Hello friends and welcome to MOOC in Enzymology.Enzymology is a specialized branch of life sciences that deals with the biochemical nature and activity of enzymes and is a subject that has relevance to students from a wide range of disciplines. In addition, it will serve as an individual reference for students pursuing their undergraduate/ post graduate degree programmes in life sciences and it could be recommended to anyone wishing to get an idea of the present day scope and applications of enzymology. The present course opens the door to all of the abundant careers in and out of the area of biological sciences including health/ medical / Environmental Sciences. The course is designed to give students an understanding of procedures involved in purification of enzymes, enzymes assays and quantitative evaluation of the influencing parameters such as concentrations of substrate / enzyme, pH, temperature and effects of inhibitors on enzyme activity.This is a course where the topics to be studied include enzyme active sites / mechanisms of enzyme action; enzyme kinetics and regulation; collision and transition state theories; Isozymes and their clinical significances /protein folding / site-directed mutagenesis and molecular structure/ function relationship etc as tools for understanding functions of enzymes. This course is rich in pedagogical features like short answer type questions / self explanatory illustrations, MCQs etc. which help students to evaluate themselves and improve upon.This course is fundamental to the acquisition of competencies to solve problems which impact daily life e.g., health and environment and also students should acquire skills to initiate a research or industrial career (food, pharmaceutics or biotechnology) in the country or abroad. The main goal of this course is to prepare students for critical analysis of scientific phenomena involving enzymes and competently work with enzyme systems in both academia and industry.Hence, the programme is designed to provide students with the knowledge and skills they need to explore the central questions in basic and applied biology.Hope that the students belonging to all the fields of life sciences may take this as a course of study in their undergraduate and post graduate programs. All the best---
Overview
Syllabus
Week 1
EnzymesNomenclature and classification of enzymes Holoenzyme, apoenzyme , cofactors, coenzyme, prosthetic groups ,metallo enzymes, monomeric and oligomeric enzymes Activation energy and transition state theory, enzyme activity, specific activity, common features of active sites, enzyme specificity: types and theories Factors affecting enzyme activity, E, S, temp and pH
Week 2
Role of NAD+ NADP+, Folic acid and Vit., B12 as biocatalystsFMN/FAD, Coenzyme A, Lipoic acid, biotin, tetrahydrofolateand metal ions as biocatalystsRole of thiamine pyrophosphate, pyridoxal phosphate as biocatalystsBiocatalysts from extreme thermophillic and hyperthermophilic archea and bacteria
Week 3 :
Enzyme substrate complex: Concept of E-S complex,binding sites, active site, specificity, kinetics of enzyme activityMichaelis- Menten equation and its derivationDifferent plots for the determination of KM and Vmax and their physiological significanceTwo substrate reactions (random, ordered and ping pong mechanisms), enzyme inhibition, types of inhibition, determination of Ki, suicide inhibitor
Week 4 :
Qualitative description of concerted and sequential modelsNegative cooperativityHalf site reactivityEnzyme regulation: Product inhibition, feedback control,covalent modification
Week 5 :
Enzyme- enzyme interaction , protein ligand bindingMeasurement analysis of binding isotherm, cooperativity, Hilland Scatchard plotsKinetics of allosteric enzymesAllosteric enzymes with special reference to aspartate transcarbomylase and posphofructokinase Week 6 :
Mechanism of enzyme action, general mechanistic principleTechniques for studying mechanism of actionFactors associated with catalytic efficiency, proximity, orientation and distortion of strainCollision and transition state theories, significance of activation energy and free energyAcid base. Nucleophilic and covalent catalysis Week 7 :
Isolation, crystallization and purification of enzymesTest of homogeneity of enzyme preparation, methods of enzyme analysisDetailed view of techniques for studying Enzyme assayChemical modification of active site groups, chymotrypsin, Lysozyme, RNase, Carboxypeptidase, GPDH, Aldolase, alcohol dehydrogenase 5 SubjectiveWeek 8 :
Zymogens and their activation (proteaseses and prothrombin)Isozymes :Multiple forms of enzymes with special reference tolactate dehydrogenaseMultienzyme complexes, RibozymesMultifunctional enzymes e.g., fatty acid synthase
Week 9 Enzyme technology: Methods for large scale production of enzymes, immobilized enzymes and their comparison with soluble enzymesMethods of immobilization of enzymes, immobilized enzyme reactorsApplication of immobilized and soluble enzymesin health and industry, application to fundamental studies of biochemistry, enzyme electrodesThermal stability and catalytic efficiency of enzyme, site directed mutagenesis and enzyme engineering-selected examples 5 SubjectiveWeek 10 :
Delivery system for protein pharmaceuticals, structure function relationship in enzymes, structural motifs and enzyme evolutionMethods of protein sequencingMethods for analysis of secondary and tertiary structure of enzymesProtein folding in vitro and in vivo
EnzymesNomenclature and classification of enzymes Holoenzyme, apoenzyme , cofactors, coenzyme, prosthetic groups ,metallo enzymes, monomeric and oligomeric enzymes Activation energy and transition state theory, enzyme activity, specific activity, common features of active sites, enzyme specificity: types and theories Factors affecting enzyme activity, E, S, temp and pH
Week 2
Role of NAD+ NADP+, Folic acid and Vit., B12 as biocatalystsFMN/FAD, Coenzyme A, Lipoic acid, biotin, tetrahydrofolateand metal ions as biocatalystsRole of thiamine pyrophosphate, pyridoxal phosphate as biocatalystsBiocatalysts from extreme thermophillic and hyperthermophilic archea and bacteria
Week 3 :
Enzyme substrate complex: Concept of E-S complex,binding sites, active site, specificity, kinetics of enzyme activityMichaelis- Menten equation and its derivationDifferent plots for the determination of KM and Vmax and their physiological significanceTwo substrate reactions (random, ordered and ping pong mechanisms), enzyme inhibition, types of inhibition, determination of Ki, suicide inhibitor
Week 4 :
Qualitative description of concerted and sequential modelsNegative cooperativityHalf site reactivityEnzyme regulation: Product inhibition, feedback control,covalent modification
Week 5 :
Enzyme- enzyme interaction , protein ligand bindingMeasurement analysis of binding isotherm, cooperativity, Hilland Scatchard plotsKinetics of allosteric enzymesAllosteric enzymes with special reference to aspartate transcarbomylase and posphofructokinase Week 6 :
Mechanism of enzyme action, general mechanistic principleTechniques for studying mechanism of actionFactors associated with catalytic efficiency, proximity, orientation and distortion of strainCollision and transition state theories, significance of activation energy and free energyAcid base. Nucleophilic and covalent catalysis Week 7 :
Isolation, crystallization and purification of enzymesTest of homogeneity of enzyme preparation, methods of enzyme analysisDetailed view of techniques for studying Enzyme assayChemical modification of active site groups, chymotrypsin, Lysozyme, RNase, Carboxypeptidase, GPDH, Aldolase, alcohol dehydrogenase 5 SubjectiveWeek 8 :
Zymogens and their activation (proteaseses and prothrombin)Isozymes :Multiple forms of enzymes with special reference tolactate dehydrogenaseMultienzyme complexes, RibozymesMultifunctional enzymes e.g., fatty acid synthase
Week 9 Enzyme technology: Methods for large scale production of enzymes, immobilized enzymes and their comparison with soluble enzymesMethods of immobilization of enzymes, immobilized enzyme reactorsApplication of immobilized and soluble enzymesin health and industry, application to fundamental studies of biochemistry, enzyme electrodesThermal stability and catalytic efficiency of enzyme, site directed mutagenesis and enzyme engineering-selected examples 5 SubjectiveWeek 10 :
Delivery system for protein pharmaceuticals, structure function relationship in enzymes, structural motifs and enzyme evolutionMethods of protein sequencingMethods for analysis of secondary and tertiary structure of enzymesProtein folding in vitro and in vivo
Taught by
Dr. Bashir Ahmad Ganai
