Enzymology Course

Lesson 1 • International Enzyme Classification System

Introduces the six major enzyme classes and the systematic numbering scheme used globally. Students can assign an enzyme to its correct class based on reaction type.

Lesson 2 • Thermodynamics of Catalysis

Explains free energy, activation energy, and transition states as they apply to enzyme-catalysed reactions. Grounds kinetic concepts introduced in later chapters.

Lesson 3 • The Active Site and Substrate Binding

Details how substrates are recognised and bound, covering lock-and-key and induced-fit models. Prepares students for mechanistic analysis in subsequent chapters.

Lesson 4 • What Enzymes Are and Do

Defines enzymes as biological catalysts and situates them within cellular metabolism. Provides the conceptual anchor for all subsequent mechanistic and kinetic content.

Lesson 1 • Multi-Substrate Reaction Mechanisms

Extends single-substrate kinetics to reactions involving two or more substrates. Students distinguish sequential and ping-pong mechanisms from kinetic patterns.

Lesson 2 • Reaction Rate Fundamentals

Covers initial velocity, substrate concentration dependence, and the concept of saturation kinetics. Establishes the experimental basis for all kinetic modelling.

Lesson 3 • Michaelis-Menten Equation Derivation

Walks through the steady-state assumption and mathematical derivation of the Michaelis-Menten equation. Students understand each parameter's physical meaning.

Lesson 4 • Graphical Analysis of Kinetic Data

Teaches linearisation methods and nonlinear regression for extracting kinetic constants from experimental data. Emphasises accuracy and pitfalls of each approach.

Lesson 1 • Uncompetitive and Mixed Inhibition

Covers inhibitors binding only the enzyme-substrate complex or both free enzyme and complex. Students distinguish these patterns using graphical and algebraic methods.

Lesson 2 • Reversible Inhibition Overview

Introduces the concept of reversible inhibition and its thermodynamic basis. Provides the framework for classifying competitive, uncompetitive, and mixed inhibition.

Lesson 3 • Irreversible and Mechanism-Based Inhibition

Analyses covalent modifiers and suicide substrates that permanently inactivate enzymes. Introduces kinact and partition ratio as key parameters.

Lesson 4 • Inhibition in Drug and Toxicology Contexts

Applies inhibition principles to pharmaceutical design and toxic compound analysis. Bridges mechanistic understanding to real-world enzyme-targeted applications.

Lesson 1 • Principles of Enzyme Regulation

Surveys the major regulatory strategies cells use to modulate enzyme activity rapidly and reversibly. Sets the stage for detailed mechanistic analysis in subsequent sections.

Lesson 2 • Allosteric Enzymes and Cooperativity

Explains how effector binding at non-active sites alters catalytic rate through conformational changes. Students apply the Hill equation to quantify cooperativity.

Lesson 3 • Covalent Modification and Zymogen Activation

Covers phosphorylation, adenylylation, and proteolytic cleavage as regulatory switches. Students trace activation cascades from zymogen to active enzyme.

Lesson 4 • MWC and KNF Allosteric Models

Compares the concerted Monod-Wyman-Changeux model with the sequential Koshland-Nemethy-Filmer model. Students evaluate which model fits experimental data.

Lesson 1 • Chromatographic Purification Strategies

Teaches ion exchange, size exclusion, hydrophobic interaction, and affinity chromatography principles. Students sequence chromatographic steps to maximise purification fold.

Lesson 2 • Assessing Purity and Yield

Explains SDS-PAGE, native PAGE, and activity staining for purity assessment alongside purification tables. Students calculate specific activity, yield, and purification fold.

Lesson 3 • Enzyme Stability and Storage

Addresses factors affecting enzyme stability during purification and long-term storage. Students formulate stabilising buffer conditions and select appropriate storage formats.

Lesson 4 • Precipitation and Bulk Fractionation

Introduces ammonium sulfate precipitation, isoelectric precipitation, and heat treatment as early purification steps. Students calculate saturation percentages and assess yield.

Lesson 1 • General Catalytic Strategies

Introduces acid-base, covalent, metal ion, and proximity-orientation catalysis as universal tools. Provides a mechanistic vocabulary applied throughout the chapter.

Lesson 2 • Oxidoreductase and Transferase Mechanisms

Examines electron transfer, hydride transfer, and group transfer reactions with cofactor involvement. Students connect cofactor chemistry to overall reaction stoichiometry.

Lesson 3 • Serine Proteases as a Model System

Uses the catalytic triad of serine proteases to illustrate concerted acid-base and covalent catalysis. Students trace each step of the acylation-deacylation cycle.

Lesson 4 • Isomerase and Ligase Mechanisms

Analyses intramolecular rearrangements and ATP-driven bond formation reactions. Students evaluate energy coupling strategies used by ligases.

Lesson 1 • Protein Data Bank and Structure Visualisation

Guides students through retrieving, visualising, and annotating enzyme structures from public databases. Students identify active site residues and ligand contacts in 3D viewers.

Lesson 2 • Molecular Docking and Virtual Screening

Covers docking algorithms, scoring functions, and their use in predicting inhibitor binding poses. Students evaluate docking results critically against experimental data.

Lesson 3 • Cryo-EM and NMR in Enzymology

Introduces cryo-electron microscopy and solution NMR as complementary structural methods. Students assess when each technique is preferred over crystallography.

Lesson 4 • X-Ray Crystallography of Enzymes

Explains crystal growth, data collection, and electron density map interpretation for enzyme structures. Students read crystallographic quality indicators such as R-factor and resolution.

Lesson 1 • Enzyme Engineering and Directed Evolution

Covers rational design, random mutagenesis, and high-throughput screening for improving enzyme properties. Students compare engineering strategies based on desired outcome.

Lesson 2 • Immobilised Enzyme Technology

Explains adsorption, covalent attachment, entrapment, and cross-linking immobilisation methods. Students evaluate stability, reusability, and activity retention trade-offs.

Lesson 3 • Bioreactor Design for Enzyme Processes

Introduces batch, fed-batch, and continuous bioreactor configurations for enzyme-catalysed production. Students calculate productivity and identify scale-up challenges.

Lesson 4 • Enzymes in Food and Agricultural Industries

Surveys amylases, proteases, lipases, and cellulases used in food processing and crop improvement. Students match enzyme properties to specific industrial process requirements.