UNIVERSITY COLLEGE OF ENGINEERING,

ANNA UNIVERSITY,
BIT CAMPUS,
TIRUCHIRAPPALLI.

GLYCOLYSIS
PRESENTED BY
CHITRA C
DEPARTMENT OF BIOTECHNOLOGY
2ND YEAR
INTRODUCTION

• Carbohydrates are the major source of energy for the living cells
• The monosaccharide glucose is the central molecule in
Carbohydrate metabolism
• Glucose is utilised as a source of energy, it is synthesized from
non-carbohydrate precursors and stored as glycogen to release
glucose as and when the need arises
CARBOHYDRATE METABOLISM

• Carbohydrate metabolism is a fundamental biochemical process

that ensures a constant supply of energy to living cells
• The most important carbohydrate is glucose, which can be
broken down via glycolysis, enter into the Krebs’s cycle and
oxidative phosphorylation to generate ATP
GLYCOLYSIS

• Extraction of energy from glucose by splitting it to two three –

carbon molecules called Pyruvates
• In cellular respiration, glycolysis is the first stage of this
process
• Glycolysis takes place in the cytosol of the cell and it has ten
steps
• Glycolysis occur in both aerobic and anaerobic organisms
NET REACTION
PHASES IN GLYCOLYSIS

•Glycolysis can be broken down to three phases

1.
Energy requiring/investment phase
2.
Splitting phase
3.
Energy generation phase
GLUCOSE TRANSPORTERS

• Glucose transporters are membrane proteins that facilitate the transport of

glucose across the membrane via facilitated diffusion
• These membrane proteins are bidirectional
• Glucose can also be transported with sodium ions via secondary active
transport pathway in renal proximal tubule and intestine
• These transport mechanisms do not require any ATP
GLUT PROTEINS

There are 4 main classes of GLUT proteins

1. GLUT I
• Found in red blood cells
• Foetus cells
• Blood brain barrier
2. GLUT II
• Found in kidney
• Liver
• Pancreas
 3. GLUT III
• Present in placenta
• Kidney
• Neuron
4.GLUT IV
• Present in muscle
• Adipose

• GLUT IV proteins are insulin dependent

ENERGY INVESTMENT PHASE
STEP 1
• A phosphate group is transferred from ATP to glucose, making glucose-6-
phosphate. It is more reactive than glucose, and the addition of phosphate
also traps glucose inside the cell since glucose with a phosphate is
impermeable to cell membrane. The enzyme Hexokinase acts only at
higher levels of glucose i.e., after meals
STEP 2

• Glucose-6-phosphate is converted to fructose-6-phosphate in the

presence of the enzyme phospho glucose isomerase and mg2+
STEP 3
• A phosphate group is transferred from ATP to fructose-6-phosphate
producing fructose-1,6-bisphosphate. It is an irreversible reaction
catalysed by the enzyme phosphofructokinase. It is a regulatory step in
glycolysis
SPLITTING PHASE
STEP 4
• The six carbon fructose-1,6-bisphosphate is split to two three
carbon compounds, glyceraldyhede 3-phosphate and dihydroxy
acetone phoshate by the enzyme aldolase
STEP 5
• The enzyme Triose phosphate isomerase catalyses the reversible
interconversion of glyceraldyhede-3-phosphate and dihydroxy acetone
phosphate. Thus two molecules of glyceraldyhede-3-phosphate are obtained
from one molecule of glucose
ENERGY GENERATION PHASE
STEP 6
• Glyceraldyhede-3-phosphate dehydrogenase converts glyceraldyhede-
3-phosphate to 1,3-bisphosphoglycerate. It results in the formation of
NADH+H^+ and a high energy compound 1,3-bisphospho glycerate.
Iodoacetate and Arsenate inhibit the enzyme GAPDH
STEP 7
• The enzyme phospho glycerate kinase acts on 1,3-bisphospho
glycerate resulting in the synthesis of ATP and formation of 3-
phospho glycerate. This step is a good example of substrate level
phosphorylation
STEP 8
• 3-phosphoglycerate is converted to 2-phosphoglycerate by
phosphoglycerate mutase. This is an isomerization reaction
STEP 9
• The high energy compound phosphoenol pyruvate is generated
from 2-phosphoglycerate by the enzyme enolase. This enzyme
requires mg2+ or mn2+ and is inhibited by fluoride
STEP 10
• The enzyme pyruvate kinase catalyses the transfer of high
energy phosphate from phosphoenol pyruvate to ADP leading to
the formation of ATP. This step is also called substrate level
phosphorylation
ANAEROBIC CONDITION

• Under anaerobic conditions , pyruvate is reduced by NADH to lactate in the presence of the
enzyme Lactate dehydrogenase
• The NADH utilized in the step is obtained from the reaction catalysed by glyceraldyhede -3-
phosphate dehydrogenase
• The formation of lactate allows the regeneration of NAD+ which can be reused by
Glyceraldyhede -3-phosphate dehydrogenase so that glycolysis proceeds even in the absence of
Oxygen to supply ATP
• Glycolysis in erythrocytes leads to lactate production
AEROBIC CONDITION

• In the presence of oxygen, pyruvate generated is converted to Acetyl- coA

• Acetyl- coA then enters into Krebs cycle and oxidative phosphorylation to
generate ATP
• Most cancer cells produce energy predominantly through Aerobic
glycolysis consisting of high level of glucose uptake and glycolysis
followed by lactic acid fermentation taking place in the cytosol even in the
presence of abundant oxygen
 PRODUCTION IN GLYCOLYSIS

• One glucose molecule yields Four ATP molecules in total

during glycolysis
• Since 2 ATP molecules are used up in the first phase of
glycolysis, there is a net gain of 2 ATP molecules
• Two NADH molecule is produced
REGULATION OF GLYCOLYSIS

• The three enzymes responsible for catalysing the irreversible

reactions regulate glycolysis
1. Hexokinase
2. Phosphofructokinase
3. Pyruvate kinase
HEXOKINASE

Allosteric regulation
• Hexokinase is activated by high amounts of glucose in the cell
• It is inhibited by Glucose-6-phosphate
Hormonal regulation
• Glucokinase is an inducible enzyme which gets induced with the help
of insulin
• Glucagon in turn try to suppress both hexokinase and glucokinase
PHOSPHOFRUCTOKINASE (PFK I)
Allosteric regulation
• The enzyme gets activated by Fructose-2,6-bisphosphate , ADP and pi
• The enzyme gets inhibited by ATP and Citrate(feed back)
Role of fructose-2,6-bisphosphate in glycolysis
• It is the most important regulatory factor for controlling PFK I and glycolysis in the
liver
• Fructose-2,6-bisphosphate is synthesized from fructose-6-phosphate by the enzyme
Phosphofructokinase II
• The generated fructose-2,6-bisphosphate stimulates Phosphofructokinase to take part
in glycolysis
• Another enzyme Fructose-2,6-bisphosphotase, inhibits fructose-2,6-bisphosphate by
converting back to fructose-6-phosphate
Allosteric regulation PFK I
Hormonal regulation

•Glucagon phosphorylates the enzyme

Phosphofructokinase II (PFKII) and Fructose-2,6-
bisphosphatase(F 2,6-bp) which activatesF 2,6-bp and
inhibits PFK II
•Insulin in turn dephosphorylates both enzymes and
results in the stimulation of PFK I to carry out glycolysis
PYRUVATE KINASE

Allosteric regulation
• The enzyme pyruvate kinase is activated by Fructose-6-phosphate
• The enzyme is inhibited by ATP and increased concentration of Acetyl- coA
Hormonal regulation
• Glucagon phosphorylates the enzyme pyruvate kinase which inhibit them
• Insulin dephosphorylates the enzyme which activates them
• The hormone glucagon inhibits hepatic glycolysis by this mechanism
CONCLUSION

• Carbohydrate metabolism
• Transport of glucose
• Glycolysis
• Steps in glycolysis
• Anaerobic and aerobic
• Regulation of glycolysis