Carbohydrate Metabolism Overview Citric Acid Cycle Overview Metabolism & Energy Summary Elmhurst College Glycolysis Summary

Pyruvic Acid - Crossroads Chemistry Depar tment Glycolysis Reactions Glycogenesis / Glycogenolysis / Gluconeogenesis Virtual ChemBook

Click for larger image Glycolysis Reactions Introduction to Glycolysis: The overall reaction of glycolysis which occurs in the cytoplasm is represented simply as: C6H12O6 + 2 NAD+ + 2 ADP + 2 P -----> 2 pyruvic acid, (CH3(C=O)COOH + 2 ATP + 2 NADH + 2 H+ At this time, concentrate on the fact that glucose with six carbons is converted into two pyruvic acid molecules with three carbons each. Only a net "visible" 2 ATP are produced from glycolysis. The 2 NADH will be considered separately late r. The major steps of glycolysis are outlined in the graphhic on the left. There ar e a variety of starting points for glycolysis; although, the most usual ones sta rt with glucose or glycogen to produce glucose-6-phosphate. The starting points for other monosaccharides, galactose and fructose, are also shown. Glycolysis - with white background for printing Overview of Metabolism Link to: Great Animation of entire Glycolysis - John Kyrk Link to: Interactive Glycolysis (move cursor over arrows) Jim Hardy, Professor of Chemistry, The University of Akron. Link to Glycolysis Aninmation 1 Link to Glycolysis Aninmation 2

Reaction 1: Phosphate Ester Synthesis Phosphate is added to the glucose at the C-6 position. The reaction is a phospha te ester synthesis using the alcohol on the glucose and a phosphate from ATP. This first reaction is endothermic and thus requires energy from a coupled react ion with ATP. ATP is used by being hydrolyzed to ADP and phosphate giving off en ergy and the phosphate for reaction with the glucose for a net loss of ATP in th e overall glycolysis pathway.

Hydrolysis: ATP + H2O --> ADP + P + energy P = PO4-3; ATP = adenine triphosphate;ADP = adenine diphosphate This reaction is catalyzed by hexokinase. Off-site chime link: Boyer Tutorial - Hexokinase Reaction 1 - Chime in new window

Reaction 2: Isomerization The glucose-6-phosphate is changed into an isomer, fructose-6-phosphate. This me ans that the number of atoms is unchanged, but their positions have changed. This works because the ring forms may open to the chain form, and then the aldeh yde group on glucose is transformed to the keone group on fructose. The ring the n closes to form the fructose-6-phosphate. This reaction is catalyzed by phosphoglucoisomerase. Off-site chime link: Phosphoglucoisomerase Reaction 2 - Chime in new window

Reaction 3: Phosphate ester synthesis This reaction is virtually identical to reaction 1 The fructosee-6-phosphate has an alcohol group on C-1 that is reacted with phosphate from ATP to make the pho sphate ester on C-1. Again this reaction is endothermic and thus requires energy from a coupled react ion with ATP. ATP is used by being hydrolyzed to ADP and phosphate giving off en ergy and the phosphate for reaction with the glucose for a net loss of ATP in th e overall glycolysis pathway. Hydrolysis: ATP + H2O --> ADP + P + energy This reaction is catalyzed by phosphofructokinase. Off-site chime link: Phosphofructokinase Link to: Rodney Boyer Animation of Phosphofructokinase Reaction 3 - Chime in new window

Reaction 4: Split Molecule in half The six carbon fructose diphophate is spit into two three-carbon compounds, an a ldehyde and a ketone. The slit is made between the C-3 and C-4 of the fructose. The ring also opens at the anomeric carbon. The product on the right is the glyceraldehyde. Technically this is called a reverse aldol condensation.

This reaction is catalyzed by aldolase. Off-site chime link: Aldolase Reaction 4 - Dihydroxyacetonephosphate Chime in new window Reaction 4 - Glyceraldehyde-3-phosphate Chime in new window

Reaction 4A: Isomerization The dihydroxyacetone phosphate must be converted to glyceraldehyde-3-phosphate t o continue the glycolysis reactions. This reaction is an isomerization between t he keone group and an aldehyde group. As a result of this reaction, all of the remaining glycolysis reactions are carr ied out a second time. The first series of reactions occurs with the first glyce raldehyde molecule from the orginal split. Then the second series of reactions o ccurs after the isomerization of the dihydroxyacetone into the glyceraldehyde. This reaction is catalyzed by triose phosphate isomerase. Off-site chime link: Triose Phosphate Isomerase (TIM) Reaction 4A - Isomerization Chime in new window

Reaction 5: Oxidation/Phosphate Ester Synthesis This reaction is first an oxidation involving the coenzyme NAD+. The aldehyde is oxidized to an acid as an intermediate through the conversion of NAD+ to NADH + H+. Then an inorganic phosphate is added in a phosphate esteer synthesis. This and all remaining reactions occur twice for each glucose-6-phosphate (six c arbons), since there are now two molecules of 3-carbons each. This reaction is catalyzed by glyceraldehyde-3-phosphate. Off-site chime link: G3P Dehydrogenase Reaction 5 - 1,3-diphosphoglycerate Chime in new window

Reaction 6: Hydrolysis of Phosphate; Synthesis of ATP One of the phosphate groups undergoes hydrolysis to form the acid and a phosphat e ion, giving off energy. This first energy producing reaction is coupled with t he next endothermic reaction making ATP. The phosphate is transferred directly t o an ADP to make ATP. This reaction is catalyzed by phosphoglycerokinase. Off-site chime link: Phosphoglycerate Kinase Reaction 6 - 3-phosphoglycerate Chime in new window

Reaction 7: Isomerization In this reaction the phosphate group moves from the 3 position to the 2 position in an isomerization reaction. This reaction is catalyzed by phosphoglycerate mutase. Off-site chime link: Phosphoglycerate Mutase Reaction 7 - 2-phosphoglycerate Chime in new window

Reaction 8: Alcohol Dehydration In this reaction, which is the dehydration of an alcohol, the -OH on C-3 and the -H on C-2 are removed to make a water molecule. At the same time a double bond forms between C-2 and C-3. This change makes the compound somewhat unstable, but energy for the final step of glycolysis. This reaction is catalyzed by enolase. Off-site chime link: Enolase Reaction 8 - phosphoenol pyruvic acid Chime in new window

Reaction 9:Phosphate Ester Hydrolysis; Synthesis of ATP This is the final reaction in glycolysis. Again one of the phosphate groups unde rgoes hydrolysis to form the acid and a phosphate ion, giving off energy. This f irst energy producing reaction is coupled with the next endothermic reaction mak ing ATP. The phosphate is transferred directly to an ADP to make ATP. This reaction is catalyzed by pyruvic kinase. Off-site chime link: Pyruvate Kinase Reaction 9 - pyruvic acid Chime in new window Conclusion: Starting with glucose-6-phosphate with 6 carbons, the final result of the glycol ysis reactions is two molecules of pyruvic acid, since reaction 5-9 are each car ried out twice. tep 1 The enzyme hexokinase phosphorylates (adds a phosphate group to) glucose in the cell's cytoplasm. In the process, a phosphate group from ATP is transferred to g lucose producing glucose 6-phosphate. Glucose (C6H12O6) + hexokinase + ATP ? ADP + Glucose 6-phosphate (C6H11O6P1) Step 2 The enzyme phosphoglucoisomerase converts glucose 6-phosphate into its isomer fr uctose 6-phosphate. Isomers have the same molecular formula, but the atoms of ea

ch molecule are arranged differently. Glucose 6-phosphate (C6H11O6P1) + Phosphoglucoisomerase ? Fructose 6-phosphate ( C6H11O6P1) Step 3 The enzyme phosphofructokinase uses another ATP molecule to transfer a phosphate group to fructose 6-phosphate to form fructose 1, 6-bisphosphate. Fructose 6-phosphate (C6H11O6P1) + phosphofructokinase + ATP ? ADP + Fructose 1, 6-bisphosphate (C6H10O6P2) Step 4 The enzyme aldolase splits fructose 1, 6-bisphosphate into two sugars that are i somers of each other. These two sugars are dihydroxyacetone phosphate and glycer aldehyde phosphate. Fructose 1, 6-bisphosphate (C6H10O6P2) + aldolase ? Dihydroxyacetone phosphate ( C3H5O3P1) + Glyceraldehyde phosphate (C3H5O3P1) Step 5 The enzyme triose phosphate isomerase rapidly inter-converts the molecules dihyd roxyacetone phosphate and glyceraldehyde phosphate. Glyceraldehyde phosphate is removed as soon as it is formed to be used in the next step of glycolysis. Dihydroxyacetone phosphate (C3H5O3P1) ? Glyceraldehyde phosphate (C3H5O3P1) Net result for steps 4 and 5: Fructose 1, 6-bisphosphate (C6H10O6P2) ? 2 molecul es of Glyceraldehyde phosphate (C3H5O3P1) Step 6 The enzyme triose phosphate dehydrogenase serves two functions in this step. Fir st the enzyme transfers a hydrogen (H-) from glyceraldehyde phosphate to the oxi dizing agent nicotinamide adenine dinucleotide (NAD+) to form NADH. Next triose phosphate dehydrogenase adds a phosphate (P) from the cytosol to the oxidized gl yceraldehyde phosphate to form 1, 3-bisphosphoglycerate. This occurs for both mo lecules of glyceraldehyde phosphate produced in step 5. A. Triose phosphate dehydrogenase + 2 H- + 2 NAD+ ? 2 NADH + 2 H+ B. Triose phosphate dehydrogenase + 2 P + 2 glyceraldehyde phosphate (C3H5O3P1) ? 2 molecules of 1,3-bisphosphoglycerate (C3H4O4P2) Step 7 The enzyme phosphoglycerokinase transfers a P from 1,3-bisphosphoglycerate to a molecule of ADP to form ATP. This happens for each molecule of 1,3-bisphosphogly cerate. The process yields two 3-phosphoglycerate molecules and two ATP molecule s. 2 molecules of 1,3-bisphoshoglycerate (C3H4O4P2) + phosphoglycerokinase + 2 ADP ? 2 molecules of 3-phosphoglycerate (C3H5O4P1) + 2 ATP Step 8 The enzyme phosphoglyceromutase relocates the P from 3-phosphoglycerate from the

third carbon to the second carbon to form 2-phosphoglycerate. 2 molecules of 3-Phosphoglycerate (C3H5O4P1) + phosphoglyceromutase ? 2 molecule s of 2-Phosphoglycerate (C3H5O4P1) Step 9 The enzyme enolase removes a molecule of water from 2-phosphoglycerate to form p hosphoenolpyruvic acid (PEP). This happens for each molecule of 2-phosphoglycera te. 2 molecules of 2-Phosphoglycerate (C3H5O4P1) + enolase ? 2 molecules of phosphoe nolpyruvic acid (PEP) (C3H3O3P1) Step 10 The enzyme pyruvate kinase transfers a P from PEP to ADP to form pyruvic acid an d ATP. This happens for each molecule of PEP. This reaction yields 2 molecules o f pyruvic acid and 2 ATP molecules. 2 molecules of PEP (C3H3O3P1) + pyruvate kinase + 2 ADP ? 2 molecules of pyruvic acid (C3H4O3) + 2 ATP Summary In summary, a single glucose molecule in glycolysis produces a total of 2 molecu les of pyruvic acid, 2 molecules of ATP, 2 molecules of NADH and 2 molecules of water. Although 2 ATP molecules are used in steps 1-3, 2 ATP molecules are generated in step 7 and 2 more in step 10. This gives a total of 4 ATP molecules produced. I f you subtract the 2 ATP molecules used in steps 1-3 from the 4 generated at the end of step 10, you end up with a net total of 2 ATP molecules produced. For a detailed view of the 10 steps, see: Details of the 10 Steps of Glycolysis.