GLYCOLYSIS

- Also known EMBDEN-MEYERHOFF PATHWAY

- Anaerobic process
- Glucose Metabolism
- FIRST STAGE of glucose metabolism.
- One molecule of glucose is converted to fructose-1,6-bisphosphate,
which gives rise to two molecules of pyruvate
- It plays a key role in the way organisms extract energy from nutrients
ONCE PYRUVATE IS FORMED, IT HAS ONE OF SEVERAL FATES

FATES OF PYRUVATE
•Anaerobic glycolysis
- happens in muscles
- overuse of muscle use
•Aerobic Oxidation
- Citric Acid Cycle (Kreb’s Cycle)
•Anaerobic alcoholic fermentation
- happens in fungi and yeast
- glucose to alcohol eg. industrial products, beer

STEP-BY-STEP GLYCOLYTIC REACTION

FIRST PHASE
• Phosphorylation of glucose to give glucose-6-phosphate
• Isomerization of glucose-6-phosphate to give fructose-6-phosphate
• Phosphorylation of fructose-6-phosphate to yield fructose-1,6-bisphosphate
• Cleavage of fructose-1,6,-bisphosphate to give
glyceraldehyde-3-phosphate and dihyroxyacetone
phosphate
• Isomerization of dihyroxyacetone phosphate to give glyceraldehyde-3-phosphate

2ND PHASE
• Oxidation of glyceraldehyde-3-phosphate
give 1,3-bisphosphoglycerate
• Transfer of a phosphate group from 1,3-bisphosphoglycerate to ADP to give 3-phosphoglycerate
• Isomerization of 3-phosphoglycerate to give 2-phosphoglycerate
• Dehydration of 2-phosphoglycerate to give
phosphoenolpyruvate
• Transfer of a phosphate group from
phosphoenolpyruvate to ADP to give pyruvate

GLUCOSE IN CYCLIC 3D will be in the circulation and enter through specific channels IN and OUT of the cell:
4 TYPES OF GLUT CHANNELS:

TYPE 1 BBB KIDNEY

BLOOD
FETUS TYPE 4 MaPa
BLOOD BRAIN BARRIER MUSCLE
FAT
TYPE 2 KLiP
KIDNEY
LIVER
PANCREAS

TYPE 3 PlaNK
PLACENTA
NEURONS
Conversion of Glucose to Glyceraldehyde-3-Phosphate
• The "Preparation Phase" of Glycolysis
• Conversion of glucose to glyceraldehyde-3-phosphate.
In the first phase of glycolysis, five reactions convert a molecule of glucose to two molecules of glyceraldehyde-3-phosphate.

STEP 1:
PHOSPHORYLATION of glucose
Substrate: GLUCOSE
Product: GLUCOSE-6-PHOSPHATE & ADP
• In step 1 of glycolysis, glucose is phosphorylated to give glucose-6-phosphate
• The reaction is endergonic, as it is driven by the free energy of hydrolysis of ATP
- ENDERGONIC - free flowing energy needs ATP
Enzymes:
Glucokinase – an enzyme in liver
Hexokinase – an enzyme in muscles
Reaction: ATP will be removed with one phosphate and will become ADP = linear glucose, phosphate will attach to the 6th carbon

STEP 2:
ISOMERIZATION
Substrate: GLUCOSE-6-PHOSPHATE (Aldehyde)
Product: FRUCTOSE-6-PHOSPHATE (Ketone)
Details: The second reaction of glycolysis is the rearrangement of glucose 6-phosphate (G6P)
into fructose 6-phosphate (F6P) by glucose phosphate isomerase (Phosphoglucose Isomerase).
PROCESS:
The second step of glycolysis involves the conversion of glucose-6-phosphate to fructose-6-phosphate (F6P).
This reaction occurs with the help of the enzyme GLUCOSEPHOSPHATE ISOMERASE (GI).
As the name of the enzyme suggests, this reaction involves an isomerization reaction.
The reaction involves the rearrangement of the carbon-oxygen bond to transform the six-membered ring
into a five-membered ring. To rearrangement takes place when the six-membered ring opens and then
closes in such a way that the first carbon becomes now external to the ring.

STEP 3:
Phosphorylation of fructose-6-phosphate
PRODUCT: fructose-1,6-bisphosphate.
Enzyme: Phosphofructokinase - key regulator enzyme

• Fructose-6-phosphate is then phosphorylated again to generate fructose-1,6-bisphosphate

• This is the second reaction to be coupled to ATP hydrolysis
•ATP will give off one phosphate and will become ADP
•Phosphate will attach to first Carbon and will yield FRUCTOSE-1,6-BISPHOSPHATE

BISPHOSPHATE: There are groups of carbons in between 2 phosphate groups.

BIPHOSPHATE: 2 groups of phosphate attached together
BLACK ARROW means IRREVERSIBLE = KEY-REGULATORY ENZYME

STEP 4:
Cleavage of fructose-1,6-bisphosphate
PRODUCT: glyceraldehyde-3-phosphate and dihyroxyacetone phosphate
ENZYME: ALDOLASE
•Fructose-1,6-bisphosphate is split into two 3-carbon fragments
- will give off 2 products
• Reaction catalyzed by aldolase
- Dihydroxyacetone phosphate (Ketone) phosphate attaches to 1st carbon
- Glyceraldehyde-3-phosphate (Aldehyde) phosphate attaches to 3rd carbon
STEP 5:
Isomerization of dihyroxyacetone phosphate
PRODUCT: glyceraldehyde-3-phosphate
ENZYME: Triosephosphate isomerase
•In step 5, dihydroxyacetone phosphate (DHAP) is converted to glyceraldehyde-3-phosphate
•These compounds are trioses.
*Glyceraldehyde phosphate is removed/used in the next step of Glycolysis.
Details:
GAP is the only molecule that continues in the glycolytic pathway.
As a result, all of the DHAP molecules produced are further acted on
by the enzyme Triosephosphate isomerase (TIM), which reorganizes the DHAP into GAP
so it can continue in glycolysis. At this point in the glycolytic pathway, we have two 3-carbon molecules
but have not yet fully converted glucose into pyruvate.

*AFTER THE FIRST PHASE, TWO GLYCERALDEHYDE-3-PHOSPHATE ARE PRODUCED

SECOND PHASE
Glyceraldehyde-3-Phosphate is Converted to Pyruvate
• The second phase of Glycolysis

STEP 6:
OXIDATION of glyceraldehyde-3-phosphate
PRODUCT: 1,3-bisphosphoglycerate
ENZYME: Glyceraldehyde-3-phosphate Dehydrogenase
• This reaction involves the addition of a phosphate group, as well as an electron transfer
•Addition of phosphate from INORGANIC PHOSPHATE (iP is stable due to H bond)
•iP will be removed with H from enzyme Gly-3-Phos Dehydrogenase (dehydration)
• The oxidizing agent, NAD+ is reduced to NADH
•Phosphate from iP will then attach to the 1st carbon

STEP 7:
Transfer of a phosphate group from 1,3-bisphosphoglycerate to ADP
PRODUCT: 3-phosphoglycerate
ENZYME: Phosphoglycerate Kinase
• 1,3-bisphosphoglycerate is converted to 3-phosphoglycerate
•This step involves another reaction in which ATP is produced by phosphorylation of ADP
• 1,3-bisphosphoglycerate transfers a phosphate
group to ADP. This is known as substrate-level phosphorylation
• Reaction is catalyzed by phosphoglycerate kinase
Co-factor: Mg2+
Details:
This reaction involves the loss of a phosphate group from the starting material. The phosphate is transferred to a molecule of ADP
that yields our first molecule of ATP. Since we actually have two molecules of 1,3 bisphoglycerate (because there were two 3-carbon
products from stage 1 of glycolysis), we actually synthesize two molecules of ATP at this step. With this synthesis of ATP, we have
canceled the first two molecules of ATP that we used, leaving us with a net of 0 ATP molecules up to this stage of glycolysis.

STEP 8:
Isomerization of 3-phosphoglycerate
PRODUCT: 2-phosphoglycerate
ENZYME: Phosphoglycerate Mutase (PGM)
•The next step involves the isomerization of 3-phosphoglycerate to 2-phosphoglycerate
• This reaction is catalyzed by phosphoglyceromutase

Details: The enzyme phosphoglyceromutase relocates the Phosphate from 3- phosphoglycerate from the 3rd carbon to the 2nd carbon
to form 2-phosphoglycerate. A mutase is an enzyme that catalyzes the transfer of a functional group from one position on a molecule
to another.
STEP 9:
Dehydration of 2-phosphoglycerate
PRODUCT: phosphoenolpyruvate
ENZYME: ENOLASE
•Next, 2-phosphoglycerate loses one molecule of water, producing phosphoenolpyruvate
• Enolase catalyzes the reaction
Details:
The enzyme enolase removes a molecule of water from 2-phosphoglycerate to form phosphoenolpyruvic acid (PEP).
*For it to be stable~ make it double bond
This step involves the conversion of 2 phosphoglycerates to phosphoenolpyruvate (PEP). The reaction is catalyzed by the enzyme
enolase. Enolase works by removing a water group or dehydrating the 2 phosphoglycerates.

STEP 10:
Transfer of a phosphate group from phosphoenolpyruvate to ADP
PRODUCT: pyruvate
ENZYME: PYRUVATE KINASE
•Phosphenolpyruvate (PEP) transfers its phosphate group to ADP, producing ATP and pyruvate
Details:
The final step of glycolysis converts phosphoenolpyruvate into pyruvate with the help of the enzyme pyruvate kinase. This reaction
involves the transfer of a phosphate group. The phosphate group attached to the 2′ carbon of the PEP is transferred to a molecule of
ADP, yielding ATP. Again, since there are two molecules of PEP, here we actually generate 2 ATP molecules.
Steps 1 and 3 = – 2ATP
Steps 7 and 10 = + 4 ATP
Net “visible” ATP produced = 2.

GLYCOLYSIS
CYTOPLASM
GLUCOSE - STARTING SUBSTRATE
2 PYRUVATE - END PRODUCT
GROSS - 4 ATP
USED - 2 ATP
NET -2ATP
2 NADH
ANAEROBIC

CONTROL POINTS
Three reactions exhibit particularly large decreases in free energy; the enzymes that catalyze these reactions are sites of allosteric control:
• Hexokinase
•Phosphofructokinase
•Pyruvate kinase