Disember 15

NITROGEN
METABOLISM
DR. AIRIANAH
airianah@ukm.edu.my

The Nitrogen cycle

N2
Atmospheric nitrogen

NO3
Nitrate

Decaying
biomass Ammonification

NH3
Ammonia

NH4+
ammonium

NO2
Nitrite

Disember 15

1. Nitrogen Fixation by
nitrogenase

Made up of 2 groups of protein

i. Fe protein
ii. MoFe protein

8H+ + 8e- + N2 + 16ATP

2NH3 + H2 + 16ADP + 16Pi

Disember 15

The reaction occurs in 2 steps

2ATP

2ADP + 2Pi

Fdred

Feox

MoFered

N2 + 8H+

Fdox

Fered

MoFeox

2NH3 + H2

Fe protein

MoFe protein

Function of ATP is not clear

However it is known to react with reduced Fe protein
(confirmation changes) and taking part in transferring e- from Fe
protein to MoFe protein

BY: NMS

Nitrification

Reaction 1 converts ammonia to the intermediate, hydroxylamine, and is

catalyzed by the enzyme ammonia monooxygenase.
Reaction 2 converts hydroxylamine to nitrite and is catalyzed by the enyzme
hydroxylamine oxidoreductase.

Chemical reaction of nitrite oxidation

Disember 15

Nitrate reductase (NR) reaction

2H+ + NO3- + 2e-

NO2- + H2O

The reaction occurs in sitosol

NO2- is very toxic and must converted to NH4+
immediately

BY: NMS

Nitrite reductase (NiR) reaction

8H+ + NO2- + 6e-

NH4+ + 2H2O

The reaction occurs in plastid (root) and

chloroplast (leaf)
NiR activity is much higher than NR
to make sure that NO2- level is as lowest as
possible

BY: NMS

Disember 15

N fixation requires:

N
Nitrogenase
ATP
Low oxygen

leghemoglobin

Summary
N : main component for protein, nucleic acid, hormone
Abundant but not available: Atmosphere contains 78% of N (N2)
Plants do not have an enzyme to break N

only prokaryotes can do that

Nitrogen enters the biosphere by the process of nitrogen fixation.
Most of N is obtained from soil in the form NO3- and NH4+
Requirements for N fixation: N, Nitrogenase, ATP, low oxygen

Adaptation to oxygen: heterocyst, leghemoglobin

Disember 15

?????

Why N fixation is important?

Why N is so abundant but yet is limited nutrient to organism?
Why N fixation so energetically costly?
What are the other sources of fixed N?
Name the transformations in N cycle?
Does plant has enzyme to break N2 bond?
Can any plant form a symbiosis with N-fixing microorganism
Why N-fixing organisms can fix N?
What is the problem for aerobic microorganism to fix N? why?
So, how this microorganism overcome the problem? How
about photosynthetic microorganism?

The Nitrogen cycle

N2
Atmospheric nitrogen

NO3
Nitrate

Decaying
biomass Ammonification

NH3
Ammonia

NH4+
ammonium

assimilation

NO2
Nitrite

Disember 15

Assimilation
Ammonia/ammonium ion generated from N2 is
assimilated into low molecular weight
metabolites such as glutamate, glutamine or
carbamoyl phosphate etc
The two most important assimilation reactions
are;
A. GDH pathway
B. GS-GOGAT pathway

A. GDH pathway
Amination of -ketoglutarate by glutamate
dehydrogenase (glutamate synthase) in lower
eukaryote

BY: NMS

Disember 15

B. GS GOGAT pathway
Alternative route in assimilation of ammonia is via
glutamine synthetase (GS) glutamate synthase
(GOGAT)

BY: NMS

glutamine is converted to glutamate

(transamination) by glutamate synthase

BY: NMS

Disember 15

GS/GOGAT pathway in plant

GDH vs GS/GOGAT pathway

Disember 15

GDH pathway is predominant when cellular

environment is rich in C sources whilst [NH3]
is low. On the other hand, GS-GOGAT
pathway is predominant in opposite
condition.
True OR False

Issues:
-

Crop rotation
Nitrogenase in plant
N-Fertilizer
Algae bloom
Effect of dinitrification

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