CH 17 Glycolysis

1. Learning outcomes:
a. Gain an understanding of the two phases of glycolysis, the prep phase and the
payoff phase. And the net high energy compounds that are produced.
b. Be able to identify the enzyme if you are given the reaction.
c. Understand what type of reaction each class of enzymes catalyzes.
d. Understand the points of control and inhibitors and the energetics.
e. Recognize oxidation reduction reactions in glycolysis and recognize the reducing
and oxidizing agents that partner with the reaction.
f. What are the fates of pyruvate that is made?
2. The overall pathway of Glycolysis
a. Glycolysis is the first stage of glucose metabolism
b. One molecule of glucose is converted to fructose-1,6-bisphosphate, which give
rise to two molecules of pyruvate.
c. It plays a key role in the way organisms extract energy from nutrients
d. One pyruvate is formed and it has one of several fates.
3. Organophosphates important in producing energy

a.
4. Fates of Pyruvate from Glycolysis
a. Glycolysis
b. Reactant : glucose, 2ATP, 2NAD​+​, 4 ADP +2P
c. Products: 2 ADP, 4 ATP, 2 NADH, 2 Pyruvate
d. Enzyme: ​Glyceraldehyde-3-phosphate dehydrogenase
i. Turns fructose-1,6 bisphosphate to 2 pyruvate
a. 2 pyruvate product from glycolysis can go through
i. Aerobic glycolysis
1. 2 NADH+2 Pyruvate→ NAD​+​+2 Lactate
2. Enzyme: Lactate dehydrogenase
ii. Aerobic Oxidation
1. Citric acid cycle
 2. Oxidative phosphorylation
a. 2 NADH, 6 O2 , 30 ADP turn into 2 NAD​+​, 30 ATP
iii. Anaerobic alcoholic Fermentation
1. 2NADH to turn 2 NAD​+
2. Producing 2CO​2​+2 ethanol
3. Enzyme :Alcohol dehydrogenase

5.
6.
7. Glycolysis steps
a. First priming reaction
i. ATP-->ADP
b. Second Priming Reaction
i. ATP-->ADP
c. Cleavage of 6-carbon sugar phosphate to the 3-carbon sugar phosphates
i. Fructose 1,6-bisphosphate---> Glyceraldehyde 3-phosphate
d. Oxidation and phosphorylation
i. (2 )Glyceraldehyde 3-phosphate +2P + 2NAD​+​ ----->1,3-
Biphosphoglycerate + 2 NADH +H+
e. First ATP- forming reaction (substrate-level phosphorylation)
i. (2) 1,3- bisphoglycerate +2 ADP----> 2 ATP +3 phosphoglycerate (2)
f. Reshapes to 2- phophoglycerate (2)
g. Loses 2- H20
h. Second ATP Forming reaction
i. Phosphophenol Pyruvate (2) +2ADP---> 2 Pyruvate + 2 ATP
8. Glycolysis and Gluconeogenesis
 a. Glucose---> 2 pyruvate Glycolysis
b. Lysis- destruction
c. Genesis- formation
d. Gucogenesis: 2 Pyruvate---> glycolysis
9. Glycolysis: The preparatory phase: Conversion of Glucose to
Glyceraldehyde-3-phosphate
a. Phosphorylation of glucose to give glucose-6-phosphate
b. Isomerization of glucose-6-phosphate to give fructose-6-phosphate
c. Phosphorylation of fructose-6-phosphate to yield fructose-1,6-bisphosphate
10. In chemistry i​ somerization​ or isomerisation is the process in which a molecule, ion or
molecular fragment is transformed into an isomer with a different chemical structur
11. Glycolysis: the preparatory phase: Conversion of Glucose to glyceraldehyde-3-phosphate
a. Cleavage of fructose-1,6,-bisphosphate to give glyceraldehyde-3-phosphate and
dihydroxyacetone phosphate
b. Isomerization of dihydroxyacetone phosphate to give glyceraldehyde-3-phosphate
12. Reactions of Glycolysis and their standard free-energy changes

a.
13. ▵G= ▵G​’°​+RTlnQ
14. Key features of the preparatory phase of glycolysis step 1 Point of control
a. First step is irreversible. This is a point of control. ▵​ G​’°​ is large and negative
and driver by ATP.
b. ​▵G​’°​= -33.9 kj/mol
 c.
d. Hexokinase I m​ uscle also hexokinase II and III
e. Hexokinase IV liver also called glucokinase
f. Kinase is an ATP dependent enzyme that transfers a phosphate from ATP
to a substrate.
15. Hexokinase

a.
i. Inactive
ii. Active
b. Glucose causes and induced fit. The conformational change occurs when
glucose and ATP binds.
16. Isozymes
a. Note that sigmoidicity for hexokinase IV and the much lower K​m​ for
hexokinase I. When blood glucose rises above 5mM, hexokinase IV
activity increases, but hexokinase I is already operating near V​max​ and
cannot respond to an increase in glucose concentration.
b. Hexokinase I is already operating near V​max​ ​ and cannot respond to an
increase in glucose concentration. Hexokinases, I, II and III have similar
kinetic properties.
c. Hexokinase vs Glucokinase
d. Hexokinase
i. Lower Km (higher affinity) lower vmax
ii. Tissue
iii. Work in low glucose level or hypoglycemia (fasted)
e. Glucokinase
f. High Km ( low affinity), higher vmax
g. Liver
h. hyperglycemia(fed)
i.

i.
ii. Hexokinase Isozymes
1. All but glucokinase phosphorylate glucose and other 6
carbon sugars
2. Glucokinase phosphorylates only glucose.
iii. Hexokinase I, II and III are inhibited by glucose-6-phosphate when it
builds up in the system. Most V versus S curves are hyperbolic and
do obey Michaelis Menton Kinetics.
iv. Hexokinase IV exhibits a sigmoidal V versus S curve. This is
puzzling because hexokinase has only one lobe. Hexokinase IV is
controlled via a regulatory protein.
v. A regulatory protein inhibits hexokinase 1V when bound. High
concentrations of fructose 6 phosphate cause hexokinase to enter
the nucleus and bind. High concentrations of glucose stimulate the
protein to release hexokinase into the cytosol.
 j.
k. When Fructose-6- phosphate is high the regulatory protein draws it into
the nucleus. When glucose is high hexokinase IV is released into the
cytosol.
17. Key features of The preparatory phase of glycolysis step 2
**​
​ ​G’​o​. ​△G​ = -2.92 kJ/mol
a. Step 2 is reversible and has a positive △ ​

b.
18. Phosphohexose isomerase two identical monomers 2 active site, His 388 and
Glu 357 for acid base reactions

a.
i. Phosphohexose isomerase
b. Isomerases catalyze the transformation of compounds into their
positional isomers.
19. Step 3 point of control
 a. Step 3 is irreversible. This is a point of control. ​△​G’​o ​is large and
negative and driven by ATP

b.
c. ​ ​△​G0​​ ’ =-14.2 kJ/mol. ​ ​△​G**​
​ =-18.8 kJ/mol
d. Phosphofructokinase is a second regulatory enzyme in glycolysis.
e. ATP is an allosteric effector; high levels inhibit the enzyme, low
levels activate it.
f. Fructose-2,6-bisphosphate is also an allosteric effector; it activates
the enzyme, without F26BP phosphofructokinase in inactive.
g. Kinase phosphorylates molecules by ATP.
20. Fructose 2,6-bisphosphate

21.
22. Phosphofructokinase
 a.
23. ATP is an allosteric effector; high levels inhibit the enzyme, low
levels activate it​.

a.
b. Step 3 point of control
i. The second priming reaction and the first commitment
ii. ATP is the donor of the second phosphate group
iii. This is an irreversible step
iv. The product, fructose 1,6-bisphosphate is committed to
become pyruvate and yield energy
v. Phosphofructokinase-1 is negatively regulated by high levels
of ATP
1. Do not burn glucose if there is plenty of ATP this
process is irreversible.
c. Key features of the preparatory phase of Glycolysis step 4
i.

.​
a. Step 4 is reversible.​ ​△G​’0​​ is positive but the reactants
are present in low concentrations. ​ ​△G​’0​​ = ​23.8 kj/mol.
△G​**​= -.23kj/mol

b.
c. Key features in step 4
i. Animal and plant aldolases employ
covalent catalysis; this is a covalent bond
forming between enzyme and substrate.
ii. Fungal and bacterial aldolases employ
metal ion catalysis.
iii. Aldolases catalyze reversible aldol
condensation. Aldolases Cleave 6 carbon
sugar.
d. Reverse of an aldol condensation
 i.
ii. Reverse of an aldol condensation
2. Aldolase

a.
3. Key feature in step 5 of the preparatory phase of
glycolysis.
a. The reaction is reversible. △G​’0​ is positive. GAP
is continuously used up. △G​’0​=+7.5 kJ/mol.
△G​**​= +2.41kJ/mol

b.
c. Aldolase creates two triose phosphates: DAP and
GAP
 d. Only GAP is the substrate for the next enzyme
(step 6)
e. DAP is converted enzymatically to GAP
f. Triose phosphate isomerase conversion
g. Isomerases catalyze the transformation of
compounds into their positional isomers.

4.
5. Triosephosphate isomerase
24. First stage Summary
a. In the first stages of glycolysis, glucose is converted to two
molecules of glyceraldehyde-3-phosphate.
b. The key intermediate in this series of reactions is
fructose-1,6-bisphosphate.The enzyme that catalyzes this reaction,
phosphofructokinase, is subject to allosteric control
c. 2 molecules of ATP were used.
d.
e. Key features of the payoff phase of glycolysis step 6

i.
ii. △G​’0​= 2(+6.3 kJ/mol). △G​**​= 2(-1.29kJ/mol)
iii. △G​’0​= 6.3kJ/mol
iv. Very high energy of hydrolysis: -49.3 kJ/mol
 f.
25. Key features of step 6
a. Oxidation of glyceraldehyde-3-phosphate and phosphorylation to
give 1,3-bisphosphoglyerate
b. Reversible reaction producing 2 NADH
c. Positive △G​’0
d. △G​’0​= 2(+6.2 kJ/mol). △G​**​= 2(-1.29 kJ/mol)

e. First energy releasing step in glycolysis

f. Oxidation of aldehyde with NAD​+ ​gives NADH
g. Phosphorylation yields an high energy reaction product
h. Glyceraldehyde-3-phosphate dehydrogenase
i. Glyceraldehyde-3- phosphate dehydrogenase
j. Dehydrogenase catalyze the removal of pair of hydrogens atom from
substrates.
 k.
l. Glyceraldehyde-3-phosphate dehydrogenase 4 monomers (there is some
allosteric regulation)
26. Step 7
a. Transfer of a phosphate from 1,3-bisphosphoglycerate to ADP to give
3-phosphoglycerate and ATP.
b. This reaction is the sum of the endergonic phosphorylation of ADP and the
exergonic hydrolysis of the mixed phosphate anhydride. This reaction is
reversible, ​ △G​’0​=2 (-18.8 kJ/mol)= -37 kJ/mol
1. △G​**​=2( +.1kJ/mol)=+.2 kJ/mol
 ii. X2
c. 1,3-bisphosphoglycerate is a high-energy compound that can donate
the phosphate group to ADP to make ATP.
d. The reaction is reversible, the reverse process transfer of phosphate
from ATP to phosphoglycerate
e. Kinases are enzymes that transfer phosphate groups from molecules like ATP to
various substrates
f. First substrate level phosphorylation.
g. In substrate level phosphorylation the source of the phosphorus is inorganic
phosphate ion, not ATP.
h. Substrate phosphorylation is phosphorylation of ADP or another nucleoside 5’
diphosphate. coupled to a dehydrogenation of an organic substrate. Steps 6 and 7
couple:
i. Glyceraldehyde-3-phosphate +​ADP​ +Pi +NAD​+​↔ 3-glycerate + ATP + NADH +
H​+
 o​
27. DG’​ = (2)(+6.2 kJ/mol) + (2) ( -18.8 kJ/mol)
28. =+12.4 kJ/mol -37.6kJ/mol = -24.6 kJ/mol
**​
29. DG​ =(2)(-1.29kJ/mol) + (2) (+0.1 kJ/mol)
30. -2.58 kJ/mol + 0.3 kJ/mol = -2.38 kJ/mol

31.
a. Phosphoglycerate kinase 2 lobes but 1 monomer
32. Step 8
a. This is a reversible isomerization reaction enzymes that shift
functional groups are called ​mutases
b. Conversion of 3- phosphoglycerate to 2-phosphoglycerate
c. The eighth step involves the isomerization of 3-phosphoglycerate to
2-phosphoglycerate
d. This reaction is catalyzed by phosphoglyceromutase
e. The reaction is reversible and ▵G​’0​ is positive
f. ▵G​’0​= 2(+4.4 kJ/mol)= +8.8 kJ/mol
g. ▵G​**​= 2(+.83kJ/mol)= 1.76 kJ/mol

h.
i. Mutases catalyze the transportation of functional groups.
j. Phosphoglycerate mutase employs covalent catalysis
k. One of the active site histidines is post-translationally modified to
phosphohistidine.
l. Phosphohistidine donates its phosphate to C2 before retrieving
another phosphate from C3.

m.
n. Notice that the phosphate from the substrate ends up bound to the
enzyme at the end of the reaction
o. The 2 negative charges in the product are fairly close now but
3-phosphoglycerate is not a good enough phosphate donor.

p.
 q. Phosphoryl transfer from C-3 of the substrate to the first active site
His. The second active-site His acts as a general acid catalyst.

33.
34. Phosphoglycerate mutase 2 monomers

35. Step 9
a. Dehydration of 2-phosphoglycerate
b. The goal here is to ​create a better phosphoryl donor
c. Loss of phosphate from 3-phosphoglycerate would merely give a primary alcohol
with no further stabilization.
d. The reaction is reversible.
e. DG’​0 ​is positive and = 2(+1.8 kJ/mol)= +3.6 kJ/mol
f. DG​**​= 2X (1.1 kJ/mol)= 2.2 kJ/mol

g.
h. 2-phosphoglycerate loses one molecule of water, producing phosphenolpyruvate.
Phosphoenolpyruvate contains a high energy bond
i. Enolase catalyzes the reaction and requires a Mg​2+​ cofactor
j. Enolase belongs to the class Lyase. Lyases catalyze removal of a group from a
molecule to form a double bond, or addition of a group to a double bond
36.
a. Enolase 2 monomer
37. Step 10 point of control
a. Pyruvate kinase catalyzes the conversion of phosphoenolpyruvate to pyruvate
b. Substrate level phosphorylation
c. The reaction is irreversible.
d. DG’​0 ​negative and = 2X-31.4 kJ/mol)=- 62.8 kJ/mol
e. DG= 2X(-23.0 kJ/mol)= -46 kJ/mol

f. X2
g. Second substrate-level phosphorylation
h. ...but loss of phosphate from phosphoenolpyruvate yields an enol that
tautomerizes into ketone
i. The tautomerization effectively lowers the concentration of the reaction product
and drives the reaction toward ATP formation. .
 i. Tautomers are isomers of a compound which differ only in the
position of the protons and electrons.

j.
38. Pyruvate Kinase is subject to regulation
a. requires divalent metals (Mg​++​ or Mn​++​) for activity
b. Under physiological conditions, the activity of pyruvate kinase is limited by
the level of Mg​++​.
c. Kinase is an ATP dependent enzyme that transfers a phosphate from ATP
to a substrate.
d. When there is plenty of ATP, the Mg ions are sequestered by ATP; this
slows down pyruvate kinase.
e. Increased concentration of metabolites in the glycolytic pathway slow
down glucose utilization

f.
g. Pyruvate kinase 3 monomers
39. Control Points in Glycolysis
a. The three reaction exhibit particularly large decrease in free energy; the
enzymes are:
i. 1. Hexokinase
1. Glucokinase exhibits sigmoidal V versus S curves. This is not
well understood.
2. Hexokinase exhibits substrate level control and follows
michaelis menten kinetics.
ii. 2. Phosphofructokinase
iii. 3. Pyruvate kinase
40. The pay off phase Glyceraldehyde-3-phosphate is converted to Pyruvate
a. Pyruvate is the end product of the second phase of glycolysis.
b. For each glucose molecule, two ATP are consumed in the preparatory
phase, giving a net yield of two ATP per molecule of glucose converted to
pyruvate
c. Keep in mind that each phosphoryl group, represented here as P, has two
negative charges (-PO​3​2-​).
i. You also regenerate 2 NADH
41. Glycolysis final stage
a. In the final stages of glycolysis, two molecules of pyruvate are produced for each
molecule of glucose that enters the pathway
b. These reactions involve electron transfer, and the net production of two ATP for
each glucose
c. There are three control points in the glycolytic pathway
42. Aerobic metabolism of pyruvate to lactate
a. •​Under anaerobic conditions, the most important pathway for the regeneration of
NAD​+​ is reduction of pyruvate to lactate
b. •​Lactate dehydrogenase (LDH) is a tetrameric isoenzyme consisting of H and M
subunits; H​4​ predominates in heart muscle, and M​4​ in skeletal muscle
c. •​Dehydrogenases catalyze the removal of pairs of hydrogen atoms from
substrates.
 1. ⃤ G= -25.5 kJ/mol

43.
1. Lactate dehydrogenase 4 monomers
44. Alcoholic fermentation in Yeast
● Two reactions lead to the production of ethanol
○ Decarboxylation of pyruvate to acetaldehyde
○ Reduction of acetaldehyde to ethanol
● Decarboxylase remove carboxyl groups and give off carbon dioxide.
● Dehydrogenases catalyze the removal of pairs of hydrogen atoms from
substrate.

●
●
○ Pyruvate Decarboxylation 4 monomers

●
○ Alcohol dehydrogenase 2 monomers
● Pyruvate summary
○ Pyruvate is converted to lactate in anaerobic tissues, such as actively
metabolizing muscle. NAD​+​ is recycled in the process.
○ In some organisms (yeast), pyruvate is converted to ethanol in a
process requiring thiamine pyrophosphate as a coenzyme.
● Energy Production in Glycolysis
○ Glycolysis is exergonic and releases 73.4kJ/mole glucose converted
to 2 moles of pyruvate.
○ Phosphorylation is also involved as 2 moles of ADP are converted to
ATP.
● Opposing pathways of glycolysis and glucogenesis in rat liver.
 ○ The reactions of glycolysis are on the left side, in red; the opposing
pathway of gluconeogenesis is on the right, in blue. The major sites
of regulation of gluconeogenesis shown here.
○ We explore gluconeogenesis in the next chapter.
Chapter 17 Homework:
1. Which reactions require ATP and which enzymes are responsible for those
reactions?
a. Phosphorylation of glucose to give glucose-6-phosphate by ​hexokinase
and glucokinase.

i.
Hexokinase reaches one half its maximum velocity in converting substrate to product
At at Km 10​-5​M
● Km is ½ Vmax

●
After meals when blood glucose may reach 10​-2​M these concentrations would swamp
the hexokinase ( 1000X its preferred concentration of glucose )and thus conversion of
the glucose to glucose-6P would be slow and blood glucose would remain elevated for
very long times.
In addition to hexokinase, the liver has another enzyme , glucokinase , that has a Km of
10​-​2​M so that even after meals it can easily convert glucose to glucose -6P as the blood
levels are right in the range of the Km of glucokinase. Since the liver is the organ
responsible for maintaining blood glucose it makes sense that it would be the source of
Glucokinase.
1000X difference in Km between glucokinase and hexokinase

b. Phosphorylation of fructose-6-phosphate to give fructose-1,6-bisphosphate

by ​phosphofructokinase.
i.

2.
3. Which reactions produce ATP and which enzymes are responsible?
a. Substrate level phosphorylation
i. Transfer of phosphate group from 1,3- bisphosphoglycerate to ADP
by phosphoglycerate kinase

a.
ii. Transfer of phosphate group from phosphoenolpyruvate to ADP by
pyruvate kinase.
 iii.
4. Which reactions require NADH and which enzymes are
responsible?
a. Reduction of pyruvate to lactate by lactate
dehydrogenase
i.

b. Reduction of acetaldehyde to ethanol by alcohol

dehydrogenase
 i.
c. Pyruvate can be converted to lactate, ethanol or
acetyl-coA.

i.
1. Citric acid cycle produces acetyl coA
5. Which reaction requires NAD​+​: and which enzyme are
responsible
 a.
i. Oxidation of glyceraldehyde-3-phosphate to give
1,3-diphosphoglycerate done by enzyme
glyceraldehyde-3-phosphate dehydrogenase.
6. What are the metabolic fates of pyruvate?
a. Pyruvate can be converted to lactate, ethanol or acetyl
CoA.
7. What is the origin of the name aldolase?
a. Aldolase catalyzes the reverse aldol condensation of
fructose-1,6-bisphosphate to
 glyceraldehyde-3-phosphate and dihydroxyacetone
phosphate.

i.
8. Why is fructose-1,6-bisphosphate the committed step?
a. Fructose-1,6-bisphosphate can only undergo the
reactions of glycolysis. The components of the pathway
up to this point can have other metabolic fates.
i. Glucose 6-phosphate and fructose 6-phosphate
can be used in other pathways.
9. Show that the reaction Glucose-->2
glyceraldehyde-3-phosphate is slightly endergonic and not
too far from equilibrium. Add the △G​0​ mol​-1​ values for the
reactions from glucose to glyceraldehyde-3-phosphate.
a. To find △G​0 ​ we need to know the △G​0 ​ for each step.
 b.

10. How does ATP act as an allosteric effector in phosphofructokinase?

a. ATP inhibits phosphofructokinase, consistent with the fact that ATP is produced
by later reactions of glycolysis.
 i. A
1. High ATP sigmoidal
2. Low ATP hyperbolic
11. Which of the enzymes are NADH-linked dehydrogenases?
a. NADH-linked dehydrogenases: glyceraldehyde-3-phosphate dehydrogenase,
lactate dehydrogenase, and alcohol dehydrogenase.
 b.
12. What is substrate level phosphorylation?
a. The free energy of hydrolysis of a substrate is the energetic driving force in
substrate-level phosphorylation. An example is the reaction
1,3-bisphosphoglycerate +ADP ---> 3 phosphoglycerate +ATP . In substrate level
phosphorylation the source of the phosphorus is inorganic phosphate ion, not
ATP. Substrate phosphorylation is phosphorylation of ADP or another nucleoside
5’ diphosphate. Coupled to a dehydrogenation of an organic substrate. Step 6
and 7 couple:
i. Glyceraldehyde-3-phosphate +ADP + Pi +NAD​+​<----> 3-glycerate + ATP
+NADH +H​+
△G​0​ = -12.2 kJ/mol
ii.

b. These are the two reactions that are substrate

phosphorylation reactions
13. Which reactions are control points in glycolysis?
a. The control points in glycolysis are the reaction catalyzed by hexokinase,
phosphofructokinase and pyruvate kinase.
i. If there is more glucose than hexokinase will activate more quickly
1. If there is more glucose 6-phosphate the reaction inhibited
ii. If there is more phosphopyruvate than pyruvate kinase will activate
more quickly
1. If pyruvate is more pyruvate kinase will be inhibited
 iii. If there is more fructose 6-phosphate phosphofructokinase is more
activated
1. If more fructose 1,6 bisphosphate is made the reaction will
be inhibiteed
b. They represent the speed of the reaction, inhibitors and activators.
14. Control points in glycolysis
a. Three reactions exhibit particularly large decreases in free energy; the
enzymes that catalyze these reactions are sites of allosteric control
i. Hexokinase
ii. Phosphofructokinase
iii. Pyruvate kinase
15. In glycolysis which molecules are inhibitors and which are activators?
a. Hexokinase I, II, III are inhibited by glucose-6-phosphate. A regulatory
protein controls response in Hexokinase IV.
b. Phosphofructokinase is inhibited by ATP and citrate.
c. Pyruvate kinase is inhibited by ATP, acetyl-CoA and alanine.
Fructose-1,6-bisphosphate.inhibits this enzyme
d. Phosphofructokinase is activated by AMP and activated by fructose-2,6-
bisphosphate. Without F26BP the enzyme is inactive. ATP inhibits
phosphofructokinase.
e. Pyruvate kinase is stimulated by AMP and fructose-1,6-bisphosphate.
16. Discuss the logic of the nature of allosteric inhibitors and activators of
glycolysis:
a. ATP is an inhibitor of several steps of glycolysis as well as other catabolic
pathways. The purpose of catabolic pathways is to produce energy , and
high levels of ATP mean the cell has sufficient energy.
b. Glucose-6-phosphate inhibits hexokinase I,II, and III. Hexokinase IV is
controlled by a regulatory protein. And is an example of product inhibition.
If glucose-6-phosphate level is high, it may indicate that sufficient glucose
is available from glycogen breakdown or that the subsequent enzymatic
steps of glycolysis are going slowly. Either way, there is no reason to
produce more glucose-6- phosphate.
c. Phosphofructokinase is activated by a special effector molecule,
fructose-2,6-bisphosphate, whose levels are controlled by hormones.it is
also inhibited by citrate which indicates that there is sufficient energy
from the citric acid cycle, probably from fat and amino acid degradation.
d. Pyruvate Kinase is also inhibited by acetyl-CoA, the presence of which
indicates that fatty acids are being used to generate energy for the citric
acid cycle. The main function of glycolysis is to feed carbon units to the
 citric acid cycle. When these carbon skeletons can form other sources,
glycolysis is inhibited to spare glucose for other purposes.
17. Show lewis dot structure for transfer of electron.

a.

b.
i. We have 2 glyceraldehyde 3-phosphate thats why 2H​+
18. What is the net gain of ATP molecules in glycolysis?
a. There is a net gain of two ATP molecules per glucose molecule consumed
in glycolysis.
Important topics Exam 4
1. Enzymes Of glycolysis: hexokinase, phosphofructokinase-1 and pyruvate kinase.
2. What is the function of
a. Kinase - K​ inase is an ATP dependent enzyme that transfers a phosphate
from ATP to a substrate.
b. Isomerase- Isomerases catalyze the transformation of compounds into
their positional isomers.
c. Mutase- M ​ utases catalyze the transportation of functional groups.
 d. Dehydrogenase- Dehydrogenase catalyzes the removal of a pair of
hydrogens atoms from substrates.
3. Basic knowledge about the substrates of glycolysis
a. Glucose forms glucose 6-phosphate by enzyme hexokinase.
b. Fructose 6-phosphate forms fructose 1,6-bisphosphate by
phosphofructokinase-1.
c. Phosphoenolpyruvate forms pyruvate by enzyme pyruvate kinase.
4. Allosteric activator and inhibitor of glycolysis
a. ATP is an inhibitor of several steps of glycolysis as well as other catabolic
pathways. The purpose of catabolic pathways is to produce energy , and
high levels of ATP mean the cell has sufficient energy.
b. Glucose-6-phosphate inhibits hexokinase I,II, and III. Hexokinase IV is
controlled by a regulatory protein. And is an example of product inhibition.
If glucose-6-phosphate level is high, it may indicate that sufficient glucose
is available from glycogen breakdown or that the subsequent enzymatic
steps of glycolysis are going slowly. Either way, there is no reason to
produce more glucose-6- phosphate.
c. Phosphofructokinase is activated by a special effector molecule,
fructose-2,6-bisphosphate, whose levels are controlled by hormones.it is
also inhibited by citrate which indicates that there is sufficient energy
from the citric acid cycle, probably from fat and amino acid degradation.
d. Pyruvate Kinase is also inhibited by acetyl-CoA, the presence of which
indicates that fatty acids are being used to generate energy for the citric
acid cycle. The main function of glycolysis is to feed carbon units to the
citric acid cycle. When these carbon skeletons can form other sources,
glycolysis is inhibited to spare glucose for other purposes.
5. Regulation of glycolysis:
6. Endergonic and Exergonic reactions
a. Endergonic delta G positive- takes in ATP
b. Exergonic delta G negative- gives ATP
i. Most of the of the exergonic reactions are substrate level
phosphorylation reaction
7. Isomerization reaction of glycolysis
a. 2 products formed, one product is isomerized to the other one to get the
final product.
i. Fructose 1,6-bisphosphate forms 2 products at equilibrium. One is
dihydroxyacetone phosphate and the other is
glyceraldehyde-3-phosphate.
 ii. dihydroxyacetone phosphate must be converted to
glyceraldehyde-3-phosphate to move the glycolysis process
forwards.
1. Glyceraldehyde-3-phosphate is the only substrate that can
continue the glycolysis pathway.
iii. Isomerase is the enzyme that is required for this reaction.
8. Control points in glycolysis- all have a large decrease in free energy; the enzymes
that catalyze these reactions are sites of allosteric control
a. Hexokinase
b. Phosphofructokinase
c. Pyruvate kinase

d.
e. Hexokinase I, II, III are inhibited by glucose-6-phosphate. A regulatory
protein controls response in Hexokinase IV.
f. Phosphofructokinase is inhibited by ATP and citrate.
g. Pyruvate kinase is inhibited by ATP, acetyl-CoA and alanine.
h. Phosphofructokinase is activated by AMP and activated by fructose-2,6-
bisphosphate. Without F26BP the enzyme is inactive. ATP inhibits
phosphofructokinase.
i. Pyruvate kinase is stimulated by AMP and fructose-1,6-bisphosphate.
9. Substrate level phosphorylation
a. Forms ATP
 b. Phosphoglycerate Kinase - responsible enzyme for transferring the
phosphate group from 1,3-bisphosphoglycerate to ADP .
c. Phosphophenol Pyruvate (2) +2ADP---> 2 Pyruvate + 2 ATP
d. Transfer of phosphate group from phosphoenolpyruvate to ADP by
pyruvate kinase.
10. Fates of Pyruvate
a. Anaerobic glycolysis ( no oxygen)- enzyme lactate dehydrogenase
b. Aerobic oxidation (oxygen)- form carbon dioxide and water
c. Anaerobic alcoholic fermentation -alcohol dehydrogenase
d. Pyruvate can be converted to lactate, ethanol or acetyl CoA.
e. Glycolysis doesn’t require oxygen
11. Irreversible and reversible reactions in glycolysis
a. All the control steps are irreversible.
b. Others are reversible
12. Glycogen-
a. Where it is created
i. ​Glycogen​ is synthesized in the liver and muscles
b. How does it convert into glucose
i. This stored form of ​glucose​ is made up of many connected
glucose​ molecules and is called ​glycogen​.
Chapter 18 Homework
1. Why is it essential that the mechanisms that activate glycogen synthesis also
deactivates glycogen phosphorylase?
a. These two pathways occur in the same cellular compartment and if both
are on the same time, a futile ATP hydrolysis cycle results. Using the same
mechanisms to turn them on/off or off/on is highly efficient. Glycogen
phosphorylase activity can be allosterically c​ ontrolled ATP and G6P
a​llosteric inhibitors, AMP allosteric activator​ as well as, controlled
through covalent modification, phosphorylation via ​hormones.
b. The activity of glycogen synthase is subject to the same type of covalent
modification as glycogen phosphorylase, however the response is
opposite. ​Glycogen synthase​ is activated by G6P. It is also controlled via
hormones.
c. Glycogen - polymer of glucose
d. Glycogen phosphorylase- enzyme that breaks down glycogen
e. Glycogen break down:
 i. N-1 - -1 glucose
ii. Glucose n - n number of glycogens
iii. Catalyzed by enzyme glycogen phosphorylase
f.

1. Breakdown and formation of glycogen

a. Glycogen synthase and UDP
2. Glycogen stored in liver or muscle cells
2. ​Why is it advantageous that breakdown of glycogen gives rise to glucose-6-phosphate
rather than to glucose?

Glucose-6-phosphate is already phosphorylated. This saves one ATP equivalent in the

early stages of glycolysis.
.

7. G6P can be used for glycogen synthesis or for glycolysis, among other fates. How
many ATP equivalents are used to store G6P as glycogen rather to use it for energy in
glycolysis? Hint: The branched structure of glycogen leads to 90% of glucose residues
being released at G1P and 10% as glucose.

It “costs” one ATP equivalent (UTP to UDP) to add a glucose residue to glycogen. In
degradation, about 90% of the glucose residues do not require ATP to produce
glucose-1-phosphate. The other 10% require ATP to phosphorylate glucose. On average,
this is another 0.1 ATP. Thus, the overall “cost” is 1.1 ATP, compared with the three ATP
that can be derived from glucose-6-phosphate by glycolysis.

21. Which steps in glycolysis are irreversible? What bearing does this have on the
reactions on which gluconeogenesis differs from glycolysis?

Three reactions of glycolysis are irreversible under physiological conditions.

They are:
· the production of pyruvate and ATP from phosphoenolpyruvate,

· ​ hosphate from fructose-6-phosphate,

the production of fructose-1,6-​bisp

· and the production of glucose-6- phosphate from glucose. These reactions

are bypassed in gluconeogenesis; the reactions of gluconeogenesis differ from
those of glycolysis at these points and are catalyzed by different enzymes.
22. What is the role of biotin in gluconeogenesis?

Biotin is the molecule to which carbon dioxide is attached to the process of being
transferred to pyruvate. The reaction produces oxaloacetate, which then undergoes
further reactions of gluconeogenesis.

23. How does the role of G6P in gluconeogenesis differ from that in glycolysis?
 In gluconeogenesis, glucose-6-phosphate is dephosphorylated to glucose (the
last step of the pathway); in glycolysis, it isomerizes to fructose-6-phosphate (an early
step in the pathway).
26. Which reactions in this chapter require ATP? Which reactions produce ATP? List the
enzymes that are required for these reactions.

Reactions that require ATP:

· formation of UDP-glucose from glucose-1-phosphate
· UTP (indirect requirement, because ATP is needed to regenerate UTP)
regeneration of UTP, and carboxylation of pyruvate to oxaloacetate.

● Glucose-1-phosphate +UTP
● →UDP-glucose +pyrophosphate

Reactions that produce ATP:

· none.
Enzymes that catalyze ATP-requiring reactions:
· UDP- glucose phosphorylase (indirect requirement),
· nucleoside phosphate kinase, and pyruvate carboxylase.
nucleoside phosphate kinase- transfer of the nucleotide diphosphate to
glucose
Enzymes that catalyze ATP-producing reactions:
· none.

28. How do glucokinase and hexokinase differ in functions?

Hexokinase can add a p​hosphate group to any of several six-carbon s​ugars, whereas
glucokinase is specific for glucose. Glucokinase has a lower affinity for glucose than
does hexokinase. Consequently, glucokinase tends to deal with an excess of glucose,
particularly in the ​liver. ​Hexokinase is the usual enzyme for phosphorylating six-carbon
sugars.

34. Why would you expect to see that the reactions of substrate cycles involve different
enzymes for different directions?

Enzymes, like all catalysts, speed up the forward and reverse reaction to the same extent.
Having different catalysts is the only way to ensure independent control over the rates of
the forward and reverse process.
39. List 3 differences in structure or function between NADH and NADPH:

· NADPH has one more phosphate group than NADH (at the 2′ position of
the ribose ring of the adenine nucleotide portion of the molecule).
· Glycolysis: ​NADH is produced in oxidative reactions that give rise to ATP.

NADPH is a reducing agent in biosynthesis (pentose phosphate shunt).

 · The enzymes that use NADH as a coenzyme are different from those that
require NADPH.

42. Show how the pentose phosphate pathway, which is connected to the
glycolytic pathway, can do the following:

(a) Make both NADPH and pentose phosphates in about equal amounts:

· By using only the oxidative reactions.

(b) Make mostly or only NADPH:

· By using the oxidative reactions, the transaldolase and transketolase
reactions, and gluconeogenesis.

(c) Make mostly or only pentose phosphates:

· By using glycolytic reactions and the transaldolase and transketolase

reactions in reverse.

46. Depict the following reactions via Lewis electron dot formulas:

Glucose-6-phosphate + NADP​+​ → 6-phosphoglucono-​d​-lactone + NADPH + H​+

The lactone is a cyclic ester that is an intermediate in the production of

6-phosphogluconate

Glucose can go to glycogen, glycolysis or PPP.

Important Topics from Chapter 18:

● Enzymes used in Gluconeogenesis:
○ Pyruvate carboxylase, Fructose 1,6-bisphosphate, glucose-6-phosphatase.
● Glycolysis vs gluconeogenesis
○ 3 different controls with different enzymes
● Glycogen breakdown and the enzyme involved
○ Glycogen phosphorylase is the enzyme that is helpful to convert glycogen
to glucose-1-phosphate .
○ glucose-1-phosphate uridylyltransferase
○ also called UDP–glucose pyrophosphorylase)
○ Transfers nucleoside diphosphate to glucose
○
○ Glucose synthase is used in the formation of glucose to glycogen
● Glucose synthesis is activated by glucose 6 phosphate
● Point of control of glycogen enzymes
● Structure and storage of glycogen
● Uridine diphosphate glucose
● Pentose Phosphate Shunt
● Oxidative and non-oxidative phases of pentose phosphate shunt
 ○ Operates mostly in cytoplasm of liver and adipose cell
■ Two oxidative processes followed by 5 non oxidative steps
■ Oxidative phase: NADPH formed in oxidative phase is used to
reduce glutathione, GSSG. reduced form of GSH which scavanges
harmful free radicals. The other product is ribose 5 phosphate,
serves as precursor for nucleotides, coenzymes and nucleic acid
■ Non oxidative phase:recyles 6 molecules of the pentose into 5
molecules of the hexose glucose 6-phosphate, allowing production
of NADPH or ribose-5-phosphate. When cells arent using ribose 5
phosphate.

Chapter 18 storage mechanisms and controls in carbohydrate mechanism

g. Learning outcomes
i. 1.​Gain an understanding of how glycogen is converted to glucose and how
glucose is converted to glycogen and the mechanisms of control.
ii. 2.​Understand the pathway of gluconeogenesis and the points of control as
they relate to control points in glycolysis.
iii. 3.​Understand the pentose phosphate pathway and how it is regulated.
iv. 4. Be able to identify the enzyme if you are given the reaction.
v. 5. Understand what type of reaction each class of enzymes catalyzes.
h.

2. Endogenous Glycogen and Starch are degrade by phosphorolysis

a. Glycogen stored in animal tissue​s (primarily liver and skeletal muscle)​, in
microorganisms, or in plant tissues ​(starch​) can be mobilized for use
within the same cell by ​a phosphorolytic reaction​ catalyzed by glycogen
phosphorylase (s​tarch phosphorylase in plant​s).
b. Phosphorylase are enzymes that catalyze the addition of a phosphate
group from an inorganic phosphate to an acceptor.
c. These enzymes catalyze an attack by P​i​ on th.joins the last two glucose residues at
a​ nonreducing ​end, generating glucose 1-phosphate and a polymer one glucose
unit shorter.
d. Phosphorolysis preserves some of the energy of the glycosidic bond in the
phosphate ester glucose 1-phosphate.
3. Feeder pathways for glycolysis
 a. Other carbohydrates undergo glycolysis when they are transformed into a
glycolytic intermediate.

b.
4. Glycogen Phosphorylase
a. P​i​ on the (𝞪1-->4) glycosidic linkage
b. Glucose 1-phosphate produced by glycogen phosphorylase is converted to
glucose 6-phosphate by p ​ hosphoglucomutase ​which catalyzes the
reversible reaction:
i. Glucose 1-phosphate ⇆ glucose 6-phosphate

ii.
 c.

d.
5. Alpha glucose and beta glucose

a.
 b.
6. Maltose

a.
7. Hemiacetal, acetal , hemiketal

a.
8. Redox reactions of simple sugars
a. •​Aldehyde groups can be oxidized to give the carboxyl group
b. •​Aldoses are ​reducing sugars​ that contain a free carbonyl group and can react
with an oxidizing agent
c. •​Ketoses can also be reducing sugars because they isomerize to aldoses
 d. •​Oxidation of a cyclic hemiacetal form gives a lactone, which is a cyclic ester
linking the carboxyl group and one of the sugar alcohols

e.
9. Formation of a Cyclic Hemiacetal

a.
i. 𝞪- D - Glucopyranose
ii. Β- D- Glucopyranose
10. Glycogen Breakdown
a. The non-reducing end of Glycogen is cleaved by phosphate to give a
a-D-glucose-1-phosphate
b. Cleavage reaction is phosphorolysis not hydrolysis
c. No ATP is involved in reaction
d. Reaction is catalyzed by glycogen phosphorylase.
11. Glycogen phosphorylase dimer
 a.
12. Phosphoglucomutase
a. Glucose 1-phosphate produced by glycogen phosphorylase is converted to
glucose 6-phosphate by this enzyme.
b. Catalyzes the reversible reaction
c. Glucose 1-phosphate⇋ glucose 6-phosphate
d. Mutases catalyze the transposition of functional groups.

e.
i. Phosphoglucomutase
f. In the second reaction, glucose, 1-phosphate is isomerized to
glucose-6-phosphate. The reaction is reversible.
g. This reaction is catalyzed by phosphoglucomutase.
 h.
13. In glycolysis it is an irreversible reaction:

a.
14. Worked example
a. Energy saving for glycogen breakdown by phosphorolysis.
b. Calculate the energy savings (in ATP molecules per glucose monomer)
achieved by breaking down glycogen by phosphorolysis rather than
hydrolysis ( ATP to ADP) to begin the process of glycolysis.
i. Solution: Phosphorolysis produces a phosphorylated glucose which
is then converted to glucose 6-phosphate- without expenditure of
the cellular energy (1 ATP) needed for formation of glucose
6-phosphate from free glucose.
c. Thus only 1 ATP is consumed per glucose monomer in the preparatory phase,
compared with 2 ATP when glycolysis starts with free glucose.
d. The cell therefore gains 3 ATP per glucose monomer (4 ATP produced in the
payoff phase minus 1 ATP used in the preparatory phase), rather than 2—a saving
of 1 ATP per glucose monomer.
e. Ingested polysaccharides and disaccharides are converted to monosaccharides by
intestinal hydrolytic enzymes, and the monosaccharides then enter intestinal cells
and are transported to the liver or other tissues.
f. •
 g. A variety of D-hexoses, including fructose, galactose, and mannose, can be
funneled into glycolysis.
h. Each is phosphorylated and converted to glucose 6-phosphate, fructose
6-phosphate, or fructose 1-phosphate.Conversion of galactose 1-phosphate to
glucose 1-phosphate also occurs.
15. Glycogen Formed from Glucose
● Synthesis requires energy
● Energy supplied by hydrolysis of UTP
● Glucose-1-phosphate reacts with UTP to make UDPG
● Glucose-1-phosphate +UTP
● →UDP-glucose +pyrophosphate
● Not exact reversal of glycogen breakdown to glucose
● glucose-1-phosphate uridylyltransferase
● also called UDP–glucose pyrophosphorylase)
● Transfers nucleoside diphosphate to glucose

●
○ Glucose-1-phosphate uridylyltransferase
○ (e coli)
■ Each subunit of ​tetramer- 4 subunits​ is dominated by an
eight-stranded mixed beta-sheet. There are two additional layers of
beta-sheet (two and three strands) and 10 alpha-helices.
○ Pyrophosphate is also formed
○ UDPG is then added to a growing chain of glycogen, catalyzed by
glycogen synthase.
 ■
● Reaction Catalyzed by Glycogen Synthase

○
● Glycogen synthase tetramer Eukaryotes

● Eukaryotes form tetramers that are regulated

2. Control of Glycogen Breakdown

❖ Glycogen phosphorylase ​is a major control point in the synthesis and breakdown
of glycogen ​it is a dimer with T and R states
❖ Glycogen phosphorylase ​activity can be allosterically controlled ATP and G6P
allosteric inhibitors, AMP allosteric activator as well as, controlled through
covalent modification,phosphorylation, Phosphorylase A is more active than B.
❖ Glycogen phosphorylase exists as an inactive monomer and tetramer the
dimer is active

➢
❖ Phosphorylase A and Phosphorylase B

➢
● Phosphorylase is most active when it has the
phosphate on it
➢ Covalent modification trumps allosteric regulation
■ The B form is less active and more allosterically controlled.
■ The A form is more active and less allosterically responsive.
❖ Control of Glycogen Breakdown
➢ Glycogen phosphorylase is also under hormonal control.
➢ During vigorous muscular activity Epinephrine triggers the formations of
cAMP. This results in an enzyme cascade shifting the b form to the a form
and producing more G1P from glycogen.
 ➢ The activity of ​glycogen synthase​ ​is subject to the same type of covalent
modification as glycogen phosphorylase, however, the response is
opposite.
➢ The A form is the active form it is unphosphorylated and Phosphorylation
produces the inactive B form. cAMP causes an enzyme cascade effect in
response to epinephrine. This time the enzyme is phosphorylated and
deactivated.
➢ Glycogen Synthase is an allosteric enzyme. It is inhibited by ATP. It is
activated by glucose-6-phosphate.
➢ The non phosphorylated form (active), glycogen synthase I is active at
even very low glucose-6-phosphate levels.
➢ The phosphorylated form (inactive) is glycogen synthase D. It is activ only
at extremely high concentrations of glucose-6-phosphate.
➢ Purified enzymes respond to allosteric effectors but true control is by
phosphorylation which is controlled by hormonal states.
➢ It is composed of 4 identical subunits in eukaryotes.
➢ .The substrates bind when the enzyme is in the R state.
➢ The phosphorylated (less active)form is allosterically activated by G6P,
which binds to the R state.
➢ The unphosphorylated (active) A form is not allosterically controlled.

➢
■ GS Eukaryotes Tetramer
Sensing the Epinephrine Signal via a G-Protein Coupled Receptor
● Adenyl Cyclase is a membrane bound protein
 ○

○
● Summary
○ Glycogen is the storage form of glucose in animals, including humans. Glycogen
releases glucose when energy demands are high.
○ Glucose polymerizes to form glycogen when the organism has no immediate need
for the energy derived from glucose breakdown
○ Glycogen metabolism is subject to several different control mechanisms,
including covalent modification and allosteric effects.
● Glucogenesis
○ Occurs mainly in liver and kidneys
○ Not the mere reversal of glycolysis for 2 reasons:
■ Energetics must change to make gluconeogenesis favorable (▵G of
glycolysis = -74 kJ/mol
■ Reciprocal regulation must turn on and the other off- this requires
something new!
○ Seven steps of glycolysis are retained:
■ Steps 2 and 4-9
○ Three steps are replaced:
 ■ Step 1,3 and 10 ( the regulated steps)
○ The new reactions provide for a spontaneous pathway (▵G negative in the
direction of glucose synthesis), and they provide new mechanisms of regulation.
● The overall pathway of Gluconeogenesis
○ Gluconeogenesis : the conversion of glucose from pyruvate
■ Gluconeogenesis is not the exact reversal of glycolysis; that is, pyruvate to
glucose does not occur by reversing the steps of glucose to pyruvate.
● Coordinated Regulation of Glycolysis and Gluconeogenesis
○ Three irreversible steps in glycolysis
■ Phosphoenolpyruvate to pyruvate + ATP
■ Fructose-6-phosphate to fructose-1,6-bisphosphate
■ Glucose to glucose-6-phosphate
● Net result of gluconeogenesis is reversal of these three steps, but
by different reactions and using different enzymes
■ Three reactions of glycolysis are so exergonic as to be essentially
irreversible: those catalyzed by hexokinase, PFK-1, and pyruvate kinase.
All three reactions have a large, negative ΔG′.
○ •​Gluconeogenesis uses detours around each of these irreversible steps; for
example, the conversion of fructose 1,6-bisphosphate to fructose 6-phosphate is
catalyzed by fructose 1,6-bisphosphatase (FBPase-1). Each of these bypass
reactions also has a large, negative ΔG′.
● Pyruvate Carboxylase
○ Pyruvate is converted to oxaloacetate
○ The reaction requires ATP and bicarbonate as substrates
○ Biotin (coenzyme) is covalently linked to an active site lysine
○ Acetyl-CoA is an allosteric activator
○ Regulation: when ATP or acetyl-CoA are high, pyruvate enters gluconeogenesis
○ Compartmentalized, takes place in mitochondria
● Oxaloacetate is an intermediate
○ In first step, Pyruvate is carboxylated to oxaloacetate
○ Requires biotin (​CO​2​ carrier​)
○ Pyruvate carboxylase ​is subject to allosteric control; it is activated by
acetyl-CoA

■
● Biotin, acetyl co-A, pyruvate carboxyalse.
● Phosphoenolpyruvate Carboxykinase PEPCK
○ Next, decarboxylation of oxaloacetate is coupled with phosphorylation by GTP to
give PEP.
 ■
■ The net reaction of carboxylation/decarboxylation is
■ Pyruvate + ATP +GTP ---> Phosphenolpyruvate + ADP + GDP + P​i
■ Overal DG’​0​ = 2.1 kJ/mol
■ Law of mass action: A small increase in oxaloacetate can drive the
reaction to the right. A small increase in PEP can drive it to the left.

○
● Oxaloacetate cannot be transported across the mitochondrial membrane. It is reduced to
malate transported to the cytosol and oxidized back to oxaloacetate.

○
● Which of the following statements concerning biotin and gluconeogenesis is false?
● a) Biotin is used to add CO​2​ ​to certain intermediates in gluconeogenesis.
● b) * CO​2​ ​is incorporated into the glucose product.
● c) Biotin is capable of binding covalently to CO​2​.
● d) Biotin helps synthesize an important precursor of phosphoenolpyruvate.
● e) ATP hydrolysis is required to attach CO​2​ ​to biotin.

★ Second Control step

★ Role of sugar phosphates

○ Other different reactions in gluconeogenesis relative to glycolysis involve
phosphate-ester bonds bound to sugar-hydroxyl groups being hydrolyzed.

■
● Fructose 1,6- bisphosphate
○ Delta G° = -16.7 kJ mol​-1
○ •​Fructose-1,6-bisphosphatas​e is an allosteric enzyme,
inhibited by AMP and activated by ATP
★ Control step 3

○
○ Role of sugar phosphates
■ Another reaction is the hydrolysis of glucose-6-phosphate to glucose and
p​i

●
-1​
○ •​Reaction also spontaneous (Delta G°’ = -13.8 kJ mol​ )
○ •​Reaction catalyzed by g​ lucose-6-phosphatase
 ● The Glycolytic Enzyme Pyruvate kinase is allosterically inhibited by ATP

★ Pyruvate can be converted to glucose and glycogen via gluconeogenesis or

★ oxidized to acetyl-CoA for energy production.
★ The first enzyme in each path is regulated allosterically; acetyl-CoA,
produced either by fatty acid oxidation or by the pyruvate dehydrogenase
complex, stimulates pyruvate carboxylase and inhibits pyruvate
dehydrogenase.
★
 ●
○ F26BP increases the affinity of PFK-1 for F6P and
decreases affinity of FBPase for G6P

1. Hexokinase is inhibited by high levels of glucose 6-phosphate. When glycolysis is

inhibited through phosphofructokinase, glucose 6-phosphate builds up, shutting down
hexokinase
2. Glucose 6-phosphatase is sequestered in the liver and kidneys. Releases glucose made
from glucose 6-phosphate. Glucose-6-phosphatase activity in controlled by the level of
glucose-6-phosphate. ​This is substrate level control.
3. Activity is not under allosteric control.
4. When levels of glucose-6-phosphate are high, glycolysis switched off.
5. Glucose is sometimes diverted through the pentose phosphate pathway
a. •​The Pentose Phosphate Pathway (PPP) is an alternative to glycolysis, and differs
in several ways
b. •​In glycolysis, ATP production is important, in PPP, this is not the case
c. •​As the name implies, five-carbon sugars, including ribose, are produced from
glucose
d. •​Oxidizing agent is NADP​+​; it is reduced to NADPH, which is a reducing agent in
biosynthesis
6. Glucose is sometimes diverted through the pentose phosphate pathway
a. •​Begins with two oxidation steps (NADP​+​) to give ribulose-5-phosphate
b. •​Following this, a series of carbon-shuffling steps occur during which three-,
four-, five-, six-, and seven-carbon monosaccharide phosphates are produced
7. Pentose Phosphate Pathway
a. The main goals are to produce NADPH for anabolic reactions, fatty acid
biosynthesis and ribose 5-phosphate for nucleotides
b. •
c. •​Two oxidative processes followed by five non-oxidative steps
d. •
e. •​Operates mostly in cytoplasm of liver and adipose cells
8.
a. General scheme of the pentose phosphate pathway
i. NADPH formed in the oxidative phase is used to reduce glutathione,
GSSG. It is the ​reduced form GSH​ which scavanges harmful free radicals
and to support reductive biosynthesis. The other product of the oxidative
phase is ribose 5-phosphate, which serves as a precursor for nucleotides,
coenzymes, and nucleic acids.
ii. In cells that are not using ribose 5-phosphate for biosynthesis, the
nonoxidative phase recycles six molecules of the pentose into five
molecules of the hexose glucose 6-phosphate, allowing continued
production of NADPH and converting glucose 6-phosphate (in six cycles)
to CO​2​.
iii. •​The carbon-shuffling reaction are catalyzed by:
iv. •​Transketolase​ ​for the transfer of two-carbon units and
v. •​Transaldolase ​for the transfer of three-carbon units
vi. Control of the pentose phosphate pathway is maintained by:
vii. •​Glucose-6-phosphate (G6P) can be channeled into either glycolysis or the
pentose phosphate pathway
viii. •​G6P channeling into glycolysis, if ATP needed
ix. •​G6P channeling into the pentose phosphate pathway, if NADPH or
ribose-5-phosphate are needed
9.
a. In a series of rearrangements of the carbon skeletons 6 five-carbon sugar
phosphates are converted to 5 six-carbon sugar phosphates, completing the cycle
and allowing continued oxidation of glucose 6-phosphate with production of
NADPH. Continued recycling leads ultimately to the conversion of glucose
6-phosphate to six CO​2​.
b.