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CARBOHYDRATE METABOLISM Cellulose – there is no hydrolase in man that can digest cellulose but it is
-Remedios P. Santos, MD important to add bulk in the diet.

Multiple Roles of Carbohydrates in All Forms of Life: DIETARY FIBERS:

1. CHO serves as energy stores, fuels and metabolic intermediates. It is the preferred source -includes cellulose, hemicelluloses, pectin, lignin.
of energy by the organism –gives 4kcal/gm, 36 or 38 moles of ATP per mole of glucose. -PECTIN and LIGNIN binds or sequesters lipid compounds such as cholesterol →
2. Ribose and deoxyribose sugars form part of the structural framework of DNA and RNA. lowers blood cholesterol levels.
3. Polysaccharides are structural elements in the cell walls of bacteria and plants, and in the -SLOWS DOWN ABSORPTION OF GLUCOSE
exoskeletons of arthropods. Cellulose is the main constituent of plant cell walls; it is the
most abundant organic compound in the biosphere. IMPORTANCE:
4. CHO are linked to many proteins and lipids forming glycoconjugates (proteoglycans, a. To normalize bowel movement
glycoproteins, glycolipids) which can serve as recognition sites for hormones, antigen -adds bulk to stool so the urge to defecate comes regularly
specificity, etc. b. Protects us against chronic constipation, colon cancer and obesity
5. CHO can generate reducing equivalents in the form of NADPH, which is essential in the -Recommended: at least 7servings/day (fruits and vegetables)
biosynthesis of important cellular components like lipids, steroids, DNA, etc. c. Good for “potential” diabetics
-(DM stages: Potential, Latent, Asymptomatic, Overt stage)
Why is Glucose considered as the most important CHO? -Delays and attenuates the postprandial rise in blood glucose, with
1. The characteristic sugar of blood and of tissue fluids is GLUCOSE. The glucose of circulating consequent reduction in insulin secretion. (because fibers slows down
blood and tissue fluids is drawn up by all cells of the body and used for the production of absorption of glucose)
energy. -Effect is beneficial to diabetic and to dieters because it reduces the
2. It is as glucose that the bulk of dietary CHO is absorbed into the bloodstream or into which rebound fall in blood glucose that stimulates appetite.
it is converted in the liver.
-in the LIVER: converts all sugar (galactose, mannose, fructose) to GLUCOSE → blood A. Digestion in the MOUTH
circulation  ENZYME: Salivary Amylase or PTYALIN
-blood sugar level refers to GLUCOSE because all the other sugars are converted to  Bonds acted upon: alpha1,4-glycosidic linkages of amylopectin and amylase
glucose  Optimum pH: 6-7 (readily inactivated at pH4.0 or less)
3. It is from glucose that all other CHO in the body can be formed.  End products: limit dextrin and maltose
-ex. RIBOSE in nucleic acids, GALACTOSE in milk, glycoproteins, proteoglycans
4. Glucose is the major fuel of the tissues of mammals and the universal fuel of the fetus. B. Digestion in the STOMACH
-3RD TRIMESTER: maximum transfer of nutrients from mother to baby.  No enzymatic digestion (due to acidity)
-If premature, the baby might suffer from malnutrition; starts supplementary feeding  Limited acid hydrolysis
earlier than a full term baby.
5. The metabolism of many of the protein amino acids proceeds by the glucose pathway and C. Digestion in the SMALL INTESTINES
some of the products of glucose metabolism are utilized by the body for the synthesis of  ENZYME: Pancreatic Amylase or AMYLOPSIN
amino acids.  Alpha-limit dextrinase
-ex. pyruvate can be converted to alanine  Sucrase
-GLUCONEOGENESIS: amino acid → glucose  Lactase (β-galactosidase)
 Maltase (α-glucosidase)
CARBOHYDRATE DIGESTION AND ABSORPTION  Isomaltase (α1,6-glucosidase)
DIGESTION OF CARBOHYDRATES
FOOD CARBOHYDRATES
60% - starches and dextrins. Starch is a glucosan or glycan (composed of glucose END PRODUCT OF CHO DIGESTION: Monosaccharides (principally GLUCOSE)
ONLY)
-HOMOPOLYSACCHARIDE; most important food source of CHO
30% - sucrose
5% - lactose
5% - other sugars

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ABSORPTION OF CARBOHYDRATES Fate of GLUCOSE-6-PHOSPHATE:

1. Conversion to GLYCOGEN for storage (10%)
Rate of monosaccharide absorption: 2. Enters GLYCOLYSIS (50%)
(MOST rapid to LEAST rapidly absorbed) 3. Conversion to fatty acids and cholesterol
Galactose → Glucose → Fructose → Mannose → Pentoses 4. Conversion to blood glucose
5. Oxidative degradation to CO2 → energy formation (KREB’S CYCLE)
Fates of Ingested Glucose: -continuation of #2; for maximum energy production
50% - converted to energy (glycolysis) 6. Degradation via HMP shunt
30-40% - converted to fats 7. Enters uronic acid pathway
10% - converted to glycogen (stored in muscles and liver)

*INSULIN: facilitates entry of glucose inside the cell. GLYCOLYSIS: “Embden-Meyerhof-Parnas Pathway”
- EXCEPT in LIVER CELLS; glucose can enter the liver cell even without insulin  Gustav Embden, Otto Meyerhof, Jacob Parnas
because liver is the most important organ for the control of blood sugar.  Major pathway for the utilization of glucose; found in all cells
 Unique pathway: can utilize oxygen if available (AEROBIC); can still function in the absence
of oxygen (ANAEROBIC)
FIRST REACTION undergone by GLUCOSE once inside the cells:  Converts GLUCOSE to PYRUVATE (AEROBIC end-product) and LACTATE (ANAEROBIC end-
product) with concomitant production of ATP
ATP ADP  Location: CYTOSOL
Glucose GLUCOSE-6-PHOSPHATE  “Basically” an EXERGONIC process (production of ATP)
Hexokinase -meaning there is a part in the pathway that is endergonic and exergonic.
Glucokinase -Exergonic (ATP production) is more than endergonic (ATP is used).

*IRREVERSIBLE reaction. FUNCTIONS of Glycolysis:

*meaning you cannot use the same enzyme for the forward and backward.  To produce energy in the form of ATP
*It is possible to convert GLUCOSE-6-PHOSPHATE → GLUCOSE using another enzyme  Intermediates formed can be converted to other substances like amino acids, fatty acids,etc.
(Glucose-6-Phosphatase, which is only found in the liver); conversion is only done in the liver.
*The reaction above is a PHOSPHORYLATION REACTION. TISSUES that depend on glycolysis as their major mechanism for ATP production:
1. RBC they lack =ANAEROBIC
Differences between HEXOKINASE and GLUCOKINASE 2. Cornea, lens, regions of the retina mitochondria GLYCOLYSIS
HEXOKINASE GLUCOKINASE 3. Kidneys medulla, testis, leukocytes and white muscle fibers
High Km for glucose -have relatively FEW mitochondria
 They consume about 40 GRAMS OF GLUCOSE PER DAY in a normal human adult.
Low Km for glucose
(so it can function ONLY when the
concentration of glucose is relatively HIGH) BRAIN: has an absolute need for glucose.
-can act even if glucose level is LOW
-cannot act if glucose level is LOW -the PYRUVATE obtained is oxidized completely to CO2 and H2O in the brain mitochondria
Inhibited by high levels of glucose-6-  Uses 120 GRAMS OF GLUCOSE PER DAY
Not inhibited by glucose-6-phosphate
phosphate (feedback inhibition)
Acts on other hexoses such as fructose, HEART MUSCLE: adapted for aerobic performance; has relatively poor glycolytic ability, and
Acts on glucose ONLY (specific) poor survival under conditions of ischemia.
mannose, etc. (general enzyme)

 Enzymes are NOT used at the same time. FIRST- Hexokinase Growing CANCER CELLS: glycolysis proceeds at a much higher rate than is required by Kreb’s
 FEEDBACK INHIBITION: the enzyme is inhibited by its own product. cycle → more pyruvate produced than can be metabolized → excessive lactate production →
 Once hexokinase is inhibited by ↑ glucose-6-phosphate, glucokinase is the only one will act. lactic acidosis.

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TWO PHASES OF GLYCOLYSIS “IN GLYCOLYSIS, all the IRREVERSIBLE STEPS are CONTROL POINTS!”
1. Energy INVESTMENT phase; PRIMING STAGE
 Endergonic CONTROL POINT:
Glucose → Fructose 1,6-bisphosphate -if you want to STIMULATE the pathway, stimulate the enzymes of the control point.
-if you want to INHIBIT the pathway, inhibit the enzymes of the control point.
2. Energy GENERATION phase; ENERGY RECOVERY PHASE
 Exergonic
 Production of ATP
I. ENERGY INVESTMENT PHASE (PRIMING STAGE)
A. SPLITTING STAGE  FIRST REACTION: (1st) Irreversible
Fructose 1,6-bisphosphate → DHAP + glyceraldehydes-3-PO4 -ENZYME: Hexokinase, Glucokinase
-use of 1ATP
B. OXIDOREDUCTION-PHOSPHORYLATION STAGE -Phosphorylation reaction
glyceraldehydes-3-PO4 → Pyruvate or Lactate
 SECOND REACTION: Reversible
-ISOMERIZATION REACTION: an aldose is converted into a ketose or vice versa
-Ex: Glucose-6-phosphate ↔ Fructose-6-phosphate
-ENZYME: Phosphoglucose Isomerase

 THIRD REACTION: (2nd) Irreversible

-use of 1ATP
-Phosphorylation reaction
-RATE LIMITNG STEP (Committed Step)
-most important CONTROL POINT of the pathway
-RATE LIMITING ENZYME: Phosphofructokinase 1 (PFK 1)

SUMMARY:
 There are 2 IRREVERSIBLE steps.
 2 ATP’s are used.
 No ATP production.

II. ENERGY GENERATION PHASE

A. SPLITTING STAGE
 FOURTH REACTION:
-Fructose-1-6-Bisphosphate (6 carbons) SPLITS into 2 sugars:
 Dihydroxyacetone phosphate (3 carbons)
 Glyceraldehyde-3-phosphate (3 carbons)
-SPLITTING ENZYME: Aldolase (aldo-cleavage?)
-Dihydroxyacetone (DHAP) phosphate and Glyceraldehyde-3-phosphate are
INTERCONVERTIBLE.
-ISOMERIZATION REACTION

GLYCOLYSIS (Embden-Meyerhof-Parnas Pathway)

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B. OXIDOREDUCTION-PHOSPHORYLATION STAGE SUMMARY:

-whatever happens to Glyceraldehyde-3-phosphate, also happens to DHAP
-energy produced in Glyceraldehyde-3-phosphate, MULTIPLY by 2.
 FIFTH REACTION:
-conversion of Glyceraldehyde-3-phosphate → 1,3 Diphosphoglycerate
-ENZYME: Glyceraldehyde-3-phosphate Dehydrogenase
-produces NADH (whenever NADH is produced, it has to go back to NAD so that the
pathway will go on; if NAD+ were not regenerated, glycolysis could not proceed beyond
glyceraldehyd-3-phosphate, which means no ATP would be generated.)
-H+ is given to Electron Transport Chain (ETC) in the mitochondria
-from the CYTOSOL, it has to go to the MITOCHONDRIA through a shuttle system.
-SHUTTLE SYSTEM: Malate-Aspartate (Complex1); Glycerol-3-phosphate (Complex2)
-MASS: 3 ATP produced x2 = 6 ATP
-G3PSS: 2 ATP produced x2 = 4 ATP
-OXIDATIVE PHOSPHORYLATION: ATP production through ETC
-Step5: ENERGY PRODUCTION THROUGH OXIDATIVE PHOSPHORYLATION
 SIXTH REACTION:
-1,3 Diphosphoglycerate → 3-phosphoglycerate
-ENZYME: Phosphoglycerate Kinase
-1 ATP produced x2 =2 ATP
-SUBSTRATE LEVEL OF PHOSPHORYLATION: energy production even without entering ETC
 2 substrate level of phosphorylation
 1st – catalyzed by Phosphoglycerate Kinase (Sixth reaction)
 2nd – catalyzed by Pyruvate Kinase (Ninth reaction)
 1 oxidative phosphorylation
 Fifth reaction
 3 irreversible steps = CONTROL POINTS!
 First reaction (catalyzed by Hexokinase, Glucokinase)
 Third reaction (catalyzed by Phosphofructokinase 1) –RATE LIMITING STEP
 Ninth reaction (catalyzed by Pyruvate Kinase)
 If becomes ANAEROBIC (No ETC), NADH will be used to convert PYRUVATE → LACTATE.
 ENZYME: Lactate Dehydrogenase
 In ANAEROBIC GLYCOLYSIS, end-product is LACTATE.
*In as much as the major function of glycolysis is production of energy, the major control
factor is the energy level of the cell.
↑energy = ↑ATP/AMP ratio = ↑ATP/ADP ratio = ↓AMP = ↓ADP
 Meaning, energy is HIGH; pathway needs to be inhibited.
 INHIBIT enzymes at CONTROL POINT.

 SEVENTH REACTION: ↓energy = ↓ATP/AMP ratio = ↓ATP/ADP ratio = ↑AMP = ↑ADP

-3-phosphoglycerate→ 2-phosphoglycerate  Meaning, energy is LOW; pathway needs to be stimulated.
-ENZYME: Phosphoglyceromutase  STIMULATE enzymes at CONTROL POINT.

 EIGHTH REACTION: RECITATION:

-2-phosphoglycerate → Phosphoenolpyruvate  If 1mol of glucose goes to AEROBIC glycolysis using the MALATE-ASPARTATE shuttle, how
-ENZYME: Enolase many ATP’s will be produced (Gross)?
-Enolase is inhibited by FLUORIDE  Fifth reaction: 6 ATP
-DEHYDRATION STEP  Sixth reaction: 2 ATP
 Ninth reaction: 2 ATP
Gross = 10 ATP (Energy generation phase)
 NINTH REACTION: (3rd) Irreversible First, Third reaction: -2 ATP (Energy investment phase)
-Phosphoenolpyruvate → Pyruvate Net = 8 ATP
-ENZYME: Pyruvate Kinase
-1 ATP produced x2 =2 ATP  Using the GLYCEROL-3-PHOSPHATE shuttle
-SUBSTRATE LEVEL OF PHOSPHORYLATION  Fifth reaction: 4 ATP
 Sixth reaction: 2 ATP
 Ninth reaction: 2 ATP
Gross = 8 ATP (Energy generation phase)
First, Third reaction: -2 ATP (Energy investment phase)
Net = 6 ATP

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RECITATION: Pyruvate Dehydrogenase Complex

 If 1mol of glucose goes to ANAEROBIC glycolysis using the MALATE-ASPARTATE shuttle, PYRUVATE ACETYL CoA
how many ATP’s will be produced (Gross)? -TPP, Lipoic Acid, FAD
 Only source of energy will be from the 2 substrate level of phosphorylation NAD+ NADH + H+
 Sixth reaction: 2 ATP
 Ninth reaction: 2 ATP
Gross = 4 ATP (Energy generation phase) PYRUVATE DEHYDROGENASE COMPLEX:
First, Third reaction: -2 ATP (Energy investment phase) ENZYMES: Prosthetic Groups
Net = 2 ATP 1. Pyruvate dehydrogenase -Thiamine Pyrophosphate (TPP)
2. Dihydrolipoyl transacetylase -Lipoamide and CoA
3. Dihydrolipoyl dehydrogenase -FAD and NAD

ENZYMATIC CONTROL OF GLYCOLYSIS: p113 5 coenzymes used:

*Inhibit/Stimulate at CONTROL POINTS 1. Thiamine Pyrophosphate (TPP)
 Most active coenzyme form of vitB1
1. HEXOKINASE 2. FAD
 Inhibited by Glucose-6-phosphate  From vitB2 (Riboflavin)
2. PHOSPHOFRUCTOKINASE 1 (PFK1) 3. CoASH
 MAJOR CONTROL ENZYME of glycolysis  From vitB5 (Pantothenic Acid)
3. PYRUVATE KINASE 4. Lipoic Acid
 From vitB3 (Niacin)
5. NAD
INHIBITORS OF THE GLYCOLYTIC PATHWAY  From vitB3 (Niacin)
1. 2-DEOXYGLUCOSE
 Inhibits HEXOKINASE  Pyruvate CANNOT be converted to Acetyl CoA without the B-complex vitamins.
2. SULHYDRYL REAGENTS (iodoacetate)
 Inhibits GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE
3. FLUORIDE INHIBITORS OF PYRUVATE DEHYDROGENASE
 Inhibits ENOLASE (catalyzes the dehydration step) 1. Aresenite and Mercuric ions
4. PENTAVALENT ARESNIC or ARESNATE -react with the –SH group of lipoic acid and inhibit pyruvate dehydrogenase
 Inhibits PYRUVATE DEHYDROGENASE, and α-KETOGLUTARATE DEHYDROGENASE 2. Thiamine Deficiency
-Pyruvate cannot be converted to Acetyl CoA → Lactic Acidosis
-nutritionally deprived alcoholics are thiamine-deficient and may develop potentially
fatal pyruvic and lactic acidosis
-Alcohol INHIBITS ABSORPTION of Thiamine.
FATES OF PYRUVIC ACID
1. Reversible reduction to LACTATE INBORN ERRORS OF METABOLISM ASSOCIATED WITH GLYCOLYSIS:
-if the condition is ANAEROBIC 1. Inherited ALDOLASE A Deficiency in Erythrocytes- can cause
2. Conversion back to GLUCOSE 2. Inherited PYRUVATE KINASE Deficiency hemolytic anemia
3. Formation of OXALOACETATE or MALATE -Glucose-6-phosphate Dehydrogenase (also causes hemolytic anemia)
4. Transamination to form ALANINE 3. Muscle PHOSPHOFRUCTOKINASE Deficiency
-Pyruvate + Amino group = Alanine -rate limiting enzyme of glycolysis → glycolysis stops → low energy production
5. Oxidative decarboxylation to Acetyl CoA -exercise capacity of patient is low, particularly on high carbohydrate diet
-MAJOR FATE OF PYRUVIC ACID 4. Inherited PYRUVATE DEHYDROGENASE Deficiency
-it will be converted to Acetyl CoA -due to defects in one or more of the components of the enzyme complex
-catalyzed by Pyruvate Dehydrogenase Complex (has 5 steps of reactions) -manifested by lactic acidosis, particularly after a glucose load because pyruvate is
channeled toward lactic acid production.

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SUMMARY: ACETYL CoA

FATE OF LACTIC ACID in ANAEROBIC Glycolysis

 Lactate can either be exported from the cell OR converted back to pyruvate.
 Much of the lactate produced in skeletal muscle cells is carried by the blood to the liver
where it is used to SYNTHESIZE GLUCOSE (Gluconeogenesis)
 “CORI CYCLE” or LACTIC ACID CYCLE
 ACETYL CoA is a CENTRAL METABOLITE
 Majority of Acetyl CoA enters the Kreb’s cycle (80%) for maximum ATP production.
 20% or less: formation of ketone bodies, cholesterol, bile acids, steroid hormones,
prostaglandins, TAG.
 ENTRY of Acetyl CoA to Kreb’s cycle REQUIRES INSULIN (has a permissive effect on Kreb’s).

CASE:
o DIABETES: LACKS INSULIN → ACETYL COA CANNOT ENTER KREB’S
→ channel more towards Ketone body production → Ketoacidosis
→ channel more towards cholesterol formation → premature atherosclerosis
GLUCONEOGENESIS  Coronary arteries → Myocardial infarction
Lactate → Glucose  Cerebral arteries → Stroke
 Retinal arteries → blindness
 Renal arteries → Renal failure

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KREB’S CYCLE (TRICARBOXYLIC ACID CYCLE)
FUNCTIONS OF KREBS in Citric Acid Cycle
1. It is the major degradative pathway for the generation of ATP
2. It provides intermediates for biosynthesis
AMPHIBOLIC PATHWAY: has both anabolic and catabolic part at the same time
CATABOLIC- degradation to form ATP
ANABOLIC- use intermediates to synthesize other substances
 ALPHA-KETOGLUTARATE: precursor of GLUTAMIC ACID
 OXALOACETATE: precursor of ASPARTIC ACID
 CITRATE: precursor of extramitochondrial acetyl CoA for FATTY ACID
biosynthesis thru the ATP-citrate lyase reaction
 SUCCINYL CoA: for the synthesis of HEME (succinyl coa + glycine)

PATHWAY:
FIRST REACTION- Oxaloacetate + Acetyl CoA → Citrate
 Enzyme: Citrate Synthase
 RATE LIMITING STEP/ Committed Step!
 MAJOR CONTROL POINT!

*RATE LIMITING STEP IS ALWAYS THE MAJOR CONROL POINT!*

Citrate → Cis-aconitic Acid → Isocitrate → Oxalosuccinate →alpha-ketoglutarate → Succinyl

CoA → Succinate → Fumarate → Malate

*MUST KNOW!
-Where NADH is being released?
-Krebs is beside ETC, so when NADH is produced → Complex1 = 3 ATP’s

3 NAD-DEPENDENT ENZYMES IN KREBS: (Oxidative Phosphorylation)

1. Isocitrate Dehydrogenase
2. Alpha-ketoglutarate Dehydrogenase
3. Malate Dehydrogenase
= Total of 9 ATP’s

Release of FADH2 (Succinate Dehydrogenase) → ETC via Complex2 = 2 ATP’s

OXIDATIVE PHOSPHORYLATION (NAD, FAD → ETC)

 TOTAL of 11 ATP’s produced.

 Hans Krebs (1937) SUBSTRATE LEVEL OF PHOSPHORYLATION

 Location: MITOCHONDRIA, beside Electron Transport Chain (ETC)  Production of ATP without the use of ETC
 Referred to as “final common pathway” of carbohydrates, fats and protein metabolism.  Succinate Thiokinase step = 1 ATP produced.

Oxidative Phosphorylation 11
Substrate level of Phosphorylation 1
1 Krebs = 12 ATP’s

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ALPHA-KETOGLUTARATE step is exactly the same as the PYRUVATE DEHYDROGENASE step. RECITATION:
 Closely resembles the pyruvate dehydrogenase complex.  If Glucose undergoes COMPLETE OXIDATION (Glycolysis + Krebs) using the MALATE-
 The same coenzymes are involved and the mechanisms are identical (TPP, FAD, Lipoic, ASPARTATE shuttle, how many ATP’s will be produced (Gross)?
CoA-SH, NAD).  Glycolysis (MAS- Aerobic) 10 ATP
 They have the SAME INHIBITOR: Arsenite Krebs 24 ATP
Pyruvate DH 6 ATP
TOTAL: Gross 40 ATP
CONTROL POINTS IN THE KREBS CYCLE: Net 38 ATP
1. CITRATE SYNTHASE
 Acetyl CoA + Oxaloacetate → Citrate  If Glucose undergoes COMPLETE OXIDATION (Glycolysis + Krebs) using the GLYCEROL-3-
 RATE LIMITING STEP PHOSPHATE shuttle, how many ATP’s will be produced (Gross)?
 INHIBITED by ATP  Glycolysis (G3P- Aerobic) 8 ATP
 Rate of the reaction is largely dependent on the availability of acetyl CoA and Krebs 24 ATP
Oxaloacetate and by the concentration of succinyl CoA which competes with acetyl Pyruvate DH 6 ATP
CoA and inhibits citrate synthase. TOTAL: Gross 38 ATP
Net 36 ATP
2. ISOCITRATE DEHYDROGENASE
 Isocitrate ↔ α-ketoglutarate  If Glucose undergoes COMPLETE OXIDATION (Glycolysis + Krebs) using the MALATE-
 INHIBITED by NADH+ ASPARTATE shuttle, how many ATP’s will be produced through OXIDATIVE
PHOSPHORYLATION alone?
3. ALPHA-KETOGLUTARATE DEHYDROGENASE  Glycolysis (G3P- Aerobic) 6 ATP
 Alpha-ketoglutarate → Succinyl CoA Krebs 22 ATP
Pyruvate DH 6 ATP
4. SUCCINATE DEHYDROGENASE TOTAL: 34 ATP
 Succinate ↔ Fumarate
 MALONATE is a competitive inhibitor of the enzyme.
ANAPHLEROTIC REACTIONS or FILLING-UP REACTIONS
 HIGH ENERGY charge → inhibits control enzymes (↓activities of citrate synthase, -replace or filling-up of intermediates
isocitrate DH, and α-ketoglutarate DH) → ↓Krebs -makes sure that intermediates are always available
 LOW ENERGY charge → stimulates control enzymes (↑activities of citrate synthase,
isocitrate DH, and α-ketoglutarate DH) → ↑Krebs 1. Formation of MALATE from PYRUVATE
Pyruvate + CO2 + NADPH + H+ ↔ MALATE + NADP
Malic enzyme
SUMMARY:
2. Glutamate + Pyruvate ↔ α-ketoglutarate + alanine
Transaminase
Aspartate + Pyruvate ↔ Oxaloacetate + alanine

3. Enzymatic carboxylation of pyruvate to form oxaloacetate

-most important anaphlerotic reaction in the liver and kidney
-MAJOR ANAPHLEROTIC REACTION
-catalyzed by PYRUVATE CARBOXYLASE
-only activated if there is Acetyl CoA to combine with Oxaloacetate (1st step in Krebs)
-occurs in 2 steps:
E-biotin + ATP + CO2 + H2O ↔ E-carboxylation + ADP + Pi
E-carboxybiotin + Pyruvate ↔ E-biotin + oxaloacetate .
ATP + CO2 + Pyruvate + H2O ↔ OXALOACETATE + ADP + Pi

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Importance of Gluconeogenesis:
-requires BIOTIN (vitB7) as coenzyme; ACETYL CoA as activator. (1st step in Krebs) 1. It meets the needs of the body for glucose when sufficient CHO is not available from the
-NOT part of glycolysis and krebs. It is an important reaction because it LINKS the two diet or glycogen reserves.
important energy-producing pathways; serves as a bridge of the two pathways.  1st- GLYCOGEN RESERVES
o GLYCOGENOLYSIS: breakdown of glycogen to glucose
o Glycogen will maintain blood sugar level for 1 DAY.
THIAMINE (VITAMIN B1) DEFICIENCY (BERI-BERI)  2nd- FATS
 In the form of THIAMINE PYROPHOSPHATE (TPP) o LIPOLYSIS → fatty acids + glycerol
 Coenzyme in PYRUVATE DH, and ALPHA-KETOGLUTARATE DH steps. o Glycerol becomes Glucose.
o Will maintain blood sugar level depending on how much fat is available.

2. Glucose is also important in maintaining the level of intermediates of the Krebs cycle
even when fatty acids are the main source of acetyl CoA in the tissues.
3. Gluconeogenesis clears LACTATE produced by muscle and erythrocytes and GLYCEROL
produced by adipose tissues.

GLUCONEOGENESIS

Manifestations:
 If THIAMINE (TPP) is deficient, PYRUVATE is channeled towards LACTIC ACID production →
LACTIC ACIDOSIS
 Cannot enter KREBS → ↓ATP (energy) production → SEVERE MUSCLE WEAKNESS
-affects skeletal muscles, and CARDIAC muscles → cardiac failure
 USUAL MANIFESTATIONS: prickling sensations at the fingertips
 MOST SEVERE FORM OF BERI-BERI: “SHOSHIN BERI-BERI”
-cardiac muscles are affected → cardiac failure

GLUCONEOGENESIS
 Reverse of Glycolysis and Krebs
 HYPERGLYCEMIC PATHWAY: ↑ blood sugar level
 Includes all pathways and mechanisms responsible for converting non-carbohydrates to
glucose or glycogen.
 Two major organs involved: LIVER and KIDNEYS
 The major gluconeogenic PRECURSORS are:
 LACTATE
 GLYCEROL (from fats)
 GLUCOGENIC AMINO ACIDS
 Reverse of GLYCOLYSIS (cytosol) and KREBS (mitochondria); starts at bottom, end in GLUCOSE
 PROPIONATE (in ruminants) –in animals  GLUCONEOGENESIS involves both CYTOSOL and MITOCHONDRIA.

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 The 3 IRREVERSIBLE steps in GLYCOLYSIS must be reversed in GLUCONEOGENESIS:

1. Phosphoenolpyruvate → Pyruvate
 No SINGLE enzyme that can reverse this step.
 Needs to enter the MITOCHONDRIA
 Pyruvate → Oxaloacetate → Malate → goes out to cytosol → Oxaloacetate
→ Phosphoenolpyruvate →→ 2-Phosphoglycerate → 3-Phosphoglycerate
→ 1-3, Diphosphoglycerate → Glyceraldehyde-3- PO4 → Fructose-1,6-
bisPO4 → Fructose-6-PO4 → Glucose-6-PO4 → GLUCOSE
 Enzyme used to revese this reaction includes PYRUVATE CARBOXYLASE
(pyruvate to oxaloacetate); another enzyme, PHOSPHOENOLPYRUVATE
CARBOXYKINASE (PEPCK) catalyzes the conversion of oxaloacetate to
phosphoenolpyruvate.
2. Fructose-6-phosphate → Fructose 1,6 bisphosphate
 Reversed by FRUCTOSE 1,6-BISPHOSPHATASE
 Enzyme is present in liver and kidney; absent in heart muscle and smooth
muscle
3. Glucose → Glucose-6-phosphate
 Reversed by GLUCOSE-6-PHOSPHATASE
 Enzyme is present in liver and kidney; absent from muscle and adipose tissues

 “BYPASS PATHWAYS”- bypassing the irreversible steps.

 All the enzymes that are used to bypass are CONTROL ENZYMES of gluconeogenesis
 Includes: Glucose-6-phosphatase; Fructose 1,6 bisphosphatase; Pyruvate Carboxylase;
and Phosphoenolpyruvate Carboxykinase (PEPCK)

GLYCEROL (from fats) to GLUCOSE

 Glycerol → Glycerol-3-PO4 → DHAP
 DHAP is the entry point of GLYCEROL in glycolysis
 DHAP + Glyceraldehyde-3- PO4 → Fructose-1,6-bisPO4
 Fructose-1,6-bisPO4 → Fructose-6-PO4 → Glucose-6-PO4 → GLUCOSE

LACTATE to GLUCOSE
 First: Lactate → Pyruvate (enzyme: Lactate Dehydrogenase)
 Pyruvate → Oxaloacetate → Malate → goes out to cytosol → Oxaloacetate →
Phosphoenolpyruvate → 2-Phosphoglycerate → 3-Phosphoglycerate → 1-3,
Diphosphoglycerate → Glyceraldehyde-3- PO4 → Fructose-1,6-bisPO4 → Fructose-6-PO4 →
Glucose-6-PO4 → GLUCOSE

ia,’12