Applied enzymes

Course Code: 4.1

Semester: 4th 

Category: Compulsory 

Hours: 2h Th + 2h Lab

ECTS : 5

 

 

LEARNING OUTCOME

Upon completion of the course, it is expected that the student will be able to:

• Knows basic principles governing Biocatalysis.
• Be able to explain the physical constants and chemical behaviour of enzymes based on their kinetic properties.
• Be able to quantify substances based on enzyme reactions.
• Know the basic classes of enzymes and be able to predict their chemical behaviour and apply it to oenological practices.
• Apply the techniques of enzyme technology to optimize wine production.

 

General skills

 

• Research, analysis and synthesis of data and information for the use of necessary techniques and technology
• Development of creative and inductive thinking
• Autonomous work and/or teamwork
• Decision making

 

COURSE CONTENT

Theoretical part 

• Introduction to Enzymology, naming and classification of enzymes.
• Description of biological catalysts, enzyme catalysis.
• Structure of enzymes, active area, specificity of enzymes.
• Enzyme activity – Factors affecting enzyme activity (pH, temperature, etc.).
• Kinetics of enzyme reactions. Michaelis-Menten equation. Linear conversions of the Michaelis-Menten equation.
• Enzyme inhibition. Reversible and irreversible enzyme inhibitors. Ways to measure the action of inhibitors.
• Hydrolysis of p-nitrophenyl-glucopyranose in the presence of the enzyme beta-glucosidase. Measurement of the enzymatic activity of beta-glucosidase.
• Basic enzymes of saccharomyces and their action – Basic enzymes of grape ripening
• Isolation and purification of enzymes from various sources
• Enzymes in analytical chemistry: Enzyme methods of analysis
• Applications of enzymes – Industrial enzymes and applications of industrial enzymes

 

Laboratory Part

• Calculation of lipase activity.
• Calculation of KM, Vmax lipase
• Determination of KM, Vmax with immobilized lipase reusable
• Determination of beta-glucosidase activity spectrometrically, during hydrolysis of p-nitrophenyl-glucopyranose.
• Effect of pH and temperature on enzyme reaction rate (hydrolysis of p-nitrophenyl-glucopyranose by beta-glucosidase)
• Determination of kinetic parameters Vmax, KM of beta-glucosidase during hydrolysis of p-nitrophenyl-glucopyranose.
• Measurement of kinetic parameters KM, Vmax of beta-glucosidase, in the presence of an inhibitor (glucose, fructose).
• Determination of ethanol by enzymatic reactions
• Quantification of malic acid by enzymatic reactions
• Determination of glucose and fructose by the method of enzymatic reactions

 

RECOMMENDED BIBLIOGRAPHY

1. Αντώνη Τρακατέλλη, «Βιοχημεία, Ένζυμα –Τεύχος Β1», Θεσσαλονίκη 1976
2. Lubert Stryer, « Βιοχημεία », Τόμος Ι, Πανεπιστημιακές Εκδόσεις Κρήτης, Ηράκλειο (1997)
3. Γ. Γεωργάτσου, Τ. Α. Γιουψάνη, Δ.Α. Κυριακίδη, «Ενζυμολογία», Εκδόσεις Ζήτη, Θεσσαλονίκη (2001)
4. Ιωάννης Κλώνης, “Ενζυμική Βιοτεχνολογία”, Παν. Εκδόσεις Κρήτης (1997)
5. Alan Fersht, “Enzyme Structure and Mechanism”, 2nd Editiοn, W.H. Freeman
6. Beutler, H. –O. (1984) “Methods of Enzymatic Analysis” (Bergmeyer, H.U., ed.) 3rd ed., Νerlag Chemie, Weinheim, Deerfield Beach/Florida, Basel
7. Boehringer Mannheim Biochemica “Methods of Enzymatic Food Analysis” (1984)
8. Xaνier Malcata F., Reyes R. H., Garcia S. H., Hill G. C., Amundsοn H.C. and Jr. “Immοbilized Lipase Reactors fοr Modification οf Fats and Οils- A Reνiew”, Νο 67, nο. 12, JAΟCS, (1990)
9. Ribéreau- Gayon, D. Dubourdieu, B. Donèche, A. Lonvaud, “Handbook of Enology”, Volume 1, “The Microbiology of Wine and Vinifications”, (2000) John Wiley & Sons, Ltd.