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Biochemistry

Buffers           

• Biological systems use buffers to maintain pH.

• Definition: A buffer is a solution that resists a significant change in pH upon addition of an acid or a base.

• Chemically: A buffer is a mixture of a weak acid and its conjugate base

• Example: Bicarbonate buffer is a mixture of carbonic acid (the weak acid) and the bicarbonate ion (the conjugate base): H2CO3 + HCO3

• All OH- or H+ ions added to a buffer are consumed and the overall [H+ ] or pH is not altered

H2CO3 + HCO3 - + H+ <- -> 2H2CO3

H2CO3 + HCO3 -  +  OH<- -> 2HCO3  - + H2O

• For any weak acid / conjugate base pair, the buffering range is its pKa +1.

 

It should be noted that around the pKa the pH of a solution does not change appreciably even when large amounts of acid or base are added. This phenomenon is known as buffering. In most biochemical studies it is important to perform experiments, that will consume H+ or OH- equivalents, in a solution of a buffering agent that has a pKa near the pH optimum for the experiment.

Most biologic fluids are buffered near neutrality. A buffer resist a pH change and consists of a conjugate acid/base pair.

Important Physiological Buffers include carbonate (H2CO3/HCO3-),

Phosphate (H2PO-4 /HPO2-4) and various protiens

Glycogen Storage Diseases are genetic enzyme deficiencies associated with excessive glycogen accumulation within cells.

  • When an enzyme defect affects mainly glycogen storage in liver, a common symptom is hypoglycemia (low blood glucose), relating to impaired mobilization of glucose for release to the blood during fasting.
  • When the defect is in muscle tissue, weakness and difficulty with exercise result from inability to increase glucose entry into Glycolysis during exercise.

Various type of Glycogen storage disease are

Type

Name

Enzyme Deficient

I

Von Geirke’s Disease

Glucose -6-phosphate

II

Pompe’s Disease

(1, 4)glucosidase

III

Cori’s Disease

Debranching Enzymes

IV

Andersen’s Disease

Branching Enzymes

V

McArdle’s Disease

Muscles Glycogen Phosphorylase

Niacin: Vitamin B3, Nicotinamide, Nicotinic Acid Niacin, or vitamin B3,

 is involved in energy production, normal enzyme function, digestion, promoting normal appetite, healthy skin, and nerves.

RDA Males: 16 mg/day; Females: 14 mg/day

Niacin Deficiency : Pellagra is the disease state that occurs as a result of severe niacin deficiency. Symptoms include cramps, nausea, mental confusion, and skin problems.

Enzyme Kinetics

Enzymes are protein catalysts that, like all catalysts, speed up the rate of a chemical reaction without being used up in the process. They achieve their effect by temporarily binding to the substrate and, in doing so, lowering the activation energy needed to convert it to a product.

The rate at which an enzyme works is influenced by several factors, e.g.,

  • the concentration of substrate molecules (the more of them available, the quicker the enzyme molecules collide and bind with them). The concentration of substrate is designated [S] and is expressed in unit of molarity.
  • the temperature. As the temperature rises, molecular motion - and hence collisions between enzyme and substrate - speed up. But as enzymes are proteins, there is an upper limit beyond which the enzyme becomes denatured and ineffective.
  • the presence of inhibitors.
    • competitive inhibitors are molecules that bind to the same site as the substrate - preventing the substrate from binding as they do so - but are not changed by the enzyme.
    • noncompetitive inhibitors are molecules that bind to some other site on the enzyme reducing its catalytic power.
  • pH. The conformation of a protein is influenced by pH and as enzyme activity is crucially dependent on its conformation, its activity is likewise affected.

The study of the rate at which an enzyme works is called enzyme kinetics.

Applications of the Henderson-Hasselbalch equation

• Calculate the ratio of CB to WA, if pH is given

• Calculate the pH, if ratio of CB to WA is known

• Calculate the pH of a weak acid solution of known concentration

• Determine the pKa of a WA-CB pair

• Calculate change in pH when strong base is added to a solution of weak acid. This is represented in a titration curve

• Calculate the pI

By rearranging the above equation we arrive at the Henderson-Hasselbalch equation:

pH = pKa + log[A-]/[HA]

It should be obvious now that the pH of a solution of any acid (for which the equilibrium constant is known, and there are numerous tables with this information) can be calculated knowing the concentration of the acid, HA, and its conjugate base [A-].

At the point of the dissociation where the concentration of the conjugate base [A-] = to that of the acid [HA]:

pH = pKa + log[1]

The log of 1 = 0. Thus, at the mid-point of a titration of a weak acid:

pKa = pH

In other words, the term pKa is that pH at which an equivalent distribution of acid and conjugate base (or base and conjugate acid) exists in solution.

 

CALCIUM

Total calcium in the human body is 1 to 1.5kg, out of which 99% is seen in bone and 1% in extracellular fluid. The main source of calcium is milk.

The daily requirement of calcium for child is 1200mg/day and for adult it is 500mg/day. During pregnancy /lactation the calcium requirement is 1500mg/day.

The absorption of calcium takes place in 1st and 2nd part of deuodenum. Calcium absorption requires carrier protein, helped by Ca2+ - dependent ATpase.

Factors responsible for increase in calcium absorption include Vitamin D, Parathyroid hormone, acidity and amino acids. Factors such as phytic acid,oxalates, malabsorption  syndromes and Phosphates decreases calcium absorption. The normal calcium level in blood is 9-11mg/dl.

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