NITROGEN METABOLISM

• Nitrogen is one of the important elements of plant life.

• It is one of the constituent of protein, nucleic acids, growth hormones,
vitamins.
• Atmosphere consists of about of 79% gaseous nitrogen .Only few
organisms can directly use the atmospheric nitrogen.
Source of nitrogen:-
1. Atmosphere
2. Soil

Nitrogen fixation:-
The conversion of molecular nitrogen into nitrogenous compounds is
known as Nitrogen fixation.
Types of nitrogen fixation:-
1. Biological nitrogen fixation:-
Certain microorganisms and blue green algae are capable of fixing the
atmosphere nitrogen. They possess certain biological mechanisms by means
of which nitrogen content of the soil and water is continuously replenished
from the atmosphere.
2. Atmospheric nitrogen fixation:-
It is brought by lighting and rainfall. Lightning causes the nitrogen to
combine with Oxygen to produce different types of oxides. The oxides
dissolve in rain water to form nitrous and nitric acid which enters into the
soil.
BIOLOGICAL NITROGEN FIXATION
Nitrogen fixation by biological means is of two types:-
1. Asymbiotic Nitrogen Fixation
2. Symbiotic Nitrogen Fixation
1.Asymbiotic Nitrogen fixation:-
• Microorganisms which lead independent life and fix atmospheric nitrogen
are known as free living diazotrophs (conversion molecular nitrogen into
usable form of plants with the help of living organisms).
• There are two groups of such microorganisms bacteria and Cyanobacteria
(Blue green algae).
Based on mode of nutrition bacteria are divided into
1. Aerobic bacteria - Azotobacter
2. Facultative anaerobic bacteria - Bacillus
3. Anaerobic bacteria - Clostridium
4. Photosynthetic bacteria - Chlorobium

• Among Cyanobacteria both heterocystous and non heterocystous forms

fix atmospheric nitrogen for example Anabaena ,Nostoc, Gloeocapsa,
Scytonema, Oscillatoria etc
Sites of N2 fixation:-
• Many cyanobacteria capable of fixing nitrogen are filamentous and contain
thick wall cell called heterocyst. These are the sites of nitrogen metabolism.
• Heterocyst lack PS II. Wall of heterocyst contain O2 binding glucolipids
which together with respiratory consumption maintain the anaerobic
condition necessary for N2 fixation.
• In other group of cyanobacteria such as Oscillatoria and bacteria where
heterocyst does not develop, N2 fixation process takes place in internally
organized cells having reduced conditions. Root nodules developed by
Rhizobium are the site of nitrogen fixation.
2. Symbiotic Nitrogen fixation:
• There are some microorganisms which establish symbiotic relationships
with different parts of plants and special structures as the site of nitrogen
fixation.
• Members of the genus Rhizobium upon infection of the appropriate
legume can cause the formation of nodules and helps in symbiotic
nitrogen fixation.
Establishment of Symbiosis:
• Establishment of Rhizobium inside the host root and development of nodules are a complex
process which follow many events such as recognition and infection of host root, differentiated of
nodules, proliferation of bacteria and conversion into bacteroids in nodules.
• The infection occurs by curling and this curling reaction of the tip of the root hair suggests that the
rhizobia around the root hair bring about their deformation.
• Nodule initiation and development are controlled by hormones secreted by both the bacteria as
well as the host which includes the indole acetic acid (IAA) and related auxin.
• The infected root hair transformed into a fine hyphae like inwardly growing tube of cellulose,
infection threat multiply and grows and penetrates into the cortex and branches into the central
portion of the developing nodule.
• The final nodule structure consists of a central core containing the rhizobia and surrounding
cortical area with vascular system.
• Doubling of the chromosomes occur in nodule and the rhizobia have peculiar morphology termed
as bacteroid which are the seat of nitrogen fixation.
 AMINO ACID SYNTHESIS
There are two methods in which amino acids can be synthesized, they are:
1. Reductive amination
2. Transamination
In both the cases glutamic acid plays an important role.
1. Reductive amination:-
• Glutamic acid is synthesized in plant cells by a reaction between ammino often originating from
reduction of nitrates and α-ketoglutaric acid (TCA cycle)
• The reaction requires a hydrogen donor and reduced coenzyme I and catalysed by the enzyme
glutamic acid dehydrogenase.
NADPH2
α-ketoglutaric acid + NH2 = α-iminoglutaric acid Glutamic acid
Glutamic dehydrogenase

Though the amination of only α-ketoglutaric acid is of real importance in the plants, there is also
some indirect evidence for the direct amination of oxaloacetic, fumaric and pyruvic acids.
Therefore four ways in which ammonia nitrogen can convert into amino acids.
α-ketoglutaric acid + NH3 Glutamic acid
oxaloacetic acid + NH3 Aspartic acid
Fumaric acid + NH3 Aspartic acid
Pyruvic acid + NH3 Alanine
2. Transamination:-
• Once glutamic acid is available, other amino acids may be formed by the transfer of its amino
group to other carbon skeletons by a process known as transamination.
• Thus transfer of amino group can take place from glutamic acid to oxaloacetic acid, another
intermediate of the metabolic giving a new amino acid, aspartic acid in the presence of the
enzyme transaminase, α-ketoglutaric acid is released in the process.
transaminase
Glutamic acid + Oxaloacetic acid α-ketoglutaric acid + Aspartic acid

transaminase
Glutamic acid + Pyruvic acid α-ketoglutaric acid + Alanine
• The transamination reaction involves the participation of pyridoxal phosphate
or pyridoxamine phosphate as a coenzyme.
• Using radioactive nitrogen it has been found that N15 is incorporated first into
glutamic acid and then into aspartic and alanine.
• Twenty or so amino acids required for protein synthesis are formed in this way.
• Amino acids present in plant cells are often of secondary origin resulting from
chemical transformation of amides and hydrolysis of proteins.