HND-311 Metabolism of Carbohydrates 2nd Notes

Glycolytic pathway
 The oxidation of glucose to pyruvate with the generation of ATP and NADH
 Present in all living organisms
Glycolysis
Glycolysis is a process where a six-carbon compound, glucose, is converted to two three-carbon
compounds (i.e. two pyruvates), which are then converted to two two-carbon compounds (acetyl-CoA)
with the liberation of carbon dioxide, and then into the Tricarboxylic acid cycle (TCA cycle), where it is
completely oxidized to carbon dioxide and water. The conversion of glucose to pyruvate is through a
series of ten reactions (Figure-1)
Steps Involved:

1. Glucose is first phosphorylated to Glucose-6-phosphate (G-6-P) by the enzyme hexokinase

(although the liver uses glucokinase). This happens immediately as glucose enters the cell and
utilizes an ATP molecule. Hexokinase is more specific than glucokinase.
2. The G-6-P is then converted to fructose-6-phosphate by phosphoglucoisomerase
3. In the third reaction, the F-6-P is converted to fructose-1,6 bisphosphate. This reaction involves
the enzyme phosphofructokinase (PFK). PFK is the regulatory enzyme for glycolysis.
4. The next phase of glycolysis involves splitting the F-1,6 bisphosphate (which is a six-carbon
compound) converted into two three-carbon compounds, known as dihydroxyacetone phosphate
(DHAP) and glyceraldehyde 3-phosphate.
5. The DHAP is then converted to glyceraldehyde 3-phosphate – so in effect there are now two
glyceraldehyde 3-phosphates, which are in turn converted to 1,3-bisphosphoglycerate by the
enzyme isomerase.
6. Later, an NADH (Nicotinamide-adenine-dinucleotide hydrogen) is formed from
NAD(Nicotinamide-adenine-dinucleotide), NADH being the reduced form of NAD.
7. The 1,3 bis-phosphoglycerate is then converted to 3-phosphoglycerate by phosphoglycerokinase
in a process that produces 2 ATP molecules from ADP (Phosphorylation).
8. The intermediate 3-phosphoglycerate being converted to 2-phosphoglycerate by the enzyme
phosphoglyceromutase and then to phosphoenolpyruvate (PEP) using enzyme enolase.
9. The final stage uses pyruvate kinase to form pyruvate and another 2 ATP molecules are made
from 2 ADP molecules (phosphorylation).
10. When oxygen is present, glycolysis produces two molecules of the three-carbon compound
pyruvate which can be used for the synthesis of other molecules or oxidized completely to carbon
dioxide and water by the citric acid cycle/kreb‘s cycle.
 Figure 1: Glycolytic Pathway

 Examination of glycolysis
 The Examination of glycolysis shows that after an initial use of two ATPs (reactions 1 and 3), a
further four ATPs are produced (remember that in reactions 5 and 9, there are two reactants from
the initial glucose molecule). So there is a net yield of two ATP. In addition, two NADH are
formed from two NAD, and these can either be re-oxidized from aerobic events in the
mitochondria (each producing three ATP)or from the conversion of pyruvic acid to lactic acid. The
production of lactic acid ensures that glycolysis continues if aerobic re-oxidation of NAD from
 NADH is not possible. The latter tends to occur during very high intensity sessions of exercise.
Regeneration of NAD is essential if glycolysis is to continue.
 The formation of lactic acid occurs when significant amount of pyruvate is formed in glycolysis.
 The ‘link’ reaction; production of acetyl-CoA:
The pyruvate formed as the end result of glycolysis passes into the mitochondria and is converted to acetyl-
CoA in the so-called ‗link‘ reaction. This reaction involves the formation of acetyl-CoA from pyruvate
with the production of carbon dioxide.
 The key regulatory enzyme is known as pyruvate dehydrogenase (PDH).
 In essence, the three-carbon pyruvate is converted to the two-carbon acetyl-CoA. The formation of acetyl-
CoA provides the crossroads between carbohydrate and fat oxidation, and so the control of PDH is seen as
an important factor in the regulation of fat and carbohydrate metabolism.
Another important factor is that NADH is formed from NAD and later this provides ATP.
  Tricarboxylic acid cycle (TCA cycle) or Krebs cycle:
 Introduction:
 The citric acid cycle (also called the tricarboxylic acid [TCA] cycle or the Krebs cycle) is a major
pathway used during aerobic conditions.
 It consists of a series of chemical reactions that take place within mitochondria.
 These enzyme-catalyzed reactions are often depicted as a circle because the product of the last
reaction of the pathway (oxaloacetate) becomes the substrate for citrate (the first reaction)— like a
chemical roundabout.
 Although the citric acid cycle is primarily a catabolic pathway, it serves other purposes. For example,
intermediate products of the citric acid cycle can ―leave‖ and enter anabolic pathways.
 Thus, the citric acid cycle is both catabolic and anabolic and is considered an amphibolic pathway
(being both catabolic and anabolic).
 Amphibolic pathways provide important intersections between catabolism and anabolism on the
metabolism ―superhighway.‖ The citric acid cycle is illustrated in Figure 2.

KEY FEATURES OF TCA CYCLE:

 The citric acid cycle begins when acetyl-CoA combines with oxaloacetate to form citrate (also known
as citric acid).
 In the process, coenzyme A (CoA) is released.
 The formation of citrate is followed by a series of chemical reactions that transfer the chemical energy
contained in acetyl-CoA to NAD+ and FAD. Carbon atoms are released at several points along the
citric acid cycle, combining with oxygen to form carbon dioxide.
 In the end, the citric acid cycle generates NADH + H+ and FADH2 , and these highly energized
compounds enter the electron transport chain for ATP production.
 Note that, in the citric acid cycle, ATP is not formed directly. Rather, it is formed from another high-
energy compound called guanosine triphosphate (GTP) via substrate phosphorylation. In all, the
oxidation of two molecules of acetyl-CoA (formed from one molecule of glucose) produces six
NADH + H+, two FADH2 , and two ATPs.
 Although small amounts of ATP are produced via substrate phosphorylation, most (90%) of the ATP
generated from glucose results from oxidative phosphorylation via the electron transport chain (Stage
4).
 In total, the complete oxidation of one molecule of glucose (via glycolysis, the citric acid cycle, and
oxidative phosphorylation) generates up to 38 ATPs, depending on the source of glucose—10 NADH
+ H+ (30 ATPs via oxidative phosphorylation), 2 FADH2 (4 ATPs via oxidative phosphorylation),
and 4 ATPs formed via substrate phosphorylation (Figure)
 TCA CYCLE:
 TCA cycle initiates when Acetyl-CoA formed during the link reaction, enters the TCA cycle, whereby
it attaches to a four-carbon compound, oxaloacetic acid (OAA).
 The combination of the four-carbon OAA and the two-carbon acetyl-CoA results in the formation of
the six-carbon, citric acid (or citrate). This cyclic sequence of reactions is found in every cell type
that possesses mitochondria
 The function of the TCA cycle is to convert the two-carbon acetyl-CoA to carbon dioxide and water
and to produce energy (ATPs). Figure-2 highlights some key events in the TCA cycle, and shows that
citrate is converted to isocitrate, then to succinyl-CoA and then to alpha-ketoglutarate. The formation
of alpha-ketoglutarate produces NADH, as does the next reaction to succinyl-CoA. The cycle is
catalyzed by a series of enzymes and yields reducing equivalents (H+)
 Succinyl-CoA then forms fumarate in a reaction that produces a Flavin-adenine-dinucleotide-
hydrogen (FADH2). Fumarate then forms malate, and then malate forms OAA, with the production of
another NADH. So, the net effect of the TCA cycle is to produce three NADH, one FADH2 and one
ATP. Each NADH results in the formation of three ATPs via oxidative phosphorylation whilst the
FADH is re-oxidized to form two ATPs via oxidative phosphorylation.
Figure-2: Tricarboxylic acid Cycle / Kreb’s Cycle / Citric Acid Cycle
 Glycogenesis
 Introduction:
 Glycogen is a homopolysaccharide formed of branched glucose units
 The primary glycosidic bond is 1-4-glycosidic linkage.
 Each branch is made of 6-12 glucose units. At the branching point, the chain is attached by 1-6
glycosidic linkage.
 Glucose is stored as glycogen predominantly in liver and muscle cells.
 Liver glycogen is about 100-120 grams approx. (about 6 % of liver weight).
 Liver glycogen: It maintains normal blood glucose concentration especially during the early stage of
fast (between meals). After 12-18 hours fasting, liver glycogen is depleted.
 Muscle glycogen: It acts as a source of energy within the muscle itself especially during muscle
contractions
 Muscle glycogen is about 300-350 grams approx. (about 1 % of total muscles weight.
 The process of glycogen formation is known as glycogenesis. For glycogenesis to occur, glucose must
be made available inside the cell and insulin (a hormone which helps regulate blood glucose levels)
elevated in the blood.
 Primarily these two products (Glucose and insulin) stimulate the glycogenesis pathway.

 Site of glycogen synthesis: Liver and Muscles.

 This typically occurs after a carbohydrate meal. Once glucose enters the cell, it gets converted into
glucose-6-phosphate (g-6-p) by the enzyme hexokinase (in muscles) and or glucokinase (in liver), the
enzyme phosphoglucomutase then converts g-6-p to g-1-p, this reaction occurs because the molecule
uridinediphosphate (UPD) can only attaches with g-1-p to make UDP-glucose.
 The UDP-glucose is then attached to the glycogen molecule by the enzyme glycogen synthase (the
key regulatory enzyme for glycogenesis is glycogen synthase).
 This is repeated until a short linear glucose polymer with α-1,4-glycosidic linkages is built up
on Glycogenin.
 Glycogen Synthase then catalyzes elongation of glycogen chains initiated by Glycogenin.
 By the action of Glycogen Synthase (key enzyme of glycogenesis) UDP-G molecules are added to
glycogen primer causing elongation of the α-1,4-branches up to 11 glucose units.
 A branching enzyme transfers a segment of glycogen molecule from the end of a glycogen chain to
the Carbon 6 hydroxyl of a glucose residue of glycogen to yield a branch with an α-1,6- linkage. The
new branches are elongated by the glycogen synthase and the process is repeated.
 In effect, this means that the glycogen molecule becomes larger by the addition of glucose molecules.
The central core of the glycogen molecule is never completely broken down, or it would not be
possible for a glucose molecule to attach.
Being a synthesis reaction, glycogenesis requires energy. One ATP is used up when the glucose molecule
enters the cell and is immediately phosphorylated to G-6-P; the other is used in the form of uridine
triphosphate (UTP), to which the G-1-P is attached to produce UDP-glucose.

 
 Glycogenolysis

INTRODUCTION
 Glycogenolysis refers to the breakdown of glycogen.
 It is the breakdown of glycogen into glucose-1-phosphate (G-1-P), which is then converted to
glucose-6-phosphate (G-6-P) before going through glycolysis (the breakdown of glucose through a
series of reactions to form pyruvic acid).
 Note that glycogenolysis feeds into glycolysis with the formation of glucose-6-phosphate.
 The process of glycogenolysis is controlled by the enzyme phosphorylase.

EXPLANATION:
The glucose formed when removed from a glycogen molecule is glucose-1-phosphate, which means that
the glucose has a phosphate group attached at the carbon-1 position. This is achieved because ATP is
involved:
The G-1-P is then converted to G-6-P, whereby the phosphate group is moved from carbon-1 to carbon-6
before undergoing glycolysis. Glycogenolysis is a process that must be capable of happening rapidly,
since it provides an important and major source of energy during prolonged physical activities. This
implies that glycogenolysis needs to be ‗switched on‘ almost instantaneously. Two major enzymes
participate in all glycogen degradation are:

 Glycogen phosphorylase
 Debranching enzyme

 Glycogen Phosphorylase (the key enzyme of Glycogenolysis)

 It adds phosphate (Pi) and releases glucose 1 phosphate. i.e.,

 It catalyzes phosphorolytic cleavage (addition of Pi) of the α-1,4-glycosidic linkages of glycogen,
releasing glucose-1-phosphate as reaction product.
 Always acts at non-reducing end, and stops at fourth glucose from α-1,4-glycosidic linkages
branch point.

 Debranching enzyme: it has 2 important function

 It transfers 3 glucose molecules to the branch by breaking the α-1,4-glycosidic linkage
 In the next stage it breaks the α-1,6-glycosidic linkages to release glucose.

As a result this reactions yields G-1-P in significant amount and some glucose molecules.
GLYCOGENESIS