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10 Steps of Glycolysis
By Regina Bailey
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Glycolysis literally means "splitting sugars." In glycolysis, glucose (a six carbon

sugar) is split into two molecules of a three-carbon sugar. Glycolysis yields two
molecules of ATP ( free energy containing molecule), two molecules of pyruvic acid
and two "high energy" electron carrying molecules of NADH. Glycolysis can occur
with or without oxygen. In the presence of oxygen, glycolysis is the first stage of
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glycolysis allows cells to make small
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Glycolysis

10 Steps of Glycolysis

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EDUCATIO

Step 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 glucose producing glucose 6phosphate .

Glucose (C 6 H 12 O 6 ) + hexokinase +

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ATP ADP + Glucose 6-phosphate (C 6 H

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O6P1)

Step 2
The enzyme phosphoglucoisomerase converts glucose 6-phosphate into its isomer
fructose 6-phosphate. Isomers have the same molecular formula , but the atoms of
each molecule are arranged differently.

Glucose 6-phosphate (C 6 H 11 O 6 P 1 ) + Phosphoglucoisomerase Fructose 6phosphate (C 6 H 11 O 6 P 1 )

Step 3
The enzyme phosphofructokinase uses another ATP molecule to transfer a
phosphate group to fructose 6-phosphate to form fructose 1, 6-bisphosphate.

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Fructose 6-phosphate (C 6 H 11 O 6 P 1 ) + phosphofructokinase + ATP ADP +

Fructose 1, 6-bisphosphate (C 6 H 10 O 6 P 2 )
Step 4

The enzyme aldolase splits fructose 1, 6-bisphosphate into two sugars that are

EDUCATIO

The enzyme aldolase splits fructose 1, 6-bisphosphate into two sugars that are
isomers of each other. These two sugars are dihydroxyacetone phosphate and
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Fructose 1, 6-bisphosphate (C 6 H 10 O 6 P
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Glyceraldehyde phosphate (C 3 H 5 O 3 P 1
)
Step 5

The enzyme triose phosphate isomerase rapidly inter-converts the molecules

dihydroxyacetone phosphate and glyceraldehyde phosphate. Glyceraldehyde
phosphate is removed as soon as it is formed to be used in the next step of
glycolysis.

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Dihydroxyacetone phosphate (C 3 H 5 O 3 P 1 ) Glyceraldehyde phosphate (C 3 H 5

O3P1)
Net result for steps 4 and 5: Fructose 1 , 6-bisphosphate (C 6 H 10 O 6 P 2 ) 2
molecules of Glyceraldehyde phosphate (C 3 H 5 O 3 P 1 )
Step 6
The enzyme triose phosphate dehydrogenase serves two functions in this step. First
the enzyme transfers a hydrogen (H - ) from glyceraldehyde phosphate to the
oxidizing agent nicotinamide adenine dinucleotide (NAD + ) to form NADH. Next
triose phosphate dehydrogenase adds a phosphate (P) from the cytosol to the
oxidized glyceraldehyde phosphate to form 1, 3-bisphosphoglycerate. This occurs
for both molecules 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 (C 3 H 5 O 3
P 1 ) 2 molecules of 1,3-bisphosphoglycerate (C 3 H 4 O 4 P 2 )
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 1,3bisphosphoglycerate. The process yields two 3-phosphoglycerate molecules and
two ATP molecules.
2 molecules of 1,3-bisphoshoglycerate (C 3 H 4 O 4 P 2 ) + phosphoglycerokinase + 2
ADP 2 molecules of 3-phosphoglycerate (C 3 H 5 O 4 P 1 ) + 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 (C 3 H 5 O 4 P 1 ) + phosphoglyceromutase 2
molecules of 2-Phosphoglycerate (C 3 H 5 O 4 P 1 )
Step 9
The enzyme enolase removes a molecule of water from 2-phosphoglycerate to form
phosphoenolpyruvic acid (PEP). This happens for each molecule of 2phosphoglycerate.
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2 molecules of 2-Phosphoglycerate (C H O P ) + enolase 2 molecules of

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2 molecules of 2-Phosphoglycerate (C 3 H 5 O 4 P 1 ) + enolase 2 molecules of

phosphoenolpyruvic acid (PEP) (C 3 H 3 O 3 P 1 )
Step 10
The enzyme pyruvate kinase transfers a P from PEP to ADP to form pyruvic acid and
ATP. This happens for each molecule of PEP. This reaction yields 2 molecules of
pyruvic acid and 2 ATP molecules.
2 molecules of PEP (C 3 H 3 O 3 P 1 ) + pyruvate kinase + 2 ADP 2 molecules of
pyruvic acid (C 3 H 4 O 3 ) + 2 ATP

Summary
In summary, a single glucose molecule in glycolysis produces a total of 2 molecules
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. If 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 .

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