Methods in Enzyme Technology
Methoden der Enzymtechnologie
8 LPWiSeEN
Voraussetzungen
Zwingend, im Wortlaut
• 12793 Purification and Characterisation of Proteins
Empfohlen, im Wortlaut
Techniques in microbiological isolation and cultivation; protein purification; analytical photometric and chromatographic methods.
Inhalte
The course consists of practical exercises for isolation and screening of enzymes from natural habitats (lignocellulose), strain cultivation for fermentative enzyme production, preparation of extra- and intracellular enzymes, enzyme conservation, characterisation of the enzymes’ application potential (factors for regulation of enzyme activity, textile bleaching, xenobiotic degradation potential, regioselective halogenation and hydroxylation). Oxidoreductases and hydrolases will be used for exemplary degradation of xenobiotic organopollutants, e.g. laccase to determine application conditions in washing processes, versatile haloperoxigenase and peroxygenase (catalysing the regiospecific hydroxylation of basic chemical structures) we will investigated and the halogenation potential will be studied. The use of enzymes in technologies for industrial waste treatment is carried out by constitutive and inducible mono- and dioxygenases of imperfect fungi.
Lernziele
For students the course with complex tasks will enhance key skills such as teamwork, communications, problem-solving and business awareness (professional methodically and social competences). The first step in the development of a new enzyme product is to find the organism that produces the optimal enzyme. Each student should isolate and determinate one fungal strain from natural sources; purify the strain and determine enzyme activities by using several screening methods. Students use well-studied microorganisms from internal strain collection for producing extracellular and intracellular oxidoreductases and hydrolases. The biocatalysts are separated, concentrated and lyophilised and individual enzyme activities, kinetic parameter and stabilities are studied. In the second part, isolated enzyme reactions - as well as bioreactions by cell preparations - will be performed in several application techniques, including analytical product identification, activities in dependence of changing incubation parameters and yields. The main objective is to demonstrate regio- and stereoselective reactions and to analyse the potential applications of oxidoreductases and hydrolases by several analytical methods.
Prüfungsleistung
• Individual practical work in the lab: preparation of experiments, performance, discussion, time management (50%) • Presentation and discussion in seminars, duration depends on the topic (20%) • Final colloquium, duration: 15 min (20%) • Final report, approx. 20 pages (10%)
Literatur (1)
- Bisswangeer H., Figura R., Möschel K., Nouaimi M: Enzymkinetik, Ligandenbindung und Enzymtechnologie. ISBN 3-8265-7175-4, 2001 Heiden S., Bock A-K., Antranikian G.: Industrielle Nutzung von Biokatalysatoren. ISBN: 3-503-04861-8, 1999 Several current publications e.g.: Kiebist, J., Schmidtke, K.-U., Zimmermann, J., Kellner, H., Jemlich, N., Ulrich, R., Zänder, D., Hofrichter, M. & Scheibner, K. (2017). A peroxygenase from Chaetomium globosum catalyzes the selective oxygenation of testosterone. ChemBioChem, Vol. 18: 563-569 (doi: 10.1002/cbic.201600677). Olmedo, A., Aranda, C., del Rio, J. C., Kiebist, J., Scheibner, K., Martínez, A. T. & Gutiérrez, A. (2016). From Alkanes to Carboxylic Acids: Terminal Oxygenation by a Fungal Peroxygenase. Angewandte Chemie International Edition, Vol. 55(40): 12248-51 (doi: 10.1002/anie.201605430). Kiebist, J., Holla, W., Heidrich J., Poraj-Kobielska, M., Sandvoss, M., Simonis, R., Gröbe, G., Atzrodt, J., Hofrichter, M. & Scheibner, K. (2015). One-pot synthesis of Human Metabolites of SAR548304 by fungal peroxygenases. Bioorganic & Medicinal Chemistry, Vol. 23(15): 4324-32 (doi:10.1016/j.bmc.2015.06.035) Friedrich, S., Gröbe, G., Kluge, M., Brinkmann, T., Hofrichter, M. & Scheibner, K. (2014). Optimization of a biocatalytic process to gain (R)-1-phenylethanol by applying the software tool Sabento for ecological assessment during the early stages of development. Journal of Molecular Catalysis B: Enzymatic, Vol. 103: 36-40. (doi:10.1016/j.molcatb.2013.10.002)
Bemerkungen
The Laboratory course takes place as a 6 week block course. The seminar is an obligatory part of the lab course.
Quelle: Modulbeschreibung der BTU · abgerufen 28.09.2026Original