Metabolism of Carbohydrates

I
LEARNING OBJECTIVES

Students should be able to:

• Recognise glucose as central compound to pathways of CHO
metabolism
• Discuss reasons why glucose is such an important fuel for most
organisms
• Define / explain glycolysis, citric acid cycle and uronic acid pathway.
• State the importance of these pathways
• The major monosaccharide obtained from the
intestinal hydrolysis of dietary carbohydrate is
glucose.
• Most tissues are partially or totally dependent on
glucose for ATP generation and for production of
precursors of other pathways
• Principal food (fuel) for the brain, kidney medulla, red
blood cells, testis, lens and cornea of the eye and
other body tissues.
• Its level in the blood (80 – 100mg/dL) represents a
balance between processes adding or removing it
from the blood.
 Adding Removing

• Absorption from • Oxidation in tissues

intestine to provide energy
• Glycogenolysis • Glycolysis
• Gluconeogenesis • Glycogenesis
• Excretion in urine at
high blood glucose
levels
Brief summary of glucose utilisation

• Oxidation….Glycolysis (Embden-Meyerhof Pathway),

HMP shunt pathway, Uronic acid pathway
• Conversion to fats (lipogenesis)
• Conversion to amino acids
• Conversion to other carbohydrates (Fructose,
Mannose, Glucosamine, Galactose, D-glucuronic acid,
Ribose, Deoxyribose, e.t.c.
• Storage as glycogen (glycogenesis).
1. Glycolysis: “Splitting of sugar”.
• It is a series of reactions that converts glucose into
pyruvate with the concomitant trapping of a portion
of the energy as ATP.
• Ancient process central to all life.
• Principal pathway for the metabolism of glucose in
the cytosol of all cells.
• Glucose (6 C) is split and oxidized to two molecules
of pyruvate (3C).
Net yield per glucose molecule:
2 ATP (substrate level phosphorylation)
2 NADH
• Overall equation:
Aerobic: Glucose + 2ADP + 2Pi + 2NAD+ 2 pyruvate + 2ATP +
2NADH + 2H+
Anaerobic: Glucose + 2ADP + Pi 2 Lactate + 2ATP + 2H2O
Note:
• Virtually all sugars (whether ingested through dietary means or
formed from catabolic reactions in the body) can be converted
to glucose which eventually undergo breakdown via glycolysis.
• Glycolysis therefore, is at the hub of CHO metabolism.
• Net generation of 2 molecules of ATP via substrate level
phosphorylation (direct transfer of high-energy phosphate
from intermediates of the pathway to ADP)
• Pyruvate is the end product of glycolysis in cells with
mitochondria and an adequate supply of oxygen.
• The pyruvate then undergo oxidative decarboxylation (aerobic
glycolysis) to acetyl coA, a major fuel for the TCA cycle
• Fate of the two Pyruvates from glycolysis
One of three possibilities :
 lactate formation
alcoholic fermentation
the obligatory step.
• Lactate Formation
Under anaerobic conditions (sprinters, race
horses) glucose is metabolized to lactate (3
carbon compound), this is energetically inefficient, but does
produce energy.
• Alcoholic fermentation
Certain yeast cells convert pyruvate to ethanol,
not common except in these cells.
• Ethanol is formed from pyruvate in yeast
and several other microorganisms. The
first step is the decarboxylation of
pyruvate. This reaction is catalyzed by
pyruvate decarboxylase, which requires
the coenzyme thiamine pyrophosphate.
• The second step is the reduction of
acetaldehyde to ethanol by NADH, in a
reaction catalyzed by alcohol
dehydrogenase. This process regenerates
NAD+.
Obligatory step
The most common fate of the two pyruvates is to
enter into the obligatory step.
• The mitochondria of the cell converts the 2
pyruvates in to acetyl coenzyme A (the
coenzyme A part is a derivative of pantothenic
acid).
• This yields no energy but does give up some
NADH and hydrogen (2 each), that later yields
some energy
Regulation of glycolysis
• PFK: This is the most important control step of glycolysis. The
enzyme, PFK is regulated by
ATP/AMP: PFK is allosterically inhibited by ATP. This inhibition is
however reversed by AMP.
Citrate: High levels of citrate inhibits PFK.
H+: PFK is inhibited by H+, hence the rate of glycolysis
decreases when the pH is very low.
• Hexokinase: HK is inhibited by Glucose 6-P.
• Pyruvate kinase: This enzyme which catalyses the 3rd
irreversible step in glycolysis is activated by fructose 1,6-bisP
and inhibited by ATP as well as alanine.
2. The Kreb’s or TCA cycle or Citric acid cycle (the
fate of the two acetyl coenzyme A’s)
• The TCA cycle occurs aerobically in the mitochondria of the cell.
• It is the final common pathway for the aerobic oxidation of
carbohydrate, lipid and protein because glucose, fatty acids, and
most amino acids are metabolized to acetyl-CoA or intermediates of
the cycle.
• It is a series of enzymatic reactions designed to split the remaining 4
carbons (2 carbons each in the 2 acetyl coA’s produced in the
obligatory step) and release them as CO2.
• The enzymes of the citric acid cycle are located in the mitochondrial
matrix, either free or attached to the inner mitochondrial
membrane.
• The cycle is the biochemical hub of the cell, oxidizing carbon fuels,
usually in the form of acetyl CoA, as well as serving as a source of
precursors for biosynthesis (it is an amphibolic pathway).
• 2 ATP’s and 16 H+ are generated.
Reactions of the Citric acid cycle:
• The initial reaction between acetyl-CoA and oxaloacetate to
form citrate is catalyzed by citrate synthase which forms a
carbon-carbon bond between the methyl carbon of acetyl-CoA
and the carbonyl carbon of oxaloacetate. The thioester bond of
the resultant citryl- CoA is hydrolyzed, releasing citrate and
CoASH.
• Citrate is isomerized to isocitrate by the enzyme aconitase.
• Isocitrate undergoes dehydrogenation catalyzed by isocitrate
dehydrogenase to form, initially, oxalosuccinate, which
remains enzyme-bound and undergoes decarboxylation to α-
ketoglutarate.
• α-Ketoglutarate undergoes oxidative decarboxylation in a
reaction catalyzed by α-ketoglutarate dehydrogenase complex
to form succinyl-CoA.
• Succinyl-CoA is converted to succinate by the enzyme
succinate thiokinase (succinyl-CoA synthetase). This
is the only example in the citric acid cycle of
substrate-level phosphorylation.
• The first dehydrogenation reaction, forming fumarate,
is catalyzed by succinate dehydrogenase.
• Fumarase (fumarate hydratase) catalyzes the
addition of water across the double bond of
fumarate, yielding malate.
• Malate is converted to oxaloacetate by malate
dehydrogenase, a reaction requiring NAD+.
Steps in the TCA cycle
Energetics of the TCA Cycle:
• 3 mols of NADH and 1 mol of FADH2 are produced for
each molecule of acetyl-CoA catabolized in one turn
of the cycle.
• Reoxidation of each NADH results in formation of 3
ATP and reoxidation of FADH2 results in formation of 2
ATP.
• 1 ATP (or GTP) is formed by substrate-level
phosphorylation catalyzed by succinate thiokinase.
• This gives a total of 12 ATP molecules per turn of the
TCA Cycle.
Consequences to impaired functioning of the TCA cycle

• Impaired functioning of the TCA cycle can occur if

oxygen is not available to accept electrons in the ETC.
The consequences include
Inability to generate ATP from fuel oxidation
• Accumulation of TCA cycle precursors e.g. inhibition
of pyruvate oxidation in the cycle results in its
reduction to lactate, which accumulates to cause
lactic acidosis.