Glycolysis, Gluconeogenesis, and the

Pentose Phosphate Pathway

Chapter 14

Lehninger
Principles of Biochemistry

5th edition

By Nelson & Cox

 Glucose •Central place in metabolism

Major fates in animals and vascular plants:

1. Good fuel (___ standard free energy).

2. Storage: starch or glycogen

(high MW polymers): 2
- does not affect cytosolic _________
could be used in future when energy 3
demand increases.
1
3. Oxidation via _______________
___________
 Oxidation of glucose
C6H12 O6 + 6O2  6CO2 + 6H2O

Glycolysis

•During this reaction some of the free energy

is conserved as _____ or _______.

Fermentation: Anaerobic breakdown of glucose and

others to obtain energy as ATP.

Most ______ form of energy obtaining process?

Glycolysis aerobic

-Breakdown of 6-C glucose into

-two molecule of the 3-C __________
-occurs in 10 steps.

- First five (1-5) are ________ phase

- energy investment

- Last five (6-10) are ________ phase

 Preparatory Phase of Glycolysis
1. Phosphorylation of Glucose

Hexokinase – ATP & Mg2+

- ______ reversible (under intracellular condition).

- Hexokinase kinase transfers terminal phosphoryl group

from _____to an acceptor.

- Hexokinase is present in ______of all organisms.

p532

negative charged sugar: cannot _____ the cell.

 Preparatory Phase of Glycolysis
2. Conversion of Glucose 6-Phosphate to Fructose 6-Phosphate

Phosphohexose isomerase – Mg2+

- Isomerization reaction

- Freely __________

If slight ______ in product, reverses reaction.
 Preparatory Phase of Glycolysis
3. Phosphorylation of Fructose 6-P to Fructose 1,6-bisphosphate


Phosphofructokinase 1 (PFK-1) – ATP & Mg++
– Essentially ___________ / first __________
step
• Fructose 1,6-bisphosphate is targeted for
glycolysis.
- It is a major point of _________ in glycolysis.
- ATP & PEP __________ regulate PFK1

p526

Fructose 6-phosphate

G´° = –14.2 kJ/mol

 Preparatory Phase of Glycolysis
4. Cleavage of Fructose 1,6-bisphosphate
Aldolase
Freely _________: yields two ________ trioses

- Lower concentration of reactants for reverse reaction.

(Products are _____________up by subsequent reaction)
p533
 Preparatory Phase of Glycolysis
5. Interconversion of Triose Phosphates
Triose phosphate isomerase
- Only G3-P (Glyceraldehyde 3-phosphate) can be ________
degraded.
- Dihydroxyacetone phosphate is readily converted into
_______.
- Freely __________ reaction but moves in forward direction
because product is readily used up.
 Payoff Phase of Glycolysis
6. Oxidation of G3-P to 1,3-Bisphosphoglycerate
Glyceraldehyde 3-phosphate dehydrogenase
- Ist step in payoff phase (energy conserving) leading to
ATP formation later on.
- Freely ________
- Acceptor of hydrogen is ________ yielding NADH.

p535

G’˚ = 6.3 kJ/mol

 Payoff Phase of Glycolysis
7. Phosphoryl transfer from 1,3-Bisphosphoglycerate to ADP

Phosphoglycerate kinase – Mg2+

• ___________ level phosphorylation.
(different from respiration-linked synthesis of ATP)
• Freely reversible, but highly _______________

p536
 Payoff Phase of Glycolysis
8. Conversion of 3-Phosphoglycerate to 2-Phosphoglycerate

Phosphoglycerate mutase – Mg2+

- Freely reversible
- Shifts phosphoryl group between C-3 and C-2
 Payoff Phase of Glycolysis
9. Dehydration of 2-Phosphoglycerate to Phosphoenolpyruvate

Enolase (dehydratase)
- Freely reversible,
- Removes _____ from 2-Phosphoglycerate to yield ___.
-

PEP

Sets up ____ substrate-level phosphorylation

 Payoff Phase of Glycolysis
10. Transfer of phosphoryl group from PEP to ADP

Pyruvate kinase (PK) – ADP & Mg2+

Yield is ________per glucose molecule.
(substrate-level phosphorylation)

PK is ____ reversible

Pyruvate
kinase
p538
Mg 2+ , K+

Products of Glycolysis?


• ________
• ________
• ________

What happens to these three products?


– ATP is ________ source.

– NADH makes more ATP if Oxygen is
_________.
– Pyruvate is further _____ if Oxygen is
available

- contains majority of the energy.


–

Anaerobic conditions produce _______ result.

Other carbohydrates:

1.Storage polysaccharides:
____________ & __________.

2. disaccharides:

_________, ______, _______ & ________.

3. monosaccharides:

__________, mannose, galactose

Feeder pathways for Glycolysis

The trick is to make:

Glycolytic _______________

 may involve one or more specific
_______.
Feeder pathways for Glycolysis
 1. Catabolism of Polysaccharides
Starch (______), and Glycogen (______) can be used
in the same _________.

•Digestive tract or sprouting seeds http://en.wikipedia.org/wiki/File:Amylose2.svg

- hydrolyzed by __________
- maltose & glucose released

•Intracellular
- transfer of glucose to Piby
___________
- glucose 1-phosphate released

Dietary glycogen/starch (similar structure) is broken

by hydrolysis through use of _________ not Pi.
ntracellular
Glycogen and Starch
Two enzymes needed

1. Glycogen Phosphorylase :
catalyzes an attack by Pi on
the ____________ bond that
joins last two glucose
residues at the
_____________ end.

- glucose 1-phosphate

This enzyme works until it

reaches a glucose that is within
_____residues of an:
_______branch point.
(glycogen & amylopectin).
 Glycogen and Starch
Debranching

• Debranching enzyme
has 2 activities
– Transferase – moves
______ glucose units
from the _____ branch
to the ______ branch


– Glucosidase – releases a
free _______ by
hydrolyzing the

-1,6-glycosidic bond
 Glycogen and Starch Detail
Glycogen
Phosphorylase
2. Phosphoglucomutase

- Converts Glucose 1-phosphate

to Glucose ___-phosphate

Glucose 1-phosphate 
Glucose ___-phosphate

- Freely ________ reaction

Hydrolysis of dietary polysaccharide:
(breakdown of starch)

•Salivary  -amylase: smaller _______________

(digestion begins in mouth).

•Pancreatic  -amylase: maltose & maltotriose

(di- and trisaccharides of glucose) and dextrins.

•Maltose and dextrins are further degraded by

enzymes
into ___________.

•Disaccharides must be hydrolyzed to

____saccharides
before entering cells.
2. Catabolism of Disaccharides
Disaccharides are hydrolyzed by specific __________

•Maltose – from breakdown of starches

Maltase – 2 ________ units are released

•Lactose – milk sugar, major source of energy for

infants in mammals.

________ (intestine) – Glucose & Galactose

•Sucrose (glucose + fructose) – cane or beet sugar

Table sugar, sweetener in all regular soft drinks

________ – Glucose & Fructose

 Lactose intolerance ?

Common among adults of most humans except

____________and parts of _______?

actase (or -galactosidase) is low or missing in many

Arabian,
Jewish,
Bantu,
Japanese,
Native Americans, and
Filipino
adults – are all _______________.
3. Catabolism of Monosaccharides

•Mannose – from breakdown of complex

________________.

•Galactose – from breakdown of _______.

•Fructose – from breakdown of sucrose.

- Main sugar in _______ and _______
 Mannose catabolism

Two enzymes convert mannose

to __________________

1.hexokinase
- Same enzyme that acts
on glucose

2. phosphomannose
isomerase

Fig 14.9
(From Lactose)
Galactose catabolism


Three enzymes convert galactose to
_______________



1. Galactokinase

2. UDP-glucose: galactose 1-
phosphate uridylyltransferase

3. UDP-glucose 4-epimerase



Defect in any of the enzymes

is cause of __________________.

Problems in Galactose Metabolism
Infants lacking: hexose 1-phosphate uridylyltransferase

- have a form of _____________

- Must _______ milk and other sources of galactose.

Adults have galactose 1-phosphate uridylyltransferase which

can metabolize galactose.

Some infants lack galactokinase– 2nd type of galactosemia



– _________ overcome as adults

Fructose catabolism

Fructose (fruits) first gets

converted to Fructose 6-
phosphate and then enters
___________.

Enzyme: ___________

This is major way of entry of

Fructose in glycolysis in _____
and _________.

But different in _______,

where most of Fructose is
catabolized.
 Fructose catabolism
in Liver



3 enzymes make glycolytic
metabolites


– ___________


– Fructose 1-Phosphate
Aldolase


– _____________
 Catabolic fates of Pyruvate

First route:
Pyruvate  _______.

Acetyl group is then oxidized

3 2 completely to CO2 by _______
1 _________________

Second route:
Reduction to lactate via lactic
acid ___________

Third route:
leads to formation of _______
via ethanol ______________
Fate of Pyruvate under Anaerobic Conditions

•Under hypoxic condition, NADH from glycolysis cannot be

______________ by O2.
•
•Non-availability of _____ as an electron
acceptor for oxidation of G3-P can
__________the Glycolysis.
Fate of Pyruvate under Anaerobic Conditions

•Under these conditions, NAD+ is regenerated from _______ by

reduction of pyruvate to _________.

Lactate formation
favored (______ –G´)

______change in
NAD+ or NADH
Fate of Pyruvate under Anaerobic Conditions

Fermentation – 2 steps-
Ist step- Pyruvate decarboxylase requires Thiamin
pyrophosphate (TPP) (co-enzyme) and Mg2+

TPP is a coenzyme derived from vitamin ____.

- Vitamin deficiencies leads to? ______
- TPP has role in cleavage of bonds adjacent to
carbonyl group.
-

IInd step - Alcohol dehydrogenase requires - _____

Freely reversible
Fate of Pyruvate under Anaerobic Conditions

Summary

Glucose + 2ADP + 2Pi 

__ lactate + 2 ATP + 2H2O

or
__ ethanol + 2 ATP + 2H2O
Anabolic fate of Pyruvate

Pyruvate can also provide

carbon skeleton for the synthesis of the
amino acid ________.
Gluconeogenesis
•Some mammalian tissue depend ________
glucose for their metabolic energy.

e.g. Human _____, nervous systems, renal medulla,

erythrocytes, embryonic tissue.

All need glucose from the blood as major/sole

source of fuel.

Brain needs ~_____ glucose/day which

is half of all the stored glucose.
Gluconeogenesis

•Body fluids carry only:

•~___g free glucose

• glycogen can provide 180-200 g of free

glucose.
- Little more then a ___-day supply.

•Glucose if not obtained from the diet, body

must make _________from the
____-carbohydrate precursors:

Gluconeogenesis
Gluconeogenesis

•Gluconeogenesis occurs in _____

- animals,
- plants,
- fungi and
- microorganisms.

•Reactions are essentially ____ in all tissues

and all species.
Gluconeogenesis

Storage of
carbohydrates

Biosynthesis of
carbohydrates

Glucogenic
molecules
Substrates of Gluconeogenesis

•Pyruvate, lactate and other non-carbohydrate

precursors can be substrates:
•most of the ____________,
•_______ and all the
•_______cycle intermediates.

• Fatty acids are _____ the substrates in animals.

• Lysine and Leucine are the only amino acids that

are ____ substrates.
Site of Gluconeogenesis in Animals
•The organ which consumesmost of the
glucose e.g. brain and muscle,
- carry out very little glucose __________.
•
•Major site of gluconeogenesis in animals:
Liver : ___%
Kidney : ___%

The synthesized glucose is released into

_____ which gets transported to the place
of need.
Gluconeogenesis Not merely _____ of Glycolysis

In some ways gluconeogenesis

is reverse of glycolysis but not
all steps are similar.

What is different?

Both are _____________ and

occur largely in _______.
Reactions specific to Gluconeogenesis

___ different reactions in

gluconeogenesis.

- Other ______ are same

as glycolysis.

Three different enzyme

catalyzed reactions are
utilized.

 start here
Reactions specific to Gluconeogenesis
1.First: Conversion of pyruvate to ____:
initiates gluconeogenesis.

Occurs in two unique steps:

A.Pyruvate carboxylase
catalyzes first converting
pyruvate to oxaloacetate.

- First pyruvate is transported

to _____________.
Reactions specific to Gluconeogenesis

Since oxaloacetate is required in cytosol but

it _________ be transported to cytosol.
-so it is converted to_________by malate dehydrogenase.

Oxaloacetate + NADH + H+  L-malate + NAD+

Malate is transported out to _________ and is

reoxidized to oxaloacetate.

Malate + NAD+  oxaloacetate + NADH + H+

Reactions specific to Gluconeogenesis

B. Then PEP carboxykinase catalyzes the conversion of

oxaloacetate to PEP in cytosol.

Reversible reaction but

effectively irreversible
due to ______ effective
concentration of PEP.
2nd specific reaction: Gluconeogenesis

Conversion of:

Fructose 1,6-bisphosphate
to

Fructose 6-P

Catalyzed by _________

- Irreversible hydrolysis of
C-1 phosphate
3rd specific reaction : Gluconeogenesis

3. Conversion of Glucose 6-P

to Glucose

catalyzed by Glucose 6-
phosphatase requires Mg2+

Glucose is transported via

_____ to brain and muscle.
Gluconeogenesis is a expensive but essential step

2 Pyruvate + __ ATP+___ GTP+ __NADH + 2 H+ + 4 H2O

 glucose + 4ADP + 2GDP + 6Pi + 2NAD+

P 556
Pentose Phosphate Pathway
- A __________for producing NADPH:
needed in synthesis of
• _________
• _________
2
-Provides a pool of ribose
5-phosphate for
synthesis of 3
•________.
1
 Pentose Phosphate Pathway: Starting point
From Glycolysis: Phosphorylation of glucose


Hexokinase – ATP & Mg2+

- ___ reversible (under intracellular condition)

- Hexokinase is present in _______of all organisms.
Pentose Phosphate Pathway
 Pentose phosphate pathway
Oxidative Phase


1. Glucose 6-P dehydrogenase–

requires _______& Mg2+

Makes 6-phosphogluconolactone

and ________ (1st molecule)



Genetic defect causes disease?

Read. Common!



2. Lactonase– Mg2+

Makes 6-phosphogluconate

 Pentose phosphate pathway
Oxidative Phase
Fig 14.21bottom

3. 6-Phosphogluconate
dehydrogenase :

- requires NADP+ & Mg2+

- Products:
- ribulose 5-phosphate
- _______ (2nd molecule)

4. Pentose phosphate isomerase:

- converts Ribulose 5-Pto its

isomer ________________
Non-oxidative phase
Pentose phosphate pathway: Non-oxidative phase
Recycles PP to G6-P
In tissue which primarily require NADPH:
Pentose-P are recycled into ____________.

-First step: epimerization of Ribulose 5-P to Xylulose 5-P

- enzyme ribose 5-phosphate epimerase

Epimers?
Non-oxidative phase Recycles PP to G6P

-Series of rearrangements of carbon skeletons:

Six , 5-C sugar phosphates are converted to
_____, 6-C sugar phosphates.