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9
CHAPTER

Carbohydrate Metabolism

INTRODUCTION Glucose is the major source of energy. There are certain tissues like erythrocytes, brain, and renal medulla, which mainly depend upon glucose as the source of energy. A continuous supply of glucose is required for the proper functioning of human body. Fasting plasma glucose level in normal individuals is 75 to 110 mgl dl. A normal individual has got the capacity to maintain the fasting glucose level with the help of various hormones. Carbohydrates are immediate source of energy. Glucose is oxidized in the body to form carbon dioxide and water. During this process, energy is extracted from it. Hence most of the tissues contain enzymes required for carbohydrate metabolism. The major metabolic pathways by which glucose is metabolized are: Glycolysis Glycogenolysis Glycogenesis Gluconeogenesis Hexose monophosphate shunt (HMP) Uronic acid pathway Galactose and fructose are also metabolized in our body by specific metabolic pathways GLYCOLYSIS Glycolysis is a pathway by which glucose is converted into pyruvate under aerobic condition, and lactate under anaerobic condition.

I I

I
i

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Textbook of Biochemistry for Nurses

Tissues

where

Glycolysis

takes Place

3. Energy

E:

RBCs depend entirely upon glycolysis for energy production. When the muscle is contracting, energy is provided by glycolysis. Glycolytic pathway takes place in almost all the tissues. Subcellular Site

Further react pyruvate as ~ pyruvate kin form ATP. So, of production

Reactions take place in the cytosol. End products formed: Under aerobic condition (presence of oxygen) pyruvate, and anaerobic condition (absence of oxygen) lactate. The glycolytic pathway can be di vided in following three phases:

1. Energy

Investment

Phase

Energy investment phase extends up to the formation of fructose 1,6-bisphosphate.

Glucose Glucokinase/Hexokinase

1.........- ATP
~ADP

Glucose-6-phosphate Phosphohexose Isomerase

Fructose-6-phosphate Phosphofructokinase

1.........- ATP
~ADP

Fructose 1,6-bisphosphate

2. Splitting

Phase

Fructose 1,6-bisphosphate formed is split into two trioses, glyceraldehyde3-phosphate and dihydroxy acetone phosphate by an enzyme called aldolase B.
Fructose 1,6-bisphosphate AldOI~

Under ae into rnitochor CoA, which i Under an called lactat equi valents fr Energetics
0

Glyceraldehyde-3-phosphate

Dihydroxyacetone phosphate ~snnotriOSee ~ pno t' i\So{\\e\c>~-

Dihydroxyacetone phosphate and glyceraldehyde-3-phosphate can be interconverted by an enzyme known as phosphotriose isomerase. Glyceraldehyde-3-phosphate enters into further glycolytic reactions. Hence, its concentration is kept low. This favors the conversion of dihydroxy acetone phosphate to glyceraldehyde-3-phosphate.

Initially, glue 6':phosphate i: one ATPeach glucose is spl One glyc. I NADH TwoATF Since tw10 ATP's are

Carbohydrate

Metabolism

99

3. Energy Extraction is. Glycolytic

Phase

Further reactions of glycolysis will convert glyceraldehyde-3-phosphate into pyruvate as shown below. During this phase, phosphoglycerate kinase and pyruvate kinase are the two enzymes that are capable of phosphorylating ADP to form ATP. So, energy is produced at the metabolic pathway level. This mechanism of production of ATP is called substrate level phosphorylation.
Glyceraldehyde-3-phosphate Glyceraldehyde-3phosphate dehydrogenase

.n)

pyruvate,

s:

f::

NAD+ NADH + H+

1'3-PhOSPf::hoglyce:~p

bisphosphate.

Phosphoglycerate kinase

ATP

3-phosphoglycerate Phosphoglycerate mutase

1
t

2-phosphoglycerate Enolase

~H20 PhOSPhoelCl pyruvate

ADP
Pyruvate kinase

ATP
Pyruvate

ceraldehydealdolase B.

Under aerobic conditions, pyruvate formed at the end of glycolysis enters into mitochondria. Inside the mitochondria, pyruvate is converted into acetyl CoA, which is further metabolized. Under anaerobic conditions, pyruvate is converted to lactate by an enzyme called lactate dehydrogenase. Lactate dehydrogenase utilizes the reducing equivalents from the NADH + H+ and reduces pyruvate to lactate. Energetics of Glycolysis

late

rhate can be
.tions. Hence,

Initially, glucose is activated to form glucose-6-phosphate, and later, fructose6-phosphate is converted to fructose 1,6-bisphosphate. Both these reactions utilize one ATP each. So, two ATP's are utilized for the activation of glucose. Activated glucose is split into two trioses: One glyceraldehyde-3-phosphate, which enters into glycolysis, and produces 1 NADH + H+, which is equivalent to 3 ATP's and Two ATP's are produced by the substrate level phosphorylation. Since two glyceraldehyde-3-phosphates 10 ATP's are formed. are formed from single glucose,

:onephosphate

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Textboo-: of Biochemistry for Nurses

Out of 10 .-\TVs, 2 are utilized during the energy investment phase. So, the net ATP's produced by aerobic glycolysis are 8. Under anaerobic conditions, NADH + H+ is utilized for the conversion of pyruvate to lactate. So, the net ATP's formed are 2. Regulation Phosphofructokinase-l is the regulated enzyme of glycolysis. Fructose 2,6-bisphosphate and ADP are allostearic activators of the enzyme. ATP and citrate are allostearic inhibitors. Pyruvate kinase activity can also be regulated. Rapoport Leubering Shunt
!.~."

I Succinate del

One of the intermediate of glycolysis, 1,3-bisphosphoglycerate can be converted into 2,3-bisphosphoglycerate. 2,3-bisphosphoglycerate has got a biological significance. It binds to hemoglobin and decreases its affinity for oxygen and increases the delivery of oxygen to the tissues. 2,3-bisphosphoglycerate is later converted to 3-phosphoglycerate, which enters glycolytic pathway. Since I,3-bisphosphoglycerate is shunted through this pathway back to glycolysis, it is called 2, 3-bisphosphoglycerate shunt (2,3-BPG Shunt). CITRIC (Kreb's ACID CYCLE Acid Cycle)
~' 1

Energetics One acetyl 0 Isocitrate ~INJ o-keroglr ~lNJ Succinate Succinate

Cycle, Tricarboxylic

Citric acid cycle takes place inside the mitochondria (Fig. I). Pyruvate, which is the end product of glycolysis, is transported into the mitochondria with the help of a transporter. Inside the mitochondria, it is converted into acetyl CoA by the action . of pyruvate dehydrogenase complex. Pyruvate dehydrogenase complex is a multienzyme complex, which requires thiamine pyrophosphate (TPP), NAD+, FAD, and lipoic acid.
NAD+ Pyruvate '-_ NADH + H+

~FAI
Malate de ~lNJ Total AT

_.... L

Acetyl CoA

Pyruvate Dehydrogenase (PDH)

Acetyl CoAcombines with oxaloacetate to form citrate. This is the first reaction of citric acid cycle. Further reactions proceed as shown in the reaction sequence. and at the end, oxaloacetate is regenerated. Reducing equivalents removed from the intermediates of TCA cycle are accepted by the coenzymes like NAD+ and FAD. leading to the formation of NADH + H+ and FADH2. These reduced forms of coenzymes donate the electrons to the electron transport chain to generate three ATP's and two ATP's respectively.

TCA cycle is produces intei produce CO2 ~-oxidation 01 enters TCA a-ketoglutarat Pyruvate ea: a-ketoglutarai cycle to yield

Carbohydrate

Metabolism

101

ise. So, the net conversion of

Reaction Sequence
Acetyl CoA ,-Oxaloacetate

I Malate dehYdrOgenasetc. I Fumarase

)f the enzyme.
.-----------'

.-1 ~

fr.1

Citrate synthase

. Citrate \[Aconita:~1 \ Fe Acomtate } ACOnitase -I

NADH + H+

MI!-a,ate NAD+

HP

I Succinate dehydrogenase I
n be converted ds to hemoglof oxygen to the :e, which enters ;h this pathway I-BPG Shunt). Energetics

Fumarate FADH2
~ FAD

I Succinate thiokinase I ~

-------'ZGDP

Succinate

GTP

lsocltrate NAD+~~-------'I
+

."

1'--'
CO
2

Fe2+

Isocitrate dehydrogenase

NADH + H+ NADH +H

Succinyl CO~-KetOglutarate

I a-Ketoglutarate

dehydrogenase

Figure 1: Citric acid cycle

One acetyl CoA, which enters TCA cycle, is oxidized to produce 12 ATP's: Isocitrate dehydrogenase --) 1 NADH + H+ a-ketoglutarate dehydrogenase --) 1 NADH + H+

3 ATP's 3 ATP's 1 ATP

= 2 ATP's

uvate, which is vith the help of A by the action : complex is a (TPP), NAD+.

Succinate thiokinase (Substrate level phosphorylation) Succinate dehydrogenase --)FADH2 Malate dehydrogenase --) 1 NADH + H+ Total AT Ps produced

= 3 ATP's = 12 ATP's

he first reaction ction sequence. of TCA eyele ) the formation nes donate the and two ATP's

TCA cycle is an amphibolic pathway. Metabolism of amino acids and fatty acids produces intermediates of TCA cycle. They are catab~lized through TCA cycle to produce CO2 and energy. For example, propionic acid is produced during the ~-oxidation of the odd chain fatty acids. Later, it is converted to succinyl CoA and enters TCA cycle. Alanine and glutamate are converted to pyruvate and a-ketoglutarate respectively. They are further oxidized by TCA cycle to yield energy. Pyruvate can be converted to acetyl CoA and oxidized by TCA cycle. a-ketoglutarate is an intermediate of TCA cycle, which can be oxidized by TCA cycle to yield energy.

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Textbook of Biochemistry for Nurses

Intermediates of TCA cycle can be utilized for the synthesis of various biologically important substances (anabolism). For example: Succinyl CoA is used for the synthesis of heme. ex-ketoglutarate is used for the synthesis of nonessential amino acid, glutamate. . Oxaloacetate is used for the synthesis of aspartate. It is required for protein synthesis. Since TCA cycle has gqt role both in anabolism as well as catabolism, it is an amphibolic pathway. When one mole of glucose is completely oxidized by glycolysis, followed by TCA cycle, it leads to the formation of 38 ATP's. From one glucose molecule, 2 pyruvate molecules and 8 ATP's are formed from glycolytic pathway. Both pymvate molecules enter mitochondria, where they are converted to acetyl CoA by pyruvate dehydrogenase complex. During this reaction, two NADH + H+ is produced, which is equivalent to 6 ATP's. AcetylCoA enters into TCA cycle and gets oxidized to CO2 and form 24 ATP's. GLUCONEOGENESIS Gluconeogenesis or neoglucogenesis is the synthesis of new glucose molecules from noncarbohydrate substances. When there is a deficiency of glucose in our body, as in the case of starvation, gluconeogenesis provides glucose to those tissues (neurons), which entirely depend upon glucose (RBCs) or are mainly dependent on glucose as a source of energy. Gluconeogenic Substrates

Pyruvate Phosphoe Fructose

Glucose-r

Reactions C; The reaction ( this reaction, 1 First, pyruvatr reaction requi energy (Fig. 2

Noncarbohydrate substances that can be converted to glucose are called gluconeogenic substrates. Lactate, glycerol, propionic acids, and various amino acids called glucogenic amino acids are gluconeogenic substrates. Lactate is formed during anaerobic glycolysis. Glycerol is produced when the fat in the adipose tissue is mobilized. The last three carbons of the odd chain fatty acids are released as propionic acir'. Proteolysis leads to the release of amino acids. Gluconeogenesis involves most of the reactions of glycolysis, but in the reverse direction, and a few reactions of TCA cycle. Simple reversal of glycolysis is impossible because of the irreversible nature of few of the reactions of glycolysis. These reactions are catalyzed by the following enzymes: Pyruvate kinase: Catalyzes the conversion of phosphoenol pyruvate to pyruvate. Phosphofructokinase 1: Catalyzes the phosphorylation offructose-6-phosphate to form fructose 1,6-bisphosphate. Glucokinase: It converts glucose to glucose-o-phosphate. Key Enzymes of Gluconeogenesis The irreversible reactions of glycolysis are reversed with the help of special enzymes called key enzymes of gluconeogenesis. These enzymes are:

co
PhoSI

Oxaloacet oxaloacetate a malate dehyd

Carbohydrate Metabolism

103

is of various bio-

o acid, glutamate.
[uired for protein

Pyruvate carboxylase Phosphoenolpyruvate carboxykinase Fructose 1,6-bisphosphatase Glucose-ti-phosphatase

Reactions Catalyzed by the Key Enzymes of Gluconeogenesis

uabolism, it is an vsis, followed by The reaction catalyzed by pyruvate kinase in glycolysis is irreversible. To reverse this reaction, the sequential action of following two different enzymes is required. First, pyruvate carboxylase converts pyruvate to oxaloacetate. It is a carboxylation reaction requiring carbon dioxide, water-soluble vitamin, biotin, magnesium, and energy (Fig. 2).

fP's are formed

idria, where they lex. During this ; ATP's. Acetyl-

Pyruvate

-7....,--------- ... ~Oxaloacetate

-I
ATP; Biotin; Mg2+

Pyruvate carboxylase

t ATP's.

icose
C

molecules

glucose in our ~to those tissues ainly dependent

-------. Oxaloacetate

e called gluco-us amino acids iroduced when f the odd chain .lcase of amino
t in the

I
Cytosol

reverse

f glycolysis is ; of glycolysis. ite to pyruvate. :e-6-phosphate

~Malate
Oxaloacetate

.J

C02~~
Phosphoenol

Malate dehydrogenase Phosphoenol pyruvate carboxykinase GTP

~ pyruvate

Figure 2: Site for the reactions of gluconeogenesis

elp of special .e:

Oxaloacetate is impermeable through the mitochondrial membrane. To transport oxaloacetate across the mitochondrial membrane, it is first converted to malate by

malate dehydrogenase.

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Textbook (~f Biochemistryfor

Oxaloacetate

Nurses
Phosphoenol pyruvate GTP '\ CO2 Lacl

Phosphoenolpyruvate carboxy kinase

1. Lact Later, pyru'

Malate is carried out of the mitochondria, and in the cytosol it is converted back to oxaloacetate by malate dehydrogenase. Later, phosphoenolpyruvatecarboxykinase (PEPCK) acts on the oxaloacetate to form phosphoenolpyruvate. PEPCK is present both in cytoplasm and mitochondria of human beings. PEPCK present inside the mitochondria can convert some amount of oxaloacetate to phosphoenol pyruvate. It is later transported to cytosol and enters further reactions of gluconeogenesis. Phosphoenol pyruvate is converted to fructose 1,6-bisphosphate by the sequential action of various enzymes of glycolysis in the reverse direction. Conversion of fructose 1,6-bisphosphate to fructose-6-phosphate is an irreversible step. It requires another key enzyme of gluconeogenesis called fructose 1,6-bis phosphatase.
Fructose 1,6-bisphosphate

2. Var They can t amino acid Alanir can be later forms oxal 3. Gly, phosphate.

---.-7~""'-"~----~~
Fructose 1,6-bisphosphatase H20 Pi

GI

Fructose-6-phosphate

Fructose-6-phosphate formed is isomerized to form glucose-6-phosphate. Glucose-6-phosphate can be converted to glucose by the action of the enzyme glucose-6- phopshatase.
Glucose-6-phosphate

----7~ ..... -"~----~~

Glucose-6-phosphatase H20 Pi

Glucose

Glucose-6-phosphatase is present in the liver and medulla of the kidney. Substances that can form pyruvate or any intermediates of glycolysis and TCA cycle can be converted to glucose by the gluconeogenic pathway. Cori's Cycle During the muscle contraction, under anaerobic conditions, glucose is converted to lactate. Lactate is transported to liver through circulation. In the liver, lactate is converted back to pyruvate. Later, by gluconeogenesis, pyruvate is converted to glucose. Glucose is transported back to the muscle. It is called Cori's cycle (Fig. 3).
Muscle Glucose Blood Liver Glucose

4. Pro Later it ge
Propi

L-Me

Succ Gluconeogenesis

Glycolysis

Lactate

----

... ---_-+
Figure 3: Con's cycle

-.

Lactate

If the B12, metl condition

Carbohydrate
,yruvate

Metabolism

105

1. Lactate is converted to pyruvate by the action of lactate dehydrogenase. Later, pyruvate is converted to glucose by gluconeogenic pathway.

Lactate

----7-,..-""'r----- ... ~
NADH + H+ NAD+

Lactate dehydrogenase

Pyruvate

Glucose

is converted iolpyruvateenolpyruvate. iman beings. .oxaloacetate ther reactions ohate by the se direction. n irreversible ctose 1,6-bis

2. Various amino acids can form pyruvate or intermediates of TCA cycle. They can be converted to glucose. These amino acids are known as glucogenic amino acids. Alanine, serine, cysteine etc. form pyruvate during their metabolism. This can be later converted to glucose. Glutamate forms a-ketoglutarate and aspartate forms oxaloacetate. Both are later converted to glucose. 3. Glycerol is converted to the intermediate of glycolysis, dihydroxy acetone phosphate. Later, it forms pyruvate that can be converted to glucose.

phosphate

Glycerol

----7-.,..-""'~---....
ATP ADP

Glycerol kinase

Glycerol-3-P

6-phosphate. , the enzyme

Glycerol-3-P dehydrogenase

NADH + H+

Dihydroxy acetone phosphate

Glucose

le kidney. /sis and TCA

acid forms succinyl CoA, which is an intermediate ofTCA cycle. Later it gets converted to glucose.
CoASH; ATP Propionic acid ------------...

4. Propionic

Propionyl CoA

converted (0 'er, lactate is converted (0 ycle (Fig. ~).

Carboxylase Racemase

Biotin; CO2; ATP Propionyl CoA

L-Methyl melonyl CoA

....~

D-Methyl melonyl CoA

Succinyl CoA

Vitamin B12 Isomerase

----..04-- --.~ Glucose

If the enzyme isomerase is defective or if there is a deficiency of vitamin B12, methyl malonic acid gets accumulated and is excreted in the urine. This condition is called methylmelonic aciduria.

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Textbook of Biochemistry for Nurses

Lactate dehydrogenase Lactate-------7-.,.-"'r-------...... NADH + H+ NAD+ CO2 GTP
2

Pyruvate

Phosphoenol pyruvate Phosphoenol pyruvate ~.

Biotin; ATP : CO2 Pyruvate carboxylase

2-phosphoglycerate

3-phosphoglycerate

l l l

C_a_r_bo_x..;y_k_in_a_.:s:::=e .;.(P_E_P_C_K~)_ CO (Xaloacetate Aspartate Succinyl CoA Glutamate a-ketoglutarate Propionic acid !., Dihydroxy acetone phosphate Glycerol Fructose-6- phosphate

<:

Fructose 2, activator of Ina we It decreases tokinase-l During is decrease increases tl

Glycogen
Glycogen i ride made

1,3-bisphosphoglycerate Glyceraldehyde-3-phosphate

and muscl
When glycogen. : decreased, the blood Glycogene Glycogen. Glycogene glycogen.l called UD

~
Fructose 1,6-bisphosphate-----------~. Fructose 1,6-bisphosphatase Glucose

-<11I.-----------1
Glucose-6-phosphatase ~Glucose-6-phosphate

Figure 4: Reactions of gluconeogenesis

Regulation of Gluconeogenesis
Insulin and glucagon are mainly involved in the regulation of gluconeogenesis. Insulin decreases the synthesis of the key enzymes of gluconeogenesis. Glucagon increases the synthesis of the key enzymes of gluconeogenesis. Insulin increases the rate of glycolysis, whereas glucagon increases the rate of gluconeogenesis and decreases the rate of glycolysis. Most of the reactions of gluconeogenesis are reversal of glycolysis. So, both these pathways cannot operate simultaneously. When gluconeogenesis is predominant, the rate of glycolysis is decreased and vice versa. Fructose 2,6-bisphosphate is formed in excess when the concentration of insulin is increased by the action of an enzyme called phosphofructokinase-2.

Fructose 1,6-bisphosphatase (Gluconeogenesis)

-"U~Fructose-6-phosphate Phosphofructokinase-2

Fructose 1,6-bisphosphate

Phosphofructokinase-1 (Glycolysis)

~
-----Fructose 2,6-bisphosphate -------'

Synth Glycogen

glycogeni
side chain glucose re is used foi

Carbohydrate

Metabolism

107

1;

ATP: CO2

late carboxylase

)
.toqlutarate sphate ycerol ate

Fructose 2,6-bisphosphate is an inhibitor of fructose 1,6-bisphosphatase and an acti vator of phosphofructokinase-I (Fig. 4). In a well-fed state, when the blood glucose level is increased, insulin is secreted. It decreases the synthesis of the enzymes of gluconeogenesis and keeps phosphofructokinase-I in an active state. During fasting or starvation, blood glucose level is decreased. So, insulin level is decreased in the blood and there is a relative increase in glucagon level, which increases the rate of gluconeogenesis and decreases glycolysis. Glycogen Metabolism

Glycogen is the storage form of carbohydrate in animals. It is a homopolysaccharide made up of glucose units. It is highly branched. Glycogen is stored in liver and muscle. When surplus amount of carbohydrate is present in the body, it is stored as glycogen. Synthesis of glycogen is called glycogenesis. If blood glucose level is decreased, the stored glycogen is broken down and glucose is released to maintain the blood glucose level. The breakdown of glycogen is called glycogenolysis. Glycogenesis and glycogenolysis constitute glycogen metabolism. Glycogenesis

late

Glycogenesis is the set of biochemical reactions leading to the formation of glycogen. During glycogenesis, glucose is activated to form its nucleotide derivative called UDP glucose.
Glucose

.oneogenesrs. oneogenesis. ieogenesis. ses the rate of

Glucokinase

Glucose-6-phosphate Phosphoglucomutase

ysis. So, both

eogenesis
IS

If

K K
,

ATP

ADP

Glucose-1-phosphate UTP UDP Glucose pyrophosphorylase

icentration of
rctokinase- 2.

PPi

UDP glucose uctoki nase-1

colysis)

10

Synthesis of glycogen requires a preexisting glycogen or a primer molecule. Glycogen primer can be synthesized on the backbone of a protein called glycogenin. This protein contains a tyrosine residue. The hydroxyl group of the side chain of tyrosine residue is linked to glucose by glycosidic linkage. Later, glucose residues are added to it, leading to the formation of glycogen primer, which is used for the synthesis of glycogen as shown below (Fig. 5).

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Textbook of Biochemistry for Nurses

Tyrosine Glycosidic linkage

Glycogenin

~t

uo!:co:e

i
l l l

)-o~

Glucose Glycogen synthase adds glucose to the growing glycogen chain by forming 0.-1 ,4-glycosidic linkage.

~'---:----')-o
Glycogen synthase adds glucose to the growing glycogen chain by forming 0.-1 ,4-glycosidic linkage. Chain length considerably increased.

Once the chain length is considerably increased, a portion of the chain is removed and added to another point by forming a.-1,6-glycosidic linkage by an enzyme called branching enzyme.

Glycogen

Subsequent addition of glucose leads to the formation of glycogen.

Figure 5: Glycogenesis

This polyme t-phosphai qlucose-B-r

GLYCOGENOLYSIS
Free glucose can be formed from glycogen during glycogenolysis only in the tissues having glucose-6-phosphatase activities. Since the muscles don't have glucose-6-phosphatase activity, they cannot form free glucose, whereas liver containing the glucose-6-phosphatase activity can convert glucose-6-phosphate to free glucose (Fig. 6). Epinej membrane converts K. Increas Active fon phosphoryl synthase is Phospl activates tl:

Regulation of Glycogen Metabolism

Glycogen synthesis takes place in a well-fed state, and glycogen breakdown in fasting condition. Insulin favours glycogenesis, and hormones like glucagon and epinephrine favour glycogenolysis.

Carbohydrate

Metabolism

109

growing :linkage. ~ Glucose-6-phosphate Glucose-6- \. phosphatase Glucose

-=.J
Glycogen Phosphorylase Glucose-1-phosphate

growing ~Iinkage.

Glycogen phosphorylase hydrolyzes the a-1 A-glycosidic linkage until four glucose residues remain on the side of the ee-l ,6- glycosidic linkage.

iased, a portion roth er point by enzyme called Three of these four residues are shifted to the other branch by forming a-1 A-glycosidic linkage.

ycosidic linkage Debranching enzyme hydrolyzes the a-1,6-glycosidic linkage and releases free glucose.

he formation of

This polymer of glucose is further acted upon by glycogen phosphorylase and forms glucose1-phosphate. Glucose-1-phosphate is isomerized to form glucose-6-phosphate. The enzyme glucose-6-phosphatase converts glucose-6-phosphate to free glucose.

Figure 6: Glycogenolysis

sis only in the es don't have whereas liver -phosphate to

breakdown in glucagon and

Epinephrine or glucagon binds to their respective receptors on the cell membrane and activate adenylate cyclase. Adenylate, cyclase in the active form converts ATP to cAMP. Increase in the cAMP concentration activates cAMP dependent protein kinase. Active form of the cAMP dependent protein kinase phosphorylates glycogen phosphorylase and glycogen synthase (Fig. 7). In the phosphorylated form, glycogen synthase is inactive. and glycogen phosphorylase is active. Phosphodiesterase is an enzyme, which converts L'AMP to S 'AMP. Insulin activates this enzyme and decreases the cAMP content of the cell. It facilitates

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Textbook of Biochemistry for Nurses

Plasma membrane Glycogen synthase (active) ATP

ATP

is broken de glucose-o-pt affected indi from glucos. able to incre Glucose and glycolys

cAMP dependent protein kinase (Inactive)

hyperuricer
~--ADP

HEXOSE ~
Glycogen synthase (inactive)

[Pentose PI
In HMP shr This pathwa adrenal corn

Glycogen phosphorylase kinase (Inactive)

Glycogen phosphorylase kinase (Active)

ATP Glycogen phosphorylase kinase (Inactive) Glycogen phosphorylase kinase (Active)

Reaction S
(

ATP'

Figure 7: Regulation of glycogen metabolism

ADP'" Glucos, NADP+' NADPH + H" s-phospn

dephosphorylation of glycogen synthase and glycogen phosphorylase. In the dephosphorylated form, glycogen synthase is active whereas glycogen phosphorylase is inactive.

Glycogen Storage Disorders

Defect or deficiency in the enzymes involved iri the metabolism of glycogen gives rise to glycogen storage disorders. There are eight different types of glycogen storage disorders (Table 1).
Table 1: Types of glycogen storage disorders

6-phos NADP+. NADPH + H+'

Type
Type Type Type Type Type Type Type Type I Il III IV V VI VII VIII

Deficiency Glucose-o-phosphatase
a-I,4-glucosidase Debranching enzyme Branching enzyme Glycogen phosphorylase Glycogen phosphorylase Phosphofructokinase Phosphorylase kinase

Name of the disease

Von Gierke's disease Pompe's disease Cori's disease Anderson's disease Me Ardle's disease Hers' disease Tauri's disease

Ribulose-!

Ribose-S Sedoheptulc Fructose-

(muscle) (liver)

Von Gierke's Disease (Glucose-ti-Phosphatase

In this disease, the enzyme glucose-o-phosphatase

Deficiency)
is defective. So, when glycogen

Fructose-t

Carbohydrate Metabolism

111

rthase (active)

is broken down, free glucose cannot be formed. Glucose gets accumulated as glucose-6-phosphate. Since free glucose is not formed during glycogenolysis, the affected individual suffers from fasting hypoglycemia. As the formation of glucose from glucose-6-phosphate is blocked, injection of glucagon or epinephrine is not able to increase the blood glucose level. Glucose-6-phosphate, which is accumulated, is diverted to HMP shunt pathway and glycolysis. This is responsible for lactic acidosis, hypertriglyceridemia and hyperuricemia seen in the type I glycogen storage disorder. HEXOSE MONOPHOSPHATE SHUNT (HMP SHUNT)

IP

rthase (inactive)

[Pentose Phosphate

Pathway (PPP)]

In HMP shunt, glucose is directly oxidized to produce NADPH + H+ and CO2. This pathway takes place in the cytosol of various tissues like liver, adipose tissue, adrenal cortex, lactating mammary gland, erythrocyte, neutrophils, testis etc. Reaction Sequence

.rylase. In the .en phosphory-

Glucose-6-phosphate NADP+~I NADPH + H 6-phosphogluconolactone Glucose-6-P dehydrogenase

A::~
I

Glucose

?;lycogengives ycogen storage

H,o~

Hydrolase

6-phosphogluconate NADP+~ NADPH + H<"

the disease ce's disease disease ease ,'s disease 's disease ~ase sease

l\..1

Dehydrogenase Epimerase -.-----------.. ~ Xylulose-5-phosphate

CO2 RibUlose-lfhosPhate

Ribose-5-phosphate

----------1
:::_-_-_-_-_ .... -_.J~---~II Erythrose-4-phosphate

Sedoheptulose-7 -phosphate Fructose-6-phosphate

4

Transaldolase

Fructose-6-phosphate .4-_--------ruG1yceraldehyde-3-phosphate

when glycogen

Figure 8: Hexose monophosphate shunt

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Textbook of Biochemistry for Nurses

Glucose is converted to glucose-6-phosphate by the action of glucokinase or hexokinase by utilizing phosphate group from the ATP. Glucose-6-phosphate is oxidized by the action of the enzyme gIucose-6-phosphate dehydrogenase to form 6-phosphogluconolactone. The enzyme glucose-6-phosphate dehydrogenase requires NADP+ as the coenzyme, and during the reaction the coenzyme is reduced (Fig. 8). In the next reaction, aQ enzyme called 6-phosphogluconolactone hydrolase hydrolyses phosphogluconolactone to 6-phosphogluconate. This product is later oxidized and decarboxylated by the action of a dehydrogenase to form ribuloseS-phosphate. During this oxidative reaction, NADP+, the coenzyme, is reduced to NADPH + H+. Some of the ribulose-S-phosphate isomerize to form ribose-S-phosphate and some are epimerized to form xylulose-S-phosphate. Ribose-S-phosphate and xylulose-S-phosphate react in the presence of an enzyme called transketolase to form sedoheptulose-7 -phosphate and glyceraldehyde-3-phosphate. The enzyme transketolase requires thiamine pyrophosphate (TPP) as the coenzyme. Later, glyceraldehyde-3-phosphate and sedoheptulose-7 -phosphate react in the presence oftransaldolase to form erythrose-4-phosphate and fructose-6- phosphate. Erythrose-4-phosphate and an another molecule of xylulose-S-phosphate react in the presence of transketolase to form fructose-6-phosphate and glyceraldehyde3-phosphate. Glyceraldehyde-3-phosphate and fructose-6-phosphate enter into glycolytic pathway. Glucose-6-phosphate is the substrate for glycolysis. It can also enter into an alternative pathway, where it gets converted to fructose-6-phosphate that enters hack into glycolysis. Hence, this pathway is called hexose monophosphate shunt. The initial reactions of the pathway where oxidation takes place is called oxidative phase. At the end of the oxidative phase, ribulose-S-phosphate is formed. It is later isomerized to ribose-S-phosphate and xylulose-S-phosphate. This phase of the metabolic pathway is called nonoxidative phase. I mportance of Hexose Monophosphate Shunt

The pen converte Provide glycoprc Glucose-6-F In glucose-r reduced. DUI oxidized stat RBCs. It darr with glucose are exposed of fava bean: be seen. METABOL
I

ATP ADP
Fructos

Glye

ATP ADP
Glyceralde

During HMP shunt, a large amount of reducing equivalents in the form of NADPH + H+ and pentose phosphate are produced. The reducing equivalents can be used for the following purposes: Reductive synthesis of fatty acids, cholesterol, steroid hormone, glutamate etc. They can be utilized by cytochrome P 450 enzyme system. To reduce the oxidized form of glutathione by the enzyme glutathione reductase. Help to keep hemoglobin in the reduced state. In neutrophil. NADPH oxidase utilizes NADPH ,lIld produces superoxide, which helps to kill bacteria. The pentose nucIeotides. sugar, ribose-S-phosphate is utilized for the formation of

Fructose

I-phosphate phosphate ir Glycera kinase, whir can enter int converted to

Carbohydrate Metabolism glucokinase or -6-phosphate is ehydrogenase -6-phosphate le reaction the .one hydrolase oroduct is later form ribulosee, is reduced to -phosphate and phosphate and 'ansketolase to e. The enzyme izyrne. hate react in the e-6- phosphate. isphate react in Iyceraldehydehate enter into enter into an late that enters osphate shunt place is called ihate is formed. ate. This phase
:0

113

The pentose sugars of dietary origin or produced inside the body can be converted to the intermediates of glycolysis. Provide pentose sugars for the synthesis of oligosaccharide chain of glycoproteins. Glucose-6- Phosphate Dehydrogenase Deficiency

In glucose-6-phosphate dehydrogenase deficiency, production of NADPH is reduced. Due to the decreased cell content of NADPH, glutathione remains in the oxidized state. This leads to the accumulation of free radicals in the cells, especially RBCs. It damages the cell membrane and gives rise to hemolysis. Usually, individuals with glucose-6-phosphate dehydrogenase deficiency are symptomless. When they are exposed to certain drugs like sulphonamides, primaquine etc. or consumption offava beans (Favism), the enzyme deficiency manifests and severe hemolysis can be seen.

METABOLISM

OF FRUCTOSE

Fructose

ATP=l1 Fructokinase ADP --------~

Fructose-1-phosphate

~
Glyceraldehyde

1
Dihydroxy acetone phosphate Kinase

ATP ~ I ADPA"""l

Glyceraldehyde-3-phosphate

lrmofNADPH ses: ormone, gluta1.

Glycolysis

Glucose

Fructose 1,6-bisphosphate

Figure 9: Fructose metebolism

me glutathione

roduces superformation of

Fructose is a ketohexose. It is phophorylated by fructokinase to form fructosel-phosphate. Fructose-l-phosphate forms glyceraldehyde and dihydroxy acetone phosphate in the presence of an enzyme called aldolase. Glyceraldehyde is converted to glyceraldehyde-3-phosphate by the action of a kinase, which utilises phosphate group from ATP. Glyceraldehyde-3-phosphate can enter into glycolytic pathway, or along with dihydroxyacetone phosphate it is converted to fructose 1,6-bisphosphate and enters gluconeogenic pathway (Fig. 9).

114

Textbook of Biochemistry for Nurses glucose beca a carbohydra normal indiv When tl circulation b glucose leve come into ac

Fructose is mainly derived from the digestion of dietary sucrose. During the metabolism of fructose, it is converted to the intermediate of glycolysis and later to pyruvate, bypassing the regulated step of glycolysis. The fructose metabolism through the glycolytic pathway is unregulated. All the fructose gets converted to acetyl CoA. Acetyl CoA pool is increased. It is the substrate for fatty acid synthesis. When the fructose content of the blood increases, blood glucose level also increases. This is responsible for the .increase in insulin content of the blood. Insulin favors fat synthesis. So, individuals who eat more sweets can become obese due to the deposition of fat in the adipose tissues.

Metabolic p
Glycoly

Essential Fructosuria
Defect in the enzyme fructokinase gives rise to essential fructosuria, fructose gets accumulated and is excreted in the urine. in which

HMP sl
Glycog: Uronic

Fructose Intolerance
Defect in the enzyme aldolase B is responsible for fructose intolerance.

Metabolic J
Glycog. Glucon A perf metabolisr fine regula hormones. Insulin hyperglycei of increasn pathways epinephriru

METABOLISM

OF GALACTOSE

Galactose is one of the constituents of milk sugar. It is converted to glucose in the liver (Fig. 10).
Galactose ATP,) ADP..

"!

Galactokinase

Galactose-1-phosphate UDP
9IUCOSe~

Galactose-1-phosphate Uridyl transferase

Glucose-1-phosphate UDP Galactose

!1

Increase il Insulin is :
number of. increases ti rate of gly suppresses

Epimerase Glycogen

UDP Glucose

Figure 10: Galactose metabolism

Galactosemia
In this disease, there is a defect in the enzyme galactose-l-phosphate tranferase. Galactose gets accumulated in the blood, so the condition galactosemia. Cataract is seen in children suffering from galactosemia. uridy I is called

Decrease' Glucagon
fasting an gluconeog state and b glycolysis glucose. S~

Blood Glucose Homeostasis

Normal fasting plasma glucose level is 70 to 110 mg/dl. Normal fasting blood glucose level is 60 to 100 mg/dl. Blood glucose level is slightly less than the plasma

Carbohydrate

Metabolism

115

se. During the lysis and later se metabolism :;con verted to acid synthesis. also increases. Insulin favors ese due to the

glucose because of the difference in the water content of RBCs and plasma. After a carbohydrate rich diet, glucose level increases slowly and is below 140 mg/dl in normal individuals. When the blood glucose level decrease, free glucose is poured into the circulation by different metabolic pathways in order to maintain the normal blood glucose level. When the blood glucose level increases, various metabolic pathways come into action to utilize the glucose or store the excess amount of glucose.

Metabolic pathways responsible for utilization and storage of glucose

. Glycolysis and citric acid cycle HMP shunt Glycogenesis Uronic acid pathway

iria, in which

Metabolic pathways responsible for the formation of glucose

nee. Glycogenolysis Gluconeogenesis A perfect balance between these two groups of pathways of carbohydrate metabolism is responsible for maintaining the blood glucose level. This fine regulation is mainly done by the hormonal regulation of blood glucose hormones. Insulin is the major hypoglycemic hormone, and glucagon is the major hyperglycemic hormone. In addition to glucagon, many other hormones are capable of increasing the blood glucose level by altering the rate of various metabolic pathways responsible for carbohydrate metabolism. These hormones are epinephrine, glucocorticoids, thyroid hormone, growth hormone etc.

glucose in the

Increase in the Level of Blood Glucose Insulin is secreted when the blood glucose level increases. Insulin increases the
number of glucose transporters on the cell membrane of the peripheral tissues and increases the rate of transport of the glucose inside the cell. Insulin increases the rate of glycolysis, glycogenesis, and HMP shunt pathway and simultaneously suppresses the rate of glycogenolysis and gluconeogenesis.

.sphate uridyl ition is called ma.

Decrease in the Level of Blood Glucose Glucagon level is increased when the blood glucose level decreases, as in case of
fasting and starvation. Glucagon increases the rate of glycogenolysis and gluconeogenesis by keeping the regulated enzymes of these pathways in the active state and by increasing their synthesis. Glucagon decreases the rate of glycogenesis, glycolysis and HMP shunt pathway. Glucagon favors those pathways, which produce glucose, so it has got an antiinsulin action.

fasting blood ian the plasma

116

Textbook of Biochemistry for Nurses

Epinephrine is s nthesized in the adrenal medulla. It helps to increase the blood glucose level. It increases the rate of glycogenolysis and gluconeogenesis. Unlike glucagon, epinephrine increases the rate of glycogenolysis, both in liver and muscle. It can stimulate the release of glucagon and can inhibit the secretion of insulin. This action of epinephrine on the release of insulin and glucagon is responsible for increasing the blood glucose level. Growth hormone is secreted in response to hypoglycemia. Growth hormone has hyperglycemic effect. Glucocorticoids and thyroxine have an effect on carbohydrate metabolism and tend to increase the blood glucose level. DIABETES MELLITUS

Lipid metal As a result 0 An alternate level of fattthe accumul If the level 0' Excretion 01 and ketaonu Since l If the blood diabetic ket Protein me. In diabetes I This leads tl Diagnosis ( Analysis o) Urine colle Benedict's t Alterna reagents are Estimation Blood is col containing oxalate acts If the j occasions, j Oral gluco

Diabetes mellitus is a disorder, which results due to the deficiency of insulin secreted by the ~-cells of pancreas, or defect in the action of insulin. In diabetes mellitus, the metabolism of carbohydrates, proteins, and lipids is altered. Signs and Symptoms Signs and symptoms of diabetes mellitus are: Polyuria (excess urine excretion) Polydypsia (increased thirst) Polyphagia (excess hunger) Diabetes mellitus may be Type I diabetes mellitus or Type 11 diabetes mellitus. Type I diabetes mellitus is insulin dependent diabetes mellitus (IDDM), and Type 11 diabetes mellitus is non-insulin diabetes mellitus (NIDDM). Type I diabetes mellitus Juvenile onset Absolute deficiency of insulin Ketoacidosis is commonly seen Alterations Type 11 diabetes mellitus Adult onset Resistance to the action of insulin Hyperosmolar coma is commonly seen proteins, and lipids

in the metabolism of carbohydrates,

Carbohydrate metabolism Insulin is required for the uptake of glucose by the peripheral tissues. Insulin favors the metabolic pathways that are involved in the utilization and storage of carbohydrates. In case of diabetes mellitus, glucose is not taken up by the peripheral tissues and its utilization is decreased. This increases the blood glucose level, and the condition is called hyperglycemia. When the blood glucose level is increased, it is excreted in urine and the condition is called glycosuria. Presence of the reducing sugar in urine can be detected by Benedict's test.

Oral glucos excess amo a load of gh Preparatioi Patient sho the test she consume IT activity of 1 elicit a mm

Carbohydrate

M etabolism

117

, increase the .oneogenesis. both in liver the secretion I glucagon is wth hormone an effect on el.

Lipid metabolism
As a result of insulin deficiency, cells will be starved of glucose (source of energy). An alternate fuel, fatty acid, is released from the adipose tissue. When the blood level of fatty acid increases, its oxidation at the liver is also increased, leading to the accumulation of acetyl CoA. Acetyl CoA is later converted into ketone bodies. If the level of ketone bodies increases in the blood, the condition is called ketonemia. Excretion of excess of ketone bodies in the urine is called ketonuria. Ketonemia and ketaonuria together is ketosis. Since ketone bodies are acidic in nature, it might decrease the pH of the blood. If the blood pH is decreased due to the accumulation of ketone bodies, it is called

diabetic ketoacidosis. cy of insulin

n. In diabetes ered.

Protein metabolism
In diabetes mellitus, protein synthesis decreases and protein breakdown increases. This leads to negative nitrogen balance.

Diagnosis of Diabetes Mellitus Analysis of urine to detect the presence of glucose

Urine collected from the individual can be tested for the reducing substance. Benedict's test is done for the detection of reducing substance in the urine. Alternatively, glucose strip containing enzyme glucose oxidase and other reagents are available to detect the presence of glucose specifically.

ietes melli tus. M), and Type

Estimation of plasma glucose

f insulin nmonly seen Blood is collected from the patients by aseptic precautions. It is stored in a container containing sodium fluoride and potassium oxalate (sugar tubes}. Potassium oxalate acts as an anticoagulant, and sodium fluoride is an inhibitor of glycolysis. If the fasting plasma glucose level is above 126 mg/dl on two separate

I lipids

occasions, it is diagnostic of diabetes mellitus. , Oral glucose tolerance test

Insulin favors Id storage of the peripheral ose level, and urine and the urine can be

Oral glucose tolerance test is done to assess the ability of the body to handle an excess amount of glucose. Plasma glucose level is measured before and after giving a load of glucose. Before performing the test, patient should be properly prepared.

Preparation of the patient

Patient should do his/her routine work. [f possible, drugs that might interfere with the test should be stopped. At least, three days prior to the test, individual. must consume minimum I SO g of carbohydrates per day. This will help to keep the activity of the enzymes of carbohydrate metabolism at the optimum level so as to elicit a maximum response.

118

Textbook of Biochemistry for Nurses

Patient should fast for about 8 to 12 hours before the test.

b) c) d)

Glycc Glyc: Diabi

Test procedure
Individual who is fasting for the past 8 to 12 hours and ready for OGTT is made to sit comfortably in the laboratory. The fasting blood sample is collected. Individual is made to drink 7S g of anhydrous glucose in about 200 ml of water. Exactly two hours after the glucose is ingested, one more blood sample is taken and it is called 2 hour post glucose load sample. The glucose present in the plasma is estimated by glucose oxidase peroxidase method. Interpretation of OGTT Result WHO Criteria Fasting plasma glucose
Normal Impaired tolerance Fasting hyperglycemia Diabetes mellitus <126 mg/dl >110 mg/dl and <126 mg/dl >110 mg/dl and <126 mg/dl >126 mg/dl

e) Oral 2. What you necqene:

Short

Answ

1. Write the (a) Galac (b) Esser 2. 3. 4. 5. 6. 7. 8. What is t What is t Describe Name thl What is ~ How will What is tl

2 hour post glucose load

<140 mg/dl > 140 mg/dl and <200 mg/dl <140 mg/dl >200 mg/dl

9. How ma anaerobi

Glycated

Hemoglobin

(HbAlc)

If blood glucose level is increased and remains high for a long time, it will glycate the proteins present in the body. The amino-terminal valine residue of the ~ chain ofhemoglobin is glycated nonenzymatically in diabetes mellitus. It is called HbAlc. Since the life span of erythrocytes is on an average 90 days, measurement of glycated hemoglobin gives an idea about the blood glucose level of an individual during the past few months. QUESTIONS Long Essay
1. Discuss in detail about the regulation of blood glucose level. 2. Discuss in detail about glycolysis. 3. What is gluconeogenesis? Mention the substrates for gluconeogenesis, and the key enzymes of gluconeogenesis. Write the reaction sequence of gluconeogenesis from lactate. 4. Describe the reactions of HMP shunt, and write the significance of this pathway. 5. Discuss in detail about citric acid cycle. 6. Discuss in detail about the glycogen metabolism. Add a note on glycogen storage disorders.

Short

Essay

1. Write short notes on the following: a) Glycogen storage disorders

Carbohydrate b) Glycogenesis

Metabolism

119

iTl'

:3

ade to
(\\0

~.:: =:::d: .idual

_ ..=.:.: ~:: _..

c) Glycogenolysis d) Diabetes mellitus e) Oral glucose tolerance test 2. What you mean by gluconeogenesis? neogenesis.

Discuss about the key enzymes of gluco-

~':::5
-,

alled : ~ ;:3 .irnated

Short Answers
1. Write the defect in: (a) Galactosemia. (b) Essential fructose intolerance.
2.

3. o :e load
4.

:: c= = rng/dl

5. 6. 7. 8.
9.

What is the significance of estimation of glycated hemoglobin? What is the biochemical basis for cataract in galactosemia? Describe Von Gierk's disease. Name the regulated enzymes of glycogen metabolism. What is ketoses? Mention any two conditions where ketoses is seen. How will you prepare the patient for oral glucose tolerance test? What is the normal blood glucose level? Metion any two causes for hyperglycemia. How many ATP's are formed when glucose is metabolized by glycolysis under anaerobic condition?

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