Digestion,

Absorption,
Glycolysis and
TCA Cycle
200 LEVEL MBB’S CBD/IBS PROGRAMME
DR. A.Z LAWAL
MEDICAL BIOCHEMISTRY DEPARTMENT
Introduction

 Dietary carbohydrates principally consist

of the polysaccharides
 Also contains disaccharides and small
amounts monosaccharides
 Liquid food materials escape digestion in
mouth but solid foods are masticated
thoroughly before they are swallowed
Digestion in the mouth

 Mastication
 Rolling/swallowing
 Saliva - Salivary amylase
 Activation - Cl
 Optimum pH 6-7
 Hydrolyzes α- (1,4) glycosidic linkage
 Action stops in stomach pH
Digestion in Stomach

 No carbohydrate splitting enzymes

 HCl may hydrolyze some dietary sucrose to equal
amounts of glucose and fructose
Digestion in Duodenum

 Pancreatic juice - pancreatic amylase

 α - amylase,
 optimum pH 7.1
 Cl- for activity
 hydrolyzes α-(1,4) glycosidic linkage
Digestion in Small Intestine
 Intestinal Juice
 pancreatic amylase:
 It hydrolyzes terminal α-(1,4), glycosidic
linkage
 liberates free glucose molecules.
 Lactase:

It is a β- glycosidase
pH - 5.4 to 6.0
Lactose - glucose and galactose.
Digestion in Small Intestine cont.
 Maltase:

hydrolyzes the α -(1,4) glycosidic linkage

liberating two glucose molecules.
pH range is 5.8 to 6.2.
 Sucrase:

PH - 5.0 to 7.0
hydrolyzes sucrose molecule
Liberates glucose and fructose.
Carbohydrate absorption
 Almost entirely from the small intestine
 Proximal jejunum is three times grater than that of
distal ileum
 Monosaccharides - blood stream
 Some disaccharides, which escape digestion, may enter
the cells of the intestinal lumen by “pinocytosis” and are
hydrolyzed within these cells.
Mechanism of Absorption
 Simple absorption (passive diffusion):
 the concentration gradient of sugar
 all monosaccharides especially fructose &
pentoses
 Facilitative diffusion
 Na+- independent glucose transporter system (GLUT5)
 mobile carrier proteins - fructose, glucose, and galactose with
their conc. gradient
 Active transport
 sodium-dependent glucose transporter system (SGLUT1)
 mobile carrier protein coupled with Na+-K+ pump.
 has 2 separate sites one for Na+ and for glucose
 Na+ ions is expelled outside the cell by Na+-K+ pump
which needs ATP and expel 3 Na+ against 2 K+.
 Exit all sugars from mucosal cell to the blood occur
by facilitative transport through GLUT2.
Pathways for transport of material
absorbed by intestine

 Water-soluble nutrients → hepatic portal system → liver

 Lipid soluble nutrients → Lymphatic vessels → thoracic
duct
GLUCOSE UPTAKE BY TISSUES
 Different protein carriers or transporters
 GLUT1 : present mainly in red cells, and retina.
 GLUT2 : liver, kidneys, pancreatic B cells, and lateral
border of small intestine, for rapid uptake and
release of glucose
 GLUT3 : mainly in brain
 GLUT4 : heart, skeletal muscles, and adipose tissues.
It is for insulin-stimulated uptake of glucose
 GLUT 5 : small intestine and testes for glucose and
fructose transport

 SGLUT 1: small intestine and kidneys, for active

transport of glucose and galactose from lumen of
small intestine and reabsorption of glucose from
glomerular filtrate in proximal renal tubules
Role of insulin in transport of glucose

 through GLUT 4 :
 Insulin produces transfer of GLUT-4 from their intracellular
pool to the outer membrane surface of these tissues
 increase GLUT-4 in the cell surface of these tissues leads to
increase glucose transport and uptake by these tissues

 Other carriers are insulin-independent

FATE OF ABSORBED SUGARS

 Hexoses or pentoses
 Pentoses are excreted in urine
 Hexoses are glucose, fructose, or
galactose
 Fructose and galactose are converted
into glucose in the liver
FATE OF ABSORBED GLUCOSE

 Oxidation
 Synthesis of other CHO substances
 Synthesis of non essential amino acids
 Excess
glucose is stored as glycogen in liver and
muscles (glycogenesis)
 More excess glucose is stored as lipid in adipose
tissue (lipogenesis)
 GLYCOLYSIS
 Glycolysis : Derived from Greek words;

Glykys = Sweet, Lysis = splitting

 Described by Embeden, Meyerhoff and Parnas,

hence called Embden Meyerhoff pathway

 Erythrocytes and nervous tissues derive its energy mainly form

glycolysis
Definition

 The glycolytic pathway represents an ancient

process in which degradation of glucose yield
pyruvic acid in the presence of oxygen (aerobic) and
lactic acid in absence of mitochondria (RBCs) and in
absence of oxygen (anaerobic) with the release of
ATP

 Glycolysis occurs in the cytoplasm of all human

cells, mainly the liver, kidney and muscle cells
Aerobic Phase

 Produces two molecules each of NADH and

pyruvate

 NADH must be re-oxidised for glycolysis to

continue
 Oxidation is carried out by dehydrogenation
 Reducing equivalent is transferred to NAD
 NADH + H+ in presence of O2 is oxidized in
electron- transport chain producing ATP
Anaerobic Phase

 Produces 2 molecules each of lactate and ATP from one

molecule of glucose
 NADH cannot be oxidized, so no ATP is produced in
electron transport chain
 But the NADH is oxidized to NAD+ by conversion of
pyruvate to Lactate, without producing ATP.
 limits the amount of energy per molecule of glucose oxidized
 Hence, to provide a given amount of energy, more glucose
must undergo glycolysis under anaerobic as compared to
aerobic
Glycolysis does occur in cells with abundant supply
of oxygen provided it contains mitochondrial.

C6H12O6 + 6O2 +32 ADP3- +32Pi2- >>>>>>>6CO2 +

6H2O +32ATP4- + 32OH-

Complete oxidation of glucose gives 32ATP/Glucose

Glucose >>>>>Lactate gives 2 ATP/Glucose

Pyruvate produced in glycolysis is oxidised to CO2

in mitochondria.
 REACTIONS OF GLYCOLYTIC
PATHWAY
 Stage I (preparatory stage)
 Uptake of Glucose by Cells
 Glucose phosphorylation- Glucokinase/Hexokinase
 Product → glycolysis, glycogenesis, glycogenolysis
gluconeogenesis, Hexosemonophosphate Shunt, uronic acid
partway
 Thus is a “committed step” in metabolic pathways
 G6P→Fructose-6-phosphate (Hexose isomerase)
 F6P → Fructose 1, 6 bisphosphate (phospho- fructokinase-1)
 Expenditure of 2 ATP molecules for two phosphorylations (-2
ATP)
 Stage II
 F1,6B → two molecules of triosephosphates (aldolase)
 The reaction is reversible
 Neither expenditure of energy nor formation ATP
  Stage III
 Energy- yielding reaction
 Aldehyde group is oxidized to an acid
 G3P → 1,3BPG
 DAP →1,3BPG (G3P Shuttle)
 The enzyme → G3p dehydrogenase-NAD+ dependant
 2 NADH → 6 ATP
 2nd reaction → 2ATP
 Net gain at this stage per molecule of glucose oxidized= +
8ATP
 Stage IV
 Recovery of the PO4 group
 3pg → 2pg (phosphoglycerate mutase)
 2pg → PEP ( Enolase )
 PEP → pyruvate ( pyruvate kinase )
 In absence of O2 re-oxidation of NADH at G3P-dehydrogenase
stage cannot take place in electron-transport chain
 the cells have limited coenzyme. Hence to continue the glycolytic
pathway NADH must be oxidized to NAD+
 Thisis achieved by re oxidation of NADH by conversion of
pyruvate to lactate (without producing ATP) by the enzyme lactate
dehydrogenase
 Occurs in cells with no mitochondria as RBCs (mature) ,or under
low O2 supply as intensive muscular exercise . In anaerobic
glycolysis per molecule of glucose oxidation 4 -2 = 2 ATP will be
produced.
 Regulation
1. Enzymatic regulation- 3 types of mechanism
 Changed in rate of enzyme synthesis:
 Induction →↑ rate of enzyme synthesis at gene expression →↑
mRNA synthesis
 Repression →↓ rate of enzyme synthesis at gene expression →↓
mRNA synthesis.
 Covalent modification by reversible phosphorylation
dephosphorylation
 Allosteric effect
 Four (4) regulatory enzymes → irreversible reactions:
 Hexokinase
 Glucokinase
 Phosphofructokinase
 Pyruvate kinase
2. Hormonal regulation
 Insulin
 Glucagons
INHIBITORS OF GLYCOLYSIS
 Arsenate
 Iodoacetate →G3Pdehydrogenase (inhibitor of
SH group)
 Fluoride→ inhibits enolase→↓↓ glycolysis in
bacteria →no production of lactic acid
produced by bacteria, which cause dental
caries. It used as anticoagulant in blood
sample used for estimation of blood
glucose→↓↓ glycolysis in RBCs.
 Clinical importance
 Formation of 2,3PG

 Diseases associated with impaired Glycolysis

 Hexokinase deficiency
 Pyruvate kinase deficiency(haemolytic anamia)
 Lactic acidosis
 When oxygen supply to a tissue is shut off, ATP levels can still
be maintained by glycolysis for at least a short period of time
 The capacity to use glycolysis as a source of energy is
particularly important during natural birth of human babies
 glycolysis with lactate as the end product is the major
mechanism of ATP production in some tissues
Oxidative Decarboxylation of Pyruvic
Acid
 Conversion of pyruvic acid into a coA derivatives
 Site: Mitochondria matrix of all tissues
 catalyzed by pyruvate dehydrogenase complex, which
composed of 3 enzymes act cooperative with each other
in presence of 5 co-enzymes: TPP, lipoic acid, FAD,
NAD+, and CoASH.
 Pyruvate
+TTP + Lipoic acid + CoA +FAD+ NAD+--→
CO2+ Acetyl-CoA + NADH + H+
 Regulation

 Product inhibition
 acetyl CoA
 elevated levels of NADH+.H
 Covalent modification
 an active non phosphorylated form and an inactive
phosphorylated form. can be interconverted by two separate
enzymes, a kinase and a phosphatase.
 Clinical important

 Inhibition → Lactic acidosis, which may be due to

a. Arsenite or mercuric ions complex the –SH
group of lipoic acid
b. Dietary deficiency of thiamine as in alcoholics
 These two factors lead to inhibition of pyruvate dehydrogenase
c. Inherited pyruvate dehydrogenase deficiency, which may be
due to defects in one or more of the components of the enzyme
complex
KREB’S CYCLE

 Aka TCA or CA Cycle

 It is the series of reactions in mitochondria, which
oxidized acetyl CoA to CO2, H2O & reduced H2 carriers
that oxidized through respiratory chains for ATP synthesis
 Site:
Mitochondria of all tissue cells
 The enzymes of the cycle are present in
mitochondrial matrix except succinate
dehydrogenase, which is tightly bound to inner
mitochondrial membrane
 Regulation of Kreb’s Cycle
 Respiratory control via the E.T.C and oxidative
phosphorylation exerts the main control
 several enzymes of TCA cycle are also important in the
regulation. Three key enzymes are:
 Citrate synthase ( allosterically inhibited by ATP and long-chain AcylCoA)
 Mitochondrial isocitrate dehydrogenase ( activated allosterically by
ADP and is inhibited by ATP and NADH )

 α-keto glutarate dehydrogenase ( allosterically inhibited by

succinylCoA, NADH-H+ and ATP)
 succinate dehydrogenase enzyme is also inhibited
by oxaloacetate (OAA) and the avability of OAA is
controlled by malate dehydrogenase, which
depends on [NADH]/[NAD+] ratio.
Functions
 Final pathway for complete oxidation of all
foodstuffs –CHO, Lipid and protein, which are
converted to acetyl CoA.
 Major source of energy for cells with Mit.
 Major source of succinylCoA, which used for :
 Perphyrine and HB synthesis.
 Ketone bodies activation.
 Converted to OAA → glucose.
 Detoxication by conjugation
Synthetic functions

 Amphibolic reactions: Some components of Kreb’s

cycle are used in synthesis of other substances as:
 In fasting state, oxaloacetic acid → glucose
 In fed state, citric acid → fatty acids
 Reactions of Kreb’s cycle → amino acid
(transamination into non essential amino acids)
 Anaplerotic reactions: Synthesis of one or more
component of Kreb’s cycle from outside the cycle
 O.A.A. ←← pyruvic acid ,and aspartic
 Fumarate ←← phenylalanine and tyrosine
 SuccinylCoA ←← valine, isoleucine, methionine,
and threonine
 α-keto glutarate ←← glutamic acid
Inhibitors of Citric Acid Cycle

 Flouro-acetate reacts with oxaloacetate forming

flourocitrate, which inhibits the aconitase enzyme
 Arsenite inhibits α-ketoglutarate dehydrogenase
 Malonate acts as competitive inhibitor for
succinate dehydrogenase.
ROLES OF VITAMINS IN CITRIC ACID
CYCLE
Four of the soluble vitamins of B complex have
important roles in citric acid cycle
 Riboflavin ( FAD)
 Niacin (NAD)
 Thiamin
 Pantothenic acid
Thank you for
listening…….Merry
xmas and Happy
New year in Advance
.