NEET MDS Lessons
Biochemistry
FACTORS AFFECTING ENZYME ACTIVITY
Velocity or rate of enzymatic reaction is assessed by the rate of change in concentration of substrate or product at a given time duration. Various factors which affect the activity of enzymes include:
1. Substrate concentration
2. Enzyme concentration
3. Product concentration
4. Temperature 5. Hydrogen ion concentration (pH)
6. Presence of activators
7. Presence of inhibitor
Effect of substrate Concentration : Reaction velocity of an enzymatic process increases with constant enzyme concentration and increase in substrate concentration.
Effect of enzyme Concentration: As there is optimal substrate concentration, rate of an enzymatic reaction or velocity (V) is directly proportional to the enzyme concentration.
Effect of product concentration In case of a reversible reaction catalyzed by a enzyme, as per the law of mass action the rate of reaction is slowed down with equilibrium. So, rate of reaction is slowed, stopped or even reversed with increase in product concentration
Effect of temperature: Velocity of enzymatic reaction increases with temperature of the medium which they are most efficient and the same is termed as optimum temperature.
Effect of pH: Many enzymes are most efficient in the region of pH 6-7, which is the pH of the cell. Outside this range, enzyme activity drops off very rapidly. Reduction in efficiency caused by changes in the pH is due to changes in the degree of ionization of the substrate and enzyme.
Highly acidic or alkaline conditions bring about a denaturation and subsequent loss of enzymatic activity
Exceptions such as pepsin (with optimum pH 1-2), alkaline phosphatase (with optimum pH 9-10) and acid phosphatase (with optimum pH 4-5)
Presence of activators Presence of certain inorganic ions increases the activity of enzymes. The best examples are chloride ions activated salivary amylase and calcium activated lipases.
Effect of Inhibitors The catalytic enzymatic reaction may be inhibited by substances which prevent the formation of a normal enzyme-substrate complex. The level of inhibition then depends entirely upon the relative concentrations of the true substrate and the inhibitor
The basic characteristics of enzymes includes
(i) Almost all the enzymes are proteins and they follow the physical and chemical reactions of proteins (ii) Enzymes are sensitive and labile to heat
(iii) Enzymes are water soluble
(iv) Enzymes could be precipitated by protein precipitating agents such as ammonium sulfate and trichloroacetic acid.
Biotin
Biotin helps release energy from carbohydrates and aids in the metabolism of fats, proteins and carbohydrates from food.
RDA The Adequate Intake (AI) for Biotin is 30 mcg/day for adult males and females
Biotin Deficiency Biotin deficiency is uncommon under normal circumstances, but symptoms include fatigue, loss of appetite, nausea, vomiting, depression, muscle pains, heart abnormalities and anemia.
Pantothenic Acid
Pantothenic Acid is involved in energy production, and aids in the formation of hormones and the metabolism of fats, proteins, and carbohydrates from food.
RDA The Adequate Intake (AI) for Pantothenic Acid is 5 mg/day for both adult males and females.
Pantothenic Acid Deficiency
Pantothenic Acid deficiency is uncommon due to its wide availability in most foods.
Protein electrophoresis is a laboratory technique used to separate proteins based on their size, charge, or other physical properties. It plays a vital role in diagnosing and monitoring various diseases, especially those involving abnormal protein production or structure.
Types of Protein Electrophoresis
1. SPE (Serum Protein Electrophoresis)
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Principle: Separation of serum proteins based on their charge.
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Major Fractions:
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Albumin
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Alpha-1 globulin
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Alpha-2 globulin
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Beta globulin
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Gamma globulin
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Clinical Applications:
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Multiple Myeloma: Detects monoclonal spike (M-protein) in gamma region.
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Chronic Infections: Polyclonal increase in gamma globulins.
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Nephrotic Syndrome: Decreased albumin, increased alpha-2 globulin.
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Liver Disease: Altered albumin and beta-gamma bridging.
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2. Hemoglobin Electrophoresis
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Principle: Separation of hemoglobin variants based on charge differences.
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Common Hemoglobins:
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HbA, HbA₂, HbF, HbS, HbC, HbE
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Clinical Applications:
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Thalassemia: Elevated HbA₂ or HbF levels.
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Sickle Cell Disease: Presence of HbS.
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Hemoglobinopathies: Differentiates variants like HbC, HbE, etc.
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3. SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis)
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Principle: Proteins are denatured and separated by molecular weight.
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SDS: Imparts uniform negative charge, eliminating charge-based separation.
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Clinical Applications:
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Protein Purification: Identifies specific proteins in research and diagnostics.
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Genetic Disorders: Detects abnormal or truncated proteins.
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Muscular Dystrophies: Analyzes dystrophin protein expression.
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Cancer Research: Studies tumor markers and oncogenic proteins.
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COENZYMES
Enzymes may be simple proteins, or complex enzymes.
A complex enzyme contains a non-protein part, called as prosthetic group (co-enzymes).
Coenzymes are heat stable low molecular weight organic compound. The combined form of protein and the co-enzyme are called as holo-enzyme. The heat labile or unstable part of the holo-enzyme is called as apo-enzyme. The apo-enzyme gives necessary three dimensional structures required for the enzymatic chemical reaction.
Co-enzymes are very essential for the biological activities of the enzyme.
Co-enzymes combine loosely with apo-enzyme and are released easily by dialysis. Most of the co-enzymes are derivatives of vitamin B complex
CLASSIFICATION OF ENZYMES
1. Oxidoreductases : Act on many chemical groupings to add or remove hydrogen atoms. e.g. Lactate dehydrogenase
2. Transferases Transfer functional groups between donor and acceptor molecules. Kinases are specialized transferases that regulate metabolism by transferring phosphate from ATP to other molecules. e.g. Aminotransferase.
3. Hydrolases Add water across a bond, hydrolyzing it. E.g. Acetyl choline esterase
4. Lyases Add water, ammonia or carbon dioxide across double bonds, or remove these elements to produce double bonds. e.g. Aldolase.
5. Isomerases Carry out many kinds of isomerization: L to D isomerizations, mutase reactions (shifts of chemical groups) and others. e.g. Triose phosphate isomerase
6. Ligases Catalyze reactions in which two chemical groups are joined (or ligated) with the use of energy from ATP. e.g. Acetyl CoA carboxylase