Section 1:

Enzyme: Glycosidase
Function: Breaks glycosidic bonds in polysaccharides using water (hydrolysis) to release
monosaccharides.
Role: Key enzyme in carbohydrate digestion.

Enzyme: α-amylase
Function: Breaks α(1→4) glycosidic bonds in starch during chewing.
Role: Begins carbohydrate digestion in the mouth by producing smaller branched
oligosaccharides.
Limitation: Cannot break α(1→6) bonds.

Pancreatic α-amylase​
Function: Continues breaking α(1→4) glycosidic bonds in the small intestine.​
Role: Picks up where salivary α-amylase left off.

Brush Border Enzymes​

Function: Final digestion of disaccharides to monosaccharides.​
Specific roles:

●​ Maltase breaks maltose → 2 glucose​

●​ Sucrase breaks sucrose → glucose + fructose​

●​ Lactase breaks lactose → glucose + galactose​

●​ Isomaltase breaks α(1→6) branches in starch

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Section 2:

Enzymes: Hexokinase and Glucokinase

Function: Phosphorylate glucose to form glucose-6-phosphate (G6P) using ATP.

Hexokinase

●​ Works in most tissues​

●​ Low Km (high affinity)​

●​ Inhibited by its product (G6P)​

Glucokinase

●​ Found in liver and pancreas​

●​ Higher Km (lower affinity, responds to high glucose)​

●​ Not inhibited by G6P

Role: First regulatory step in glycolysis, trapping glucose inside cells as G6P.

Enzyme: Phosphofructokinase-1 (PFK-1)

Function: Catalyzes the phosphorylation of fructose-6-phosphate to

fructose-1,6-bisphosphate using ATP.

Role: Committed and second regulatory step of glycolysis.

Allosteric activation: AMP (signals low energy)

Fructose-2,6-bisphosphate (most potent activator, levels increase in response to insulin

via PFK-2)

Allosteric inhibition: ATP (signals high energy)

Citrate (signals abundant metabolic intermediates)

Insulin effect:

Insulin indirectly activates PFK-1 by increasing levels of fructose-2,6-bisphosphate,

enhancing glycolysis in fed states.

Enzyme: Pyruvate kinase

Function: Converts phosphoenolpyruvate (PEP) to pyruvate, generating 2 ATP molecules

(one per PEP).​
Role: Final step of glycolysis.

Activated by: Fructose-1,6-bisphosphate​

Produces: 2 pyruvate + 2 ATP

Enzyme: Glyceraldehyde-3-phosphate dehydrogenase (G3P dehydrogenase)

Function: Catalyzes the oxidation of glyceraldehyde-3-phosphate (G3P) to

1,3-bisphosphoglycerate.​
Role: Produces NADH (2 per glucose) and adds an inorganic phosphate (Pi), not from
ATP.

Key point: This step is the first energy-generating step of glycolysis, though it doesn't
yet produce ATP—it sets up for it.

Enzyme: Phosphoglycerate kinase

Function: Transfers a phosphate group from 1,3-bisphosphoglycerate to ADP, forming

3-phosphoglycerate and ATP.

Role: This is the first substrate-level phosphorylation step in glycolysis, producing 2 ATP
per glucose, which repays the 2 ATP invested earlier.

Enzyme: Pyruvate kinase

Function: Transfers a phosphate from phosphoenolpyruvate (PEP) to ADP, forming 2 ATP

and 2 pyruvate per glucose.​
Role: Final step of glycolysis and the third irreversible regulatory step.

Activated by: Fructose-1,6-bisphosphate (feed-forward activation)​

This ensures coordination with earlier glycolytic steps and prevents intermediate
buildup.

Step 1 – Hexokinase / Glucokinase​

Inhibitors: Glucose-6-phosphate (for hexokinase), Glucagon​
Activators: Insulin

Step 3 (Committed Step) – Phosphofructokinase-1 (PFK-1)​

Inhibitors: ATP, Citrate, Glucagon​
Activators: AMP, Fructose-2,6-bisphosphate, Insulin​
Note: PFK-1 is allosterically regulated

Step 10 – Pyruvate Kinase​

Inhibitor: Glucagon​
Activators: Fructose-1,6-bisphosphate, Insulin​
Note: Feed-forward activation by fructose-1,6-bisphosphate

____________________________________________________________
Section 3:

Enzyme: Pyruvate Dehydrogenase Complex (PDH Complex)​

Function: Converts pyruvate into acetyl-CoA via oxidative decarboxylation, producing
NADH and CO₂.

Allosteric Regulation:

Inhibited by (product inhibition):

●​ Acetyl-CoA​

●​ NADH​

●​ ATP​

Activated by:

●​ Pyruvate​

●​ ADP​

●​ Ca²⁺ (especially during muscle contraction)​

Covalent Regulation (Phosphorylation):

1.​ PDH Kinase​

○​ Inactivates PDH by phosphorylation​

○​ Activated by: ATP, NADH, Acetyl-CoA​

○​ Inhibited by: ADP, NAD⁺, Pyruvate​

2.​ PDH Phosphatase​

○​ Activates PDH by dephosphorylation​

○​ Activated by: Ca²⁺

Enzyme: Citrate synthase

Function: Catalyzes the first step of the TCA cycle by combining acetyl-CoA and
oxaloacetate to form citrate, releasing CoA-SH.
Role: Entry point for carbon from glycolysis (via acetyl-CoA) into the TCA cycle. This
step is irreversible and highly regulated.

Inhibited by: Citrate (product inhibition)

Enzyme: Isocitrate dehydrogenase

Function: Catalyzes the oxidative decarboxylation of isocitrate to α-ketoglutarate,

producing CO₂ and NADH.

Role: This is a rate-limiting and regulatory step of the TCA cycle.

Inhibited by:

●​ ATP​

●​ NADH​

Activated by:

●​ ADP (signals low energy)​

●​ Ca²⁺ (especially in muscle cells during contraction)

Enzyme: α-Ketoglutarate dehydrogenase

Function: Converts α-ketoglutarate to succinyl-CoA, producing CO₂ and NADH.

Role: This is another irreversible and regulatory step in the TCA cycle.

Inhibited by:

●​ NADH​

●​ Succinyl-CoA (product inhibition)​

Activated by:

●​ Ca²⁺ (especially during muscle activity)

Section 4, part 1
Enzyme: Pyruvate carboxylase

Function: Converts pyruvate to oxaloacetate in the mitochondria. This is the first step of
gluconeogenesis and requires biotin and CO₂.

Role: An anaplerotic reaction that replenishes oxaloacetate and helps initiate

gluconeogenesis.

Activated by:

●​ Acetyl-CoA (signals fat breakdown and the need for glucose production)​

Inhibited by:

●​ ADP (signals low energy, so gluconeogenesis is suppressed)

Enzyme: PEP-carboxykinase (PEPCK)

Function: Converts oxaloacetate to phosphoenolpyruvate (PEP) using GTP in

gluconeogenesis.

Role: This is a key step in bypassing pyruvate kinase (from glycolysis) and moving
oxaloacetate into the gluconeogenic pathway.

Inhibited by:

●​ ADP (indicates low energy, so gluconeogenesis is suppressed)

Enzyme: Fructose-1,6-bisphosphatase

Function: Converts fructose-1,6-bisphosphate to fructose-6-phosphate in

gluconeogenesis.

Role: A key regulatory and irreversible step in gluconeogenesis; it bypasses

phosphofructokinase-1 (PFK-1) from glycolysis.

Activated by:

●​ ATP​

●​ Citrate​
Inhibited by:

●​ AMP​

●​ Fructose-2,6-bisphosphate (the most potent inhibitor)​

This regulation ensures gluconeogenesis is favored during high-energy states and

suppressed during low-energy conditions.

Enzyme: Pyruvate carboxylase

Function: Converts pyruvate to oxaloacetate in the mitochondria during

gluconeogenesis. Uses biotin as a coenzyme and consumes ATP.

Mechanism:

1.​ ATP hydrolysis forms a biotin-CO₂ intermediate​

2.​ CO₂ is transferred to pyruvate to form oxaloacetate​

Activated by:

●​ Acetyl-CoA (signals fat breakdown and need for gluconeogenesis)​

Inhibited by:

●​ ADP (indicates low energy; gluconeogenesis is suppressed)​

Summary: When acetyl-CoA is high (from β-oxidation of fats), pyruvate is diverted away
from the TCA cycle and toward glucose production via gluconeogenesis.
Enzyme: PEP-carboxykinase (Phosphoenolpyruvate carboxykinase)

Function: Converts oxaloacetate (OAA) into phosphoenolpyruvate (PEP) in the cytosol

by decarboxylation and phosphorylation during gluconeogenesis.

Energy source:​
Uses GTP (not ATP), which is hydrolyzed to GDP during the reaction. This makes the
reaction independent of ATP levels—advantageous when ATP is low and
gluconeogenesis is active.

Inhibited by:

●​ ADP, which signals low energy and suppresses gluconeogenesis.​

Key point:​
This is a crucial regulatory step in gluconeogenesis. It helps bypass the irreversible
pyruvate kinase step from glycolysis.

Enzyme: Fructose 1,6-bisphosphatase

Function: Removes a phosphate group from fructose 1,6-bisphosphate, forming fructose

6-phosphate during gluconeogenesis. This step bypasses the irreversible PFK-1 step
from glycolysis and is crucial for regulating glucose production.

Location: Liver and kidney cells.

Regulation:

●​ Inhibited by:​

○​ AMP (signals low energy)​

○​ Fructose 2,6-bisphosphate (major hormonal regulator favoring glycolysis)​

●​ Activated by:​

○​ Citrate (signals abundant energy and substrates)​

○​ ATP (indicates high energy status)​

This enzyme is one of the key regulatory checkpoints in gluconeogenesis.

Enzyme: Glucose 6-phosphatase​

Function: It removes a phosphate group from glucose 6-phosphate to produce free

glucose and inorganic phosphate (Pi).​

Location: This reaction takes place in the lumen of the endoplasmic reticulum (ER). The
enzyme is located in the ER membrane.​

Context: This step bypasses hexokinase/glucokinase from glycolysis, which is the

enzyme that initially traps glucose in the cell during glucose breakdown.​

Transport: Glucose 6-phosphate is transported into the ER, dephosphorylated by

glucose 6-phosphatase, and then glucose is shuttled back into the cytoplasm and
potentially released into the blood.

Section 4: part 2:

Enzyme name: Glycogen phosphorylase – regulatory enzyme for glycogen breakdown.

Activated by:

– Glucagon (signals low blood glucose)

– Ca²⁺ and AMP in muscle (indicate energy demand) → allosteric activators

Inhibited by:

– Glucose-6-phosphate and ATP (enough energy) → allosteric inhibitors

– Insulin (signals fed state, promotes storage)

Enzyme name: Glycogen synthase – key enzyme for glycogen synthesis.

Activated by:​
– Glucose-6-phosphate (glucose is available)​
– ATP (energy is high)​
– Insulin (fed state, promotes storage)

Inhibited by:​
– Glucagon (fasting state, stops storage)

Enzyme name: Glycogen synthase​

Function: Forms α(1→4) glycosidic bonds (linear chain)

Enzyme name: Branching enzyme (glycosyl α(1→4):α(1→6) transferase)​

Function: Creates α(1→6) bonds (branch points)

Enzyme name: Glycogenin

Function: Primer that starts glycogen synthesis by attaching glucose to itself (on
tyrosine)

→ Needed because glycogen synthase cannot start a chain on free glucose

Enzyme name: Glycogen synthase

Function: Extends the chain by forming α(1→4) bonds using UDP-glucose

Enzyme name: UDP-glucose pyrophosphorylase

Function: Converts glucose-1-phosphate + UTP → UDP-glucose (activated glucose

donor)

Energy used: UTP (acts like ATP here)

Enzyme name: Glycogenin

Function: Accepts glucose on its tyrosine OH group to begin the chain

→ Once a short chain forms, glycogen synthase takes over

Enzyme name: Glycogen synthase

Function: Adds glucose from UDP-glucose to existing glycogen chain

Enzyme name: Glycogen synthase

Function: Adds glucose at non-reducing ends by forming α(1→4) bonds

Enzyme name: Branching enzyme (amylo-α(1,4)→α(1,6)-transglycosylase)

Function: Cuts 6–8 glucose units and forms a branch by making an α(1→6) bond

Enzyme name: Glycogen phosphorylase

Function: Breaks α(1→4) glycosidic bonds to release glucose-1-phosphate

Note: Stops 4 glucose units before a branch point

Enzyme name: Debranching enzyme (has two activities)

1.​ Activity: Glycosyl (4→4) transferase​

Function: Moves 3–4 glucose units from the branch to the main chain (α1→4)​

2.​ Activity: α(1→6) glucosidase​

Function: Cleaves the single remaining glucose at the branch point​
Product: Free glucose (not phosphorylated)

Enzyme name: Glycogen synthase​

Allosteric activation:​
– Glucose-6-phosphate​
– ATP

Hormonal activation:​
– Insulin → increases glycogen synthesis

Enzyme name: Glycogen phosphorylase​

Allosteric inhibition:​
– Glucose-6-phosphate​
– ATP​
– Glucose (in liver)

Allosteric activation (muscle only):​

– Ca²⁺​
– AMP​
(both increase during muscle contraction)

Hormonal activation:​
– Glucagon → increases glycogen degradation

Section 5:

Enzyme name: Glucose-6-phosphate dehydrogenase (G6PD)​

Function: Catalyzes the rate-limiting step in the pentose phosphate pathway:​
→ Oxidation of glucose-6-phosphate to 6-phosphogluconolactone

Inhibited by: NADPH (competitive inhibitor — feedback regulation)​

Upregulated by: Insulin (increases G6PD expression)

This step is irreversible and crucial for producing NADPH, used in biosynthesis and
antioxidant defense.

Enzyme name: Glucose-6-phosphate dehydrogenase (G6PD)

Function: Converts glucose-6-phosphate → 6-phosphogluconolactone

Produces: 1 NADPH

Regulation:

– Inhibited by NADPH (competitive)

– Activated by insulin (increases G6PD expression)

Enzyme name: Lactonase

converts 6-phosphogluconolactone → 6-phosphogluconate

Enzyme name 6-Phosphogluconate dehydrogenase

converts 6-phosphogluconate → ribulose-5-phosphate

Produces: 2nd NADPH + CO₂

Enzyme name: Ribose-5-phosphate isomerase

Function: Converts ribulose-5-phosphate ↔ ribose-5-phosphate (for nucleotide

synthesis)

Enzyme name: Glutathione peroxidase​

Function: Reduces H₂O₂ → 2 H₂O using 2 glutathione (GSH) molecules​
→ GSH is oxidized to form GSSG (a disulfide)

Enzyme name: Glutathione reductase​

Function: Regenerates GSH from GSSG using NADPH​
→ NADPH is oxidized to NADP⁺

Enzyme name: NADPH oxidase​

Function: Transfers electrons from NADPH to O₂ → forms superoxide (O₂⁻)

Enzyme name: Superoxide dismutase​

Function: Converts superoxide → hydrogen peroxide (H₂O₂)

Enzyme name: Myeloperoxidase​

Function: Uses H₂O₂ and Cl⁻ to produce hypochlorous acid (HOCl) → a powerful
antimicrobial agent (like bleach)