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Lecturer(s)
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Kanďár Roman, prof. Mgr. Ph.D.
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Štěpánková Šárka, Mgr. Ph.D.
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Course content
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History of enzymology. Structure of enzymes (holoenzyme, apoenzyme, cofactor). Multiple forms of enzymes (isoenzymes, polymeric enzymes), multienzymes (multienzyme complexes, polypeptides). Enzyme nomenclature (trivial, semisystematic, systematic). Classification of enzymes. Enzyme activity (catalytic, specific). Conventional activity units. Methods for determination of enzyme activity (optical, electrochemical). Mechanism of enzyme catalysis (thermodynamics and kinetics of conversion of substrate to product). Formation of complex enzyme-substrate. Energetic profile of noncatalysed and catalysed reaction, possibilities to affect of activation energy. Enzyme kinetics. The effect of reaction conditions and effectors on the rate of enzyme reaction. Kinetics of enzyme reaction in steady state. Kinetics of uni-substrate reactions (Michaelis-Menten equation). Kinetics of multi-substrate reactions. Software for enzyme konetics. Enzyme inhibition. Inhibition by substrate, product, inhibitor. Reversible/irreversible inhibitors. Types of full inhibition (competitive, noncompetitive, uncompetitive, mixed). Cooperativity in enzyme action. Homotropic and heterotropic, cooperativity, allosteric effect. Isolation of enzymes. Animal, plant, microbial sources of enzymes. Releasing of intracellular enzymes from cells. Choice of separative techniques. Purification procedures. Study of enzyme structure. Crystallization of enzymes. X-ray crystallography. Enzymes in biochemical analytics. Determination of clinical important enzymes. Determination of analytes using solute enzymes. Immobilization of enzymes. Applications. Enzyme biosensors. Enzyme immunoanalysis. Enzymatic markers and methods of preparation of enzyme conjugates. Enzymes as markers of gene expression. Enzyme engineering. Enzymes in practice (medicine, food industry, biodetergents, organic synthesis).
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing)
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Learning outcomes
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The aim of this subject is to intensify knowledge of enzymology gained in subject General Biochemistry and Clinical Biochemistry.
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Prerequisites
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Students will get basic information about properties of enzymes, their structure, partition, nomenclature, mechanism of catalytic action, enzyme kinetics and practical applications of enzymes.
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Assessment methods and criteria
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Written examination
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Recommended literature
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Elektronické podklady (k dispozici na studijním webu univerzity STAG).
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Nelson DL, Cox MM. Lehninger Principles of Biochemistry. New York: Freeman WH, 2004.
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Palmer T, Bonner PL. Enzymes: Biochemistry, Biotechnology and Clinical Chemistry. Woodhead Publishing: Horwood, 2007.
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Voet, Donald. Biochemie. Praha: Victoria Publishing, 1995. ISBN 80-85605-44-9.
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