Asst. Prof. Dr.

Muhammad Haroon
MD, ECEA, MPH (JHSPH)
Lecturer at Kandahar Medical faculty & MPH
Former Biochemistry Guest lecturer at SMS
medical college, India.
Email:[email protected]
 Overview

l Alternate pathway for oxidation of glucose

l Other Names:
– Pentose Phosphate Pathway
– Phosphogluconate Oxidative Pathway
– Direct Oxidative Pathway
l Purpose:
– Production of NADPH
– Production of Ribose
– No ATP generated & consumed
2 HMP-Shunt
 Overview Cont.….......

l Site:
– Liver
– Adipose Tissue
– Adrenal Cortex
– Erythrocytes
– Gonads
– Thyroid
– Lactating Mammary Glands
l Location within cells:
3 – Cytoplasm HMP-Shunt
 Phases & Products of HMP Shunt
Pathway

l Phases:
– Oxidative Phase (Irreversible)
– Non-oxidative Phase (Reversible)
l Oxidation of one molecule of Glucose-6 phosphate
in HMP shunt produces:
– One molecule of CO2
– Two molecules of NADPH
– One molecule of pentose phosphate

4 HMP-Shunt
 Oxidative Phases

5 HMP-Shunt
 Introduction

l This portion of pathway involves:

– Three irreversible reactions

l Glucose -6-Phosphate leads to the formation of:

– Ribulose 5-phosphate
– CO2
– Two molecules of NADPH

6 HMP-Shunt
 Introduction Cont.….......

l This portion is important:

– For NADPH-dependent Biosynthesis of fatty Acids in:
q Liver & Adipose Tissue
q Lactating Mammary Glands
– For NADPH-dependent Biosynthesis of Steroid
Hormones in:
q Testes & Ovaries
q Placenta
– For NADPH-dependent Reduction of Glutathione in:
q RBCs
7 HMP-Shunt
 Reaction-1: Oxidation of Glucose-6-
Phosphate to 6-Phosphoglucanolactone

l Reaction:
– Glucose-6-phosphate is oxidized to 6-phosphoglucono
lactone
– NADP+ is reduced to NADP+H
l Catalyzed by:
– Glucose-6-phosphate dehydrogenase (G6PD)
l (G6PD) enzyme is:
– Irreversible (Most regulated)
– Inhibited by NADPH (Competitively)
8 – Activated by Insulin HMP-Shunt
 Reaction-2: Hydration of 6
Phosphoglucanolactone

l Reaction:
– A molecule of water is added to 6-
phosphogluconolactone to generate 6-phosphogluconate

l Catalyzed by:
– Gluconolactone hydrolase

9 HMP-Shunt
 Reaction 1 & Reaction 2

Glucose-6-phosphate 6-Phospho- O O-
Dehydrogenase glucono- 1C
6CH 2OPO 32- 6 2-
lactonase
+ CH 2OPO 3
NADPH + H HC OH
H
5 O OH NADP
+
H
5 O H2O H+ 2
H H HO 3CH
4 OH H 1 4
OH H O
1 HC OH
4
OH H OH
3 2 3 2 HC OH
5
H OH H OH CH 2OPO 32-
6
glucose-6-phosphate 6-phoshogluconolactone 6-phosphogluconate

10 HMP-Shunt
 Reaction-3: Decarboxylation of 6-
phosphogluconate

l Reaction:
– Oxidative decarboxylation of 6-phosphogluconate to
Ribulose-5 Phosphate
– Second NADP+ is reduced to NADP+H
– One molecule of CO2 is produced
l Catalyzed by:
– 6-phosphogluconate Dehydrogenase

11 HMP-Shunt
 Reaction-3: Decarboxylation of 6-
phosphogluconate Cont.….......

O O- Phosphogluconate
1C Dehydrogenase
HC OH CH OH
2 NADP + NADPH + H+ 1 2
HO 3CH C O
2
HC OH HC OH
4 3
HC OH CO 2 HC OH
5 4
CH 2OPO 32- CH 2OPO 32-
6 5
6-phosphogluconate ribulose-5-phosphate
12 HMP-Shunt
 Non-Oxidative Phases

13 HMP-Shunt
 Introduction

l Concerned with the Reversible interconversion of:

– Three, four, five and seven carbon monosaccharides
l Sites:
– Rapidly dividing cells
q Bone marrow
q Skin
q Intestinal mucosa
q virtually in all tissues
l Importance:
14 – Pentose production
HMP-Shunt
 Reaction-4: Isomerization or
Epimerization of Ribulose-5 Phosphate

l Reaction 4a:
– Some of the ribulose-5-phosphate molecules are
isomerized to ribose-5- phosphate
l Catalyzed by:
– Ribose-5-Phosphate ketoisomerase
l Reaction 4b:
– Some of the ribulose-5-phosphate molecules are
epimerized to xylulose-5-phosphate
l Catalyzed by:
15 – Ribulose-5-phosphate epimerase HMP-Shunt
 Reaction-4: Isomerization or Epimerization of
Ribulose-5 Phosphate Cont.….......

CH2OH

C O
Epimerase
HO C H

CH2OH H C OH

C O CH2OPO32-
xylulose-5-
H C OH phosphate
H C OH HC O

CH2OPO32- H C OH
ribulose-5- H C OH
phosphate Isomerase
H C OH

CH2OPO32-
ribose-5-
16 phosphate HMP-Shunt
 Reaction-5: Transketolase Reaction

l Reaction:
– Two-carbon unit (with keto group) from xylulose-5-
phosphate will be transferred to ribose-5-phosphate
l It will form:
– Sedoheptulose-7-phosphate (7 Carbon sugar)
– Glyceraldehyde-3-Phosphate
l Catalyzed by:
– Transketolase (Thymine & Mg+ Dependent Enzyme)

17 HMP-Shunt
 Reaction-5: Transketolase Reaction
Cont.….......

CH 2OH
Transketolase
C O

CH 2OH HC O HO C H

C O H C OH H C OH

HO C H H C OH HC O H C OH

H C OH
+ H C OH H C OH
+ H C OH

CH 2OPO32- CH 2OPO32- CH 2OPO32- CH 2OPO32-

xylulose- ribose- glyceraldehyde- sedoheptulose-

5-phosphate 5-phosphate 3-phosphate 7-phosphate
18 HMP-Shunt
 Reaction-6: Trans aldolase
Reaction

l Reaction:
– Three-carbon unit from sedoheptulose 7-phosphate will
be transferred to glyceraldehyde 3-phosphate
l It will form:
– Fructose 6-phosphate
– Erythrose 4-phosphate
l Catalyzed by:
– Trans-aldolase

19 HMP-Shunt
 Reaction-6: Trans aldolase
Reaction Cont.….......

CH2OH
Transaldolase
C O H2C OH

HO CH C O

HC OH HC O HO CH

HC OH HC O HC OH HC OH

HC OH
+ HC OH HC OH + HC OH

H2C OPO32- H2C OPO32- H2C OPO32- H2C OPO32-

sedoheptulose- glyceraldehyde- erythrose- fructose-

7-phosphate 3-phosphate 4-phosphate 6-phosphate
20 HMP-Shunt
 Reaction-7: Transketolase Reaction

l Reaction:
– Two-carbon unit from xylulose-5-phosphate will be
transferred to Erythrose-4-phosphate
l It will form:
– Fructose -6-Phosphate
– Glyceraldehyde-3-Phosphate
l Catalyzed by:
– Transketolase

21 HMP-Shunt
 Reaction-7: Transketolase Reaction
Cont.….......

22 HMP-Shunt
 Uses of NADPH & Pentoses

23 HMP-Shunt
 Introduction

l NADPH is required for:

– Reductive biosynthesis—fatty acids, cholesterol and
steroid hormones
– Free radical scavenging
– Maintain RBC membrane integrity by keeping GSH in
reduced state
– Prevention of met-hemoglobin formation
– Detoxification by hydroxylation
– Bactericidal activity of macrophages
24 HMP-Shunt
 Reduced Glutathione

l In erythrocytes, glutathione is oxidized:

– To detoxify hydrogen peroxide and free radicals

l NADPH is required for:

– Restoring reduced glutathione

l This prevents:
– Premature destruction of erythrocytes

25 HMP-Shunt
 Reduced Glutathione Cont.….......

26 HMP-Shunt
 Special functions of NADPH in
RBCs

l Maintains the concentration of reduced glutathione

– Mentioned earlier

l Prevention of Methemoglobin (Fe3+):

– NADPH necessary to keep the ferrous iron (Fe2+) of
hemoglobin in the reduced state

27 HMP-Shunt
 Special functions of NADPH in
RBCs Cont.….......

28 HMP-Shunt
 Macrophage Bactericidal Activity

l NADPH is required for:

– Production of reactive oxygen species (ROS) (superoxide
anion radical) by macrophages to kill bacteria

29 HMP-Shunt
 Use of Pentose

l In the HMP shunt, hexoses are converted into:

– Pentoses
l The most important being:
– Ribose 5-phosphate
l This Pentoses are useful for the synthesis of:
– Nucleic acids (RNA and DNA)
– Many nucleotides such as ATP, NAD+, FAD and CoA

30 HMP-Shunt
 Glucose-6-Phosphate Dehydrogenase
Deficiency

31 HMP-Shunt
 Introduction

l Most common enzyme deficiency

l Cause:
– Genetic Defects
l Inheritance:
– X-linked recessive trait
l Clinical manifestations:
– Hemolytic Anemia
– Methemoglobinemia

32 HMP-Shunt
 Exacerbating Factors

l Oxidant Drugs: (3As)

– Antibiotic (sulfamethoxazole and chloramphenicol)
– Ant—malarial (Primaquine)
– Anti-Pyretic (Acetanilide)
l Favism:
– Ingestion of Fava Bean
l Infections:
– Most common

33 HMP-Shunt
 G6PD Deficiency and Resistance to
Malaria

l The enzyme deficiency offers resistance to:

– Plasmodium infection (P. Falciparum)

l Cause of Resistance:
– Reduced glutathione is necessary for parasite survival
– Which is not available in adequate amounts

34 HMP-Shunt
 Diagnosis & Treatment

l Biochemical Diagnosis:
– Detecting reduced activity of G6PD in RBC
l Pathologic Diagnosis:
– Visualization of Heinz bodies
l Treatment:
– Avoiding oxidative stress
– Symptomatic treatment of hemolysis

35 HMP-Shunt
 Diagnosis & Treatment Cont.….......

36 HMP-Shunt
 References
l Rodwell, V. W., Botham, K. M., Kennelly, P. J., Weil, P. A., & Bender, D. A.
(2015). Harper's illustrated biochemistry (30th ed.). New York, N.Y.: McGraw-
Hill Education LLC.

l Harvey, Richard A., Ph. D. (2017). Lippincott's illustrated reviews: Biochemistry.

Philadelphia :Wolters Kluwer Health

l Lehninger, A. L., Nelson, D. L., & Cox, M. M. (2000). Lehninger principles of

biochemistry. New York: Worth Publishers. Chicago (Author-Date, 15th ed.) L

l U. Satyanarayana, Biochemistry, Books and Allied (P) Ltd., Calcutta, Fourth

Edition

37 l The images are retrieved from: www.google.com/images

HMP-Shunt
 Thank You

38 HMP-Shunt