Amity Institute Of Biotechnology

Amity Institute Of Biotechnology

M.Sc.(BT)
Enzymes and Enzyme Technology(GCMB718)
Dr. Benu Kumar
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Multi-Enzyme System
Pyruvate dehydrogenase Complex(PDH)

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Introduction:
• Many enzymes in living cells catalyse chains of reaction in a
sequential order either in a biosynthetic or a catabolic pathway. These
enzymes bound in the complex in a spatial organization form a multi-
enzyme system.
• Multi-enzyme complexes or multifunctional enzymes form the
structures where the active sites of the enzymes in the subsequent
sequence are close to each other-called as substrate or metabolic
channeling
Metabolic channeling:
• increase the catalytic efficiency, as the diffusion of substrates and
products in the bulk solvent is minimized,
• increase the frequency of enzyme-substrate collisions
• minimizes the time required by substrates to diffuse between
successive active sites
• Reduces the probability of side reactions 3
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PDH –Pyruvate dehydrogenase multi-enzyme complex

allows entry of metabolites into citric acid cycle(TCA)
Consists Of:

Three different enzymes Five Cofactors

(Non-covalently associated)

E1: pyruvate dehydrogenase +TPP

E2: dihydrolipoyl transacetylase + lipoamide

E3: dihydrolipoyl dehydrogenase + FAD

catalyse

+ NADH

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Five Sequential steps + Coenzyme-A
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Pyruvate
dehydrogenase
regulates entry
into the citric
acid cycle of
metabolites
leaving
glycolysis

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• Reaction Sequence
1) Pyruvate is decarboxylated to hydroxyethyl, requires TPP to stabilise
the intermediate.

2) hydroxyethyl oxidised to acetyl, collected by lipoamide of E 2, which

gets reduced.

3) lipoamide of E2, passes acetyl to coenzyme A  acetyl CoA.

4) lipoamide of E2, gets re-oxidised, gives its electrons to FAD

5) E3 which passes electrons to NAD  NADH

• The whole process takes place with the substrate bound to the 7
enzyme,
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• The enzyme complex is about 300 A in diameter as

observed by electron microscopy. It has a polyhedral
structure, with each of the subunits appearing
approximately spherical. The complex is held together
by noncovalent forces and may easily undergo
dissociation

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• PDH controlled by covalent modification and product

inhibition

Phosphatase Kinase activated by

activated by insulin NADH and acetyl-CoA
(high [glc]) and Ca2

Ca2+ and glucose are allosteric positive effectors of pyruvate dehydrogenase

phosphatase, not the PDH complex. They do not act on PDH complex directly, but
on the phosphatase that turns on PDH complex
End products inhibition of PDH is mediated by NADH and Acetyl –CoA that
activates PD kinase and make the complex inactive(Phosphorylation makes the
enzyme inactive)-An example of covalent modification 9
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Overall Reaction

high
NAD +
CoA energy
bond
(2C)
pyruvate AcCoA CoA

(3C)

CO2
irreversible
NADH
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Summary:
1.Pyruvate dehydrogenase complex is an example of a
multi-enzyme complex comprising of three enzymes-
Pyruvate Dehydrogenase (E1), Dihydro- Lipoyl Trans
Acetylase (E2), Dihydro- Lipoyl Dehydrogenase(E3)

2. It regulates entry into the citric acid cycle of metabolites

leaving glycolysis

3.Regulation of PDH complex: by covalent modification

De/Phosphorylation of E1 and product inhibition of E2 and
E3
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