GLYCOLYTIC PATHWAY D-Glucose is a major fuel for most organisms.

DGlucose metabolism occupies the center position for all metabolic pathways. Glucose contains a great deal of potential energy. The complete oxidation of glucose yields −2,840 kJ/mol of energy. Glucose + 6O2 6CO2 + 6H2O Definition: Breakdown of glucose into pyruvate in aerobic condition or lactate in anaerobic condition. splitting"),a molecule of glucose is degraded in a series of enzyrnecatalyzed reactions to yield two molecules of the three-carbon compound pyruvate. During the sequential reactions of glycolysis, some of the free energy released from glucose is conserved in the form of ATP and NADH Source: Dietary sources and stored glycogen Site: Cytosol of all cells. The glycolytic pathway is often referred to as the Embfen- Meyerhof pathway in honor of the two of the three biochemical pioneers. It is the pathway by which glucose is converted via fructose-1,6-bisphosphate to pyruvate with the generation of 2 mol of ATP per mole of glucose. Under aerobic conditions, the pyruvate formed by glycolysis is further oxidized by the citric acid cycle (Chapter 21) and oxidative phosphorylation (Chapter 22) to CO2 and water. Under anaerobic conditions, however, the pyruvate is instead converted to a reduced end product, which is lactate in muscle (homolactic fermentation; a fermentation is an anaerobic biological reaction process) and ethanol _ CO2 in yeast (alcoholic fermentation). Glycolysis has two phases



IMPORTANCE OF HEXOKINASE In glycolysis pathway, glucose is first phosphorylated to glucose-6-phosphate by an enzyme hexokinase which is present in almost all the tissues. It catalyses the phosphorylation of various hexoses like fructose, mannose has low Km (about 0.1 mM) for substrates. Glucokinase which is also an additional enzyme helps in phosphorylation of glucose to glucose 6 phosphate present in liver. It has high Km for glucose (10 mM). Hexokinase has a higher affinity for glucose than glucokinase. Its functions are to supply glucose to the tissues even in low blood glucose concentration. It can catalyze the phosphorylation of other hexoses but at a slower rate than glucose. Hexokinase requires Mg2+ for activity. The true substrate for this enzyme is not ATP4- but MgATP2-. Hexokinase is the poster enzyme for the induced fit mechanism where substrate binding induces conformational changes in the enzyme. Hexokinase is found in all cells of all organisms.

Isozymes of hexokinase are found in yeast and mammals. Glucokinase is an isozyme of hexokinase sometimes called Hexokinase IV (other times called Hexokinase D). Glucokinase is found in hepatocytes (liver cells). The hexokinase isozyme found in skeletal muscle has a Km for glucose of 0.1 mM. The skeletal muscle maintains steady state concentration of glucose around 4 mM. This isozyme is allosterically inhibited by high concentrations of glucose-6-phosphate. The isozyme glucokinase catalyzes the same reaction but has a very high Km for glucose of 10.0 mM. This enzyme is not allosterically regulated by high glucose-6-phosphate concentrations. The high Km, means that glucokinase only becomes metabolically important when glucose levels are high. This enzyme produces glucose-6-phosphate which is then stored by the liver in the form of glycogen. Glucokinase is an inducible enzyme. The amount of this enzyme present in hepatocyte cells is regulated by insulin. 2.REGULATION OF GLYCOLYSIS
The three enzymes namely hexokinase,phosphofructo kinase and pyruvate kinase catalyzing the irreversible reactions regulate glycolysis.

• Glucokinase is a molecular sensor of high glucose levels Four hexokinase genes have been identified in humans (hexokinase I-IV), all of which are capable of converting glucose to glucose-6-P at the expense of ATP hydrolysis (step 1 of glycolysis). hexokinase I (reaction 1), which has a high affinity for substrate (Km for glucose is ~0.1mM), is expressed in all tissues, phosphorylates a variety of hexose sugars, and is inhibited by the product of the reaction, glucose-6-P. In contrast, hexokinase IV, also known as glucokinase, has a low affinity for substrate (Km for glucose is ~10mM), is highly specific for glucose, is expressed primarily in liver and pancreatic cells and is not inhibited by glucose-6-P. This difference in tissue expression and glucose affinity between hexokinase and glucokinase plays an important role in controlling blood glucose levels, which ultimately controls rates of glycolytic flux in all cells by limiting substrate availability. Since blood glucose levels are maintained around 5mM, significant levels of glucose phosphorylation by glucokinase only occur under conditions of high glucose, such as after consuming a carbohydrate-rich meal. Moreover, since glucokinase is not inhibited by glucose-6-P, it is able to continue functioning even if flux through glycolysis cannot keep up with product formation. The role of glucokinase in liver cells is to trap the extra glucose that is available from the diet so that it can be stored as glycogen for an energy source later. Allosteric control of phosphofructokinase activity

Of all the enzymes in glycolysis, phosphofructokinase is the best characterized because of its vital role in controlling flux through the pathway. There are actually two phosphofructokinase isozymes (distinct genes that encode proteins with similar functions), phosophofructokinase-1 (PFK-1) which catalyzes reaction 3 in glycolysis, and phosphofructokinase-2 (PFK-2) a bifunctional enzyme that catalyzes the synthesis of fructose-2,6-bisophosphate (F-2,6-BP), a potent allosteric regulator of PFK1 activity .
Rate limiting step: Phosphofructukinase (PFK) is the most important regulatory enzyme in glycolysis which catalyses the rate limiting committed step. Allosteric inhibitors: Phosphofructokinase is an allosteric enzyme regulated by allosteric effectors.ATP; citrate and H+ ions (low pH) are the most important allosteric inhibitor. Allosteric activators: Fructose 2, 6 bisphosphate, ADP, AMP and Pi are the allosteric activators. Fructose 2, 6 bis phosphate is the most positive allosteric effector of phosphofructokinase-1. It relieves inhibition of phosphofructokinase-1 by ATP and increases affinity for fructose 6 phosphate. Fructose 2, 6 bis phosphate is formed by phosphorylation of fructose 6 phosphate by phosphofructokinase-2 which is also responsible for its breakdown since it contains fructose 2,6 bis phosphatase activity. In the superfluity of glucose, the concentration of fructose 2,6 bis phosphate increases stimulating glycolysis by activating phosphofructokinase-1 and inhibiting fructose 1,6 bis phosphatase. In glucose shortage, gluconeogenesis is stimulated by a decrease in the concentration of fructose 2, 6 bis phosphate which deactivates phosphofructokinase- 1 and deinhibits fructose 1,6 bis phosphatase.

Pyruvate kinase also regulates glycolysis. This enzyme is inhibited by ATP and

activated by Fructose 1,6bisphosphate.pyruvate kinase is active in dephosphorylated state and inactivated in phosphorylated state. Inactivation of pyruvate kinase by phosphorylation is brought about by cAMP-dependent proteinkinase.the hormone glucagon inhibits hepatic glycolysis by this mechanism.