Nitrification:

Nitrification is a two steps process of oxidation of

ammonium first to nitrite (nitritation) and then nitrate (nitratation) carried
out by highly specified specialized autotrophic bacteria. So, nitrification is
an oxidative process.
In the first step, nitrosomonas (and nitrosococcus & to
some extent by nitrosogoea, nitrosocystic) participates in the oxidation of
ammonium to nitrite, whereas the second step, i.e. the oxidation of nitrite to
nitrate is carried out by Nitrobactor. This type of nitrification is also known
as autotrophic nitrification.
However, recent literature reviews show that apart from
autotrophic nitrification, the heterotrophic nitrification, carried out by
heterotrophic bacteria and fungi, also takes place in soil. In this case, there is
no formation of nitrite, i.e. ammonium is directly converted to nitrate. The
heterotrophic process of nitrification is of little agronomic importance since
only small amount of nitrate is formed through this process.
The first step of reaction in nitrification is,
Nitrosomas
+
NH4 + 1 ½ O2 NO2- + H2O + 2H+ +66Kcal
Bacteria

This step is a direct oxidation not involving any

cytochrome system or auxiliary carriers.
The second step of oxidation of nitrite to nitrate by
nitrobactor undoubtedly involves a cytochrome (respiratory enzyme) and
their oxidation cannot take place by the addition of atmospheric oxygen to
nitrate. It can be expressed in following way:

NO2-.H2O + 2Cy+.Fe++ NO3- +2Cy+Fe2+ +2H+

2Cy+.Fe++ +2H+ + ½ O2 2Cy+.Fe+++ + H2O

The overall autotrophic process of nitrification can be

shown simply as:
NH3 H+ NH4+ NO2- NO3-
H+ nitrosomas nitrobactor
Pathway of nitrification:
The six electron transfer accompanying the oxidation of
NH4 (oxidation state of -3) to NO2- (oxidation state of +3) by nitrosomas
+

suggests of least two intermediates, the most likely candidates being

hydroxylamine (NH2OH) and nitroxyl (NOH).
NH4+ +1/2 O2 NH2OH NOH +1/2 O2 NO-2
-H+ -H+
Ammonium Hydroxyl amine Nitroxyl Nitrite

Energy is released by the reaction (65Kcal/mol) is used

by the organisms for carrying out its life activities.
Several studies have indicated that N2O is a by-product
+
of NH4 oxidation. This gas may arise by chemical dismutation of nitroxyl
(NOH) and through the action of nitric reductase.

NH4+ 2e-1
NH2OH 2e- [NOH] 2e-
NO2-
Chemical
Dismutation Denitrifying
Nitrite reductase

N2O

The oxidation of NO2- to NO3- by nitrobactor involves a

two-electron change in the oxidation state of N (from +3 to +5) with release
of 17.8 Kcal/mole of energy:
NO2- + ½ O2 2e-
NO3-
Nitrite (+3) nitrate (+5)

NO intermediates are suspect. The reaction is facilitated

-
by an NO oxidase system with the elements being carried to O2 via a
2
cytochrome system, with generation of ATP.

Why nitrite does not usually accumulate in soil? If do, then under what
condition nitrite accumulate in soil?
NO2- does not usually accumulate in soil but when it
does, it can adversely affect plants and microorganisms.
Ammonium also does not accumulate in soil. But, under
same condition, it does .
----- Temperature is very low
----- Water saturated soil condition
When NH4+ accumulates in soil , a portion of it is utilized
by microorganisms and plants. Another portion undergoes volatilization.
According to J.P Robertson nitrification is NH4+ limited;
the amount of NH4+ accumulated, is immediately transformed to nitrate
(NO3-) by nitrifiers.
From this, we can say that microbial bacteria, although
they are small in number, they are efficient (Nitrosomonas, nitrobactor,
nitrosococcus, etc. bacteria convert NH4+ to NO3-). But when pH is above
9.0, then some nitrite can accumulate in soil.
Thus, we can conclude that nitrifiers are much more
efficient than ammonifiers.

Which organism is more active in the transformation of nitrite to nitrate?

Bacteria
Bacteria---- 1500 μg/ml NO2—N or NO3--N
Fungi------ 30 μg/ml NO3—N or NO3—N
Chemical composition and functional group of humic substances:
Elemental composition – C,O2, N, H mainly
In humic acid rich in carbon 41-57%, H2 and n contents
O2--------33-46%
N2------2-5%
In fulvic acid lower carbon content, H2 & N2 content is also
lower.
O2------44-54%
N2-----07-2.6%
Functional groups:
1. Carboxyl groups (acid groups) :R-COOH
2. Hydroxyl groups (total hydroxyl, phenolic OH-groups and
alcoholic OH-group)-R-OH-R-O-
3. Carboxyl groups (ketonic or quinoid)- R=O-R-O-