Carbohydrate Metabolism

Dr. Milind Dudhane

Glycolysis
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After being absorbed from the intestinal tract the monosaccharides are carried by the portal circulation directly to the liver.  In liver most of the D-Glucose is phosphorylated to Glucose -6- phosphate which can not diffuse back out of the cell as plasma membrane is impermeable to the glucose -6phosphate.  Remaning glucose passes into systemic blood supply. Other dietary monosaccharides D-fructose & Dgalactose are phosphorylated & may be converted to glucose in the liver.
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Glucose -6- phosphate is an intermediate in several metabolic pathways that uses glucose in liver depe Glycogen nding upon supply & demand.
Blood Sugar

Glycogenesis

Glucose-6-phosphate Pentose phosphatePathway Glycolysis

Glycogen

Pyruvate Acetyl CoA CO2 + H2O

Pentose phosphate, NADPH

Cholesterol

Fatty acid

TG
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Glycolysis

(Embden Meyerhof pathway)

Definition : Glycolysis is the sequence that converts glucose into pyruvate in presence of oxygen(aerobic) or lactate in absence of oxygen (anarerobic) with the production of ATP Location : Cytosol of cell Reactions of Glycolysis: The breakdown of glucose to two moles of pyruvate is brought about by sequential action of 10 enzymes which can be devided into two phases.
First phase or energy requiring phase or preparative phase First phase or energy requiring phase
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Phases of Glycolytic pathway

Glucose
ATP

First phase:Energy requiring phase

ATP

Fructose1,6 Bisphosphate

2- triose phosphate
2 NADH 6 ATP

Second phase:Energy releasing phase

2 ATP 2 ATP

2 Pyruvate
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First phase or energy requiring phase or preparative phase

1. Glucose is phosphorylated to Glu-6-phosphate by enzyme Hexokinase &ATP is required as phosphate donar.[ Hexokinase occur in different isoenzyme forms type I to IV] Brain And Kidney Skeletal Muscles Adipose tissues Liver Type I Type II Type I & II All types from I to IV
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2. Conversion of glucose-6-phosphate to fructose-6-phosphate by enzyme phosphohexose isomerase is freely reversible reaction. 3. fructose-6-phosphate is phophorylated to fructose-1-6-bisphosphate by phosphofructokinase-I. phosphofructokinase-I is both allosteric and inducible enzyme which is rate limiting & regulatory enzyme of glycolysis.

4. Fructose-1-6-bisphosphate is cleaved by enzyme aldolase to two triose phosphates, glyceraldehyde-3-phosphate & dihydroxy acetone phosphate(DHAP). Several tissue specific isoenzymes of aldolase exists.Aldolase A occurs in most tissues & aldolase B occurs in liver & kidney. 5. DHAP is isomerized to glyceraldehyde-3phosphate by the enzyme phosphotriose isomerase,so that for every molecule of Glucose entering glycolysis,2 moles of glyceraldehyde-3phosphate are formed.
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Second phase or energy releasing phase reactions 1. Oxidation of glyceraldehyde-3-phosphate to 1,3 bisphosphoglycerate by glyceraldehyde-3phosphate dH, an NAD dependent, reversible reaction. NADH++ H+ are reoxidized by ETC to generate 3 ATP molecules. 2. 1,3-bisphosphoglycerate to 3phosphoglycerate catalyzed by phosphoglycerate kinase. This is the first step in glycolysis that generates ATP, by substrate level phosphorylation(SLP).
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Since two molecules of triose phosphate are formed per molecule of glucose 2 ATPs are generated at this stage. Arsenate can uncouple oxidation & phosphorylation at this step. 3. Phosphoglycerate mutase catalyze the reversible reaction 3-phosphoglycerate to 2phosphoglycerate. 4. Enolase converts 2-phosphoglycerate to phosphoenol pyruvate. Enolase in inhibited by fluoride.

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5. Conversion of phosphoenol pyruvate to pyruvate is brought about by Pyruvate kinase(an allosteric enzyme)ATP is generated by substrate level phosphorylation. (2 ATPs per molecule of glucose). Under aerobic condition pyruvate is taken up into mitochondria and after conversion to acetyl Co A is oxidized to CO2 by TCA.

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Hexokinase Glucose Glucose-6- P Phospho hexose isomerase ATP Mg2+ ADP Fructose-6- P
ATP Mg2+ ADP

Phospho fructokinase

Fructose-1-6-bisphosphate Aldolase Dihydroxy acetone P Glyceraldehyde-3- P Phospho triose isomerase Glyceraldehyde 3NAD phosphate dH NADH 1,3-bisphosphoglycerate Phospho glycerate kinase 3-Phosphoglycerate
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3-Phosphoglycerate Phospho glycerate Mutase 2 -Phosphoglycerate Enolase H2O Phosphoenolpyruvate

ADP Mg2+ ATP

Pyruvate kinase PDH Acetyl CoA

Pyruvate

Anaerobic Oxidation

Lactate
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Conversion of Pyruvate to Acetyl CoA

Conversion of Pyruvate to Acetyl CoA occurs in mitochondria & hence pyruvate must be transpoted into mitochondria by special pyruvate transport. Within the mitochondria pyruvate is oxidatively decarboxylated to acetyl Coa by multienzyme complex,consisting . Enzymes : Pyruvate decarboxylate Dihydrolipoate transacetylase & Dihydrolipoate dH Coenzymes: TPP, Lipoate, CoASH,FAD & NAD
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Oxidative decarboxylation of Pyruvate to by Pyruvate dH complex

O II CH3 C COO Pyruvate

NAD+ PDH

NADH + H+ CO2

TPP, Lipoate, CoA-SH, FAD

O II CH3 C SCoA Acetyl CoA

Significance : Conversion of Pyruvate to Acetyl CoA is a central step linking glycolitic pathway with TCA. Acetyl CoA is also an important intermediate of lipid metabolism, Cholesterol biosynthesis and acetylation reactions.
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Oxidative decarboxylation of Pyruvate to by Pyruvate dH complex CO2

-OH ethylthiamine pyrpphosphate Pyruvate Pyruvate Pyruvate Decarboxylase decarboxylase Oxidized lipoate NAD FADH Dihydrolipoate dH FAD NADH + H+
2

Thiamine pyrpphosphate

Dihydrolipoate Transacetylas e S-Acetyl lipoate Reduce d lipoate CoASH CH3

C SCoA II O Acetyl CoA

3 ATP ETC
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Metabolic fates of Acetyl CoA Xenobiotic metabolism Acetyl Choline

N-Acetylglutamate in urea biosynthesis Acetylation of amino sugar Glycoprotein synthesis

Ketone bodies

Acetyl CoA
Catabolism TCA cycle

N-Acetylneuraminic acid ganglioside synthesis

Cholesterol Synthesis

ATP CO2 + H2O

Fatty acid Synthesis

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Anaerobic Glycolysis :
If anaerobic condition prevail the reoxidation of NADH(formed in glyceraldehyde-3-phosphate step) by transfer of reducing equivalents through the respiratory chain to oxygen is prevented. But are reoxidized by conversion of pyruvate to lactate By LDH. Tissues that function under anaerobic condition produce lactate,e.g. skeletal muscle, smoth muscle & erythrocytes. In erythrocytes even under aerobic conditions, glycolysis terminates in lactate because of absence of Mitochondria.
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The liver, kidney & heart usually take up lactate & oxidize it under hypoxic condition. This allows other active muscle cells to utilize glucose. During vigorous exercise the rate of formation of pyruvate exceeds its utilization. Moreover rate of formation of NADH is greater than its utilization. This accumulated NADH is to oxidized to NAD in order to supply enough NAD for glycolysis in skeletal muscle & erythrocytes.

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Significance of Anaerobic Glycolysis : The reoxidation of NADH via lactate formation allows glycolysis to proceed in the absence of oxygen by regenerating sufficient NAD for another cycle of reaction catalyzed by glyceraldehyde-3-phosphate dH Anaerobic glycolysis is an emergency source of ATP.

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Energetics of Glycolysis: The overall reaction of aerobic glycolysis alone using either free glucose, fructose or galactose yielding pyruvate generates: 2 molecules of NADH 4 molecules of ATP at SLP But 2 molecules of ATP per mole of hexose are consumed. The NET gain of 2 moles of each NADH & ATP . The NADH thus formed must be transported to mitochondria so that it can be utilized there.

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Mitochondrial membrane is impermeable to both NADH & NAD & hence the shuttle systems namely. Glycerol phosphate & Malate aspartate shuttle which yield 2 &3 moles of ATP respectively per mole of NADH. Further oxidation of Pyruvate to CO2 & H2O yields 38 ATPs in citric acid cycle.

In anaerobic glycolysis on the other hand only 2 moles of ATP & NO NADH are produced.
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Consumption of ATP in Glycolysis

Reaction Reaction Catalyzed by
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Gluc to Gluc-6Hexokinase, phosphate Glucokinase Fruct-6-phosphate Phosphofructokinase to Fruct-1,6bisphosphate

No. of ATP consumed/m ole Glucose -1 -1 Total = -2

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Production of ATP in Glycolysis

Reaction No.of ATP of ATP formed/Gl Production u molecule Glyc-3-P to 1,3 Glycaldehyde- ETC bisphosphoglycerat 3-phsphate +6 e dH
Mechanism

Reaction catalyzed by

1,3 bisphosphoglycerate to 3-hosphoglycerate Phosphoenol pyruvate to pyruvate

Phosphoglycer SLP ate kinase Pyruvate kinase SLP Total

So the NET production is 10 2 = 8 ATP

+2 +2
production 10

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Rapoport Lubering Cycle OR Bisphosphoglycerate Shunt : Mature erythrocytes contain no mitochondria. So they are totally dependent upon glycolysis for ATP production. Erythrocytes metabolizes excessive amount of glucose in the glycolytic pathway to maintain the structural integrity of the erythrocyte membrane,resulting in generation of more ATPs.Those are in excess & can not be used by erythrocytes.This may inhibit glycolysis by inhibiting phosphofructokinase-I

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In Rapoport Lubering cycle ATP production by SLP from 1,3 BPG is bypassed in RBCs. 1,3 BPG is converted to 2,3 BPG by an enzyme bisphosphoglycerate mutase. 2,3 BPG is converted to 3-phosphoglycerate by 2,3 bisphosphoglycerate phosphatase with loss of energy in the form of heat. Due to lackof mitochondria in RBCs glycolysis occur anaerobically producing lactate.

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Rapoport Lubering cycle

Glucose

Glyceraldehyde-3-phosphate Glyceraldehyde-3-phosphate dH
NAD Pi NADH+ H+ ADP ATP

1,3-bisphosphoglycerate

Bisphosphoglycerate mutase 2,3-bisphosphoglycerate 2,3Bisphosphoglycerate phosphatase

3-phosphoglycerate Pi Pyruvate

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Significance of Rapoport Lubering cycle : It prevents accumulation of ATP not needed by the erythrocyte It supplies 2,3-BPG in oxygen transport which is required for Hb function.2,3-BPG regulatesthe binding & release of oxygen from Hb. 2,3 BPG accounts for 16% of noncarbonate buffer value.

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Krebs Cycle, TCA Cycle

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TCA CYCLE
(CITRIC ACID OR KREB CYCLE OR COMMON

METABOLIC PATHWAY)

The TCA cycle is a series of cyclic reaction that catalyse the oxidation of acetyl Co A to CO2 and water liberating reducing equivalents.

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SITE : The TCA cycle occurs inside the mitochondria as the enzymes required for the cycle are located in the mitochondria matrix, either free or attached to the inner surface of inner mitochondria membrane. The enzyme of TCA cycle are located in mitochondria matrix, in close proximately to the electron transport chain.

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TCA CYCLE - THE CENTRAL METABOLIC PATHWAY The citric acid cycle is the final common oxidative pathway for carbohydrates, fats and amino acid. The cycle not only supplies energy but also provides many intermediates required for the synthesis of amino acids glucose etc. Krebs cycle is the most important central pathway.

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TCA Cycle is an open cycle Krebs cycle is the a cyclic process. It should not be viewed as closed circle many compounds enter the cycle and leaves. TCA cycle is comparable to a heavy traffic circle in a national highway with many connecting roads. Reactions of TCA cycle Oxidative decarboxylation of pyruvate to acetyl COA by pyruvate dehydrogenase complex is discussed above. The TCA cycle connecting glycolysis.

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Steps The various steps of the cycle are as follows Step-1 citrate synthase Acetyl CoA + Oxaloacetate Citrate
H2O CoASH

Formation of citrate Acetyl CoA condense with oxaloacetate to from citrate. It is catalyzed by condensing enzyme citrate synthase. the condensation product citryl CoA is hydrolysed to yield citrate and CoA.

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Step -2
Citrate is Isomerized to Isocitrate Citrate is converted to isocitrate by the enzyme aconitase. the enzyme contain iron in in the from of iron sulfur protein. The conversion takes place in 2 stage First, there occur dehydration of citrate to cis aconitate. Next, Cis aconitate undergoes rehydration to form isocitrate.
H2O H2O

Citrate

Cis-aconitase

Isocitrate

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STEP-3 Formation of - ketoglutarate Isocitrate is converted to - ketoglutarate in 2 stage by the enzyme isocitrate dehydrogenase. First, There occurs dehydrogenation of isocitrate to from oxalosuccinate. NAD acts as on hydrogen acceptor. Next , Oxalosuccinate undergoes decarboxylation to form -ketoglutarate. Isocotrate + NAD Oxalosuccinate + NADH + H+ CO2 Oxalosuccinate - ketogutarate

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Step -4 Conversion of - ketoglutarate to succinyl CoA -Ketoglutarate undergoes oxidative decarboxylation to produce succinyl CoA. The enzyme is - ketoglutarate dehydrogenase complex and the reaction is irreversible. the enzyme requires NAD ,FAD ,thiamine pyrophosphate, lipoate and co-enzyme A as cofactors similar to PDH complex

- ketoglutarate dH complex

- ketoglutarate + CoA + NAD

Succinyl CoA+ NADH + H+ CO2

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Step 5 Formation of succinate Succinyl Co A is converted to succinate by the enzyme succinate thiokinase requires & GDP converted GTP.
Succinyl CoA + GDP Succinate + GTP + CoA

The enzyme nucleoside diphosphate kinase converts GTP in to ATP.

Succinyl thiokinase GTP+ ADP ATP + GDP

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Step -6 Conversion of succinate to formula : Succinate undergoes dehydrogenation to from fumarate The enzyme is concerned is succinate dehydrogenase which is found to inner surface of inner mitochondrial membrane unlike other enzymes of the cycle. Succinate + FAD Fumarate + FADH2 The step is inhibited competitively by malonate or oxaloacetate.

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Step -7 Formation of Malate A molecule of water is added to fumarate to form malate. The enzyme required is fumarase which is specific informing :- Isomer of malate. Fumarase Fumarate + H2O L-Malate

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Step - 8 Conversion of malate to oxaloacetate Malate undergoes dehydrogenation to form oxaloacetate the enzyme is malate dehydrogenase NAD as the hydrogen acceptor. Malate dH L-Malate + NAD Oxaloacetate + NADH

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ENERGETICS OF GLUCOSE OXIDATION When a molecule of glucose undergoes, glycolysis 2 molecules of pyruvate or located are produced pyruvate is oxidatively decarboxylated to acetyl Co A. Which enters the citric acid cycle and gets completely oxidized to and glycolysis and citric acid cycle.
C6H12O6 + 6O2 +38 ATP +38 Pi 6 CO2 +6H2O +38 ATP

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The enzyme of glucose metabolism responsible for generating ATP. When a molecule of glucose is burnt in a calorie meter. In the living system energy is trapped leading to the synthesis of 38 ATP which is equivalent to 1159 KJ. The 48% of the energy in glucose combustion is actually captured for ATP generation.

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Regulation of TCA The cellular demands of ATP are crucial in controlling the rate of citric acid cycle. The regulation is brought about either by enzymes or the levels of ADP. Their enzymes Citrate synthase. In inhibited by ATP , NADH, acyl CoA and succinyl CoA Isocitrate dehydrogenase Is activated by ADP and inhibited by ATP and NADH. and - ketoglutarate dehydrogenase. Is inhibited by succinyl CoA and NADH.

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Amphibolic Nature of the TCA The citric acid cycle is the final common oxidative pathway in the living cell. The cycle provides various intermediates for the synthesis of many compounds needed by the body. Krebs cycle is the both catabolic anabolic nature hence regarded as amphibolic.

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The important synthetic reaction connected with TCA cycle are given : 1. Oxaloacetate and - Ketoglutarate, respectively serves as precursors for the synthesis of aspartate, glutamate which is turn are required for the synthesis of other non essential amino acid, Purines & Pyrimidines. 2. Succinyl CoA is used for the synthesis of porphyrins and heme. 3. Mitochondrial citrate is transported to the cytosol were it is cleaved to provide acetyl CoA for the biosynthesis of fatty acids.

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Anaplersis or Anaplerotic Reaction The synthesis reaction described above deplete the intermediates of citric acid cycle. The reaction concerned to replenish or fill up the intermediates of citric acid cycle are called anaplerotic reaction or anaplerosis. The important synthetic pathways that draw the intermediates of TCA cycle and the anaplerotic reaction are described. 1. Pyruvate carboxylase catalyses the conversion of pyruvate to oxaloacetate. This is an ATP dependent carboxylation reaction.

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1. Pyruvate carboxylase catalyses the conversion of pyruvate to oxaloacetate. This is an ATP dependent carboxylation reaction. 2. Pyruvate is converted to malate by NADP+ dependent malate dehydrogenase. 3. Transamination is process where in an amino acid transfers its amino groups to a keto acid and itself gets converted to a keto acid. 4. - Ketoglutarate can also be synthesized form glutamate by glutamate dehydrogenase action. The formation of - ketoglutarate and oxaloacetate occurs by this mechanism.

Pentose phosphate pathway HMP Shunt Pathway

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This is aerobic pathway for the oxidation of glucose in the liver , adipose tissue in addition to the Embdonmeyorhop pathway for glycolysis.It is anabolic in nature , since it is concerned with the biosynthesis of NAPDH and pentose's . HMP shunt -a unique multifunctional pathway: The enzyme of this pathway are present in extra mitochondrial portion of the cell . This pathway is active in adipose tissue , adrenal cortex, liver , thyroid , erythrocytes.

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Glu-6-P
NADP++ H2O

Glu-6-P
NADP++ H2O

Glu-6-P
NADP++ H2O

Glu-6-P dH

Mg2+ or Ca2+ NADP++ H+ NADP++ H+

P
NADP++ H+

H
6-Phosphogluconolactone 6-Phospho gluconola Mg2+ or Ca2+ ctone hydratase 6-Phosphogluconate 6-Phospho gluconate dH
NADP+ H2O

6-Phosphogluconolactone 6-Phosphogluconolactone

A S
6-Phosphogluconate
NADP+

6-Phosphogluconate
NADP+

NADP + H+ CO2

NADP + H+ CO2

NADP + H+ CO2

Ribulose-5-P

Ribulose-5-P

Ribulose-5-P
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Ribulose-5-P Ribulose-5-P Ribulose-5-P Ketoisomerase 3-epimerase Xylulose-5-P Ribose-5-P Transketolase TPP

Ribulose-5-P Ribulose-5-P 3-epimerase Xylulose-5-P

P H A

Glyceraldehyde-3-P Transaldolase

Sedoheptulose-7-P

S E
Fructose-6-P Erythrose-4-P

II

Fructose-6-P Glucose-6-P Glucose-6-P

Glyceraldehyde-3-P 1/2 Glucose-6-P

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 The pathway starts with glucose 6-phosphate .As such no ATP is directly utilize or produced in HMP pathway. It is unique and multifunctional , since their are several interconvertible substance produced which proceed in different direction in the metabolic reaction.
REACTIONS :

Reactions of HMP shunt is divided in to 2 phases: Oxidative phase Non-oxidative phase

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1. Oxidative phase : Glucose 6-phosphate dehydrogenase (G6PD)is an NADP dependent enzyme that converts glucose 6-phosphate to 6-phospogluconate.the latter is then hydrolysed by the gluconoactone hydrolase to 2. 6-phospogluconate.The next reaction involving the synthesis of NADPH is catalysed by 6phosphogluconate which then undergoes decaboxylation to give ribulose 5-phosphate.

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2.Non-oxidative phase : The non-oxidative reaction are concerned with the interconvertion of 3,4,5,&7carbon monosaccharide . Ribulose 5-phosphate is acted upon by an epimer to produce xylulose 5-phospate while ribose 5-phosphate ketoisomerase converts ribulose5-phosphate to ribose 5-phosphate.

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 Significance of HMP shunt HMP shunt is unique in generating two important products Pentose & NADPH needed for the biosynthetic reactions & other function: A)Importance of Pentose In the HMP shunt hexos are converted into pentose, the most important being ribose 5phosphate.The pentose or its derivative are useful for the synthesis of nucleic acid & many nucleotides such as ATP ,NAD+, FAD,& CoA.
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B) Importance of NADPH 1) NADPH is required for the reductive biosynthesis of fatty acid & steroids, hence HMP shunt is more active in the tissue concerned with lipogenesis . Example adipose tissue, liver. 2)NADPH is used in the synthesis of certain ammino acid involving the enzyme glutamate dehydrogenase.

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3)Microsomal cytochrome P450 system(in liver)brings about the detoxification of drugs & foreign compounds by hydroxylation reactions involving NADPH. 4)Phagocytosis is the engulfment of foreign particles including microorganism , carried out by W.B.C.The process requires the supply of NADPH.

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5)Special function of NADPH in RBC: NADPH produced in erythrocytes has special function to perform. It maintain the concentration of reduced glutathione, which is essentially required to preserve the integrity of RBC membrane. 6)NADPH is also necessary to keep the ferrous iron (Fe2+) of hemoglobin in the reduced state so that accumulation of methemoglobin (Fe3+) is prevented .

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Glucose 6- Phosphate dehydrogenase deficiency :

G6PD deficiency is more severe in RBCs. HMP shunt is the only means of providing NADPH in erythrocytes . Decreased activity of G6PD impairs the synthesis of NADPH in RBCs . This results in the accumulation of methamoglobin & peroxides in erythrocytes leading to hemolysis .

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The G6PD deficiency develop Hemolytic Anemia . If they are admired with drugs such as primaquine (antimalaria), sulfamethaxoazole (antibiotic), produce hemolytic jaundice in patients.

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Wernicke Korsakoff Syndrome :

This is a genetic disorder associated with HMP shunt An alteration in Transketolase activity reduce affinity with thiamine pyrophosphate is the biochemical lesion. The symptoms include the mental disorder, loss of memory & partial paralysis. The symptoms are manifested whose diet are vitamin deficient .

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URONIC Acid Pathway

This is an alternative oxidative pathway for glucose and is known as glucuronic acid pathway. 1.uronic acid pathway is concerned with the production of glucuronic acid (involved in detoxification)/pentose & vitamin C. 2.In uronic acid pathway the free sugar or sugar acids are involved

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Formation & Importance of UDP- Glucuronate

Glucose 6-phosphate is first converted to glucose1-phosphate UDP-glucose is then synthesized by the enzyme UDP-Glucose Pyrophosporylase. UDP- glucuronate is the metabolically active form of glucuronate which is utilized for conjugation with many substance like bilirubin , steroid hormones & certain drugs

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Several insoluble compounds are converted to soluble ones through conjugation & further the drugs are detoxified. UDP- glucuronate is also require for the synthesis of glycosaminoglycans & proteoglycan. Conversion of UDP- Glucuronate to L- Gluonate UDP- glucuronate loses its moiety in a hydrolytic reaction & releases D glucuronate which is reduced to L gluonate by an NADPH dependent reaction.

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 Effect of drugs on Uronic acid pathway Administration of drugs (barbital , chlorobutanol) significantly increase the uronic acid pathway to achive more synthesis of glucouronate from glucose.Certain drugs (aminopyrine , antipyrine)were found to enhance the synthesis of ascorbic acid in rats.

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Gluconeogenesis
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Definition :The formation of Glucose or Glycogen from non-carbohydrate precursors is called gluconeogenesis. The major non-carbohydrate substrates for gluconeogenesis are Lactate, Glycerol, Glucogenic amino acids Propionate & Intermediates of citric acid cycle

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Significance : Blood sugar level is maintained via gluconeogenesis. D-glucose is absolutely necessary for the tissues such as brain, erythrocytes, kidney & eyes. Glu-6-phosphatase is absent in muscle. Therefore, during exercise and starvation, the large amount of lactate produced by glycolysis & glycerol generated by lipolysis of TGs are used up by gluconeogenesis.
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Location : In animals, gluconeogenesis takes place mainly in the liver. (85 95 %). In the cortex of kidneys during periods of fasting, starvation, or intense exercise about 50 % The pathway can begin in the mitochondria or cytoplasm, depending on the substrate being used.

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Characteristics of Gluconeogenesis :
Gluconeogenesis involves Glycolysis, Citric acid cycle plus some special reactions. Glycolysis & Gluconeogenesis share the same pathway but in opposite direction. Seven reactions of glycolysis are common. However three reactions are irreversible involving following enzymes. Pyruvate carboxylase (Mitochondrial) Phosphoenol pyruvate carboxykinase Fructose-1,6 bisphosphatase and Glucose -6-phosphatase
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Pi Glucose-6-phosphatase H2O

Glucose

Glucose-6-phosphate

ATP Glucokinase Hexokinase ADP

Pi Fructose-6-phosphate Fructose-1,6-bisphosphatase H2O Fructose-1,6-bisphosphate

ATP Phosphofructokinase ADP DHAP

Glyceraldehyde-3-phosphate Contd
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Glyceraldehyde-3-phosphate
NAD NADH +H+

DHAP
NADH +H+ Glycerol-3-phosphate dH NAD

1,3-Bisphosphoglycerate

Glycerol-3-phosphate
ADP

Glycerol kinase
ATP

3-phosphoglycerate

Glycerol

2-phosphoglycerate

Phosphoenol Pyruvate
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Phosphoenolpyruvate CO2 ADP CYTOSOL GDP Phosphoenolpyruvate Pyruvate kinase GTP carboxykinase ATP Pyruvate Lactate Oxaloacetate NADH+H+ NAD+
Pyruvate 1
NADH+H+ ATP,CO2

..

Pyruvate Carboxylase
Oxaloacetate 2 -KetoGlutarate 3

MDH
NAD NADH+H+ NAD

ADP+Pi

Malate MITO.

Malate

5 Fumarate

Succinyl CoA 4 Propionate

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The Nos. allotted in above Pathway are for Gluconeogenic amino acids are ( Gluconeogenesis from amino acids) 1.Pyruvate : Alanine, Glycine, Serine Cysteine,
Threonine & Tryptophan

2.Oxaloacetate : Aspartate & Arginine 3. Ketoglutarate : Arginine, Glutamate, Glutamine,

Histidine, Prolein

4.Succinyl CoA: 5.Fumarate :

Isoleucine, Methionine, Valine Phenylalanine, Tyrosine

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Reactions of gluconeogenesis : Only those reactions which are not common to glycolysis are : Reaction 1 : Carboxylation of pyruvate Pyruvate is converted to oxaloacetate with the help of CO2, ATP & pyruvate carboxylase.Biotin, Mg++, Mn++ are required. There is an absolute requirement of acetyl CoA. This reaction occurs in mitochondrial matrix. Oxaloacetate produced in mitochondria cannot cross the membrane. It is first reduced to malate, which then crosses mito. Membrane where it is re-oxidized to Oxaloacetate .
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Reaction 2 : Conversion of Oxaloacetate to phosphoenol pyruvate : The enzyme required is phosphoenolpyruvate carboxykinase, it requires Mn++ & GTP. CO2 & GDP are formed. The reaction occurs in cytosol. Reversal of reactions of glycolysis now occurs until fructose 1-6 bisphosphate. Reaction 3 : Dephosphorylation of fructose 1-6 bisphophate : Fructose 1-6 bisphosphatase, the major regulatory enzyme then forms fructose-6phosphate. The enzyme is activated by citrate & inhibited by AMP and fructose2-6 bisphosphate
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Reaction 4 : Dephosphorylation of glucose 6 phosphate: This reaction gives free Glucose & inorganic phosphate. It is catalyzed by glucose 6 phosphatase which is present only in liver, kidney & epithelial cells of small intestine. The overall summary of Gluconeogenesis for conversion of pyruvate to glucose. 2 Pyruvate + 4 ATP +2 GTP + 2 NADH+ 2H+ + 6H2O -Glucose + 2 NAD + 4 ADP + 2 GDP + 6 Pi + 6 H+
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Gluconeogenesis from Glycerol : Glycerol liberated by fat hydrolysis in adipose tissues. In liver & kidneys glycerol is activated to Glycerol-3- P by the enzyme Glycerokinase (Which present in liver & kidneys and not in adipose tissues).Glycerol-3phosphate is then converted to DHAP which is the intermediate of glycolysis by the enzyme Gly.-3-P dH.

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Gluconeogenesis from Propionate : Oxidation of odd chain fatty acid & breakdown of methionine & isoleucine yields propionyl CoA. This is carboxylated to form methyl malonyl CoA which requires Biotin & ATP by propionyl CoA carboxylase. It further forms Succinyl CoA, the intermediate of TCA

PropyonylCoA Methyl malonyl CoA Succinyl CoA

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Gluconeogenesis from Lactate : Lactate produced during anaerobic oxidation of glucose in muscles. Due to absence of enzymes Glucose-6-phosphatase & Fructose 1,6 bisphosphatase, Lactate can not be used in muscle &to be transported to liver for Gluconeogenesis. LDH Pyruvate + NADH + H+ Lactate + NAD

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Regulation of Gluconeogenesis : The hormone Glucagon & the availability of substrate mainly regulates gluconeogenesis. Glucagon stimulates gluconeogenesis by two mechanisms. 1. Active form of pyruvate kinase is converted to its inactive form. Decreased pyruvate kinase results in reduced conversion of phosphoenol pyruvate to pyruvate & former is diverted for synthesis of glucose. 2. Glucagon reduces the concentration of fructose 2,6 bisphosphate. This compound allosterically inhibit phosphofructokinase and activates fructose 1,6 bisphosphatase. 94

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Availability of substrate : Glucogenic amino acids have stimulating effect on gluconeogenesis. In conditions like diabetes mellitus amino acids are mobilized from muscle protein for the purpose of gluconeogenesis. During starvation acetyl CoA accumulates in liver due to lypolysis. Acetyl CoA allosterically activate pyruvate carboxylase resulting in enhanced glucose production.

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Glycogen Metabolism
Glycogenesis & Glycogenolyss

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Glycogenesis
Glycogen is storage form of glucose in animals. It is stored mostly in liver (6-8 %) & muscle(1-2 %). Due to more muscle mass, the quantity of glycogen in muscle(250 g) is about three times higher than that in the liver (75 g).

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Glycogenesis: Glycogenesis is the formation of Glycogen from Glucose. Glycogen is synthesized depending on the demand for glucose and ATP (energy) If both are present in relatively high amounts, then the excess of insulin promotes the glucose conversion into glycogen for storage in liver and muscle cells (In fed state).

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 In the synthesis of glycogen, one ATP is required per glucose incorporated into the polymeric branched structure of glycogen. Glucose-6-phosphate is synthesized directly from glucose or as the end product of gluconeogenesis. The synthesis of Glycogen from glucose takes place in cytosol and requires ATP, UTP & glucose.

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Reactions of Glycogenesis: Glucose is phosphorylated to Glu-6-phosphate catalyzed by hexokinase in muscle & glucokinase in liver. Glu-6-phosphate is converted to Glu-1phosphate by phosphoglucomutase. Glu-1-phosphate reacts with uridine tri phosphate (UTP) to form uridine di phosphate glucose(UDPGlc) by the enzyme UDP-glucose phosphorylase.

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By the action of glycogen synthase the C1 of activated glucose of UDP-Glu form glycosidic bond with C4 of terminal residue of glycogen primer to 1-4 glycosidic linkage. When the chain has been lenthened to a minimum of 11 residues the branching enzyme amylo-1,4 to 1-6 transglucosidase transfers a part of the 1,4 chain minimum lenth of 6 glucose residue to neighbouring chainto form -1,6 linkage as a branching point.
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Shifting

New 1,6 bond

Glycogen Primer

Branching enzyme

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Glycogenolysis
.

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 The conversion of glycogen to glucose -6phosphate and glucose, which occurs in Muscle & liver respectively. Glycogenolysis is not reverse of glycogenesis but is a separate pathway. Reactions of Glycogenolysis The enzyme glycogen phophorylase cleaves -1,4 linkage yielding glu-1-phosphate & residual Glycogen molecule. This continues til about 4 glucose residue remain on either side of branch point. The four unit chain is then transferred to one branch by glucan transferase. The debranching enzyme splits -1,6 linkage
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-1,4 bond Glucagon Debranching enzyme

Phosphorylase
Pi Glu-1-phosphate

Transferase

Free Glucose

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Next, glu-1-phosphate is converted to glu-1-phosphate by the phosphoglucomutase. In the liver but not in muscle, glu-6-phosphatase removes phosphate to form glucose, which difuse from cell into blood. Regulation of Glycogenesis & Glycogenolysis Glycogen synthase-a, an active or DEPHOSPHORYLATED form Glycogen synthase-b, an inactive or PHOSPHORYLATED form

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Regulation of Glycogenesis & Glycogenolysis Glycogen

Glu-6-P ATP _ + Insulin _

GlycogenPhosphorylase +

cAMP

Glycogen synthase + Glu-6-phosphate

Glucagon, Epinephrine

Ca++ ,AMP

Glucose-1-phosphate

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 In skeletal muscle glycogen is degraded to glucose 6-phosphate, which is then converted into pyruvate and used in ATP production during glycolysis and the Krebs cycle . However, pyruvate can also be converted, in the liver, to glucose; thus muscle glycogen is indirectly a source of blood glucose.

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Reaction cascade for the control of Glycogen Synthesis

.

Glucagon(Liver) ,Epinephrine(Muscle)

ATP

Insulin

+
Inactive adenylate cyclase Active adenylate cyclase

+
Phosphodiesterase

c-AMP

5 -AMP

Active c-AMP dependent Protein kinase ATP

Inactive c-AMP dependent Protein kinase

ADP

Insulin

Active Glycogen Inactive Glycogen synthase a Glu-6-Phsphate synthase b

+ +

H2O Glycogen synthesis

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Pi Insulin

Protein phosphatase - 1

Reaction cascade for the control of Glycogen Breakdown

Glucagon(Liver) ,Epinephrine(Muscle)

+
Inactive adenylate cyclase Active adenylate cyclase
Inactive c-AMP dependent Protein kinase Active c-AMP dependent Protein kinase ATP Ca2+ ADP

Inactive phosphorylase kinase

Ca2+

Active phosphorylase kinase H2O

Pi Protein phosphatase 1

ATP

Insulin

+
c-AMP

+
Phosphodiesterase

5 -AMP

+
GlycogenPhosphorylase a active

Glu-6-P

+
Protein phospha tase

ATP

Insulin GlycogenPhosphorylase b Inactive

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Uronic acid pathway (Glucuronic acid cycle) Definition : A pathway in liver for conversion of Glucose to Glucuronic acid & pentoses as referred to as the uronic acid pathway. Reactions of pathway: Glucose is phosphorylated to Glu-1-phosphate by phosphoglucomutase. with Glu-1-1P then reacts withUTP to form Uridine diphosphate Glu (UDP-Glc) an active nucleotide by UDP- glucose phosphorylase. UDP-glucose is oxidized at carbon 6 to glucuronate via UDP-glucuronate by UDPglucose dH ( NAD dependent)
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 Glucuronate is reduced at carbon 1 to L-gulonate by NADPH dependent enzyme gulonate dH. L-gulonate is precursor of ascorbate ( Vit C) in those animals capable of synthesizing this vitamin. In humans & other primates due to absence of enzyme L-gulonolactone oxidase. L-gulonate is oxidized and decarboxylated to the pentose, L-xylulose by the enzyme L-gulonate decarboxylase. L-xylulose is reduced to xylitol catalysed by NADPH dependent L-xylulose dH.

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 Xylitol is oxidised to D-xylitol by NAD dependent D-xylulose dH. D-xylulose is phophorylated by ATP in the presence of Xylulose kinase to yield xylulose-5phosphate, which is further metabolized in pentose phosphate pathway & lead to formation of glucose.

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 Uronic acid pathway (Glucuronic acid cycle) -D-Glucose-6-phosphate Phosphoglucomutase Glucose-1-phosphate UDP-glu-pyro UTP phosphorylase
UDP

Pentose phosphate pathway

.
D-Xylulose-5- phosphate
ADP

D-xylulose kinase
ATP

Mg+ +

Uridine diphosphate glucose (UDP-Glc) +

2NAD + H2O

D-Xylulose D-xylulose dH
NADH + H+ NAD+

UDP-glucose-dH

2NADH + 2H+

Uridine diphosphate glucose (UDP-Glc) H2O

UDP

Xylitol
NADH + H+

NADPH + H+
Block in NADP essential pentosuria

NAD+ H+

CO2

D-Glucuronate
NADPH +

L-Xylulose L-Gulonate-dH

Gulonate-dH L-Gulonate

NADP

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Thanks

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