Bahir Dar University

Department of Industrial Chemistry

Biochemistry (Ichem.3052)
Chapter 8: Carbohydrate Metabolism

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 Structure of Carbohydrate

Isomers - compounds that have the same chemical formula but have
different structures

- For example, fructose, glucose, mannose, and galactose ,having

a formula of C6H12O6

Epimers- when two monosaccharides differ in configuration around

only one specific carbon atom

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C1 and C4 are an epimers
and an isomer of glucose

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 Digestion of Carbohydrates

-Carbohydrates digestion are in mouth and intestinal lumen.

-Enzymes needed for degradation of most dietary carbohydrates
(that break oligosaccharides and polysaccharides).
- Hydrolysis of glycosidic bonds is catalyzed by a family of
glycosidases that degrade carbohydrates into their reducing sugar
components

-Hydrolysis of glycosidic bond

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 Digestion of carbohydrates begins in the mouth

-Polysaccharides of animal (glycogen) and plant origin (starch,

composed of amylose and amylopectin).
- Salivary α-amylase acts on dietary starch in breaking some α(1-4)
bonds.
-Humans are unable to digest cellulose because, we do not have β(1-
4)-endoglucosidase
-Further digestion of carbohydrates by pancreatic enzymes occurs
in the small intestine
-acidic in small intestine are neutralized by bicarbonate secreted
by the pancreas, and pancreatic α-amylase continues the process of
starch digestion. 5
-The final digestive processes occur at the mucosal lining of the small
intestine
-Several disaccharidases (lactase (β-galactosidase), sucrase , maltase
and isomaltase ) produce monosaccharides.

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 Glycogen Metabolism

- Excess glucose is converted to polymeric forms for storage—

glycogen in vertebrates and many microorganisms.

- Glycogen in muscle provide a quick source of energy for either

aerobic or anaerobic metabolism.

-Muscle glycogen can be exhausted in less than an hour during

vigorous activity.

-Liver glycogen serves as a reservoir of glucose for other tissues

when dietary glucose is not available.

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 Glycogen breakdown by Glycogen Phosphorylase

-Glycogen enter to glycolytic pathway through the action of three

enzymes:
- Glycogen Phosphorylase,

-Glycogen debranching enzyme, and

- Phosphoglucomutase

Glycogen Phosphorylase- Catalyzes the reaction in which an (α1-> 4)

glycosidic linkage between two glucose residues at a non reducing end

of glycogen undergoes attack by inorganic phosphate (Pi), removing

the terminal glucose residue as -D-glucose 1-phosphate

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This process is repetitive; the enzyme removes successive glucose
residues until it reaches the fourth glucose unit from a branch point
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-Glucose residues near a branch
Are removed in a two-step
process that requires a
bifunctional “debranching
enzyme.”

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Glucose 1-phosphate to glucose 6-phosphate by phosphoglucomutase

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 steps

1. The enzyme donates its phosphoryl group (green) to glucose

1-phosphate, producing glucose 1,6-bisphosphate.
2. The phosphoryl group at C-1 of glucose 1,6-bisphosphate (red) is
transferred back to the enzyme, re-forming the phosphoenzyme and
producing glucose 6-phosphate.

- Glucose 6-phosphate goes to glycolytic pathways

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 Metabolism of Monosaccharides and Disaccharides

Fructose Metabolism
-Fructose is found as a free monosaccharide in high-fructose corn
syrup, in many fruits, and in honey.
-In contrast to glucose, fructose does not promote the secretion of
insulin.
- For fructose to enter the pathways of intermediary metabolism, it
must first be phosphorylated.
-Fructokinase provides the primary mechanism for fructose
phosphorylation
- Found in liver , kidney, and the small intestinal mucosa
- Converts fructose to fructose 1-phosphate and ATP used as
phosphate donor 13
-Fructose 1-phosphate is not converted
to fructose 1,6-bisphosphate as fructose
6-phosphate, but is cleaved by aldolase B
(fructose1-phosphate aldolase) to
dihydroxyacetone phosphate (DHAP)
and glyceraldehyde.

Glycolysis pathway
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-Both aldolase A(glycolysis pathway) and aldolase B cleave fructose
1,6-bisphosphate produced during glycolysis to DHAP and
glyceraldehyde3-phosphate
-DHAP can directly enter glycolysis or gluconeogenesis, whereas
glyceraldehyde can be metabolized by a number of pathways.

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 Galactose Metabolism

-Major dietary source of galactose is lactose (galactosyl β-1 ,4-

glucose) obtained from milk and its products.
-Some galactose can also be obtained by lysosomal degradation of
complex carbohydrates, such as glycoproteins, and glycolipids,.
-The entry of galactose into cells is not insulin-dependent.

Phosphorylation of galactose
-Galactose phosphorylated before it is further metabolized.
-The enzyme galactokinase produces galactose 1-phosphate and
ATP is used as phosphate donor.

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- Formation of UDP-galactose (Uridine diphosphate galactose
-Galactose 1-phosphate cannot enter the glycolytic pathway unless
it is first converted to UDP-galactose.
-Galactose 1-phosphate transfer to UDP-galactose . The enzyme
galactose 1-phosphate uridyltransferase catalyzes the reaction .

-Useof UDP-galactose as a carbon source for glycolysis or

gluconeogenesis
- In order for UDP-galactose to enter the mainstream of glucose
metabolism, it must first be converted to its C-4 epimer UDP-
glucose, by UDP-hexose 4-epimerase.

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 Lactose synthesis

- Lactose is a disaccharide that consists of a molecule of β-galactose

attached by β(1->4) linkage to glucose. Therefore, lactose is
galactosyl β(1 ->4)-glucose .
- Lactose(milk sugar) is produced by the mammary glands of most
mammals. Therefore, milk and other dairy products are sources.

- Lactose is synthesized in the endoplasmic reticulum by lactose

synthase, (UDP- galactose:glucose galactosyltransferase), which
transfers galactose from UDP-galactose to glucose.

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 Glycolysis

-The glycolytic pathway is employed by all tissues for the breakdown

of glucose to provide energy (in the form of ATP) and intermediates
for other metabolic pathways.
-Glycolysis is at the hub of carbohydrate metabolism because
virtually all sugars—whether arising from the diet or from
catabolic reactions in the body –can ultimately be converted to
glucose .

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-Pyruvate is the end product of glycolysis in cells with mitochondria
and an adequate supply of oxygen. In aerobic glycolysis , oxygen is
required to reoxidize the NADH formed during the oxidation of
glyceraldehyde 3-phosphate.
-Alternatively, glucose can be converted to pyruvate, which is
reduced by NADH to form lactate.
- This conversion of glucose to lactate is called anaerobic glycolysis
because it can occur without the participation of oxygen. Anaerobic
glycolysis allows the continued production of ATP in tissues that lack
mitochondria (for example, red blood cells) or in cells deprived of
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sufficient oxygen.
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 Reactions of Glycolysis

-The conversion of glucose to pyruvate occurs in two stages .

-The first five reactions of glycolysis correspond to an energy
investment phase in which the phosphorylated forms of
intermediates are synthesized at the expense of ATP.

-The subsequent reactions of glycolysis constitute an energy

generation phase in which a net of two molecules of ATP are formed
by substrate level phosphorylation per glucose molecule
metabolized.
-Two molecules of NADH are formed when pyruvate is produced
(aerobic glycolysis), whereas NADH is reconverted to NAD+
when lactate is the end product (anaerobic glycolysis). 24
 Energy investment Phase

- Phosphorylation of glucose 25
 Energy Generative Phase
- Conversion of glyceraldehyde 3-phosphate to pyruvate

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 Overall reactions of glycolysis
1. Phosphorylation of glucose
- Phosphorylation of glucose to glucose 6-phosphate is catalyzed by
hexokinase
2. Isomerization of glucose 6-phosphate

- Isomerization of glucose 6-phosphate to fructose 6-phosphate is

catalyzed by phosphoglucose isomerase
3. Phosphorylation of fructose 6-phosphate

The irreversible phosphorylation reaction catalyzed by

phosphoructokinase- (PFK-1) is the most important control point
and the rate-limiting step of glycolysis
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4. Cleavage of fructose 1,6-bisphosphate
Aldolase A cleaves fructose1,6-bisphosphate to dihydroxyacetone
phosphate and glyceraldehyde 3-phosphate . The reaction is
reversible and not regulated.
Aldolase B in the liver and kidney also cleaves fructose 1,6-
bisphosphate.
5.Isomerization of dihydroxyacetone phosphate
- Triose phosphate isomerase interconverts dihydroxyacetone
phosphate to glyceraldehyde 3-phosphate.
- Dihydroxyacetone phosphate must be isomerised to
glyceraldehyde3-phosphate for further metabolism by the glycolytic
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pathway.
 6. Oxidation of glyceraldehyde 3-phosphate
-The conversion of glyceraldehyde 3-phosphate to 1,3-bisphospho-
glycerate (1,3-BPG ) by glyceraldehyde 3-phosphate dehydrogenase
-It is the first oxidation-reduction reaction of glycolysis .
- The NADH formed by this reaction must be reoxidized to NAD+
for glycolysis to continue
7. Synthesis of 3-phosphoglycerate producing ATP

- 1,3-BPG converted to 3-phosphoglycerate, by phosphoglycerate

kinase.
- The high-energy phosphate group of 1,3-BPG is used to synthesize
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ATP from ADP .
8. Shift of the phosphate group from carbon 3 to carbon 2

- The shift of the phosphate group from carbon 3 to carbon 2 of

phosphoglycerate by phosphoglycerate mutase and it is freely
reversible

9. Dehydration of 2-phosphoglycerate

- Dehydration of 2-phosphoglycerate by enolase, resulting in the

formation of phosphoenolpyruvate (PEP)
- It contains a high energy - enol phosphate.

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 10. Formation of pyruvate producing ATP
- Conversion of PEP to pyruvate is catalyzed by pyruvate kinase , it is
irreversible reaction .
- The pyruvate kinase reaction favors the formation of ATP
- Glycolytic enzyme deficiencies cause for anemia. Anemia is a
consequence of the reduced rate of glycolysis, leading to decreased
ATP production. Resulting alterations in the red blood cell membrane
, lead to changes in the shape of the cell and, ultimately, to
phagocytosis

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11. Reduction of pyruvate to lactate
Lactate is formed by the action of lactate dehydrogenase,
- Lactate is the final product of anaerobic glycolysis in eukaryotic
cells like red blood cells, lens and cornea of the eye, kidney
medulla, testes, and leukocytes.

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 FATES OF PYRUVATE
A. Oxidative decarboxylation of pyruvate
- Pyruvate dehydrogenase irreversibly converts pyruvate, the end
product of glycolysis, into acetyl CoA.
- acetyl CoA a major fuel for the tricarboxylic acid cycle and the
building block for fatty acid synthesis
B. Carboxylation of pyruvate to oxaloacetate
- Carboxylation of pyruvate to oxaloacetate (OAA) by
pyruvatecarboxylase.
- This reaction is important because it replenishes the citric acid cycle
intermediates, and provides substrate for gluconeogenesis

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C. Reduction of pyruvate to ethanol (microorganisms)
- The conversion of pyruvate to ethanol.
- The decarboxylation of pyruvate by pyruvate decarboxylase
occurs in yeast and certain microorganisms. The enzyme requires
thiamine pyrophosphate as a coenzyme.

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 Energy Yield of Glycolysis

- Production of ATP during glycolysis, the end products, pyruvate or

lactate, still contain most of the energy originally contained in
glucose.
1. . Aerobic glycolysis:

- Two molecules of ATP and Two molecules of NADH are

produced per molecule of glucose .
- Ongoing aerobic glycolysis requires the oxidation of most of this
NADH by the electron transport chain, producing approximately
three ATP for each NADH molecule entering the chain.

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2. Anaerobic glycolysis:
- Two molecules of ATP and two molecules of lactate are generated
from a molecule of glucose.
- No net production or consumption of NADH.
Anaerobic glycolysis: is a valuable source of energy under
conditions of
1) when the oxygen supply is limited, as in muscle during intensive
exercise:
2) For tissues with few or no mitochondria, such as the medulla of
the kidney, mature erythrocytes, leukocytes, and cells of the lens,
cornea, and testes. 36
 Tricarboxylic Acid Cycle

- Tricarboxylic acid cycle (TCA cycle ) , also called the Krebs cycle or
Citric acid cycle

-TCA cycle is the major energy-yielding metabolic pathway in cells,

providing the reduced coenzymes that will be oxidized by
the electron transport chain to yield adenosine triphosphate (ATP).
- Oxaloacetate, a key intermediate in the cycle, is the main precursor
for gluconeogenesis in the fasting state.

-The cycle take place in the mitochondria and it is an aerobic

pathway, O2 is required as the final electron acceptor.
-The cycle functions in the formation of glucose from the carbon
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skeletons of some amino acids.
 Reactions of the TCA cycle

- In the TCA cycle, oxaloacetate is first condensed with an a acetyl

CoA, and then is regenerated as the cycle is completed.

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A. Oxidative decarboxylation of pyruvate
-Pyruvate is converted to acetyl CoA by the pyruvate
dehydrogenase complex, which is a multienzyme complex .
- The pyruvate dehydrogenase complex is not part of the TCA cycle
, but is a major source of acetyl CoA.
B. Synthesis of citrate from acetyl CoA and oxaloacetate
- Condensation of acetyl CoA and oxaloacetate to form citrate is
catalyzed by citrate synthase
C. Isomerization of citrate
- Citrate is isomerized to isocitrate by aconitase

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D. Oxidation and decarboxylation of isocitrate
-Isocitrate dehydrogenase catalyzes the irreversible oxidative
decarboxylation of isocitrate, yielding the first of three NADH
molecules produced by the cycle, and the first release of CO2 .

E. Oxidative decarboxylation of α-ketoglutarate

- Conversion of α-ketoglutarate to succinyl CoA is catalyzed by the
α-ketoglutarate dehydrogenase complex.

F. Cleavage of succinyl CoA

- Succinate thiokinase (also called succinyl CoA synthetase)
cleaves the high-energy thioester bond of succinyl CoA.
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G. Oxidation of succinate
-Succinate is oxidized to fumarate by succinate dehydrogenase,
producing the reduced coenzyme FADH.
H. Hydration of fumarate
- Fumarate is hydrated to malate in a freely reversible reaction
catalyzed by fumarase (also called fumarate hydratase) .

I. Oxidation of malate
-Malate is oxidized to oxaloacetate by malate dehydrogenase.

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 Energy produced by the TCA cycle

-Two carbon atoms enter the cycle as acetyl CoA and leave as CO2.
- The cycle does not involve net consumption or production of
oxaloacetate or of any other intermediate

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 Electron transport chain and Oxidative phosphorylation.

Electron transport chain

-Energy-rich molecules, such as glucose, are metabolized by a series
of oxidation reactions ultimately yielding CO2and water.

-The metabolic intermediates of these reactions donate electrons to

specific coenzymes_ nicotinamide adenine dinucleotide (NAD+ )
and flavin adenine dinucleotide (FAD) to form the energy-rich
reduced coenzymes, NADH and FADH2 . These reduced coenzymes
can, in turn, each donate a pair of electrons to a specialized set of
electron carriers collectively called the electron transport chain .

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-The electron transport chain is present in the inner mitochondrial
membrane and is the common pathway by which electrons derived
from different fuels of the body flow to oxygen.

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 - Part of energy can be captured and stored by the production of
ATP from ADP and inorganic phosphate (Pi). This process is called
oxidative phosphorylation .

- Electron transport and ATP synthesis by oxidative phosphorylation

proceed continuously in all tissues that contain mitochondria
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 Reactions of the electron transport chain

1. Formation of NADH : NAD+ is reduced to NADH by dehydrogenases

that remove two hydrogen atoms from their substrate.
- Both electrons but only one proton (that is a hydride ion, :H ) are
transferred to the NAD+, forming NADH Plus a free proton H+
2. NADH dehydrogenase: The free proton plus the hydride ion
carried by NADH are next transferred to NADH dehydrogenase, an
enzyme complex I embedded in the inner mitochondrial membrane.
This complex has a tightly bound molecule of flavin mononucleotide
(FMN, a coenzyme structurally related to FAD, )that accepts the two
hydrogen atoms (2e- ,2H+), becoming FMNH2.
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NADH dehydrogenase also contains several iron atoms paired with
sulfur atoms to make iron-sulfur centers, necessary for the transfer of
the hydrogen atoms to the next member of the chain, ubiquinone
(known as coenzyme Q).

3. Coenzyme Q: It can accept hydrogen atoms both from FMNH2

produced by NADH dehydrogenase, and from FADH2 (Complex
II), which is produced by succinate dehydrogenase and , acyl CoA
dehydrogenase .

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4. Cytochromes:
Cytochromes contain a heme group made of a porphyrin ring
containing an atom of iron. Cytochrome iron atom is reversibly
converted from its Ferric ( Fe 3+) to its ferrous (Fe2+) form as a
normal part of its function as a reversible carrier of electrons.

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 Release of free energy during electron transport

Free energy is released as electrons are transferred along the

electron transport chain from an electron donor to an electron
acceptor .
The (electrons can be transferred in the forms, as hydride ions
(:H-) to NAD+, as hydrogen atoms (.H) to FMN, coenzyme Q,
and FAD, or as electrons ( .e-) to cytochromes .

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- The transfer of electrons down the electron transport chain is
energetically favored because NADH is a strong electron donor and
molecular oxygen is an avid electron acceptor. However, the flow of
electrons from NADH to oxygen does not directly result in ATP
synthesis .

ATP synthesis
After protons have been transferred to the cytosolic side of the inner
mitochondrial membrane, they re-enter the mitochondrial matrix by
passing through a channel in the ATP synthase complex v, resulting
in the synthesis of ATP from ADP + Pi .
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The enzyme complex ATP synthase

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 Hexose Monophosphate Pathway

- Hexose monophosphate pathway also called pentose phosphate

pathway or 6-phosphogluconate pathway, occurs in the cytosol of
the cell.
-Glucose 6-phosphate is entering to the oxidative part of the
pathway and released as CO2 and two NADPH per glucose 6-
phosphate molecule.
-The pathway provides a major portion of the body's NADPH.
-No ATP is directly consumed or produced in the cycle

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-It consists of irreversible oxidative reactions and reversible
sugar-phosphate interconversions.

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 1. Irreversible oxidative reactions
-Formation of ribulose 5-phosphate, CO2 and two molecules of
NADPH for each molecule of glucose 6-phosphate oxidized

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 -Glucose 6-phosphate dehydrogenase (G6PD) catalyzes glucose 6-
phosphate to 6-phosphogluconolactone.

-6-Phosphogluconolactone is hydrolyzed by 6-phosphogluconolactone

hydrolase.
-The subsequent oxidative decarboxylation of 6-phosphogluconate is
catalyzed by 6-phosphogluconate dehydrogenase.
-This irreversible reaction produces a pentose sugar-phosphate
(ribulose 5-phosphate), CO2 , and a second molecule of NADPH

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 Reversible nonoxidative reactions

-The nonoxidative reactions of the pentose phosphate pathway

occur in all cell types synthesizing nucleotides and nucleic acids.

-These reversible reactions permit ribulose 5-phosphate to be

converted either to ribose 5-phosphate (needed for nucleotide
synthesis), or to intermediates of glycolysis (fructose 6-phosphate
and glyceraldehyde 3-phosphate ).

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 NADPH
- NADPH is a high-energy molecule . The coenzyme NADP+ differs
from NAD+ only by the presence of a phosphate group (-PO4=)
on one of the ribose units.
-Electrons of NADPH are destined for use in reductive biosynthesis,
rather than for transfer to oxygen as is the case with NADH .
-Metabolic transformations of the pentose phosphate pathway, part of
the energy of glucose 6-phosphate is conserved in NADPH.

Structure of NADPH

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 Gluconeogenesis

Gluconeogenic precursors are molecules that can be used to produce

a net synthesis of glucose.
Substrates for gluconeogenesis
-All the intermediates of glycolysis and the citric acid cycle.
-Glycerol, lactate, and the α-keto acids are the most important
gluconeogenic precursors .

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Reactions of gluconeogenesis

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 Reactions unique to gluconeogenesis

- Glycolytic reactions used in the synthesis of glucose from lactate or

pyruvate.
A. Carboxylation of pyruvate

- Pyruvate is first carboxylated by pyruvate carboxylase to

oxaloacetate (OAA)
B. Transport of oxaloacetate to the cytosol
-OAA is unable to directly cross the inner mitochondrial
membrane; it must first be reduced to malate by mitochondrial
malate dehydrogenase.

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-Malate can be transported from the mitochondria to the cytosol,
where it is reoxidized to oxaloacetate by cytosolic malate
dehydrogenase
C. Decarboxylation of cytosolic oxaloacetate
- Oxaloacetate is decarboxylated and phosphorylated in the cytosol
by PEP-carboxykinase
D. Dephosphorylation of fructose 1,6-bisphosphate
- Hydrolysis of fructose 1,6-bisphosphate by fructose 1,6-
bisphosphatase and provides an energetically favorable pathway for
the formation of fructose 6-phosphate.

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 E. Dephosphorylation of glucose 6-phosphate
- Hydrolysis of glucose 6-phosphate by glucose 6- phosphatase and
provides an energetically favorable pathway for the formation of free
glucose.
- Liver and kidney are the only organs that release free glucose
from glucose 6-phosphate.

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 Regulation of gluconeogenesis

- The regulation of gluconeogenesis is determined primarily by

circulating level of glucagon, and by the availability of
gluconeogenic substrates.

Glucagon
Pancreatic islet hormone stimulates gluconeogenesis by three
mechanisms
1. Changes in allosteric effectors: Glucagon lowers the level of
fructose 2,6-bisphosphate, resulting in activation of 1,6-bis fructose
phosphatase and inhibition of phosphofructokinase .

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2.Covalent modification of enzyme activity: Glucagon and cAMP-
dependent protein kinase activity, stimulates the conversion of
pyruvate kinase to its inactive (phosphorylated) form. This decreases
the conversion of PEP to pyruvate, which has the effect of diverting
PEP to the synthesis of glucose.

3. Induction of enzyme synthesis: Glucagon increases the

transcription of the PEP carboxykinase gene, thereby increasing the
availability of this enzyme's activity as levels of its substrate rise
during fasting.

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 Substrate availability

The availability of gluconeogenic precursors, particularly

glucogenic amino acids, significantly influences the rate of
hepatic glucose synthesis. Decreased levels of insulin favor
mobilization of amino acids from muscle protein, and provide
the carbon skeletons for gluconeogenesis.

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THANK YOU FOR YOUR ATTENTION!

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