NUR8103Lecture- Carbohydrate Metabolism I

Glycolysis
 Series of reactions that takes place in the cytoplasm of all prokaryotes and eukaryotes
 Converts one molecule of glucose into two molecules of pyruvate
 2 ATPs are needed for early glycolysis pathway, 4 ATPs are generated, giving a net yield of 2 ATPs
per glucose degraded.
 Roles of glycolysis
1. to produce energy
2. to produce intermediates for biosynthetic pathways
Pathway
Control point
1. Glucose is phosphorylated by ATP to form glucose 6-phosphate and ADP.
Enzyme: hexokinase

2. Phosphoglucoisomerase catalyze the conversion of glucose 6-phosphate to fructose 6-phosphate

Control point
3. Phosphofructokinase (PFK) catalyzed the phosphorylation reaction of fructose 6-phosphate by
ATP to form fructose 1,6-bisphosphate and ADP.

4. Aldolase splits fructose 1,6-bisphosphate (6-carbon molecule) into two 3-carbon

molecules-glyceraldehyde 3-phosphate and dihydroxyacetone phosphate

5. Glyceraldehyde 3-phosphate is the only molecule that can be used in glycolysis.

Dihydroxyacetone phosphate can be converted to glyceraldehyde 3-phosphate by
triose phosphate isomerase. Dehydrogenase (oxidation reduction)
6. The energy for creating this new high-energy phosphate bond comes from the oxidation of the
aldehyde group of the glyceraldehyde 3-phosphate.

7. Phosphoglycerate kinase catalyzes the transfer of the phosphoryl group from the 1,3-BPG to ADP,
generating ATP and 3-phosphoglycerate.

8. The reaction is a movement of the phosphate group to a different carbon atom within the same
molecule. Catalyzed by phosphoglycerate mutase.
9. Converts the low-energy phosphate ester bond of 2-phosphoglycerate into the high-energy
phosphate bond of phosphoenolpyruvate (PEP) catalyzed by enolase

Control point
10. Pyruvate kinase catalyzes the physiologically irreversible transfer of the phosphoryl group from
PEP (2-adp to 2-atp)

Fates of Pyruvate
 Aerobic conditions: pyruvate converted by pyruvate dehydrogenase to acetyl CoA which enter
the citric acid cycle
 Anaerobic conditions: pyruvate is converted to lactate by lactate dehydrogenase (LDH)
 Yeast and other organisms carry out alcoholic fermentation
 Converts pyruvate to acetaldehyde and then to ethanol
 Regenerating NAD+ that allows glycolysis to continue

Fructose Metabolism
 Fructose can be metabolized by two routes.
1. Adipose tissue and muscle, fructose is converted to fructose 6-phosphate by fructose
hexokinase then enters glycolysis.
2. Liver, most of the enzyme present is glucokinase, fructose is metabolized by the fructose
1-phosphate pathway.
Galactose Metabolism
 Galactose enters glycolysis via the galactose–glucose interconversion galactose pathway
 The lack of galactose 1-phosphate uridylyl transferase, leads to the disease galactosemia
 Galactosemia, toxic substances accumulate such as galactitol, formed by the reduction of
galactose

Regulation
 Main control step is catalyzed by PFK
 Hexokinase and pyruvate glycolysis kinase are additional control sites
 PFK is allosterically inhibited by ATP, but this inhibition is relieved by AMP
 Build up of fructose 6-phosphate stimulates the formation of fructose 2,6-bisphosphate
that in turn stimulates PFK
 Phosphofructokinase 2; PFK2 and fructose bisphosphatase 2; FBPase2 are also regulated
hormonally by glucagon that causes glycolysis to slow down when the blood glucose level falls

 PFK is also inhibited by H+ ions, preventing excessive formation of lactate under anaerobic
conditions
 Hexokinase is inhibited by glucose 6-phosphate which builds up after PFK is inhibited.
 Pyruvate kinase is activated by fructose 1,6-bisphosphate but allosterically inhibited by ATP and
alanine; regulated hormonally by glucagon.

Citric Acid Cycle

 Kreb cycle/ Tricarboxylic acid cycle
 Oxidizes pyruvate (from glycolysis) to CO2 and H2O, with the production of energy
 Acetyl CoA from fatty acid breakdown and amino acid degradation products are oxidized
 Role: producing precursors for biosynthetic pathways
 Location: mitochondria of eukaryotes and the cytosol of prokaryotes

8 Stages
1. Production of citrate from oxaloacetate and acetyl CoA catalyzed by citrate synthase.
2. Isomerization of citrate to isocitrate catalyzed by aconitase. Citrate is chiral (walang chiral
carbon) only one stereoisomer can enter krebs cycle
 3. Oxidation of isocitrate to α-ketoglutarate catalyzed by isocitrate dehydrogenase; the reaction
requires NAD+
4. Oxidation of α-ketoglutarate to succinyl CoA catalyzed by the α-ketoglutarate dehydrogenase
complex; the reaction requires NAD+ (oxidative decarboxylation reaction)
5. Conversion of succinyl CoA to succinate by succinyl CoA synthetase; the reaction requires
inorganic phosphate and GDP or ADP
6. Oxidation of succinate to fumarate catalyzed by succinate dehydrogenase; the reaction
involves FAD.
7. Hydration of fumarate to malate by fumarase. Produce L-malate…
8. Oxidation of malate to oxaloacetate by malate dehydrogenase; the reaction requires NAD+

Energy yield
 For each turn of the cycle, 12 ATPs molecules are produced
 1 ATP directly from the cycle
 11 ATPs from the re-oxidation of the three NADH and one FADH2 molecules produced by
oxidative phosphorylation.
Regulation
 regulated at the steps catalyzed by citrate synthase, isocitrate dehydrogenase and
α-ketoglutarate dehydrogenase

 Feedback inhibition by ATP, citrate, NADH and succinyl CoA,

 Stimulation of isocitrate dehydrogenase by ADP
 Pyruvate dehydrogenase is inhibited by acetyl CoA and NADH
 Pyruvate dehydrogenaseis inactivated by phosphorylation
 High ratio of NADH/NAD+, acetyl CoA/CoA or ATP/ADP stimulates phosphorylation of pyruvate
dehydrogenase
 Pyruvate inhibits the kinase
 Dephosphorylation by a phosphatase reactivates pyruvate dehydrogenase

Electron Transport Chain

 In eukaryotes, occurs in the inner membrane of mitochondria
 In prokaryotes, occurs in the plasma membrane
 NADH from glycolysis are shuttled into the mitochondrial matrix
 These NADH are re-oxidized to NAD+, while forming H+ ions and electrons
 NADH from citric acid cycle delivers more H+(protons) and electrons, these are translocated via
co-enzymes across the membrane
 Translocation is from matrix into the intermembrane space increasing the proton gradient in the
intermembrane space
 Protons from the intermembrane enter once again, the mitochondrial matrix through ATP
synthase
 ATP continues to be made as protons move through ATP synthase enzyme
 Oxygen is the final electron acceptor, combining with H+ to produce H2O
 Anaerobic conditions, NADH acts as reducing agent producing lactic acid from pyruvic acid
 HW-NUR8103Lecture- Carbohydrate Metabolism I

Glycolysis
After digestion of carbohydrates, glucose enters the cell membrane and into the cytoplasm, glycolysis
takes place where glucose-6-phosphate (a 6C-molecule) is split into two 3C molecules of gly-3-P,
producing 2 pyruvate. The two pyruvates are then converted to two acetyl-CoA and enter the
mitochondria where the citric acid occurs.

1. What reaction is involved in the so called, energy-investing reactions? What chemical compound is
needed to initiate the reaction?

2. Cite the steps in the glycolysis pathway where these energy-investing reactions occur.
There are 4 pathways involved. It starts off with glucose that has 6 carbon compounds. Second, the
glucose converts into glucose-6 phosphate by an enzyme which is hexokinase. Third, glucose-6
phosphate converts into fructose-6-phosphate by isomerase. Fourth, the phosphofructokinase
catalyzed the phosphorylation reaction of fructose 6-phosphate by ATP to form fructose 1,6-
bisphosphate and ADP.

3. The conversion from glu-6-P to fru-6-P involves isomerization, why?

Isomerization means the chemical process of converting a compound into any of its isomeric forms.
Isomerization is considered a significant reaction due to its numerous applications in biomass
valorization.

4. What stages in the glycolysis pathway are considered energy harvesting reactions? What are the
products of these reactions that will prove its energy yield?
For the first part, energy production starts at the conversion of 1,3 biphosphoglycerate to 3
phospho-glycerate by phosphoglycerate kinase. While this conversion occurs, 1 molecule of ADP
also converts to 1 molecule of ATP. Second is the conversion of 3 phospho-glycerate into 2
phospho-glycerate by phosphoglycerate mutase. Third is the conversion of 2 phospho-glycerate
into phosphoenol pyruvate by enolase. The final step is the conversion of phosphoenol pyruvate
into pyruvate by a pyruvate kinase wherein 2 ADP also convert to 2 ATP.

5. The conversion from gly3-P to 1,3-diP uses NAD + and a dehydrogenase enzyme. What is the role
of NAD+? What reactions therefore are involved in these conversions?
Glycolysis oxidizes glyceraldehyde 3-phosphate. It extracts the high energy electrons that
collected by the electron carrier NAD+ which produces NADH. The reaction involved in this step
(6th step of glycolysis) depends on the presence or availability of NAD +. NADH is continuously
oxidized into NAD+ to keep the process in motion. The second half of Glycolysis stops or slows
down if NAD+ is not present.
https://bio.libretexts.org/Bookshelves/Introductory_and_General_Biology/Book
%3A_General_Biology_(Boundless)/7%3A_Cellular_Respiration/7.2%3A_Glycolysis/7.2C
%3A_The_Energy-Releasing_Steps_of_Glycolysis

6. The conversion from 1,3-di-P-glycerate to 3-phosphoglyceric acid uses ADP. Why?

The conversion from 1,3-di-P-glycerate to 3-phosphoglyceric acid is done by the enzyme
phosphoglycerate kinase, involving the transfer of the phosphoryl group from the 1,3-BPG to ADP
which produces ATP and 3-phosphoglycerate. ADP is used to generate ATP in the seventh step of
glycolysis.

7. In a reaction where the enzyme involved is phosphoglycerate mutase, what happens to the
substrate, 3-phosphoglycerate?
Phosphoglycerate is an enzyme that catalyze the movement of a phosphate group from C-3 to
C-2, which contributes to the conversion of 3-phosphoglycerate to 2-phosphoglycerate. The
reaction is simply a movement of the phosphate group to a different carbon atom within the
same molecule.

8. The formation of pyruvic acid (a keto-acid) involves what reaction? Account for the total ATP
produced from glycolysis.
The formation of pyruvic acid involves pyruvate kinase wherein it catalyzes the physiologically
irreversible transfer of the phosphoryl group from PEP. Furthermore, in the process of
glycolysis, glucose breaks down into pyruvate and energy. The total amount of ATP derived in
the process of glycolysis is 2 ATP.
 Citric Acid Cycle
9. What does amphibolic pathway mean? What three molecules produced during the citric acid cycle
are indirect or direct source of high-energy compounds?
Amphibolic pathway is a biochemical pathway. They contain enzymes which engages in both anabolic
and catabolic pathways. The citric acid cycle is a type of amphibolic pathway. Amphibolic pathways
breaks down carbohydrates to smaller molecules and produces energy (ATP). Molecules
NADH, FADH2, and GTP produced in the citric acid cycle are indirect or direct sources of high-energy
molecules.

10. Which steps of aerobic metabolism of pyruvate through the citric acid cycle are control points?
11-17. Two moles of pyruvic acids produced per molecule of glucose from glycolysis in the cytoplasm,
enters mitochondria, successively.

11. In the initial reaction, considering the products in the conversion of pyruvic acid to acetyl-CoA,
aside from the condensation reaction of pyruvic acid with CoA-SH, what is the other reaction
involved?
In the process of oxidative decarboxylation, transition reaction converts 2 molecules of 3-carbon
pyruvate to 2 molecules of the 2-carbon molecule of acetyl-CoA and 2 molecules of CO 2.
https://bio.libretexts.org/Bookshelves/Microbiology/Book
%3A_Microbiology_(Kaiser)/Unit_7%3A_Microbial_Genetics_and_Microbial_Metabolism/18%3A_
Microbial_Metabolism/18.3%3A_Aerobic_Respiration#:~:text=and%20facultative
%20anaerobes.-,Aerobic%20respiration%20involves%20four%20stages%3A%20glycolysis%2C
%20a%20transition%20reaction%20that,electron%20transport%20chain%20and
%20chemiosmosis.

12. What reactions between acetyl-CoA and oxaloacetic acetic acid are involved in the formation of
citric acid?
The citric acid cycle is catalyzed by the citrate synthase, in this reaction acetyl-CoA and oxaloacetic
acetic acid is joined together to form a citric acid. Acetyl group CH3COO transfers from CoA to
oxaloacetic acid at the ketone group, afterwards it changes into alcohol.
http://chemistry.elmhurst.edu/vchembook/611citricrx.html

13. In the 2nd and 3rd steps, what are the reactions involving the formation of isocitric acid?
In the 2nd step the reaction involved in the formation of isocitric acid is catalyzed by aconitase
wherein water molecule is removed from the citric acid and is placed back on in another location.
In the 3rd step the oxidation of isocitrate to α-ketoglutarate is catalyzed by isocitrate
dehydrogenase. During this process NAD+ is reduced to form NADH.

14. Compare the number of carbons between isocitric and a-ketoglutaric acid.
Isocitric acid or isocitrate has a 6-carbon compound whereas a-ketoglutaric acid has 5 carbon
compound.
https://www.toppr.com/ask/question/match-the-compounds-given-in-column-i-with-the-number-of-carbon-
atoms-present-in/
15. What are the reactions involved in the formation of succinyl-CoA? Compare the structure of
a-ketoglutaric acid and succinyl CoA.
The conversion of succinyl CoA to succinate is catalyzed by the enzyme succinyl CoA synthetase.
The reaction requires inorganic phosphate and GDP or ADP. During this process guanosine
triphosphate is synthesized. Synthesis of GTP occurs when a free phosphate group is added to a
GDP molecule. A free phosphate now attacks the succinyl CoA molecule which releases the CoA.
As the phosphate attaches to the molecule, it is transferred to GDP to form GTP which results
the molecule succinate.
https://www.sparknotes.com/biology/cellrespiration/citricacidcycle/section2/page/2/

16. In the 8th step, identify the change of functional group of the reactant and the product.
oxaloacetate is regenerated by oxidation of malate. The coenzyme which is NAD+ causes the
transfer of 2 hydrogens and electrons to NADH + H+.
http://chemistry.elmhurst.edu/vchembook/611citricrx.html
 17. Account the sum of ATP produced from glycolysis and citric acid cycle.
The ATP produced from glycolysis and citric acid cycle yields a total of 30-32 ATP
36 daw

Electron Transport Chain

18-20. The inner membrane has specialized protein channels coenzymes where H + and electrons pass
through and translocated, from the inner to the intermembrane space.

18. Show the reaction what happens to NADH from citric cycle as it moves to co-enzymes and
delivers H+ and electrons.
- NADH oxidizes and transfers more H+(protons) and electrons to the electron transport
chain.
19. As H+ ions remain in the intermembrane space, electrons are continuously passed on through
series of co-enzymes. Which acts as the final acceptor of electrons?
- Oxygen acts as the final acceptor of electrons by removing the de-energized electrons from
the chain. Oxygen binds to H+ ions in the matrix to form water. This is done to maintain the
electrochemical gradient.

20. As H+ increases in the intermembrane space, the proton gradient also increases. What is the
effect of this increase in proton gradient to H + ions?
H+ ions will move its electrochemical gradient down and diffuse back into matrix. The process of
the said diffusion is called chemiosmosis. Hydrogen ions will move through the ATP synthase
and trigger the molecular rotation of enzyme which will synthesize ATP.
https://ib.bioninja.com.au/higher-level/topic-8-metabolism-cell/untitled/electron-transport-
chain.html

extra
Citric acid cycle
https://www.khanacademy.org/science/biology/cellular-respiration-and-
fermentation/pyruvate-oxidation-and-the-citric-acid-cycle/a/the-citric-acid-cycle

REVIEWER FOR QUIZ ON TUESDAY

https://microbenotes.com/krebs-cycle/
https://www.youtube.com/watch?v=A1nJRoPGkRs&feature=share – GLYCOLYSIS

recorded sessions
https://drive.google.com/drive/folders/1s0aZeCkysbQXXBPmT_2hlzyLU1o84Y5b