Nitrogen cycle/nitrogen fixation

Nitrogen is an important constituent of biological molecules. The availability of N can affect plant growth and thus primary Production. Microbes are intimately involved in this process.


But N fixation is a critically important process in the environment and in agriculture .Nitrogen is a very stable and common molecule. A lot of energy is required to break the NN bond. Most organic nitrogen is recycled from the more easily available forms nitrate and NH4+.

nitrate is used as an electron acceptor. Pseudomonas denitrificans 2NO3. . eutrophication. Nitric and nitrous oxide can be released into the atmosphere causing potential problems.+2H+  N2 + 6H2O The electrons come from metabolism of carbohydrates etc. NO2. Note they will grow aerobically using O2 as an electron acceptor if it is available (THIS IS NOT FERMENTATION). It is beneficial in waste water treatment. producing N2. removing nitrate.+ 10e.Denitrification Dissimilatory (anaerobic process). NO3. NO  N2O  N2 Thought to be an enzyme for each stage. thus reducing algal growth (blooms).

Ammonia at neutral and acid pH. Nitrosomas and Nitrobacter together (example of syntrophism). aerated soils by two nitrifying bacteria. Manure and sewage promote nitrification. Occurs in well drained. This (nitrapyrin) increases efficiency of the fertiliser and reduces run off water pollution. Nitrate is rapidly absorbed by plants but as it is very soluble it is easily leached out by rain. chemicals are added to inhibit nitrification. .Nitrification It is oxidation of ammonia to nitrate (via nitrite). so it is not always of benefit. Anhydrous ammonia is used as a fertiliser. is cationic and is absorbed by clay minerals.

generating few ATP molecules the bacteria grow slowly. Nitrosomas NH4 + 3/2O2.Nitrification is a two stage process The process in energetically fairly inefficient.NO2.+2H+ + energy Nitrobacter NO2.+ 3/2O2 – NO3.+ energy The energy generated is used to fix CO2 .

Assimililatory is is the conversion of nitrate (or NH3) to NH3 and then to nitrogenous compounds like amino acids. .

and only a small number of organisms can do it. N2 is very stable and there is a large a reservoir of N in the atmosphere. all prokaryotes.Nitrogen Fixation This reaction is very important. Both free living and in symbiotic associations. It requires a lot of energy to break the triple bond. .

clover etc) Frankia with woody shrubs Anabaena with azolla (fern) in paddy fields . Cyanobacteria (some) Free living anaerobes Closridium. Azomonas. Rhodobacter etc Symbiotic Rhizobium and Bradyrhizobium with legumes (Soya.Examples of nitrogen fixers Free living aerobes Azotobacter. peas.

. FeS forms a cage. Two molecules of FeMo-co per molecule. The formula is MoFe7S9. Made up of two proteins – dinitrogenase and dinitrogen reductase. Both contain Fe and DR also has Mo. 940 kj compared to 493kj for O2. In DR the Fe and Mo are contained in a cofactor FeMo-co and the reduction of N2 occurs here. 6 electrons are required to reduce N2 to 2NH3. This reduction process is catalysed by Nitrogenase.Biochemistry of Nitrogen fixation Nitrogen has a triple bond and this requires a lot of energy to break it.

FeMo co complex .

or O2 is removed by special chemicals. nitrogenase is protected from O2 either by rapid respiration. 16-24 molecules ATP are used. production O2 retarding slime or production of special cells (heterocysts). . In aerobic bacteria. As it is a highly reducing process. For every molecule of N2 fixed. Both enzymes are rapidly and irreversibly inactivated by O2.Properties of Nitrogen fixation Nitrogen fixation is inhibited by O2.

Electron Flow Ferredoxin. They transfer electrons to dinitrogen reductase. Only 6 electrons used in the useful reduction. which can back react withN2H2. another two are wasted to make H2. ATP is hydrolysed and the two proteins disassociate to begin another cycle of reduction. which is then able to interact with dinitrogenase and transfer electrons to it. flavodoxin or low potential iron-sulphur protein are the electron donors. . dintrogen reductase binds two ATP. For each cycle of e.transfer.


Steps in Nitrogen Fixation .

Assay Nitrogenase activity by acetylene to ethylene Artificial substrate HCCH  H2C=CH2 .

Soybean Root Nodules .


Leguminous plants are at an advantage in poor soils. N2 alone. Bound: free O2 is 10. they need the plant. The bacteria and plant form this iron containing compound which binds O2.000:1 . These bacteria are unable to fix. In the nodule O2 levels are controlled by leghaemolobin.

3 Travel to main root 4 Formation deformed cells.Stages in Root Nodule Formation 1 Recognition and attachment of bacterium to root hair 2 Invasion of root hair. by formation of an infection thread. bacteroids to get to N fixing state 5 Formation of Nodule .

the infection thread. responding well to plant secretions. inducing formation of a cellulosic tube. Bacterial cells become swollen into bacteroids. Invasion of the root hair is via the tip as a result of the action of bacterial encoded nod factors. When the plant dies. Root cells adjacent to this thread also become infected. or in groups by plant cell membrane to become symbiosome. nodule deteriorate. Bacteria multiply rapidly in the root. but some dormant rods always there which proliferate on the products released from the dying nodules.Rhizobia grow well in the rhizopsphere. Nod factors stimulate plant cell division. The fixed N is released to the soil . become surrounded singly. Only then does nit fix take place. Rhicadhesin on the surface of bacterium may bind calcium complexes on the root hair surface. bacteroid cannot divide.





Stem nodulating legumes in tropics – Sesbania (water plant). Soils get leached because of high microbial activity. .

.Azolla pinnata (left) 1cm. Anabaena from crushed leaves Of Azolla.