Syllabus topics (RTMNU)
- Lipids: Definition, Properties and role of fatty acids, oils and waxes
- Degradation of fats (β-Oxidation and Glyoxylic acid cycle)
- Enzymology: Definition, Characteristics (Properties), and classification of enzymes
- Basic concepts of enzymology: Holoenzyme, Apoenzyme, Prosthetic group, Co-enzyme, Co-factor, Active site and Isoenzyme
- Mechanism of enzyme action: Enzyme-substrate complex theory, Lock and key model, Induced fit model
- Enzyme inhibitors: Definition, Competitive and non-competitive inhibition
Lipids: Definition, Properties and Role of Fatty Acids, Oils and Waxes
Definition: Lipids are a group of organic compounds that are insoluble in water but soluble in organic solvents such as ether, chloroform and benzene. They are esters of fatty acids with alcohols (mainly glycerol).
Fatty acids are long-chain carboxylic acids (R–COOH), usually with an even number of carbon atoms (C12–C24).
- Saturated — no double bond, e.g. palmitic acid (C16), stearic acid (C18); solid at room temperature.
- Unsaturated — one or more C=C double bonds, e.g. oleic (1), linoleic (2), linolenic acid (3); liquid at room temperature.
Oils and fats are triglycerides (triacylglycerols): one glycerol + three fatty acids joined by ester bonds. Oils (liquid, rich in unsaturated fatty acids) are common in seeds — groundnut, sunflower, mustard, castor.
Waxes are esters of long-chain fatty acids with long-chain monohydric alcohols, e.g. cutin and suberin-associated waxes, carnauba wax on palm leaves.
Properties: insoluble in water; lower density than water; saponification (hydrolysis with alkali gives soap + glycerol); hydrogenation of unsaturated oils gives solid fat (vanaspati); rancidity on oxidation; high energy content (about 9 kcal/g).
Role in plants:
- Stored food in seeds (energy reserve) — used during germination.
- Structural part of membranes (phospholipids, glycolipids).
- Waxes form the cuticle and reduce water loss (transpiration) and protect from pathogens.
- Source of hormones and pigments (carotenoids are lipid-derived).
Degradation of Fats: β-Oxidation and Glyoxylate (Glyoxylic Acid) Cycle
First, lipase hydrolyses a triglyceride → glycerol + 3 fatty acids. Glycerol enters glycolysis (as dihydroxyacetone phosphate).
β-Oxidation (in glyoxysomes in germinating oily seeds, also mitochondria): the fatty acid is broken down two carbons at a time from the carboxyl end; oxidation happens at the β-carbon (C-3).
- Activation: fatty acid + CoA + ATP → fatty acyl-CoA (acyl-CoA synthetase).
- Oxidation (dehydrogenation): acyl-CoA → trans-Δ2-enoyl-CoA; FAD → FADH2.
- Hydration: + H2O → L-3-hydroxyacyl-CoA.
- Oxidation: → 3-ketoacyl-CoA; NAD+ → NADH.
- Thiolysis: + CoA → acetyl-CoA + acyl-CoA shorter by 2 C.
The cycle repeats. Palmitic acid (C16) gives 8 acetyl-CoA after 7 rounds.
Glyoxylate cycle (in glyoxysomes of germinating fatty seeds like castor and groundnut): converts acetyl-CoA into succinate and then sugar. It bypasses the two CO2-releasing steps of Krebs cycle using two special enzymes — isocitrate lyase (isocitrate → succinate + glyoxylate) and malate synthase (glyoxylate + acetyl-CoA → malate).
Net: 2 acetyl-CoA → succinate. Succinate → mitochondria → oxaloacetate → phosphoenolpyruvate → glucose (gluconeogenesis) → sucrose transported to the growing seedling.
Enzymology: Definition, Characteristics and Classification of Enzymes
Definition: Enzymes are biological catalysts, mostly proteins, that speed up biochemical reactions without being used up. (Ribozymes are RNA enzymes.)
Characteristics: (1) protein in nature; (2) high catalytic efficiency — small amount needed; (3) specificity for substrate; (4) lower activation energy; (5) do not change equilibrium; (6) sensitive to temperature (optimum around 25–40 °C, denatured by heat); (7) sensitive to pH; (8) can be inhibited; (9) reversible action in many cases.
IUB classification (6 classes):
| Class | Reaction | Example |
|---|---|---|
| 1. Oxidoreductases | Oxidation–reduction | Dehydrogenases, catalase |
| 2. Transferases | Transfer of groups | Hexokinase, transaminase |
| 3. Hydrolases | Hydrolysis | Amylase, lipase, protease |
| 4. Lyases | Removal of groups without hydrolysis | Aldolase, decarboxylase |
| 5. Isomerases | Rearrangement within molecule | Phosphohexose isomerase |
| 6. Ligases | Joining two molecules using ATP | DNA ligase, PEP carboxykinase |
Basic Concepts: Holoenzyme, Apoenzyme, Prosthetic Group, Co-enzyme, Co-factor, Active Site and Isoenzyme
- Apoenzyme: protein part of an enzyme; inactive alone.
- Co-factor: non-protein part needed for activity. It may be a metal ion (Mg2+, Zn2+, Fe2+) or an organic molecule.
- Holoenzyme = apoenzyme + co-factor (complete active enzyme).
- Prosthetic group: co-factor tightly (often covalently) bound, e.g. haem in catalase, FAD in succinate dehydrogenase.
- Co-enzyme: organic co-factor loosely bound, often a vitamin derivative, e.g. NAD+, NADP+, CoA.
- Active site: small pocket on the enzyme where the substrate binds and the reaction takes place.
- Isoenzymes (isozymes): different molecular forms of an enzyme that catalyse the same reaction, e.g. lactate dehydrogenase (5 forms), peroxidase isozymes in plants.
Mechanism of Enzyme Action: E–S Complex, Lock and Key, Induced Fit
Enzyme–substrate complex theory (Michaelis and Menten): E + S ⇌ ES → E + P. The enzyme binds the substrate to form a temporary ES complex, lowers the activation energy, and releases the product; the enzyme is free again.
Lock and key model (Emil Fischer, 1894): the active site is rigid and exactly fits the substrate, like a key in a lock. Explains specificity, but not flexibility.
Induced fit model (Koshland, 1958): the active site is flexible; binding of substrate changes the shape of the enzyme so that it fits closely, like a glove on a hand. Explains broad specificity and inhibition better.
Enzyme Inhibitors: Competitive and Non-competitive Inhibition
Inhibitor: a substance that reduces or stops enzyme activity.
| Basis | Competitive | Non-competitive |
|---|---|---|
| Structure of inhibitor | Similar to substrate | Not similar |
| Binding site | Active site | Another site (allosteric) |
| Effect of more substrate | Inhibition removed | Not removed |
| Km / Vmax | Km increases, Vmax same | Km same, Vmax decreases |
| Example | Malonate inhibits succinate dehydrogenase | Cyanide, heavy metals (Hg2+, Ag+) |
Likely exam questions
- (Long, 8 marks) Describe β-oxidation of fatty acids and the glyoxylate cycle in germinating oily seeds.
- (Long, 8 marks) Define enzymes. Give their characteristics and IUB classification, and explain the mechanism of enzyme action.
- (Short, 4 marks) Distinguish between competitive and non-competitive inhibition.
- (Short, 4 marks) Explain apoenzyme, co-enzyme and prosthetic group.
- (Short, 4 marks) Write the role of waxes in plants.