752338 Practical course in enzyme technology

Type
Practical course
Semester hours
3
Lecturer (assistant)
Giorgianni, Angela , Ludwig, Roland
Organisation
Offered in
Sommersemester 2025
Languages of instruction
Englisch
Content

Please also read the course information on BOKUlearn!
A detailed schedule will be provided on BOKUlearn.
The practical part will take place in the food biotechnology laboratory in Muthgasse 11, 1st floor (Monday to Thursday from 11:00 to 17:00).

Pyranose 2-oxidase, an FAD-dependent carbohydrate oxidase can be employed for the production of rare carbohydrates. A potential drawback is its narrow substrate specificity. For the production of technologically interesting sugars (e.g. tagatose from galactose) a series of P2O variants were developed. Two of these mutant enzymes (variants) will be compared in this practical course. Time schedule: We will start with biomass (E.coli containing recombinant pyranose 2-oxidase in the cytosol), disrupt it mechanically, purify the enzyme by a chromatographic step (IMAC), and determine the technologically important constants (pH optima, Km and kcat values for different sugars and electron acceptors). Based on the obtained results we will calculate batch conversion processes (for different carbohydrates, with or without cofactor regeneration by laccase) and will run laboratory scale batch conversions in bioreactors. Product analysis will be done by HPLC and results used to evaluate our determined catalytic constants and calculations.

The purpose of this intense practical course is to enable students from the master studies in Food Technology and Biotechnology to learn and practice fundamentals of biocatalysis by a practical approach. The course demands the application of your theoretical knowledge of biochemical principles on given practical problems. The influence of various physical parameters on enzyme activity, and thus on the productivity of enzymatic processes will be examined. Through the use of coupled enzyme systems for the regeneration of the required coenzyme, the redox reaction selected can be judged on the basis of its economic aspects. Exercises will be supplemented by the relevant analytical methods and the demonstration of alternative processes and advanced techniques for enzyme recovery, biocatalytic processes and product processing.
Previous knowledge expected

Participants must be able to perform elementary practical routines like pipetting, buffer calculation and preparation, centrifugation and weighing.
Theoretical knowledge is covered by the provided course script (on BOKUlearn), which has to be worked through before the course starts.
Objective (expected results of study and acquired competences)

After the positive completion of the course students can:

1) Apply different biochemical methods for cell disruption (homogenisation, sonication,...), protein purification (chromatography) and kinetic characterization (pH profiles, determination of Km and Vmax) of enzymes and point out, if necessary, other or modified procedures.
2) Plan and calculate enzymatic processes featruring a single enzyme
3) Identify and evaluate problems in enzymatic processes and improve these processes
4) Identify critical process steps and argue consequences for the process
5) Compile the obtained results in a scientific publication based on criteria discussed in class

Additionally, you have the opportunity to:
Apply your accumulated biochemical knowledge on practical tasks in the course
Work in a small team, to prioritise and delegate tasks and to organise the team work
You can find more details like the schedule or information about exams on the course-page in BOKUonline.