BIO 202 Biochemistry II

by
Seyhun YURDUGÜL

Lecture 3
Carbohydrate Metabolism I
Glycolysis
 CONTENT OUTLINE
• Brief information on the carbohydrates
• Glycolysis
• Pathways by chart illustration
• Linker pathways on brief
• Regulation
Carbohydrates meets with our body

• Dietary carbohydrate from which humans

gain energy
• enter the body in complex forms,
• such as: disaccharides
• and the polymers:
• a)starch (amylose and amylopectin)
• b) glycogen.
 Carbohydrates meets with our body
• polymer cellulose:
• also consumed but not digested.
• The first step in the metabolism of digestible
carbohydrate:
• the conversion of the higher polymers to
simpler, soluble forms:
• that can be transported across the intestinal
wall and delivered to the tissues.
Carbohydrates meets with our body

• The breakdown of polymeric sugars

begins in the mouth.
• Saliva has a slightly acidic pH of 6.8;
• and contains lingual amylase that begins
the digestion of carbohydrates.
Carbohydrates meets with our body

• The action of lingual amylase:

• limited to the area of the mouth and the
esophagus;
 Carbohydrates
• virtually inactivated by the much
stronger acid pH of the stomach.
• Once the food has arrived in the stomach,
• acid hydrolysis contributes to its
degradation;
• specific gastric proteases
• and lipases aid this process for proteins
and fats, respectively.
 Carbohydrates
• The mixture of gastric secretions, saliva, and food,
known collectively as chyme, moves to the
intestine
• The resultant glucose and other simple
carbohydrates:
• transported across the intestinal wall to the
hepatic portal vein
 Carbohydrates
• and then to liver parenchymal cells and other
tissues.
• there converted to fatty acids,
• amino acids,
• and glycogen,
• or else oxidized by the various catabolic
pathways of cells.
 Structurally
• Starch (in plants)
• and glycogen (in animals)
• compose of one single unit: glucose
• So how we can degrade glucose?
 Glycolysis
• the sequence of reactions that converts
glucose into pyruvate;
• with the concomitant production of a
relatively small amount of ATP.
 Glycolysis
• Glycolysis can be carried out anaerobically
(in the absence of oxygen)
• and an especially important pathway for
organisms that can ferment sugars.
 Glycolysis
• is the pathway utilized by yeast to produce the
alcohol found in beer.
• serves as a source of raw materials for the
synthesis of other compounds.
• e.g. 3 phosphoglycerate can be converted into
serine,
• while pyruvate can be aerobically degraded by the
Krebs or TCA cycle;
• to produce much larger amounts of ATP.
 Glycolysis
• Oxidation of glucose
• oxidized to either lactate
• or pyruvate.
• Under aerobic conditions,
• the dominant product in most tissues is pyruvate
• and the pathway:
• known as aerobic glycolysis.
 Glycolysis
• When oxygen is depleted,
• as for instance during prolonged vigorous
exercise,
• the dominant glycolytic product in many
tissues:
• lactate
• and the process: anaerobic glycolysis.
The Energy Derived from
Glucose Oxidation
• Aerobic glycolysis of glucose to pyruvate,
• requires two equivalents of ATP
• to activate the process,
• with the subsequent production of four equivalents
of ATP
• and two equivalents of NADH.
The Energy Derived from Glucose
Oxidation

• Thus, conversion of one mole of glucose

• to two moles of pyruvate :
• accompanied by the net production of two moles each of
ATP and NADH.
• Glucose + 2 ADP + 2 NAD+ + 2 Pi -----> 2 Pyruvate + 2
ATP + 2 NADH + 2 H+
The Energy Derived from Glucose
Oxidation
• The NADH generated during glycolysis:
• used to fuel mitochondrial ATP synthesis via
oxidative phosphorylation,
• producing either two or three equivalents of ATP
depending upon whether the glycerol phosphate
shuttle
• or the malate-aspartate shuttle:
• used to transport the electrons from cytoplasmic
NADH into the mitochondria.
The Energy Derived from Glucose
Oxidation

• The net yield from the oxidation of 1 mole

of glucose to 2 moles of pyruvate:
• either 6 or 8 moles of ATP.
The Energy Derived from Glucose
Oxidation
• Complete oxidation of the 2 moles of
pyruvate, through the TCA cycle:
• yields an additional 30 moles of ATP;
• the total yield:
• either 36 or 38 moles of ATP from the
complete oxidation of 1 mole of glucose to
CO2 and H2O.
The Individual Reactions of
Glycolysis
• The pathway of glycolysis:
• can be seen as consisting of 2 separate phases.
• The first is the chemical priming phase requiring
energy in the form of ATP,
• and the second is considered the energy-yielding
phase.
The Individual Reactions of
Glycolysis

• In the first phase, 2 equivalents of ATP are used to

convert glucose to:
• fructose 1,6-bisphosphate (F 1,6 BP).
• In the second phase F 1,6 BP is degraded to
pyruvate,
• with the production of 4 equivalents of ATP
• and 2 equivalents of NADH.
 The Hexokinase Reaction:
First Step of Glycolysis

– The ATP-dependent phosphorylation of

glucose to form glucose 6-phosphate (G6P):
– the first reaction of glycolysis,
– catalyzed by tissue-specific isoenzymes known
as hexokinases.
 The Hexokinase Reaction:
First Step of Glycolysis

The phosphorylation accomplishes two goals:

☻ First, the hexokinase reaction converts:
☻ nonionic glucose into an anion that is trapped in the
cell,
☻ since cells lack transport systems for phosphorylated
sugars.
☻Second, the otherwise biologically inert glucose
becomes activated into a labile form;
☻ capable of being further metabolized.
 The Hexokinase Reaction:
First Step of Glycolysis

☻ Four mammalian isozymes of hexokinase are

known
☻(Types I - IV),
☻ with the Type IV isozyme often referred to as
glucokinase.
☻ Glucokinase is the form of the enzyme found
in hepatocytes.
 Explanation of the previous figure
• Comparison of the activities of hexokinase and
glucokinase.
• The Km for hexokinase is significantly lower
(0.1mM) than that of glucokinase (10mM).
• This difference ensures that non-hepatic tissues
(which contain hexokinase) rapidly and;
• efficiently trap blood glucose within their cells by
converting it to glucose-6-phosphate.
 Liver and glycolysis
• One major function of the liver:
• to deliver glucose to the blood;
• by a glucose phosphorylating enzyme
(glucokinase);
• whose Km for glucose :
• sufficiently higher than the normal
circulating concentration of glucose (5mM).
 Liver and glycolysis
• This feature of hepatic glucokinase:
• allows the liver to buffer blood glucose.
• After meals,
• when postprandial blood glucose levels are
high,
• liver glucokinase :
• significantly active,
• which causes the liver preferentially to trap
• and to store circulating glucose.
 Other glucose supplies in the body
• When blood glucose falls to very low levels,
• tissues such as liver and kidney which contain
glucokinases but are not highly dependent on glucose:
• do not continue to use the glucose supplies that remain
available.
• At the same time, tissues such as the brain, which are
critically dependent on glucose,
• continue to scavenge blood glucose using their low Km
hexokinases,
• and as a consequence their viability is protected.
 An important glucose provider
pathway: Gluconeogenesis
• Under various conditions of glucose deficiency,
• such as long periods between meals,
• the liver:
• stimulated to supply the blood with glucose through
the pathway of gluconeogenesis.
• The levels of glucose produced during
gluconeogenesis :
• insufficient to activate glucokinase,
• allowing the glucose to pass out of hepatocytes;
• and into the blood.
 Regulation of hexokinase and
glucokinase
• The regulation activities of them:
• different.
• Hexokinases I, II, and III : allosterically
inhibited by product (G6P) accumulation,
• whereas glucokinases are not.
 Regulation of hexokinase and
glucokinase
• The latter further insures liver accumulation
of glucose stores during times of glucose
excess,
• while favoring peripheral glucose
utilization;
• when glucose is required to supply energy
to peripheral tissues.
Phosphohexose Isomerase:
The Second Reaction
• The second reaction of glycolysis :
• an isomerization,
• in which G6P is converted to fructose 6-
phosphate (F6P).
• The enzyme catalyzing this reaction:
• phosphohexose isomerase
• also known as phosphoglucose isomerase.
Phosphohexose Isomerase:

• The reaction : freely reversible at normal cellular

concentrations of the two hexose phosphates
• and thus catalyzes this interconversion during
glycolytic carbon flow
• and during gluconeogenesis
 6-Phosphofructo-1-Kinase
(Phosphofructokinase-1, PFK-1):

• The next reaction of glycolysis

• involves the utilization of a second ATP:
• to convert F6P to fructose 1,6-bisphosphate
(F1,6BP).
• catalyzed by 6-phosphofructo-1-kinase,
• better known as phosphofructokinase-1 or PFK-
1.
 6-Phosphofructo-1-Kinase
(Phosphofructokinase-1, PFK-1):
• This reaction is not readily reversible
• because of its large positive free energy:
• ΔG = +5.4 kcal/mol in the reverse direction.
• Nevertheless, fructose units readily flow in the
reverse (gluconeogenic) direction;
• because of the presence of the hydrolytic enzyme,
• fructose-1,6-bisphosphatase (F-1,6-BPase).
 6-Phosphofructo-1-Kinase
(Phosphofructokinase-1, PFK-1):
• The presence of these two enzymes in the same
cell compartment:
• provides an example of a metabolic futile cycle,
• which if unregulated would rapidly deplete cell
energy stores.
 6-Phosphofructo-1-Kinase
(Phosphofructokinase-1, PFK-1):

• However, the activity of these two enzymes is so

highly regulated that :
• PFK-1: considered to be the rate-limiting enzyme
of glycolysis
• and F-1,6-BPase : considered to be the rate-
limiting enzyme in gluconeogenesis.
 Aldolase:

• catalyses the hydrolysis of F1,6BP:

• into two 3-carbon products:
• dihydroxyacetone phosphate (DHAP)
• and glyceraldehyde 3-phosphate (G3P).
• aldolase reaction proceeds readily in the reverse
direction,
• being utilized for both glycolysis and
gluconeogenesis
 Triose Phosphate Isomerase:
• The two products of the aldolase reaction
equilibrate readily in a reaction;
• catalyzed by triose phosphate isomerase.
• Succeeding reactions of glycolysis utilize
G3P as a substrate;
• thus, the aldolase reaction:
• pulled in the glycolytic direction by mass
action principals.
Glyceraldehyde-3-Phosphate
Dehydrogenase:

• The second phase of glucose catabolism:

• features the energy-yielding glycolytic reactions
that produce ATP and NADH.
Glyceraldehyde-3-Phosphate
Dehydrogenase:
• In the first of these reactions,
• glyceraldehyde-3-P dehydrogenase (G3PDH):
• catalyzes the NAD+-dependent oxidation of G3P
to 1,3-bisphosphoglycerate (1,3BPG)
• and NADH.
• The G3PDH reaction : reversible,
• and the same enzyme catalyzes the reverse
reaction during gluconeogenesis.
Phosphoglycerate Kinase:

• The high-energy phosphate of 1,3-BPG:

• used to form ATP and 3-phosphoglycerate
(3PG);
• by the enzyme phosphoglycerate kinase.
• the only reaction of glycolysis or gluconeogenesis
that involves ATP
• and yet is reversible under normal cell
conditions.
 Phosphoglycerate Kinase:

• Associated with the phosphoglycerate kinase

pathway:
• an important reaction of erythrocytes,
• the formation of 2,3-bisphosphoglycerate,
2,3BPG (see Figure below):
• by the enzyme bisphosphoglycerate mutase.
• 2,3BPG is an important regulator of
hemoglobin's affinity for oxygen.
Phosphoglycerate Kinase:
• Note that 2,3-bisphosphoglycerate phosphatase
degrades 2,3BPG to 3-phosphoglycerate,
• a normal intermediate of glycolysis.
• The 2,3BPG shunt thus:
• operates with the expenditure of 1 equivalent of
ATP per triose passed through the shunt.
• The process: not reversible under physiological
conditions.
Phosphoglycerate Mutase and
Enolase:
• The remaining reactions of glycolysis:
• are aimed at converting the relatively low energy
phosphoacyl-ester of 3PG;
• to a high-energy form
• and harvesting the phosphate as ATP.
Phosphoglycerate Mutase and
Enolase:
• The 3PG is first:
• converted to 2PG by phosphoglycerate mutase
• and the 2PG conversion
• to phosphoenoylpyruvate (PEP):
• catalyzed by enolase
 Pyruvate Kinase:

• The final reaction of aerobic glycolysis

• catalyzed by the highly regulated enzyme
pyruvate kinase (PK).
• In this strongly exergonic reaction,
• the high-energy phosphate of PEP :
• conserved as ATP.
 Pyruvate Kinase:
• The loss of phosphate by PEP:
• leads to the production of pyruvate in an
unstable enol form,
• which spontaneously tautomerizes to the more
stable, keto form of pyruvate:
• contributes a large proportion of the free energy
of hydrolysis of PEP.
 Anaerobic Glycolysis

• Under aerobic conditions,

• pyruvate in most cells:
• further metabolized via the TCA cycle.
• Under anaerobic conditions
• and in erythrocytes under aerobic conditions,
• pyruvate:
• converted to lactate by the enzyme lactate
dehydrogenase (LDH),
• and the lactate: transported out of the cell into the
circulation.
 Anaerobic Glycolysis
• The conversion of pyruvate to lactate,
• under anaerobic conditions, provides the cell with
a mechanism for the oxidation of NADH
(produced during the G3PDH reaction):
• to NAD+;
• which occurs during the LDH catalyzed reaction.
• This reduction is required since NAD+:
• a necessary substrate for G3PDH;
• without which glycolysis will cease.
 Anaerobic Glycolysis
• Normally, during aerobic glycolysis:
• the electrons of cytoplasmic NADH:
• transferred to mitochondrial carriers of the
oxidative phosphorylation pathway;
• generating a continuous pool of cytoplasmic
NAD+.
 Anaerobic Glycolysis

• Aerobic glycolysis :
• generates substantially more ATP per mole of
glucose oxidized than;
• does anaerobic glycolysis.
• The utility of anaerobic glycolysis;
• to a muscle cell when it needs large amounts of
energy stems from the fact that:
• the rate of ATP production from glycolysis is
approximately 100X faster than;
• from oxidative phosphorylation.
 Anaerobic Glycolysis
• During exertion muscle cells:
• do not need to energize anabolic reaction
pathways.
• The requirement to generate the maximum amount
of ATP,
• for muscle contraction,
• in the shortest time frame.
• This is why muscle cells derive almost all of the
ATP consumed during exertion;
• from anaerobic glycolysis.
 LITERATURE CITED
• Devlin,T.M. Textbook of Biochemistry with
Clinical Correlations,Fifth Edition,Wiley-Liss
Publications,New York, USA, 2002.
• Lehninger, A. Principles of Biochemistry, Second
edition, Worth Publishers Co., New York, USA,
1993.
• Matthews, C.K. and van Holde, K.E.,
Biochemistry, Second edition, Benjamin /
Cummings Publishing Company Inc., San
Francisco, 1996.