Lecture 2 PHBC622

Carbohydrate
Metabolism
(Aerobic Metabolism)

May 23, 2023 Dr. Mohamed Z Gad 1

 Learning Objectives
By the end of this session you should be able to:
• Illustrate the conversion of pyruvate to acetyl CoA and
its regulation.
• Describe the reactions and key features of aerobic
metabolism.
• Mention the regulation of TCA cycle and energy
generated.
• Describe
May 23, 2023
citrate metabolism.
Dr. Mohamed Z Gad 2
 Aerobic Metabolism
The Krebs cycle, also called Citric acid
cycle or Tricarboxylic acid cycle (TCA
cycle), is a series of biochemical reactions
that are responsible for the complete
oxidation of organic substrates to form Sir Hans Adolf Krebs
CO2 and H2O. (1900-1981)
In presence of O2, pyruvate generated from glycolysis is
completely oxidized to CO2 and H2O and the high-energy electrons
that are removed from TCA cycle are transferred to NAD+ and
FAD+ to form NADH and FADH2. Energy is captured in the form of
ATP during a complex process called oxidative phosphorylation.
Before entering the TCA cycle, pyruvate should be converted to
acetyl CoA.
May 23, 2023 Dr. Mohamed Z Gad 3
Conversion of
Pyruvate to Acetyl
CoA
After its transport into the
1
mitochondrial matrix,
pyruvate is converted to
acetyl CoA in a series of
reactions catalysed by a
2
multienzyme complex called 5
Pyruvate dehydrogenase
complex [PDH complex].
PDH reaction requires 5 4 3
coenzymes: TPP, Lipoic
acid, CoA, FAD & NAD.
The net reaction is:
Pyruvate + NAD
May 23, 2023
+
+ CoA-SH acetyl-CoA
Dr. Mohamed Z Gad + NADH + CO2 + H4
+
Steps of the PDH Reaction:
1 Pyruvate decarboxylase (E1) activity of PDH catalyses the decarboxylation
of pyruvate. An intermediate, hydroxyethyl-TPP (HETPP), is formed thereafter
with the coenzyme thiamine pyrophosphate (TPP). 2 HETPP is converted to
acetyl CoA by dihydrolipoyl transacetylase (E2). Lipoic acid plays a crucial
role in this transformation. It bound to the enzyme, reacts with HETPP to form
an acetylated lipoic acid and free TPP. 3 The acetyl group is then transferred to
the –SH group of Coenzyme A. 4 Subsequently, the reduced lipoic acid is
reoxidized by dihydrolipoyl dehydrogenase (E3) in presence of FAD. 5 The
FADH2 is reoxidized by NAD+.
Regulation of PDH complex:
By two mechanisms:
1. Product inhibition: The enzyme complex is inhibited directly by high
concentrations of the reaction products acetyl CoA and NADH.
2. Covalent modification: Acetyl CoA and NADH also activate a kinase,
kinase
which converts the active PDH complex to an inactive phosphorylated
form. The PDH complex is reactivated by a dephosphorylation reaction
catalysed by a phosphoprotein
May 23, 2023 phosphatase.
phosphatase
Dr. Mohamed Z Gad This enzyme is activated
5
when the mitochondrial ATP is low.
 Reactions of the TCA Cycle
In order to follow the passage of acetyl-CoA through the
cycle, the 2 carbon atoms of the acetyl radical are shown
labelled on the carboxyl carbon (using the designation [*]
and on the methyl carbon (using the designation )

May 23, 2023 Dr. Mohamed Z Gad 6

 Key Points in TCA Cycle
1) Two forms of isocitrate dehydrogenase in mammals:
NAD-requiring : in mitochondria
NADP–requiring: in both mitochondrial matrix and cytoplasm, generates
NADPH not NADH
2) Oxidative decarboxylation of -ketoglutarate is similar to PDH reaction.
Similarities: requirement of same cofactors (TPP, CoASH, Lipoic acid , NAD,
FAD) & inhibition by same allosteric effectors
3) Succinyl CoA synthase requires GDP -> GTP
4) Succinate dehydrogenase requires FAD -> FADH2
5) All TCA cycle enzymes are in mitochondrial matrix EXCEPT succinate
dehydrogenase, in inner mitochondrial membrane
Outcome of TCA Cycle
TCA cycle begins with the condensation of acetyl CoA with oxaloacetate to form
citrate, which is then reconverted to oxaloacetate. During this process, 2 molecules
of CO2, 3 molecules of NADH (3x3ATP=9ATP), 1 molecule of FADH2 (2ATP), and 1
molecule of GTP (1ATP) are produced. Thus: For each TCA cycle turn, 1 acetyl CoA
May 23, 2023 Dr. Mohamed Z Gad 7
produces 12 ATP .. TRY TO FIND ON THE CYCLE !!!!
 The Amphibolic Nature of TCA Cycle
TCA cycle functions in both anabolic and catabolic ways:
Catabolic Nature: acetyl groups are oxidized to form CO2 and energy in the
form of NADH & FADH2.

Anabolic Nature: several TCA cycle intermediates serve as precursors in

biosynthetic pathways,
e.g. oxaloacetate: in gluconeogenesis and a.a synthesis, -ketoglutarate: role in
a.a synthesis., succinyl CoA: in synthesis of porphyrins & ketone bodies, acetyl
CoA: synthesis of fatty acids and cholesterol.

TCA cycle intermediates are replenished by several anaplerotic reactions

e.g. synthesis of succinyl CoA from certain a.a & transamination reactions that
generate -ketoglutarate and oxaloacetate from a.a.
One of the most important anaplerotic reactions is the pyruvate carboxylase
reaction, activated when concentration of acetyl CoA is high.
ATP ADP
May 23, 2023 Dr. Mohamed Z Gad 8
-OOC-CO-CH3 + CO2 -OOC-CH2-CO-COO-
 Regulation of the TCA Cycle

By modulation of key enzymes & availability of certain substrates and

coenzymes such as NAD+, FAD & ADP.
Enzyme Activator Inhibitor

Pyruvate dehydrogenase Low mitochondrial ATP High acetyl CoA, ATP

& NADH
Pyruvate carboxylase High acetyl CoA
Citrate synthase Substrate availability High succinyl CoA,
citrate , ATP & NADH

Isocitrate dehydrogenase High ADP & NAD+ High ATP & NADH

- ketoglutarate Decrease ATP High succinyl CoA &

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dehydrogenase NADH
 ATP Generated from Complete Oxidation of
Glucose

Complete oxidation

Aerobic = 38 or 36 ATP
(Aerobic Glycolysis = 6 or 8 + TCA cycle = 30
(Pyruvate dehydrogenase = 6 , TCA= 2 x 12 = 24)

Anaerobic = 2 ATP (anaerobic glycolysis)

May 23, 2023 Dr. Mohamed Z Gad 10

Citrate Metabolism
Citrate transport from
mitochondria to cytosol is used
as a mechanism for transferring
acetyl CoA out of mitochondria.

Citrate in cytoplasm has several

functions:
1) Gives acetyl CoA -> FA
synthesis.
2) Gives oxaloacetate ->
biosynthesis reactions e.g. to
malate -> by malic enzyme
(source of NADPH) ->
pyruvate.
3) Allosteric activator of 1st
reaction of FA synthesis.
4) An inhibitor of PFK-1.
Movement
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of large no. of citrate into cytoplasm indicates that the 11
Dr. Mohamed Z Gad
cells'
current requirement for energy is low (no need for further Krebs cycle)
 ‫كلمات باقية‬

‫العدل جنة المظلوم ‪ ..‬و جحيم الظالم‪.‬‬ ‫•‬

‫إني ألهاب الرجل صامتا ‪ ..‬حتي يتكلم‪.‬‬ ‫•‬

‫الخليفة عمر بن الخطاب‬

References for Further Readings:
Latest editions of:
•Biochemistry by L. Stryer, Freeman & Company New York ….
•Harper’s Biochemistry by R.K. Murray, D.K. Granner, P.A.
Mayes & V.W. Rodwell. Biochemistry. Appleton & Lange, New
York, Connecticut, California.
•Biochemistry by T. Mckee & J. Mckee, Wm. C. Brown
Publishers, London, Madrid, Tokyo ….
•Biochemistry by I.D.K. Halkerston, Second Edition, John Wiley
& Sons Inc., Williams E Wilkins, USA.
• Textbook of Biochemistry With clinical Correlations by T.M.
Devlin, Wiley-Liss Publication, New York, Toronto ….
•Biochemistry Illustrated by P.N. Campbell & A.D. Smith,
Longman Group UK Ltd, UK.
•Principles Of Biochemistry by G.L. Zubay, W.W. Parson & D.E.
May 23, 2023 Dr. Mohamed Z Gad 13
Vance, Wm. C. Brown Publishers, Iowa, Melbourne, Ox for d.
May 23, 2023 Dr. Mohamed Z Gad 14