C8112 Enzyme biotechology

Faculty of Science
Spring 2021
Extent and Intensity
2/0/0. 4 credit(s). Type of Completion: zk (examination).
Teacher(s)
prof. RNDr. Igor Kučera, DrSc. (lecturer)
Guaranteed by
prof. RNDr. Igor Kučera, DrSc.
Department of Biochemistry – Chemistry Section – Faculty of Science
Supplier department: Department of Biochemistry – Chemistry Section – Faculty of Science
Timetable
Mon 1. 3. to Fri 14. 5. Tue 14:00–15:50 online_BCH2
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
fields of study / plans the course is directly associated with
Course objectives
Enzyme biotechnology deals with the use of enzymes as biocatalysts to manufacture bulk and high added-value products such as food, fine chemicals, and pharmaceuticals. Enzymes are also used to improve services, as in textile washing and environmental management, or for analytical and diagnostic purposes. The course will provide students with the basic principles of enzymology. An overview of procedures for industrial scale protein production will be presented, including downstream processing steps, and enzyme optimization through enzyme discovery and protein engineering. A number of case studies illustrating the use of enzymes will be discussed in detail. The subject also introduces students to some commercial aspects, such as patent practice and current trends in enzyme market.
Learning outcomes
By the end of this course, students will be able to: • -demonstrate a familiarity with the basic classes of enzymes, and their potential for use in biotechnology • -design an enzymatic assay and estimate kinetic parameters from raw kinetic data • -design, carry out and evaluate enzymatic transformations • -describe methods for selection and optimization of industrial enzymes using genetic and biochemical techniques • -summarize current processes involved in industrial enzyme production, from protein production to purification and formulation • -understand soluble or immobilized enzyme technologies for the production of industrial and medical products • -discuss various application of enzymes that can benefit human life
Syllabus
  • 1) Introduction to enzymology. General terminology, nomenclature and classification of enzymes. Structure of enzymes and their active sites. The role of cofactors. Origin of catalytic enhancement. Catalytic flexibility and selectivity/specificity. Advantages and disadvantages of the use of enzymes as catalysts. • 2) Enzyme assays. Expression and measurement of enzyme activity. Essential conditions for enzyme assays (substrate concentration, buffer, temperature). Progress curve and initial velocity. Continuous and stopped assays. Assays for high-throughput screening. Instrumental aspects of measurement. • 3) Enzyme kinetics in homogeneous systems. Kinetics of single substrate reactions; estimation of Michelis-Menten parameters. Alternative substrates. Multisubstrate reactions. Enzyme inhibition: reversible (competitive, non-competitive and un-competitive) and irreversible inhibition. Substrate and product inhibition. Allostery and sigmoidal kinetics. pH and temperature effects on enzymes. Deactivation kinetics. • 4) Enzyme kinetics in heterogeneous systems. Mass transfer limitations in enzyme catalysis. Enzymes in non-conventional media. • 5) Enzyme discovery and protein engineering. Enrichment cultivation, metagenome approach, in silico screening. Directed (molecular) evolution, rational design, computational design. • 6) Large-scale enzyme production and purification. Enzyme sources. Techniques of microorganism cultivation. Possibilities to improve enzyme yield. Downstream processing steps: recovery, concentration, purification, formulation. • 7) Enzyme immobilization. Carriers (inorganic, organic and synthetic polymers), binding methods (adsorption, covalent binding, cross-linking and entrapment). Properties of immobilized enzymes. Applications of immobilized enzymes in bioreactors and biosensors. • 8) Biotransformation applications of enzymes. Synthesis of antibiotics, production of therapeutics and fine chemicals. • 9) Applications of enzymes in food and beverage industry • 10) The use of enzymes in the household care industry, textile and leather industry. Enzymes in bioremediation. Biofuel production from biomass. • 11) Medical applications of enzymes as diagnostic, analytical and therapeutic agents. Enzymes for molecular biology. • 12) Enzyme market, enzyme companies and marketed enzymes. Academic and patent literature on enzymes. Concluding remarks.
Literature
  • ILLANES, Andrés, Lorena WILSON and Carlos VERA. Problem solving in enzyme biocatalysis. Chichester: Wiley-Blackwell, 2014, xxiii, 318. ISBN 9781118341711. info
  • BUCHHOLZ, Klaus, Volker KASCHE and Uwe BORNSCHEUER. Biocatalysts and enzyme technology. Second, completely revised a. Weinheim: Wiley-VCH, 2012, xx, 606. ISBN 9783527329892. info
  • CORNISH-BOWDEN, Athel. Fundamentals of enzyme kinetics. 4th, completely revised and. Weinheim: Wiley-Blackwell, 2012, xviii, 492. ISBN 9783527330744. info
  • BISSWANGER, Hans. Practical enzymology. 2nd, completely rev. ed. Weinheim: Wiley-Blackwell, 2011, xvi, 360. ISBN 9783527320769. info
  • DEVASENA, T. Enzymology. Oxford: Oxford University Press, 2010, xvi, 492. ISBN 9780198064435. info
  • Handbook of enzyme biotechnology. Edited by Alan Wiseman. 2nd ed. Chichester: E. Horwood, 1985, 457 s. ISBN 0853124205. info
Teaching methods
Lectures with PowerPoint presentations that are available online as pdf files.
Assessment methods
Written examination (20 questions, 8 correct answers is needed to pass)
Language of instruction
Czech
Further Comments
Study Materials
The course is taught annually.
The course is also listed under the following terms Spring 2020, Spring 2022, Spring 2023, Spring 2024, Spring 2025.
  • Enrolment Statistics (Spring 2021, recent)
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