Carbohydrate Metabolism-II

Instructor: Asst. Prof. Dr. Akıle Tuncal

akile.tuncal@kstu.edu.tr
 Gluconeogenesis

• Gluconeogenesis is the term used to include all mechanisms and

pathways responsible for converting non-carbohydrates to
glucose or glycogen.

• The major substrates are the glucogenic amino acids; lactate,

pyruvate; glycerol; and propionate.

• Gluconeogenesis occurs in the liver and kidney, the only organs

with a full complement of the necessary enzymes.
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 Gluconeogenesis
The Cori cycle:

• Lactate produced in glycolysis during muscle exertion is transported to

the liver, for resynthesis of glucose by gluconeogenesis.

• Transport of glucose back to muscle for synthesis of glycogen, and its

reutilization in glycolysis, completes the cycle.
 Gluconeogenesis
• In all organisms a three-carbon acyl-CoA, propionyl-CoA, is generated either from
the breakdown of some amino acids or from the oxidation of fatty acids with odd
numbers of carbon atoms.

• Propionyl-CoA enters gluconeogenesis via its conversion to succinyl-CoA and then

to oxaloacetate.

• The process involves a coenzyme derived from vitamin B12.

Z Cori
cycle

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 Biomedical Importance
• The biosynthesis of glucose is an absolute necessity of all mammals, because
the brain and nervous system, as well as erythrocytes, testes, renal medulla,
and embryonic tissue, require glucose from the blood as their sole or major
fuel source.
• The human brain alone requires 120 g of glucose each day. Below a critical
blood glucose concentration (normal = 65-110 mg/dL or 3.6-6 mM), brain
dysfunction can occur which can lead to coma and death.
• Even when fat may be supplying most of the caloric requirements of an
organism, there is always a certain basal requirement for glucose e.g. in
skeletal muscle under anaerobic conditions.
• Glucose is precursor of lactose in the mammary gland.
• Gluconeogenic mechanisms are used to clear lactate (from muscle and
erythrocytes) and glycerol (adipose tissue) from blood.

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 Gluconeogenesis
Synthesis and use of glucose in the human body:

• Liver and kidney cortex are the primary

gluconeogenic tissues.

• Brain, skeletal muscle, kidney medulla,

erythrocytes, and testes use glucose as their sole
or primary energy source, but they lack the
enzymatic machinery to synthesize it.

• Synthesis of glucose from non-carbohydrate

precursors is essential for maintenance of blood
glucose levels within acceptable limits.
 Gluconeogenesis
Synthesis and use of glucose in the human body:

 When dietary intake of carbohydrate is reduced and blood glucose

concentration declines, hormones trigger accelerated glucose synthesis
from non-carbohydrate sources.

 Lactate, pyruvate, glycerol (a catabolic product of triacylglycerols), and

certain amino acids represent the important non-carbohydrate sources.

 The process of producing glucose from such compounds is termed

gluconeogenesis.

 The liver is the major site of this activity, although under certain
circumstances, such as prolonged starvation, the kidneys become
increasingly important in gluconeogenesis.
 Gluconeogenesis

Synthesis and use of glucose in the human body:

 The glucose formed by the liver and the kidney is mostly returned to the
blood to maintain blood glucose levels.

 Note that intermediates of the TCA cycle can be converted to glucose, but
fatty acids cannot. They are metabolized to acetyl-CoA, which cannot be
converted to glucose.

 Gluconeogenesis essentially reverses the glycolytic pathway. It synthesizes

glucose and consumes ATP and NAD+ rather than producing ATP and NADH.
 Gluconeogenesis
• Gluconeogenesis uses specific enzymes to bypass
three irreversible reactions of glycolysis.

Reactions of glycolysis and gluconeogenesis:

• Irreversible reactions of glycolysis are shown in dark

purple.

• The opposed reactions in gluconeogenesis, which

bypass these steps, are shown in dark blue.

• Pale arrows identify reversible reactions used in both

pathways.
 Gluconeogenesis
Bypass 1: Conversion of Pyruvate to Phosphoenolpyruvate

• The bypass of pyruvate kinase begins in the mitochondrion and

involves two reactions.

• Pyruvate carboxylase catalyzes the ATP- and biotin-dependent

conversion of pyruvate to oxaloacetate.

• The enzyme requires acetyl-CoA as an allosteric activator:

 Gluconeogenesis
• To be used for gluconeogenesis, oxaloacetate must move out of the
mitochondrion to the cytosol, where the remainder of the pathway occurs.

• However, the mitochondrial membrane does not have an effective

transporter for oxaloacetate.

• Therefore, oxaloacetate is reduced by mitochondrial malate dehydrogenase

to malate, which is transported into the cytosol by exchange for
orthophosphate and then reoxidized by cytosolic malate dehydrogenase.

• Once in the cytosol, oxaloacetate is acted on by phosphoenolpyruvate

carboxykinase (PEPCK) to give phosphoenolpyruvate:
 Gluconeogenesis
• Bypass 2: Conversion of Fructose-1,6-bisphosphate to Fructose-6-phosphate

o The PFK reaction of glycolysis is essentially irreversible, but only because it is driven
by phosphate transfer from ATP.

o A bypass reaction in gluconeogenesis involves a simple hydrolytic reaction,

catalyzed by fructose-1,6- bisphosphatase.

• Bypass 3: Conversion of Glucose-6-phosphate to Glucose

o Glucose-6-phosphate cannot be converted to glucose by reverse action of

hexokinase or glucokinase because of the high positive DGo of that reaction;
phosphate transfer from ATP makes that reaction virtually irreversible.

o Another enzyme specific to gluconeogenesis, glucose-6-phosphatase, also

involves a simple hydrolysis.
Gluconeogenesis
Outline of pathways for glucose synthesis
from the major gluconeogenic precursors:

• Note that both glucose and lactate are

carried in the blood.

• The equivalent of 11 high-energy

phosphates are consumed per mole of
glucose synthesized by gluconeogenesis.

• The most important gluconeogenic

precursors are lactate, alanine, glycerol,
and propionate.

• Animals cannot undergo net conversion of

fat to carbohydrate.
Gluconeogenesis from Glycerol

• Occurs in liver, kidney, lactating mammary gland, heart and intestine.

• Glycerol is one of the major precursors for gluconeogenesis.

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Gluconeogenesis from Propionic Acid

• PA from methionine, isoleucine, oxidation of odd number fatty acids, cholesterol conversion
into bile acids, and, fermentation of fibers in large intestine and rumen of herbivorous animals.

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 Regulation of Gluconeogenesis
• Glucose-6-phosphatase, fructose-1,6-diphosphatase, phosphoenol pyruvate carboxykinase, and pyruvate
carboxylase are the key regulatory enzymes.

1. Hormonal Control

2. Metabolic Control

a. ATP/ADP Ratio

b. Acetyl-CoA

c. Citrate

d. Fructose-2,6-Biphosphate

e. NADH/NAD+ Ratio

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Gluconeogenesis

Copyright © 2013 Pearson Canada Inc.

 Polysaccharide Metabolism

 In animal metabolism, two primary sources of glucose are

derived from polysaccharides:

1. Digestion of dietary polysaccharides, chiefly starch from

plant foodstuffs and glycogen from meat.

2. Mobilization of the animal’s own glycogen reserves.

Glycogenolysis
• Degradation of Glycogen.

• The degradative pathway that mobilizes stored glycogen in liver and skeletal muscle
is not a reversal of the synthetic reactions.

• Instead, a separate set of cytosolic enzymes is required.

• When glycogen is degraded, the primary product is glucose 1-phosphate, obtained

by breaking α (1→4) glycosidic bonds.

• In addition, free glucose is released from each α (1→6)-linked glucosyl residue.

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 Glycogen Metabolism in Muscle and Liver

Glycogenolysis

 The principal glycogen stores in vertebrates are in skeletal muscle and liver.

 Breakdown of these stores into usable energy, or mobilization of glycogen, involves

sequential phosphorolytic cleavages of bonds, catalyzed by glycogen
phosphorylase.

 In plants, starch is similarly mobilized by the action of starch phosphorylase .

 Both reactions release glucose-1-phosphate from non-reducing ends of the glucose

polymer:
 Polysaccharide Metabolism

Cleavage of a glycosidic bond by hydrolysis

or Phosphorolysis:

This formal diagram shows how the elements

of water or phosphoric acid, respectively, are
added across a glycosidic bond.
 Polysaccharide Metabolism

 Dietary polysaccharides are metabolized by

hydrolysis to monosaccharides.

 Intracellular carbohydrate stores, as glycogen,

are mobilized as phosphorylated
monosaccharides by phosphorolysis.

 a-Amylase in saliva cleaves bonds a(14)

between the maltose units of amylopectin (or
glycogen).

 However, it cannot cleave a(16) glycosidic

bonds in the branched polymer, and a limit
dextrin accumulates unless a(16)-
glucosidase (debranching enzyme) is present.
 Glycogen Metabolism in Muscle and Liver
The debranching process in glycogen catabolism:

(a) A glycogen chain following activity by phosphorylase, which

cleaves off glucose residues to within four residues of the branch
point.

(b) The glycogen chain following transferase activity by the

debranching enzyme. The three remaining glucose residues with
linkage have been transferred to a nearby nonreducing end.

(c) The glycogen chain following a(16)-glucosidase activity by the

debranching enzyme, which has removed the last remaining
glucose residue of the branch.

• Phosphorylase will cleave off all but four glucose units of the
newly elongated branch, beginning the debranching process
again.
Copyright © 2013 Pearson Canada Inc.
 Glycogen Metabolism in Muscle and Liver

 At times of heightened glycogenolytic activity, the formation of increased amounts of

glucose-1-phosphate shifts the glucose phosphate isomerase reaction toward production
of the 6-phosphate isomer.

 The glucose-6- phosphate can enter into the oxidative pathway for glucose (glycolysis) or
become free glucose (in the liver or kidney).

 The conversion of glucose-6-phosphate to free glucose requires the action of glucose-6-

phosphatase.

 This enzyme is not expressed in muscle cells or adipocytes.

 Therefore, free glucose can be formed only from liver or kidney glycogen and transported
through the bloodstream to other tissues for oxidation.
 .

• Regulation of glycogen synthesis and degradation is accomplished on

two levels.

• First, glycogen synthase and glycogen phosphorylase are allosterically

controlled.

• Second, the pathways of glycogen synthesis and degradation are

hormonally regulated.

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Figure Allosteric regulation of glycogen synthesis and degradation. 27
 Glycogen Metabolism in Muscle and Liver
Glycogenesis

 This pathway is particularly important in hepatocytes because the liver is a major site
of glycogen synthesis and storage.

 Glycogen accounts for as much as 7% of the weight of the liver.

 Liver glycogen can be broken down to glucose and re-enter the bloodstream.

 The other major site of glycogen storage is skeletal muscle.

 In human skeletal muscle, glycogen generally accounts for a little less than 1% of the
weight of the tissue.

 The glycogen stores in muscle are an energy source within that muscle fiber and
cannot directly contribute to blood glucose levels.
 Glycogenesis
• is the Synthesis of Glycogen from α-D Glucose.

• occurs in the cytosol, and requires ATP and UTP.

• Site- Though every cell can form glycogen it chiefly occurs in liver and
skeletal muscle.

• Even though energy rich fat is abundant in the body, skeletal muscle
prefers to store glucose (energy) as glycogen because;
– Fat can not be oxidized under anaerobic condition.

– Skeletal muscle is unable to mobilize fat rapidly.

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 Glycogenesis

3
2

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 Reaction Sequence of Glycogenesis
• The glycogen synthesis occurs in four main reactions.
• Glycogenesis begins with glucose-6-phosphate an intermediate of
glycolysis.
1. the conversion of glucose-6- phosphate to glucose-1-phosphate.
The reaction is catalyzed by phosphoglucomutase.

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2. Synthesis of UDP-glucose
• UDP-glucose is synthesized from glucose 1-phosphate and UTP
by UDP-glucose pyrophosphorylase .

• α-D-Glucose attached to uridine diphosphate (UDP) is the

source of all the glucosyl residues that are added to the
growing glycogen molecule.

UDPG is active
sugar. It transfers
glucose residues to an
acceptor.

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 3. Synthesis of a primer to initiate glycogen synthesis

 Glycogen synthase is responsible for making the α(1→4) linkages in

glycogen.

 This enzyme cannot initiate chain synthesis using free glucose as an

acceptor of glucose from UDP-glucose.

 Instead, it can only elongate already existing chains.

 In the absence of a glycogen fragment, a protein, called glycogenin, can

serve as an acceptor of glucose residues from UDP-glucose .

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 ,

• The side chain hydroxyl group of a specific tyrosine serves as the

site at which the initial glucosyl unit is attached. The reaction is
catalyzed by glycogenin itself (autoglucosylation).

• Glycogenin then catalyzes the transfer of the next few molecules

of glucose from UDP-glucose, producing a short, α(1→4)-linked
glucosyl chain.

• This short chain serves as a primer that is able to be elongated by

glycogen synthase.

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4. Elongation of glycogen chains by glycogen synthase

• Elongation of a glycogen chain involves the transfer of glucose from

UDP-glucose to the nonreducing end of the growing chain, forming a
new glycosidic bond .

• The enzyme responsible for making the α(1→4) linkages in glycogen

is glycogen synthase.

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 Formation of branches in glycogen
• Still we have linear (unbranched) molecule of glucosyl residues attached by
α(1→4) linkages.

• But glycogen has branches located, on average, eight glucosyl residues apart,
resulting in a highly branched, tree-like structure

• Branches are made by the action of the “branching enzyme,” amylo-α (1→4) → α
(1→6)-transglucosidase.

• This enzyme transfers a chain of six to eight glucosyl residues from the
nonreducing end of the glycogen chain, breaking an α(1→4) bond to another
residue on the chain, and attaches it by an α(1→6) linkage.

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 Glycogen Metabolism in Muscle and Liver

Glycogenesis
Enzymes active in liver and
muscle?
• UDP-glucose is the metabolically
activated form of glucose for glycogen
synthesis.

• Glycogen biosynthesis requires glycogen

synthase for polymerization and a
transglycosylase to create branches.
 Glycogen Metabolism in Muscle and Liver

The glycogen synthase reaction:

Coordinated Regulation of Glycogen Metabolism
 The branching process in glycogen synthesis.

 Branching is brought about by the action of amylo-

(1,41,6)-transglycosylase.
THANK YOU!!!...=)

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