In addition to the grade)
ECTS
7,5 ECTS
Type of assessment
Oral examination, 20 min
Evaluation of project report supplemented with individual oral presentation and examination in curriculum (no preparation time).

Weight: Oral presentation of project report and subsequent examination in the report and curriculum are 100% of the final grade.
Aid
All aids allowed
Marking scale
7-point grading scale
Censorship form
No external censorship
Several internal examiners
Criteria for exam assessment

See Learning Outcome

Single subject courses (day)

  • Category
  • Hours
  • Lectures
  • 24
  • Preparation
  • 109
  • Theory exercises
  • 8
  • Practical exercises
  • 64
  • Exam
  • 1
  • English
  • 206