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Lecture 8 PDF

Nitrogenase is the enzyme complex that catalyzes nitrogen fixation. It consists of two protein components, the molybdenum-iron protein and iron protein. Nitrogenase is oxygen labile and must be protected from oxygen by structures like leghemoglobin in legume nodules or heterocysts in cyanobacteria. The enzyme requires metals and utilizes a complex cofactor to reduce nitrogen gas into ammonia.

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217 views

Lecture 8 PDF

Nitrogenase is the enzyme complex that catalyzes nitrogen fixation. It consists of two protein components, the molybdenum-iron protein and iron protein. Nitrogenase is oxygen labile and must be protected from oxygen by structures like leghemoglobin in legume nodules or heterocysts in cyanobacteria. The enzyme requires metals and utilizes a complex cofactor to reduce nitrogen gas into ammonia.

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Enzyme Nitrogenase and its

component
Nitrogenase
• Nitrogen fixation is catalyzed by an enzyme
complex known as nitrogenase
• Found only in diazotrophic prokaryotes, because
they have the genes coding for this enzyme
• Nitrogenase is rapidly and irreversibly inactivated
by molecular oxygen
• It is very sensitive to oxygen and must be
protected from oxygen-inactivation within the
cell
N2 + 8H+ + 8e- + 16ATP 2NH3 + H2 + 16ADP + 16 Pi
Nitrogenase
• In legume nodules, the protection against
oxygen is provided by leghaemoglobin (oxygen
binding protein).
• Whereas in many cyanobacteria it is achieved
by a special structure called the heterocyst.
Heterocyst have thickened, multilayered cell
walls that restricts the diffusion of oxygen.
Also characterized by a high respiratory
activity that maintains a low intracellular
oxygen
Nitrogenase
nitrogenase consists of two major protein
components
1. Dintrogenase (MoFe-protein)
2. Dinitrogenase reductase (Fe-protein)
Nitrogenase
1. Dintrogenase (MoFe-protein)
a. It contains molybdenum(Mo) and iron (Fe)
b. Fe and Mo are contained in a cofactor called MoFe-
cofactor or MoFe-co
c. Contains two molecules of Mo ions in the form of an iron-
molybdenum-sulphur cofactor. Also contains Fe4S4
clusters
d. Molecular weight is 220,000
e. Composition is MoFe7S8-homocitrate
f. Present in two copies per molecule of nitrogenase
g. Actual site of reduction of N2
Nitrogenase
2. Dintrogenase reductase (Fe-protein)
a. Contains Fe only
b. Molecular weight – 64,000
c. Has 4 iron atoms
d. Rapidly and irreversibly inactivated by O2
e. In the beginning of the process of nitrogen
fixation, the ferredoxin first reduces Fe-protein,
which in turn reduces to MoFe-protein
Nitrogenase
1. Nitrogenase, is an equilibrium mixture of
MoFe-protein and Fe-protein
2. In which Fe-protein’s two subunits are
arranged like a pair butterfly wings with Fe-S
cluster between the wings like the ‘Head’ of
the butterfly
3. The enzyme complex requires Mg2+ ions for
its activation
Nitrogenase enzyme complex unit
Nitrogenase
• Factors affecting nitrogenase activity
1. Oxygen sensitivity
2. Temperature
3. pH
4. Soil moisture
Nitrogenase
• Organisms that synthesize nitrogenase
1. Free-living diazotrophs, e.g.
a. Cyanobacteria (by means of differentiated
heterocysts)
b. Azotobacteraceae
2. Symbiotic diazotrophs, e.g.
a. Rhizobia
b. Frankia

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