BCH301: METABOLISM OF CARBOHYDRATES

DEGRADATION AND DIGESTION OF CARBOHYDRATES

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 THE GLYCOLYTIC PATHWAY (GLYCOLYSIS)

•The glycolytic pathway (Embden–Meyerhof pathway) is a ten-step enzyme-

catalyzed reactions in which a molecule of glucose is oxidized to yield 2
molecules of pyruvate, a 3-carbon compound, with energy conserved as ATP &
NADH.

•It takes place in the cytosol of the cell.

•The first 5 reactions of the pathway constitute the preparatory phase, where 2
molecules of ATP are invested to convert glucose to fructose 1,6-bisphosphate.

•The bond between C-3 and C-4 of the fructose 1,6-bisphosphate is then
broken to form 2 molecules of glyceraldehyde 3-phosphate.
•The last 5 reactions of the pathway constitute the payoff phase, where each
of the 2 molecules of the glyceraldehyde 3-phosphate derived from glucose is
oxidized to pyruvate.

•For each molecule of glyceraldehyde 3-phosphate oxidized to pyruvate, one

NADH & two ATP are formed.

•Thus, 4 ATP are produced for a molecule of glucose oxidized to pyruvate under
aerobic condition.

•When the 2 ATP used up in the preparatory phase are deducted, the net ATP
produced by the glycolytic pathway under aerobic condition is 2.

•The overall equation for the glycolytic pathway under aerobic condition is:

Glucose + 2NAD+ + 2ADP + 2Pi → 2 pyruvate + 2NADH + 2H+ + 2ATP + 2H2O

Reactions of The First (Preparatory) Phase of Glycolysis

Reaction 1: Phosphorylation of glucose at the C-6 to form glucose-6-phosphate

(G-6-P), catalyzed by hexokinase (glucokinase), which requires magnesium ion
(Mg2+) as a cofactor

•This is the first priming reaction of glycolysis, in which 1 ATP (energy) is

consumed as the phosphoryl donor to make the formation of G-6-P
thermodynamically favorable.

Reaction 2: Isomerization of G-6-P to fructose-6-phosphate (F-6-P), catalyzed

by phosphoglucoisomerase (glucose phosphate isomerase), which requires
Mg2+.
Reaction 3: Phosphorylation of F-6-P at the C-1 to form fructose-1,6-
bisphosphate (F-1,6-bisP), catalyzed by phosphofructokinase-1 (PFK-1), which
requires Mg2+

•This is the second priming reaction of glycolysis, in which 1 ATP is consumed

as the phosphoryl donor.

•This 3rd reaction is irreversible under cellular conditions; it is the first

committed (rate-limiting) step in the glycolytic pathway and the major point of
regulation in glycolysis.

•The activity of PFK-1 is increased whenever the cellular ATP level is depleted
or when ADP and AMP (ATP breakdown products) are in excess.

•In some organisms, fructose-2,6-bisphosphate is a potent allosteric activator

of PFK-1.

•The enzyme is inhibited whenever the cellular ATP level is high.

Reaction 4: Cleavage of F-1,6-bisP between the C-3 and C-4 to yield two triose
phosphates (glyceraldehyde-3-phosphate and dihydroxyacetone phosphate
[DHAP]), catalyzed by fructose-1,6-bisphosphate aldolase

Reaction 5: Inter-conversion of the triose phosphates

•Of the 2 triose phosphates formed in the 4th reaction, only glyceraldehyde 3-
phosphate, can be directly degraded in the subsequent steps of glycolysis.

•Hence, dihydroxyacetone phosphate is rapidly and reversibly isomerised to

glyceraldehyde-3-phosphate, catalyzed by triose phosphate isomerase.

•This reaction completes the first (preparatory) phase of glycolysis, with each
glucose that passes through being converted to 2 molecules of glyceraldehyde-
3-phosphate.
Reactions of The Second (Payoff) Phase of Glycolysis

•In this phase the 2 molecules of glyceraldehyde-3-phosphate produced in the

1st phase are converted to 2 molecules of pyruvate, with the formation of four
molecules of ATP from ADP.

Reaction 6: Oxidation of glyceraldehyde-3-phosphate to 1,3-

bisphosphoglycerate, catalyzed by glyceraldehyde-3-phosphate dehydrogenase

•This reaction is the first of the 2 energy-conserving reactions of glycolysis that

eventually lead to the formation of ATP.

Reaction 7: Transfer of the high-energy phosphoryl group from the carboxyl

group of 1,3-bisphosphoglycerate to ADP, to form ATP and 3-phosphoglycerate,
catalysed by phosphoglycerate kinase

•This is the first ATP-synthesizing reaction of glycolysis.

•The formation of ATP by transfer of phosphoryl group from a substrate such as
1,3-bisphosphoglycerate is referred to as a substrate-level phosphorylation, to
differentiate this mechanism from respiration-linked phosphorylation.

Reaction 8: Conversion of 3-phosphoglycerate to 2-phosphoglycerate,

catalysed by phosphoglycerate mutase, which requires Mg2+.

Reaction 9: Dehydration of 2-phosphoglycerate to phosphoenolpyruvate (PEP),

catalyzed by enolase.

•This is 2nd energy-conserving reaction of glycolysis that eventually lead to the

formation of ATP, as the PEP formed in this reaction has high phosphoryl group
transfer potential.
Reaction 10: Transfer of the phosphoryl group from phosphoenolpyruvate to
ADP, forming pyruvate, catalyzed by pyruvate kinase, which requires K+ and
either Mg2+ or Mn2+

•This is the 2nd ATP-synthesizing reaction of glycolysis (also a substrate-level

phosphorylation).

•The pyruvate formed first appears in its enol form, then tautomerizes rapidly
and non-enzymatically to its keto form, which predominates at pH 7.

•Pyruvate kinase is activated by AMP and fructose-1,6-bisphosphate, and

inhibited by ATP, acetyl-CoA and alanine.
Figure: The glycolytic pathway
FEEDER PATHWAYS FOR GLYCOLYSIS

•Beside glucose, many other carbohydrates including polysaccharides (glycogen

and starch); the disaccharides (maltose, lactose, trehalose, and sucrose); and
the monosaccharides (fructose, mannose, and galactose) enter into the
glycolytic pathway for degradation.

•These carbohydrates are first transformed into one of the intermediates of

glycolysis.
Glycogen and Starch

•Glycogen in animal tissues and starch in plants can be mobilized for use within
the same cell by a phosphorolytic reaction catalyzed by glycogen
phosphorylase and starch phosphorylase, respectively.

•This phosphorolytic cleavage occurs on the (1→4) glycosidic linkage that joins
the last two glucose residues at a non-reducing end, generating glucose 1-
phosphate and a polymer one glucose unit shorter .

•Glycogen phosphorylase (or starch phosphorylase) acts repetitively until it

approaches an (1→6) branch point, where its action stops.

•Then a de-branching enzyme removes the branches.

•Glucose 1-phosphate produced by glycogen phosphorylase is converted to

glucose 6-phosphate by phosphoglucomutase, which enter the glycolytic
pathway.
•Dietary polysaccharides and disaccharides are hydrolysed to their constituent
monosaccharides, which then enter the glycolytic pathway.

•Dietary starch is first hydrolysed (digested) to oligosaccharides by salivary-

amylase (ptyalin) in the mouth.

•Starch hydrolysis continues in the stomach, where pancreatic-amylase yields

mainly maltose, maltotriose and dextrins.

•Maltose and dextrins are further hydrolysed to monosaccharides by enzymes

present in the intestinal brush border.
•Intestinal disaccharides are hydrolyzed to monosaccharides by enzymes
present in the outer surface of the intestinal epithelial cells.

•The D-hexoses, including fructose, galactose and mannose, can enter the
glycolytic pathway, after being phosphorylated to either glucose 6-phosphate
or fructose 6-phosphate.
Fig. Entry of glycogen, starch, disaccharides & hexoses into the preparatory
phase of glycolysis
 METABOLIC FATE OF PYRUVATE UNDER AEROBIC & UNDER ANAEROBIC
CONDITIONS

•Under aerobic condition, pyruvate is oxidized to acetyl-CoA and CO2, by an

irreversible oxidative decarboxylation catalyzed by the pyruvate dehydrogenase
complex.

•The acetyl-CoA enters the citric acid (TCA) cycle and is oxidized to CO2 and
H2O.

•The pyruvate dehydrogenase complex consists of 3 different enzymes and

requires five coenzymes.

•The enzymes are pyruvate dehydrogenase (E1), dihydrolipoyl transacetylase

(E2), and dihydrolipoyl dehydrogenase (E3).
•The coenzymes are thiamine pyrophosphate (TPP), flavin adenine dinucleotide
(FAD), coenzyme A (CoA), nicotinamide adenine dinucleotide (NAD), and
lipoate.

•The overall reaction of oxidative decarboxylation of pyruvate to acetyl-CoA is

as follows:
•The mechanism of pyruvate dehydrogenase complex-catalyzed oxidative
decarboxylation of pyruvate to acetyl-CoA is shown below.

Fig. Mechanism of pyruvate dehydrogenase complex-catalyzed oxidative

decarboxylation of pyruvate to acetyl-CoA
•Under anaerobic (hypoxic) conditions, the pyruvate produced in glycolysis has
two different fates.

•In many organisms, pyruvate is reduced to lactate to regenerate NAD + , a

reaction catalyzed by lactate dehydrogenase.

•The reduction of pyruvate to lactate under anaerobic conditions is referred to

as lactate fermentation.
•In other organisms, such as yeast, pyruvate is reduced to ethanol and CO2 to
regenerate NAD+, in a two-step reaction catalyzed by pyruvate decarboxylase
and alcohol dehydrogenase.

•In the first reaction, pyruvate is irreversibly decarboxylated to acetaldehyde,

catalyzed by pyruvate decarboxylase, which requires Mg2+ (cofactor) and
thiamine pyrophosphate (coenzyme).

•In the second reaction, acetaldehyde is reduced to ethanol, catalyzed by

alcohol dehydrogenase, with the reducing power furnished by NADH derived
from the dehydrogenation of glyceraldehyde 3-phosphate.

•The reduction of pyruvate to ethanol under anaerobic conditions is referred to

as alcohol fermentation.

•The overall equation of these reactions is:

Glucose + 2ADP + 2Pi → 2 ethanol + 2CO2 + 2ATP + 2H2O

 THE PENTOSE PHOSPHATE PATHWAY

•The pentose phosphate pathway, also known as the hexose monophosphate

shunt, or the phosphogluconate pathway, is an alternative route for the
metabolism of glucose that begins from glucose 6-phosphate.

•The reactions of the pentose phosphate pathway occur in the cytosol in two
phases.

•The first phase of the pentose phosphate pathway comprises two oxidative
reactions that convert glucose 6-phosphate to ribulose 5-phosphate and
reduce NADP to NADPH.

•The second phase consists of non-oxidative reactions that convert pentose

phosphates to glucose 6-phosphate, which begins the cycle again.
•The NADPH formed in the oxidative phase is used to reduce glutathione, GSSG
and to support reductive biosynthesis.

•The ribose 5-phosphate produced in the oxidative phase serves as a

precursor for nucleotides, nucleic acids and coenzymes.
Fig. General scheme of the pentose phosphate pathway
Fig. Oxidative phase of the pentose
phosphate pathway
Figure. Non-oxidative phase of the pentose phosphate pathway
 TRICARBOXYLIC ACID (TCA) CYCLE

•The TCA cycle, discovered by Hans Krebs in 1937, is also known as citric acid
cycle or Krebs cycle.

•It is a central catabolic pathway, taking place in the mitochondria of

eukaryotes, in which the acetyl-CoA produced from pyruvate, during glycolysis
under aerobic condition, is oxidized to CO2.

•Fatty acids and amino acids are also oxidized to CO2 via the TCA cycle.

•In the process, metabolic energy is captured in the form of electron carriers
(NADH and FADH2)

•During aerobic metabolism, these electrons are transferred to O 2 and the

energy of electron flow is trapped as ATP.

•The TCA cycle has eight steps as shown in the next slide.
Fig. The TCA Cycle
AMPHIBOLIC NATURE OF THE TCA CYCLE

•The TCA cycle is amphibolic in aerobic organisms, as its intermediates serve in

both catabolic and anabolic processes.

•It provides carbon skeletons (precursors) for anabolic pathways such as

gluconeogenesis, fatty acid synthesis, and inter-conversion of amino acids, in
addition to its role in oxidative catabolism of carbohydrates, fatty acids, and
amino acids.

Examples:

•α-Ketoglutarate and oxaloacetate can serve as precursors of the amino acids

aspartate and glutamate by simple transamination.
•Then, through aspartate and glutamate, the carbons of oxaloacetate and α-
ketoglutarate are used to biosynthesize other amino acids, as well as purine
and pyrimidine nucleotides.

•Oxaloacetate can also be converted to glucose in gluconeogenesis; while

succinyl-CoA is a central intermediate in the biosynthesis of the porphyrin ring
of heme groups, which serve as oxygen carriers in hemoglobin and myoglobin,
and electron carriers in cytochromes.
Regulation of the TCA Cycle

•The overall rate of the TCA cycle is regulated by the rate of conversion of
pyruvate to acetyl-CoA, catalyzed by pyruvate dehydrogenase (PDH) complex,
and by the reactions catalyzed by citrate synthase, isocitrate dehydrogenase,
and α-ketoglutarate dehydrogenase.

•The activities of these enzymes depend largely on the concentrations of

substrates and products such that the end products (ATP and NADH) inhibit
them, while the substrates (NAD+ and ADP) stimulate them.

•The production of acetyl-CoA for the citric acid cycle by the PDH complex is
allosterically inhibited by metabolites that indicate a sufficiency of metabolic
energy (ATP, acetyl-CoA, NADH, and fatty acids) and stimulated by metabolites
that indicate a reduced energy supply (AMP, NAD+, CoA).

•On the other hand, succinyl-CoA is an intra-cycle regulator that inhibits citrate
synthase and α-ketoglutarate dehydrogenase.
THE GLYOXYLATE CYCLE

•The glyoxylate cycle is used by plants, yeast, and many bacteria to convert
acetyl-CoA to oxaloacetate.

•The glyoxylate cycle takes place in the glyoxysomes.

•Thus, these organisms can use fatty acids as the starting material for glucose
synthesis (gluconeogenesis).

•However, this is not possible in mammals because the pyruvate

dehydrogenase reaction is irreversible and cells have no other pathway to
convert acetyl-CoA to pyruvate.
•The glyoxylate cycle serves as important source of glucose during the
germination of seedlings, before photosynthesis can serve as a source of
glucose.

•In plants and many invertebrates, the glyoxylate cycle serves as a mechanism
for converting acetate to carbohydrate.

Reactions of the glyoxylate cycle

•Acetyl-CoA condenses with oxaloacetate to form citrate, catalyzed by citrate

synthase.

•The citrate is converted to isocitrate, catalyzed by aconitase.

•The isocitrate is cleaved to succinate and glyoxylate, catalyzed by isocitrate

lyase.
•The glyoxylate then condenses with a second molecule of acetyl-CoA to yield
malate, in a reaction catalyzed by malate synthase.

•The malate is subsequently oxidized to oxaloacetate, which can condense with

another molecule of acetyl-CoA to start another turn of the cycle.

Note that:

•Each turn of the glyoxylate cycle uses up two molecules of acetyl-CoA and
produces one molecule of succinate, which is then available for biosynthetic
purposes.

•The succinate may be converted through fumarate and malate into

oxaloacetate.
•The oxaloacetate can then be converted to phosphoenolpyruvate (PEP) by PEP
carboxykinase.

•The PEP can be converted to glucose by gluconeogenesis.

•The glyoxylate cycle bypasses the two oxidative decarboxylation steps of the
citric acid cycle, making possible the net formation of succinate, oxaloacetate,
and other cycle intermediates from acetyl-CoA.

•The two oxidative decarboxylation steps of the citric acid bypassed are the
oxidation of isocitrate to α-ketoglutarate by isocitrate dehydrogenase, and
oxidation of α-Ketoglutarate to succinyl-CoA by α-ketoglutarate dehydrogenase
complex.
Fig. The Glyoxylate cycle