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BIOCHEMISTRY

GLUCONEOGENESIS
Dr. Catherine L. Co-Reportoso | October 31, 2018 LE3 TRANS2

OUTLINE  Glucose: 6 carbons


I. Introduction and Review IV. Major Substrates of  Pyruvate: 3 carbons
A. Molecular Structures Gluconeogenesis  Acetyl-coA: 23 carbons
B. Glycolysis A. Lactate → Acetyl: 2 carbons
C. Kreb’s Cycle B. Glucogenic AAs → CoA: 21 carbons
II. Gluconeogenesis C. Glycerol
A. Definition D. Propionyl-coA Oxidation and Reduction
B. Steps, Reactions and V. Glucose-Forming Pathways
Products A. Cori Cycle/Glucose-  Common features of almost all metabolic reactions
C. Glycolysis vs. Lactate Pathway  Oxidation of a molecule leads to reduction of another
Gluconeogenesis B. Cahill Cycle/Glucose- o Oxidation: Loss of H or gain of O
III. Key Features of Alanine Pathway o Reduction: Gain of H or loss of O
Gluconeogenesis VI. Regulation of  1° OH can be oxidized to aldehyde then carboxylate or
A. Pyruvate to Gluconeogenesis reverse
Phosphoenolpyruvate A. Reciprocal Regulation  2° alcohol can be oxidized to ketone
(PEP) B. Rate of Enzyme  Brought by reducing or oxidizing agents
B. Fructose 1,6- Synthesis o Reducing agents undergo oxidation
Biphosphate to C. Covalent Modification o Oxidizing agents undergo reduction
Fructose-6-phosphate D. Allosteric Effectors
C. Glucose-6-phosphate VII. Physiological Importance Example reaction: Conversion of Malate to Oxaloacetate
to Glucose of Gluconeogenesis
VIII. Guide Questions
IX. References
LEARNING OBJECTIVES
 Review Gycolysis and Tricarboxylic Acid Cycle
 Describe the physiological significance of gluconeogenesis
 Compare and contrast the steps of glycolysis and
gluconeogenesis
 Discuss the key features of gluconeogenesis
 Describe the pathway in terms of its substrates: lactate,
Figure 3. Conversion of Malate to Oxaloacetate
glucogenic amino acids, glycerol, propionyl CoA
 Differentiate between Cori cycle and Glucose-Alanine cycle
 C2 of malate is oxidized to ketone forming oxaloacetate
 Discuss the regulation of the pathway during fasting
 NAD+ is the oxidizing agent: reduced to form NADH
I. INTRODUCTION AND REVIEW
B. GLYCOLYSIS
A. MOLECULAR STRUCTURES

Figure 1. Structure of Glucose and Pyruvate

Figure 2.Structure of Acetyl-coA

No. of Carbons Figure 4. Overview of Glycolysis [2021 B Trans]

L.E. # 3 - Trans # 2 Group F: Ramirez, R.J., Rapisora, Raymundo, Regalado, Reyes, A.A. 1 of 10
 Breakdown of 1 mol glucose o 1 FADH2 x 1.5
o Aerobic - 2 mol pyruvate, yields 5-7 ATP depending on o 1 ATP (GTP)
the shuttle system used  Site for entry of most amino acids for gluconeogenesis
o Anaerobic - 2 mol lactate, yields 2 ATP
 Glycolysis is linear pathway consisting of 10 total steps Recall:
 ATP is consumed in the 1st and 3rd reactions within the first  Anaplerotic reactions – replenish TCA intermediates
phase (energy investment phase) o Pyruvate carboxylase (pyruvate to oxaloacetate)
 ATP is produced in the 7th (Phosphoglycerate kinase) and  Cataplerotic reactions - utilizes TCA intermediates
10th reactions (Pyruvate kinase) within the second phase o Helps remove buildup of excess molecules
(energy generation phase). o Intermediates can be used for biosynthesis of other
 Function in fuel metabolism via provision of ATP macromolecules
 Steps 1,3, and 10 are irreversible o Malate to aspartate during Malate-Aspartate shuttle
o Step 1 (Hexokinase / Glucokinase): dedicates glucose [2021 A Trans]
to the cell for metabolism
o Step 3 (PFK-1): rate-limiting step II. GLUCONEOGENESIS
 Step 7 is a reversible step GLUCONEOGENESIS
 Activated during fed state [2021 A Trans] A. DEFINITION
 First step
o Glucose  Glucose 6-Phosphate  The process of synthesizing glucose or glycogen from
o Enzyme: Hexokinase/Glucokinase (Glucokinase has noncarbohydrate precursors [Harper]
higher Km than hexokinase for glucose)  Synthesis of glucose from non-carbohydrate compounds
o Phosphate: comes from ATP (Investment phase)  Converts 2 moles of pyruvate to 1 mole of glucose
 Substrate level of phosphorylation: Phosphoglycerate  Also involves TCA cycle intermediates especially
kinase and pyruvate kinase because they have two high oxaloacetate
phosphate energy compounds. [Lecturer]  Gluconeogenic flux is regulated primarily by fructose-2,6
 1, 3 Biphosphoglycerate and Phosphoenol pyruvate donate biphosphate
phosphate to ADP to become ATP. They have higher - ∆G.  Energetically expensive (Consumes 6 ATP when
[Lecturer]
converting 2 mol of pyruvate to 1 glucose)
 The reverse of Gluconeogenesis. Because they share the
same enzymes on the reversible reactions. In the irreversible Functions
reactions, these are the steps bypassed in gluconeogenesis:  Enables maintenance of blood glucose levels long after all
o Reactions catalyzed by Pyruvate kinase, PFK1 and dietary glucose has been absorbed and completely
Glucokinase/Hexokinase oxidized and glucose stored as glycogen has been used up
[Devlin]
C. TRICARBOXYLIC ACID CYCLE  Maintain level of intermediates of citric acid cycle [2021 A]
T  Recycles waste products of metabolite [2021A]:
→ Lactate from: Muscle and Erythrocytes
→ Glycerol from: adipose tissues

Locations
 Major gluconeogenic tissues: liver and kidney
o Overnight fast
 Liver: 90%
 Kidney: 10%
o Prolonged fasting
 Kidney: 40%
 Already occurs in lesser extent in the liver 4 hours after meal
o Not major source compared to glycogenolysis
 Renal gluconeogenesis occurs about 2 days of fasting
 Gluconeogenesis is highly active during 2-5 days of fasting
After 5 days, the body the amino acids are conserved
while the body undergoes ketogenesis [2021A]
 Occurs largely in the CYTOSOL

Major substrates
 Glucogenic Amino Acids
 Lactate
 Glycerol
 Propionate [Harper]
B. STEPS, REACTIONS AND PRODUCTS
 7 of the 10 enzymatic reactions of gluconeogenesis are the
reverse of glycolytic reactions [Lehninger].
→These reactions have a 𝚫G near 0
Figure 5. Diagram of Kreb’s Cycle [Quora]
→The 3 irreversible steps are bypassed by a separate set of
 Final common pathway for oxidation of CHO, lipids, and enzymes that are also exergonic:
proteins with Acetyl CoA as substrate.  Carboxylation of pyruvate to PEP
 Occurs in the mitochondria  Conversion of Fructose-1,6-bisphosphate to fructose-
 Occurs at inner mitochondria and accounts for the two-thirds 6-phosphate
of ATP produced in fuel oxidation.  Glucose-6-phosphate to Glucose
 10 ATP per cycle
o 3 NADH x 2.5
Biochemistry Gluconeogenesis 2 of 10
11. F6P will be isomerized to glucose-6-phosphate (G6P) using
phosphohexose isomerase
12. G6P will be dephosphorylated by glucose-6-phosphatase to
yield one glucose molecule

*the steps unique to gluconeogenesis are steps 1, 3, 10, and 12


Net Reactions and Products
2 Pyruvate + 2 NADH + 4 ATP + 2 GTP + 6H2O + 2H+ → Glucose
+ 2 NAD + 2 GDP + 4 ADP + 2 GDP + 6 Pi
 Major product: glucose
 2 mol pyruvate → 1 mol glucose
 Uses 4 mol ATP, 2 mol GTP and 2 mol NADH
 Produces 4 mol of ADP, 2 mol of GDP and 2 mol of NAD
 consumes 6 net ATP
C. GLYCOLYSIS VS. GLUCONEOGENESIS
Table 1. Comparison of Glycolysis and Gluconeogenesis
Glycolysis Gluconeogenesis
Catabolic Anabolic
-catabolizes glucose and -Synthesizes sugars and
other carbohydrates polysaccharides
Glucose  Pyruvate Pyruvate  Glucose
Rate Limiting Enzymes: Rate Limiting Enzymes:
hexokinase, Pyruvate carboxylase,
phosphofructokinase, PEPCK, Fructose-1,6-
pyruvate kinase biphosphatase, Glucose-6-
phosphatase
Muscles and tissues Liver and kidneys (prolonged
starvation)
2 ATP used 6 ATP used
Generates 4 ATP Generates 1 glucose
Net: 2 ATP and glucose
.
III. KEY FEATURES OF GLUCONEOGENESIS

Figure 6. Steps of Glycolysis and Gluconeogenesis [Lecturer’s PPT]

Steps

1. Pyruvate is carboxylated to oxaloacetate by the mitochondrial


pyruvate carboxylase
Consumes ATP
Coenzyme: Biotin
a. Biotin binds CO2 from bicarbonate as carboxybiotin prior
to addition of CO2 to pyruvate [Harper]
2. Resultant oxaloacetate is reduced to malate, exported from
mitochondrion into the cytosol where it is oxidized back to
oxaloacetate (malate-asparate shuttle)
3. Oxaloacetate is decarboxylated and phosphorylated by
phosphoenolpyruvate carbokinase (PEPCK) to PEP
a. Uses GTP as the phosphate donor Figure 7. Enzyme difference of glycolysis and gluconeogenesis [Lecturer’s PPT]
4. PEP is converted to 2-phosphoglycerate via enolase
5. 2-PG is isomerized to form 3-Phosphoglycerate via the  The 1st, 3rd, and 10th reactions of glycolysis are irreversible
enzyme phosphoglycerate mutase →Reactions with kinase enzymes are irreversible except for
6. 3-PG is phosphorylated at C1 with the enzyme 
phosphoglycerate kinase (step 7)
phosphoglycerate kinase to form 1,3 Bisphosphoglycerate
 Four enzymes are unique to gluconeogenesis:
7. 1,3 BPG is reduced to Glyceraldehyde-3-phosphate (GADP)
o Glucose-6-phosphatase
by Glyceraldehyde phosphate dehydrogenase
o Fructose-1,6-Bisphosphatase
8. GADP is isomerized and converted to dihydroxyacetone
o PEP carboxykinase (PEPCK)
phosphate (DHAP) using the enzyme triose phosphate
o Pyruvate carboxylase
isomerase
9. Since there are two GADP from the previous reaction, the other Table 2. Enzymes bypassed in glycolysis vs. enzymes unique to
mol of GADP remains as GADP, only one mol will be used in gluconeogenesis
this reaction GLYCOLYSIS GLUCONEOGENESIS
a. Both GADP and DHAP will be catalyzed by the aldolase Enzyme bypassed Unique enzyme
enzyme to form Fructose-1,6-bisphosphate (F-1,6-BP) Pyruvate kinase Pyruvate carboxylase
10. F-1,6-BP will by cleaved by F-1,6-bisphosphatase to form PEP carboxykinase
fructose-6-phosphate (F6P) Phosphofructokinase-1 Fructose-1,6-bisphosphatase
Hexokinase/glucokinase Glucose-6-phosphatase
Biochemistry Gluconeogenesis 3 of 10

Table 3. Energy expenditure of gluconeogenesis


Enzyme Energy required
Pyruvate carboxylase 2 ATP*
PEP carboxykinase 2 GTP (= 2 ATP)
Phosphoglycerate kinase 2 ATP
TOTAL 6 ATP Figure 10. Main route [2021A Trans]

*Remember that 2 mol of pyruvate are needed to generate 1 mol  Alternative route
of glucose • OAA is reduced to malate in the mitochondria
 catalyzed by malate dehydrogenase
A. PYRUVATE TO PHOSPHOENOL PYRUVATE (PEP)
• Malate is transferred to the cytosol
• Malate is then oxidized back to OAA in the cytosol
 catalyzed by malate dehydrogenase
• Cytosolic PEPCK will catalyze the conversion of OAA to
PEP

Figure 11. Alternative route [2021A Trans]


B. FRUCTOSE-1,6-BISPHOSPHATE TO FRUCTOSE-6-
PHOSPHATE
Figure 8. Route for conversion of pyruvate to PEP [Lecturer’s PPT]  Removal of phosphate group from C1
 Catalyzed by fructose-1,6-bisphosphatase
 Accomplished in two steps: o Present in liver, kidney, and skeletal muscle [Harper’s,
• Pyruvate  Oxaloacetate (OAA)
2012]
 Catalyzed by pyruvate carboxylase
• OAA  PEP C. GLUCOSE-6-PHOSPHATE TO GLUCOSE
 Catalyzed by PEP carboxykinase (PEPCK)  Catalyzed by glucose-6-phosphatase
o Present in liver and kidney [Harper’s, 2012]
Conversion of pyruvate to oxaloacetate (OAA) o Absent from muscle (therefore, muscle cannot export
 Carboxylation reaction glucose into the bloodstream) [Harper’s, 2012]
 Catalyzed by pyruvate carboxylase Note: Recall that Glucose-6-phosphatase is also an enzyme
o Solely a mitochondrial enzyme present in glycogenolysis. GSD-1 (Von Gierke Disease), which
o Biotin-dependent is due to the deficiency of this enzyme, leads to more severe
o Allosterically activated by acetyl CoA hypoglycemia since it affects both glycogenolysis and
o ATP is needed gluconeogenesis [Lecturer]
 Endergonic reaction (from 3C to 4C)
OF GLUCSIS
 ATP is a carrier of carbon
IV. MAJOR SUBSTRATES OF GLUCONEOGENESIS
 Carbon in bicarbonate cannot be directly transferred to
pyruvate A. LACTATE
 Occurs in mitochondria of liver and kidney cells where
pyruvate carboxylase is located
→This is why gluconeogenesis only takes place in the liver
and kidneys [2021A Trans]

Figure 9. Conversion of pyruvate to OAA [2021A Trans]


NTK: All carboxylase enzymes require biotin except for AIR Figure 12. Conversion of Lactate to Pyruvate
carboxylase in nucleotide metabolism [Lecturer] Lactate
 formed through anaerobic glycolysis in skeletal muscles
Conversion of OAA to phosphoenol pyruvate (PEP) and erythrocytes.
 Catalyzed by PEPCK  In anaerobic glycolysis there is a net yield of 2 ATP,
 GTP is needed and acts as the phosphate donor NADH is not used in oxidative phosphorylation and is
 Release of CO2 in the process (left with 3 carbon atoms) used to convert pyruvate to lactate by Lactate
 Main route Dehydrogenase (reversible step). [Lecturer]
• Decarboxylation and phosphorylation of OAA to PEP  predominant source of carbon atoms for glucose
happens in the mitochondria synthesis by gluconeogenesis [2021A Trans]
• Mitochondrial PEPCK will catalyze the reaction  Transported to the liver and kidney where it reforms
glucose which becomes available via circulation for
oxidation in tissues. [Harper]

Oxidation of Lactate to Pyruvate


Enzyme: Lactate dehydrogenase

Biochemistry Gluconeogenesis 4 of 10
 Common enzyme during anaerobic glycolysis o In the liver, alanine undergoes transamination via alanine
 Catalyzes the reversible reaction of converting lactate to pyruvic aminotransferase to produce pyruvate
acid o Amino group is donated to -ketoglutarate and becomes
 Redox reaction glumatate
 NAD+ is converted to NADH • Cys, Gly, Thr, Ser and Trp also enters pyruvate [Lecturer’s PPT]
 The cycling of lactate and glucose between liver and peripheral • Pyruvate enters gluconeogenesis
tissues is called the Cori Cycle (Glucose Lactate Cycle)
Entry through TCA
B. GLUCOGENIC AMINO ACIDS  All glucogenic AAs that participates in TCA cycle will enter
 Amino Acids that can be converted into glucose through gluconeogenesis via oxaloacetate
gluconeogenesis [2021 B Trans]  Consumes 4 ATP
 Carbon atoms of AAs derived from proteins can be catabolized o Pyruvate carboxylase reaction is skipped
to pyruvate or to intermediates of the citric acid cycle [Lehninger] o Since 2 moles of Oxaloacetate are also used for
 Alanine and Glutamine are important glucogenic AAs in gluconeogenesis 2 ATP is deducted
mammals that transport amino groups from extrahepatic tissues  Anaplerotic reactions are the reactions that lead to net
to the liver. synthesis of TCA cycle intermediates and support
 Amino group is first removed in the liver mitochondria and the gluconeogenesis as they provide for net synthesis of
remaining carbon skeletons are funneled into gluconeogenesis Oxaloacetate.
[Lehninger]
o Examples are reactions catalyzed by pyruvate carboxylase
 All amino acids except Leucine and Lysine can supply carbon and glutamate dehydrogenase
for net synthesis of glucose by gluconeogenesis [Devlin]  Reactions catalyzed by glumatate-oxaloacetate
aminotransferase are not anaplerotic as net synthesis of a TCA
cycle intermediate is not accomplished.
 Glucogenic AAs enter through 4 intermediates:
-ketoglutarate
→Succinyl CoA
→Fumarate
→Oxaloacetate
C. GLYCEROL
 Can be derived from the degradation of Triacylglycerides
 (TAG) by lipases.
 Hydrolysis of TAGs yield three fatty acids and glycerol [Devlin]
 Results to the formation of the glycerol molecule along with
 three molecules of fatty acid chains.
 Glycerol is converted to an intermediate of glycolysis,
Dihdroxyacetone phosphate [Lecturer]
 Glycerol enters the gluconeogenesis while the remaining fatty
 acids undergo beta oxidation to produce acetyl-CoA
 Consumes: 2 ATP if all carbon of glucose is derived from
glycerol

Figure 13. Glucogenic Amino Acids According to Site of Entry [Lehninger]
*Are also ketogenic amino acids

• Glucogenic amino acids result to net synthesis of either


pyruvate or oxaloacetate
• Both glucogenic and ketogenic AAs also yield ketone body
acetoacetate or acetyl CoA (readily converted to
acetoacetate)
• No pathway exists that can convert these two products to
pyruvate or oxaloacetate. Acetyl CoA can’t be used for net
synthesis of glucose as reaction of pyruvate dehydrogenase
is irreversible. [Devlin]

Figure 15. Production of Glycerol and Free Fatty Acids by Degradation of


Triglycerides by Lipases [2021 A Trans]

Phosphorylation of Glycerol
 Carbon 3 of glycerol is phosphorylated
 Enzyme: Glycerol kinase
 Produces Glycerol-3-phosphate
 Requires ATP as source of phosphate group
 Adipocytes lack glycerol kinase thus they cannot directly
phosphorylate glycerol [Lehninger]
→ Glycerol produced in adipocytes is transported to the liver.

Figure 14. Entry point of Different Glucogenic AAs [Lecturer’s PPT]


Entry through Pyruvate
• Consumes 6 ATP [2021 A Trans]
• Alanine -most important Glucogenic AA
Biochemistry Gluconeogenesis 5 of 10
2. Isomerization of D- Methylmalonyl-CoA to L-
Methylmalonyl-CoA (enantiomers)
 Substrate: D- Methylmalonyl-CoA
 Enzyme: Methylmalonyl-CoA epimerase
 Cofactor: Biotin
Figure 16. Phosphorylation of Glycerol by Glycerol kinase  Product: L-methylmalonyl-coA
 Mutation of the gene encoding for the enzyme may cause
Oxidation of Glycerol-3-P to DHAP deficiency leading to mild to moderate
methylmalonyl aciduria.
 Substrate: Glycerol-3-Phosphate
 Enzyme: Glycerol-3-Phosphate Dehydrogenase
 Cofactor: NAD+ 3. Isomerization of L- Methylmalonyl-CoA to Succinyl-CoA
 Products: Dihydroxyacetone Phosphate (DHAP) + NADH (2o Carbon to 3o Carbon)
 DHAP is readily converted to Glyceraldehyde-3-Phosphate or  Substrate: L-methylmalonyl-coA
produces Fructose-1,6-Bisphosphate which would continue in  Enzyme: methylmalonyl-CoA mutase
gluconeogenic pathway [2021 A Trans]  Cofactor: Vitamin B12 (cyanocobalamin)
 Product: Succinyl-CoA
 Succinyl-CoA can be utilized in the the TCA cycle
forming Oxaloacetate which can then continue in the
gluconeogenesis process (6 ATP)

• Glucogenic amino acids result to net synthesis of either


pyruvate or oxaloacetate
• Both glucogenic and ketogenic AAs also yield ketone body
acetoacetate or acetyl CoA (readily converted to
Figure 17. Oxidation of G-3-P to DHAP by Glycerol-3-Phosphate acetoacetate)
Dehydrogenase • No pathway exists that can convert these two products to
pyruvate or oxaloacetate. Acetyl CoA can’t be used for net
D. PROPIONYL-COA
synthesis of glucose as reaction of pyruvate dehydrogenase
 Propionyl-CoA
is irreversible. [Devlin]
o Produced during degradation of odd-numbered fatty acid
M
chains
V.GLUCOSE FORMING PATHWAYS IN LIVER AND MUSCLE
o good precursor for gluconeogenesis as it yields
oxaloacetate by an anaplerotic pathway  Consists of two pathways
o Also produced during catabolism of valine and isoleucine → Glucose-Lactate Pathway/Cori Cycle
and conversion of cholesterol into bile acids [Devlin] → Glucose-Alanine Pathway/Cahill Cycle
 Converted to Succinyl-CoA in a three-step process.  Liver acts to maintain blood glucose concentration and also
→followed by conversion to oxaloacetate, which is an converts non-carbohydrate metabolites such as lactate, glycerol
intermediate of gluconeogenesis [Lecturer] and amino acids to glucose [Harper]
 Both cycles depend on gluconeogenesis in the liver followed by
delivery of glucose and its use in a peripheral tissue
 Provide continuous supply of glucose to tissues that require it
as their primary energy source [Devlin]
 Peripheral tissues release either alanine or lactate as end
product of glucose metabolism

Figure 18. Conversion of Propionyl-CoA to Succinyl-CoA [2021 A Trans]

1. Carboxylation of Propionyl-CoA to D-Methylmalonyl-CoA

 Substrate: Propionyl-coA and HCO3-


 Enzyme: Propionyl-CoA carboxylase
 Cofactor: Biotin
 Product: D-methylmalonyl-coA Figure 19. Diagram of Glucose-Lactate and Glucose-Alanine Cycle
 Requires ATP [2021 A Trans]
Differences Between the Two Pathways

Biochemistry Gluconeogenesis 6 of 10
 The Cori Cycle uses Lactate as a Substrate and undergoes  2 moles of ATP are produced along with 2 pyruvate and 2
anaerobic glycolysis to produce 2 ATP NADH
→Occurs during exercise when there is high production of  NADH produced will not go to ETC and instead will be
lactate when O2 supply is depleted thus cannot proceed to ETC. oxidized by LDH to convert Pyruvate to Lactate
→Reoxidizes NADH to NAD+  Lactate is then transported to the major gluconeogenic
→Less ATP production in muscle (only 2 ATP) organs [2021 A Trans]
 In the liver, lactate undergoes gluconeogenesis via pyruvate
there is a consumption of 6 ATP. During Gluconeogenic Step
 The Cahill Cycle utilizes Alanine produced in the muscles  Hepatic LDH oxidizes lactate to reform pyruvate which transfers
through the transamination of pyruvate. free electron to NAD forming NADH
→ In contrast to the Cori cyle, it undergoes aerobic
 Pyruvate then proceeds to steps of gluconeogenesis
glycolysis which is more energy efficient for muscles
[Devlin]
 Presents liver with amino nitrogen (ammonium) which must be
disposed as urea [Devlin]
→ In the liver, the urea cycle consumes 4 ATP while
gluconeogenesis consumes 6 ATP, total consumption of 10
ATP

Table 4. Comparison of Cori Cycle and Cahill Cycle

Cori Cycle Cahill Cycle


Glycolytic Anaerobic Aerobic
Process Utilized
Enzyme Lactate Alanine
dehydrogenase Transaminase
Substrate for Lactate Alanine
Figure 20. The Cori Cycle Utilizes LDH to reversibly convert Lactate to
Gluconeogenesis Pyruvate
More Common Fed-State Fasting-state
State Used  Cori cycle utilizes 6 ATPs and produces 2 ATPs
ATP Production 2 ATP 5/7 ATP
(depending on
shuttle used)
ATP Required 6 ATP 10 ATP

 In tissues with mitochondria, reducing equivalents are


transported into mitochondria via the malate-aspartate shuttle
or glycerol-phosphate shuttle for synthesis of ATP via oxidative
phosphorylation.
 In the Malate-Aspartate shuttle NADH remains as NADH as it
enters complex I while in the G-P Shuttle, NADH becomes Figure 21. Diagram of ATP consumption and production in Cori Cycle
FADH2 which enters complex II [Lecturer]
B. CAHILL CYCLE
 As a result, 5 to 7 molecules of ATP can be formed per glucose
molecule in peripheral tissues that participate in alanine cycle  Usually observed in the Fasting State
[Devlin]  Consists of two main processes, Aerobic Glycolysis and
 7 ATP (Malate-Aspartate) and 5 ATP (Glycerol-Phosphate Transamination.
Shuttle) [2021 B Trans]  In Fasting State, there is considerable output of alanine from
skeletal muscle in excess of the amount of muscle protein
A. CORI CYCLE
catabolized [Harper]
 Lactate molecules from RBC and muscle from aerobic
 Formed by transamination of pyruvate produced by glycolysis
glycolysis are transported to the liver and kidneys to generate
of muscle glycogen and is exported to the liver and after
glucose [2021 A Trans]
transamination back to pyruvate, becomes substrate for
 After vigorous exercise, lactate produced by anaerobic
gluconeogenesis [Harper]
glycolysis in skeletal muscle returns to the liver and is converted
 Provides an indirect way of utilizing muscle glycogen to
to glucose which movies back to muscle and is converted to
maintain blood glucose in fasting state [Harper]
glycogen [Lehninger]
 Produces more ATP molecules (5-7 ATP in the muscles), it
 Cycle that serves as bridge connecting anaerobic glycolysis of
also uses more ATP to produce glucose
the muscle and RBC to gluconeogenesis in the liver and
o Amount of ATP depends on shuttle used, Glycerol-
kidneys.
Phosphate or Malate-Aspartate.
 More active in well-fed state due to glycolytic nature of RBCs as
 In the muscles:
well as white muscle fibers [2021 A Trans]
o Glucose in muscles undergoes glycolysis producing 5-7
 Inhibition of cycle may lead to lactic acid accumulation causing
ATP
acidosis
 5 ATP in Skeletal Muscles
 Energy expensive as it uses 6 ATP to produce Glucose from  7 ATP in Cardiac and smooth muscles
Lactate. Pyruvate undergoes transamination using the enzyme
o
 Enzyme: Lactate Dehydrogenase (LDH) Alanine Transferase with Glutamate as main source of
→ NADH generated by glycolysis is used to reduce pyruvate to amino group [2021 A Trans]
lactate [Devlin] o Produced Alanine is transported to the liver.
→ Redox reaction where pyruvate is reduced, lactate is  In the liver:
oxidized. o Alanine is deaminated with the amino part used to
produce urea (requires 4 mol of ATP per mol of urea)
During Anaerobic Glycolysis
Biochemistry Gluconeogenesis 7 of 10
o Remaining carbon skeleton is used to generate the  When one pathway is active, the other is inactive. Applicable in
needed glucose for gluconeogenesis vice versa. [2021A Trans]
o After removal of amino groups of alanine and glutamine
in the liver mitochondria, the carbon skeleton remaining
(pyruvate and -ketoglutarate) are funneled into
gluconeogenesis [Lehninger]
 Once converted to each intermediate, Gluogenic AAs, -
ketoglutarate, Succinyl-CoA and fumarate will become
oxaloacetate first [Lecturer]

During Aerobic Glycolysis


 2 mol of ATP are produced along with 2 pyruvate and 2
NADH [2021 A Trans]
 NADH produced will continue to the ETC (aerobic process) to
produce ATP

During Transamination
 Close relationship between urea synthesis and glucose
synthesis as gluconeogenesis from AAs imposes nitrogen
load on liver [Devlin]
 Muscles degrade amino acids for energy needs and the
resulting nitrogen is transaminated to pyruvate to form alanine
 Alanine undergoes transamination again to form pyruvate
→ Alanine transaminase (ALT) converts glutamate into -
ketoglutarate and vice versa
→ ALT is formerly known as serum-glutamate pyruvate
transaminase (SGPT) [Lecturer]
 Resulting alanine is shuttled to the liver where nitrogen enters
the urea cycle (4 ATP) and pyruvate is utilized to make
glucose (6 ATP)
 The cycle utilizes 10 ATPs and produces 5/7 ATPs Figure 23. Reciprocal Regulation of Gluconeogenesis [Lecturer’s PPT]
 ATP Produced: C. COVALENT MODIFICATION
→ 2 ATP + 2 NADH  Due to reversible phosphorylation
→2 NADH x 1.5 =3 ATP (Glycerol-Phosphate Shuttle)  Rapid
→ 2 NADH x 2.5 =5 ATP (Malate-Aspartate Shuttle)  Glucagon and Epinephrine
o Inhibits glycolysis and stimulates gluconeogenesis
o Increases concentration of cAMP
o Activates cAMP-dependent protein kinase that leads to
inactivation of pyruvate kinase
o Affects the concentration of fructose 2,6-bisphosphate
 Glucagon other functions[Lippincott]:
o Changes in allosteric effectors - lowers the level of
fructose 2,6-bisphosphate, resulting in activation of fructose
1,6-bis-phosphatase and inhibition of phosphofructokinase-
1, thus, favoring gluconeogenesis over glycolysis
o Induction of enzyme synthesis - increases the
transcription of the gene for PEP-carboxykinase, thereby
increasing the availability of this enzyme as levels of its
substrate rise during fasting
Figure 22. Conversion of Alanine to -ketoglutarate and vice versa o Covalent modification - Glucagon binds its G protein-
[Lecturer’s PPT] coupled receptor and, via an elevation in cyclic AMP
(cAMP) level and cAMP-dependent protein kinase activity,
stimulates the conversion of hepatic pyruvate kinase to its
inactive (phosphorylated) form. This decreases the
conversion of PEP to pyruvate, which has the effect of
diverting PEP to the synthesis of glucose.

Figure 23. Diagram of the Cahill Cycle [2021 A Trans]

VI.REGULATION OF GLUCONEOGENESIS
A. RECIPROCAL REGULATION
 Glycolysis and gluconeogenesis share the same pathway but in
opposite directions. [Harper’s]
 Both pathways are reciprocally regulated

Biochemistry Gluconeogenesis 8 of 10
Figure 24. Covalent modification of Glucagon [2021 A Trans] Phosphofructokinase-1
B. RATE OF ENZYME SYNTHESIS  Inhibited by citrate
 Induction and repression of key enzymes take hours.  Activated by AMP
 Enzymes involved catalyze physiologically irreversible non-  90% inhibited at normal levels of ATP
equilibrium reactions.  when ATP is converted to ADP for energy, AMP increases
 Enzymes of gluconeogenesis have low activity when there is  AMP acts as an indicator for the energy status of the cell
super-fluidity of glucose.  AMP increases glycogenolysis activity by activation glycogen
 Enzymes that induce gluconeogenesis, glucocorticoids and phosphorylase
glucagon-stimulated cAMP, are antagonized by the presence of  Inhibition of PFK-1 by ATP = increase in glucose-6-phsophate
insulin.  Increased G6P = inhibits glucose uptake and action of
hexokinase
Table 5. Regulatory and Adaptive Enzymes Associated with Carbohydrate
Metabolism (Table 19-1 in Harper’s) Fructose 2,6-Bisphosphate
Inducer Repressor Activator Inhibitor  Potent positive allosteric activator of PFK-1, by increasing
Glycogenolysis, glycolysis, and pyruvate carboxylation – The PFK-1 affinity for fructose-6-phosphate
following enzymes increases in Carbohydrate feeding and  Inhibitor of fructose 1,6-bisphosphate by increasing fructose
decreases in Fasting & Diabetes 1,6-bisphosphate’s Km
Glycogen Insulin, Glucagon
Synthase glucose-6-
phosphate

Hexokinase Glucose-
6-
Phosphate

Glucokinase Insulin Glucagon


Phosphofru Insulin Glucagon 5’AMP, Citrate,
ktokinase-1 fructose-6- ATP,
phosphate glucagon
, fructose
2,6
bisphosph
ate, Pi

Pyruvate Insulin, Glucagon Fructose ATP,


Kinase fructose 1,6- alanine,
bisphosph glucagon,
ate, insulin norepine
phrine
Pyruvate CoA, Acetyl
dehydroge NAD+, CoA,
nase insulin, NADH+,
ADP, ATP
pyruvate (fatty
acids,
ketone
bodies)
Gluconeogenesis – The following enzymes decreases in
Carbohydrate feeding and increases in Fasting & Diabetes
Pyruvate Glucocortic Insulin Acetyl ADP
Carboxylase oids, CoA
glucagon,
epinephrine
Phosphoeno Glucocortic Insulin Glucagon
lpyruvate - oids, Figure 25. Function of Fructose 2,6-Bisphosphate in Glycolysis and
carboxykina glucagon, in Gluconeogenesis [2021 A Trans]
se epinephrine
Table 6. Summary of Covalent and Allosteric regulators.[Lecturer’s PPT
Glucose-6- Glucocortic Insulin
phosphate oids,
glucagon,
epinephrine

D. ALLOSTERIC EFFECTORS
Acetyl-CoA
 An allosteric activator of pyruvate carboxylase
 Together with pyruvate carboxylase, catalyzes the synthesis of
oxaloacetate from pyruvate
 As acetyl-CoA is formed from pyruvate, this ensures production
of oxaloacetate
 Activation of pyruvate carboxylase = inhibition of pyruvate
dehydrogenase

Biochemistry Gluconeogenesis 9 of 10
VII. PHYSIOLOGICAL IMPORTANCE OF 4. An increased NAD/NADH ratio in alcohol metabolism
GLUCONEOGENESIS inhibits gluconeogenesis by
 Some of the organs in the body requires continues supply of a. Favoring malate Oxaloacteate
glucose b. Favoring pyruvate  lactate
o These organs are supplied by a non-carbohydrate source c. Inhibit DHAP  glycerol 3P
in the process of gluconeogenesis. [Lecturer] d. Inhibit alanine  pyruvate
o These organs are the brain, heart, RBC, renal medulla,
cornea and testis [2021 A Trans] 5. Glycerol from TAG degradation enters gluconeogenesis
 Four gluconeogenic sources: via which intermediate?
o Lactate a. DHAP
o Glucogenic amino acids b. Glyceraldehyde 3P
o Glycerol c. Fructose 1,6 bisP
o Propionyl CoA d. Pyruvate
 Two Major Gluconeogenic Organs:
o Liver 6. Which reaction in gluconeogenesis utilizes energy?
o Kidneys a. Oxaloacetate  PEP
b. Glucose 6P  Glucose
• Only two because of Glucose 6 - phosphatase (the
c. Fructose 1,6bisP  Fructose 6P
enzyme that converts glucose 6 phosphate to glucose).
d. 1,3Phosphoglycerate Glyceraldehyde 3P
Glucose 6 - phosphatase is only found in the endoplasmic
reticulum of liver and kidneys [Lecturer]
7. In the glucose synthesis via Cori cycle and Glu-Ala cycle
 Source of glucose during fasting [2021B Trans] respectively, what is the correct number of ATP pair used?
o 10-18 hours of fasting: glycogenolysis a. 4 & 6
o Prolonged fasting: Gluconeogenesis b. 6 & 4
 CHO Metabolism pathways during fed state [2021B Trans]: c. 6 & 10
o Glycolysis d. 10 & 6
o Glycogenesis
Effect of Alcohol Intake Answers: 1. c., 2. a. 3. b., 4. b. 5. a., 6. a., 7. c.
IX. REFERENCES
 Hypoglycemia
o caused by the consumption of alcohol and drinking Devlin, T. (2011). Textbook of Biochemistry with Clinical Correlations (7th ed).
United States, John Wiley & Sons, Inc.
alcohol after strenuous exercise. Rodwell, V. et al, (2003). Harper’s Illustrated BIochemistry (30th ed). McGraw Hill
o results from the inhibitory effects of alcohol on hepatic Education.
gluconeogenesis and occurs under circumstances of Nelson, D. and Cox, M. (2008) Lehininger Principles of Biochemistry (5th ed) New
York, W.H. Freeman Company
hepatic glycogen depletion [Devlin]. Harvey R., and Ferrier D., (2014) Lippincot’s Illustrated Reviews (6th ed).
 During Alcoholic intoxication: Philadelphia, Wolters Kluwer/Lippincott Williams and Wilkins
o There’s an increase in NADH/NAD ratio that inhibits Dr. Reportoso’s Lecture PPT
2021 A and 2021 B Trans
gluconeogenesis
o NADH is used by lactate dehydrogenase to convert
pyruvate to lactate
o Triacylglycerols are increased because of NADH/NAD
ratio.
 The DHAP will be converted to glycerol 3 –
phosphate and later to glycerol.
 Glycerol is the backbone of TAGs.

VIII.REVIEW QUESTIONS
1. Glucogenic amino acids enter which Krebs cycle
intermediate for glucose synthesis?
a. Citrate
b. Malate
c. Oxaloacetate
d. Succinate

2. Only the liver and kidneys can synthesize glucose from


non-carbohydrate source because of the presence of which
enzyme?
a. Glucose 6-phosphatase
b. Fructose 1,6 Bisphosphatase
c. PEP carboxykinase
d. Pyruvate carboxylase

3. Catherine Matibay, 35 y/o female, had been rescued alive


from a collapsed building 24 hours after an earthquake.
Which of the following molecules is the source of her
metabolic fuel for gluconeogenesis?
a. Acetyl CoA
b. Glycerol
c. Leucine
d. Palmitate

Biochemistry Gluconeogenesis 10 of 10

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