752338 Practical course in enzyme technology


Type
Practical course
Semester hours
3
Lecturer (assistant)
Ludwig, Roland
Organisation
Institute of Food Technology
Offered in
Sommersemester 2018
Languages of instruction
Englisch

Content

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 cytosolic recombinant pyranose 2-oxidase), disrupt it mechanically, purify the enzyme by one 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.
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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 the Moodle pages), which has to be worked through before the course starts.
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Objective (expected results of study and acquired competences)

The purpose of this intense training course is to enable students from Foodtechnology and Biotechnology to learn and practice fundamentials of biocatalysis by their practical work. 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 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.
After the course you will be able to:
1) plan and calculate simple enzymatic processes
2) evaluate problems in biocatalytic processes and deal with them
3) apply several biochemical methods for cell disruption, purification and kinetic characterisation of enzymes and if neccessary, modify them
4) evaluate the importance of indivitual process steps for the whole process
Additionally, you will have been trained to:
5) apply your so far accumulated biochemical knowledge on practical tasks
6) work in a small team, to prioritise and delegate tasks and to organise the team work
7) Compile the obtained results in a scientific publication
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You can find more details like the schedule or information about exams on the course-page in BOKUonline.