Detoxicol
SDLS 2008
File Created on 8/31/2004 03:30 PM
Transcriber(s): Audrey, Mark longus, Henry, Lance
Editors: JC Tayco, Mark Lomboy, Pre Ausan, Lulu Velazco
The CITRIC ACID CYCLE
Known as: TRICARBOXYLIC ACID CYCLE (TCA
CYCLE, KREBS CYCLE).
Also a major pathway for the inter conversion
of the metabolites arising from the
transamination and deamination
amino acids.
 oxidize acetyl residues(as Acetyl CoA) and
reduce coenzymes that upon reoxidation are
linked to the formation of ATP.
 final common pathway for the aerobic
oxidation of Carbohydrates, Lipid and
Protein, because glucose, fattyacids and
most amino acids are metabolised to Acetyl-
CoA or intermediates of the cycle. (able to
donate substrate to the TCA cycle).
Found in: MITOCHONDRIAL MATRIX (Location of
enzymes of TCA, also the site of enzymes
of the respiratory chain.
FUNCTIONS:
1.Catabolic
 Transfers electron pairs (potential energy)
from Acetyl Coenzyme A to NAD and FAD.
CH3CO-CoA + 3NAD + FAD + GDP + Pi
2CO2 + 3NADH + FADH2 + GTP + CoA
9ATP= Oxidative Phosphorylation
1ATP= Substrate level Phosphorylation
2. Anabolic
Specific metabolites used for
fatty acid synthesis
- gluconeogenesis
- synthesis of heme
- amino acid synthesis
*collectively known as Amphibolic(functions in both
oxidative and synthetic process)
ANAPLEROTIC REACTIONS
 are “filling-up” reactions
 replenishment of intermediates to ensure
continuity of cycle.
 not part of TCA cycle, it only produce
intermediates for TCA cycle.
1. pyruvate carboxylase – in the
liver and kidney.
- most important anaplerotic reactions.
Pyruvate + HCO3 + ATP  OXAA + ADP +
Pi
Medicine for the intoxicated
Topic: TCA Cycle
Lecturer: Dr. Dalisay Chionglo-Sy
No. of pages: 5
2 . glutamate dehydrogenase – in the
liver
GLUT + NADP + H 2O  α -
KETOGLUTARATE + NADPH + NH4
of
PEP carboxykinase – in heart and skeletal
muscle.
PEP + CO2 + GDP  OAA + GTP
Malic enzyme - in many tissues
PYRUVATE + HCO3 + NADPH 
MALATE + NADP
TCA PRODUCTS PER REVOLUTION
 Release of 2 moles of CO2
 Regeneration of one mole OXAA
 Generation of 9 moles ATP (via
oxidative phosphorylation)
 Synthesis of one high energy
phosphate (as GTP)
TCA CYCLE REGULATION
Citrate synthase
- acetyl CoA + OXAA = citrate
- allosteric inhibitor: ATP
- increases Km for acetyl CoA
Isocitrate dehydrogenase
- converts isocitrate to α -ketoglutarate
- allosteric activator: ADP
- inhibitors: ATP,NADH
3. α -ketoglutarate
- converts α -ketoglutarate to succinyl
Coa
- inhibitors: SUCCINYL COENZYME A and
NADH
Important Terms:
• Oxidation of Acetyl-CoA- Coenzymes are
reduced and subsequently reoxidized in the
respiratory chain, linked to the formation of
ATP
-Aerobic process
• Citrate synthase- enzymes that catalyze the
initial reaction between Acetyl-CoA and
Oxaloacetate to form citrate.
• Aconitase- Aconitase Hydratase
 • Lyase-enzyme that isomerizes citrate to
isocitrate.
• Isocitrate Dehydrogenase- enzyme that
catalyzes the dehydrogenation of Isocitrate
to form Oxalosuccinate.
• α -Ketoglutarate dehydrogenase complex-
requires the same cofactors as the pyruvate
dehydrogenase complex-thiamine
diphosphate, lipoate, NAD+, FAD and CoA-
and results in the formation of succinyl-CoA.
• Arsenite- inhibits the reactions(oxidative
decarboxylation); causing substrate Alpha-
Ketoglutarate to accumulate.
• Succinate thiokinase- succinyl-CoA synthase
-converts Succinyl-CoA to Succinate.
*Converted to Oxaloacetate by malate
Dehydrogenase, a reaction requiring NAD+.
B-Vitamins in Citric Acid cycle:
1. Riboflavin- in the form of flavin adenine
dinucleotide.
-cofactor in the Alpha- Ketoglutarate
dehydrogenase complex and in succinate
dehydrogenase.
2. Niacin-in the form of nicotinamide adenine
dinucleotide(NAD).
-Coenzyme for the three dehydrogenases in
the cycle- Isocitrate dehydrogenase, Alpha-
Ketoglutarate dehydrogenase and Malate
dehydrogenase.
3. Thiamine- as thiamine diphosphate
-enzyme for decarboxylation in the Alpha-
Ketoglutarate dehydrogenase reaction.
4. Pantothenic Acid-part of coenzyme A
-cofactor attached to active carboxylic acid
residues such as Acetyl-CoA are Succinyl-
CoA.
Individual Reactions of the Citric Acid Cycle:
 Conversion of Pyruvate to Acetyl-CoA
 Pyruvate Dehydrogenase Complex
-Enzyme
-Responsible for conversion of Pyruvate
to Carbon Dioxide and Acetyl portion
of Acetyl-CoA.
 CoA Molecule- has –Sh group at one end
where Acetyl group is attached.
 Acetyl-CoA- is a Thiolester with a sulfur
atom, replacing oxygen of the usual
carboxylic ester.
 Thiolester- high energy compound
 CoA is a Thiol(Sulfur[THIO] analog of an
alcohol)
*Hydrolysis of esters release energy to drive
reactions.
Enzymes that makeup the Pyruvate
Dehydrogenase Complex:
 Pyruvate Dehydrogenase
 Dihydrolipoyl Transacetylase
 Dihydrolipoyl Dehydrogenase
 Lipoic Acid-compound that has di-
sulfide group in its oxidized form
and two sulfhydryl groups in its
reduced form.
-is a Vitamin
-formation of Thiolester
linkage with the acetyl group before it
is transferred to Acetyl-CoA.
REACTION SEQUENCE:
First step in the reaction sequence that converts
Pyruvate to Carbon Dioxide and Acetyl-CoA.
• is catalyzed by Pyruvate Dehydrogenase
• requires Thiamine Pyrophosphate is a
coenzyme (metabolite of vitamin B1)
• coenzyme not covalently bonded to the
enzyme, held together by noncovalent
interactions.
• Mg2+ required
• in Pyruvate Dehydrogenase reaction a –Keto
acid, pyruvate,loses Carbon Dioxide; the
remaining two carbon unit becomes
covalently bonded to TPP.
Second step of the reaction is catalyzed by
Dihydrolipoyl Transacetylase.
• Dihydrolipoyl Transacetylase requires Liopic
Acid as a coenzyme.
• Lipoic acid is covalently bonded to the
enzyme.
• The two carbon unit that originally came
from pyruvate is transferred from the
Thiamine Pyrophosphate to the Lipoic acid,
in the process a hydroxyl group is oxidized
to produce an Acetyl group.
• Disulfide group of Lipoic acid is the oxidizing
agent, which itself reduced, the product is a
Thiolester.
• Acetyl group is noncovalently bonded to the
lipoic acid by a thiolester linkage.
Third step of the reaction Is also catalyzed by
Dihydrolipoyl Transacetylase.
• a molecule of CoA-Sh attacks the Thiolester
linkage, the acetyl groupis transferred to it.
• Acetyl group remains bound to the
Thiolester linkage, this time as Acetyl-CoA
rather than Esterified to Lipoic acid.
• the reaction of pyruvate and CoA-SH has non
reach the stage of the products,CO2 and
Acetyl-CoA, but the lipoic acid coenzyme is
in a required form.
Fourth step- the enzyme Dihydrolipoyl
Dehydrogenase reoxidezes the reduced lipoic acid
from sulfhydryl to the disulfide form.
THE CITRIC ACID CYCLE PROPER:
Step 1:Formation of Citrate
 Reaction of Acetyl-CoA and Oxaloacetate to
form citrate and CoA-SH(Reaction is called
condensation).
 condensation is followed by the hydrolysis of
citryl-CoA to give citrate and CoA-SH.
 reaction catalyzed by citrate
synthase(Condensing enzyme).
 Citrate Synthase-Allosteric enzyme
 Inhibitors= ATP, NADH and Succinyl-CoA.
=is a feedback control mechanism.
Step 2: Isomerization of citrate to isocitrate
 catalyzed by Aconitase, is the isomerization
of citrate to isocitrate.
 requires Fe2+
 Isocitrate has four possible isomers, only one
produced in this reaction.
 removal of H2O molecule from citrate to
produce cis-Aconitase, and then H2O is added *The
back to the cis-Aconitase to give isocitrate.
 There is some evidence that citrate is
completed to the Fe(II).
 In the reaction mechanism, a phosphate
group covalently bonded to the enzyme
is directly transferred to GDP.
 The energy required for the
phosphorylation of GTP is provided by
the Hydrolysis of Succinyl-CoA to Produce
Succinate and CoA.
enzyme nucleoside di-phosphate kinase
catalyze the transfer of a phosphate
group from GTP to ADP to give GDP and
ATP.

Step 3: Formation of Alpha –Ketoglutarate and

CO2 first oxidation.
 is the Decarboxylation of citrate to isocitrate Step 6: Formation of Fumarate-FAD-Linked
to Alpha-Ketoglutarate and CO2.
Oxidation
 This reaction is the first of the two oxidative
 Succinate is Oxidized to fumarate, A
Decarboxylation of CTA cycle; enzyme that
reaction that is catalyzed by the enzyme
catalyze it is Isocitrate Dehydrogenase.
Succinate Dehydrogenase(Integral
Steps:
protein of the inner mitochondrial
-First, Isocitrate is to oxidezed to oxalo
membrane).
succinate.
-Second, Oxalosuccinate is decarboxylated
and the CO2 and Alpha-Ketoglutarate are
released.
-first of the reactions in which NADH
is produced.
*Inhibited by ATP and NADH, and activated by ADP
and NAD+

Step 7:Formation of L-Malate

 catalyzed by the enzyme Fumarase.
 water is added across the double bond of
Fumarate in the hydration reaction to
give Malate.
Step 4: Formation of Succinyl-CoA are formed
from Alpha-Ketoglutarate and CoA.
 Carbondioxide and Succinyl-CoA are formed
from Alpha-KetoGlutarate and CoA.
 Catalyzed by as enzyme as enzyme system
called Alpha-Ketoglutarate Dehydrogenase
complex.

Akala mo may greatings noH!

(wrong spelling pa eh dapat ito: Grit-engs) –ed.
moi
Kala mo may chisms!
Masydo na kyong CHISMOSA!(smiling face)
FINAL STAGES OF CITRIC ACID CYCLE - SUCCINATE
IS CONVERTED TO OXALOACETATE(Conversion of
methylene group to a Carbonyl group)
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Step 5: Formation of Succinate MM.
*LOOK FOR
 the thiolesterbond of succinyl-CoA is
hydrolyzed to produced succinate and
First come, first served
CoA-SH, an accompanying reaction is the
Phosphorylation of GDP to GTP.
basis! Hurry! Avail now!
 is catalyzed by Succinyl-CoA Synthase.