nitrogen fixation nitrogen fixation nitrogen fixation is a chemical process by which molecular nitrogen (n2), which has a strong triple covalent bond, is converted into ammonia (nh3) or related nitrogenous compounds, typically in soil or aquatic systems but also in industry. the nitrogen in air is molecular dinitrogen, a relatively nonreactive molecule that is metabolically useless to all but a few microorganisms. biological nitrogen fixation or diazotrophy is an important microbe-mediated process that converts dinitrogen (n2) gas to ammonia (nh3) using the nitrogenase protein complex (nif). nitrogen fixation is essential to life because fixed inorganic nitrogen compounds are required for the biosynthesis of all nitrogen-containing organic compounds, such as amino acids and proteins, nucleoside triphosphates and nucleic acids. as part of the nitrogen cycle, it is essential for agriculture and the manufacture of fertilizer. it is also, indirectly, relevant to the manufacture of all nitrogen chemical compounds, which include some explosives, pharmaceuticals, and dyes. nitrogen fixation is carried out naturally in soil by microorganisms termed diazotrophs that include bacteria, such as azotobacter, and archaea. some nitrogen-fixing bacteria have symbiotic relationships with plant groups, especially legumes. looser non-symbiotic relationships between diazotrophs and plants are often referred to as associative, as seen in nitrogen fixation on rice roots. nitrogen fixation occurs between some termites and fungi. it occurs naturally in the air by means of nox production by lightning. all biological reactions involving the process of nitrogen fixation are catalyzed by enzymes called nitrogenases. these enzymes contain iron, often with a second metal, usually molybdenum but sometimes vanadium. history schematic representation of the nitrogen cycle. abiotic nitrogen fixation has been omitted. biological nitrogen fixation was discovered by jean-baptiste boussingault in 1838. later, in 1880, the process by which it happens was discovered by german agronomist hermann hellriegel and hermann wilfarth and was fully described by dutch microbiologist martinus beijerinck. "the protracted investigations of the relation of plants to the acquisition of nitrogen begun by saussure, ville, lawes and gilbert and others culminated in the discovery of symbiotic fixation by hellriegel and wilfarth in 1887." "experiments by bossingault in 1855 and pugh, gilbert & lawes in 1887 had shown that nitrogen did not enter the plant directly. the discovery of the role of nitrogen fixing bacteria by herman hellriegel and herman wilfarth in 1886-8 would open a new era of soil science." in 1901 beijerinck showed that azotobacter chroococcum was able to fix atmospheric nitrogen. this was the first species of the azotobacter genus, so-named by him. it is also the first known diazotroph, species that use diatomic nitrogen as a step in the complete nitrogen cycle. biological biological nitrogen fixation (bnf) occurs when atmospheric nitrogen is converted to ammonia by a nitrogenase enzyme. the overall reaction for bnf is: n 2 + 16 atp + 16 h 2 o + 8 e − + 8 h + ⟶ 2 nh 3 + h 2 + 16 adp + 16 p i _ the process is coupled to the hydrolysis of 16 equivalents of atp and is accompanied by the co-formation of one equivalent of h2. the conversion of n2 into ammonia occurs at a metal cluster called femoco, an abbreviation for the iron-molybdenum cofactor. the mechanism proceeds via a series of protonation and reduction steps wherein the femoco active site hydrogenates the n2 substrate. in free-living diazotrophs, nitrogenase-generated ammonia is assimilated into glutamate through the glutamine synthetase/glutamate synthase pathway. the microbial nif genes required for nitrogen fixation are widely distributed in diverse environments. for example, decomposing wood, which generally has a low nitrogen content, has been shown to host a diazotrophic community. the bacteria enrich the wood substrate with nitrogen through fixation, thus enabling deadwood decomposition by fungi. nitrogenases are rapidly degraded by oxygen. for this reason, many bacteria cease production of the enzyme in the presence of oxygen. many nitrogen-fixing organisms exist only in anaerobic conditions, respiring to draw down oxygen levels, or binding the oxygen with a protein such as leghemoglobin. importance of nitrogen part of a series onbiogeochemical cycles water cycle water cycle deep water cycle carbon cycle global atmospheric terrestrial oceanic sequestration carbon sink deep carbon cycle soil carbon mycorrhizal fungi boreal forests nutrient cycle hydrogen cycle nitrogen cycle human impact nitrification nitrogen and lichens fixation assimilation oxygen cycle phosphorus cycle assimilation sulfur cycle assimilation rock cycle calcium cycle silica cycle carbonate–silicate cycle marine cycle marine biogeochemical cycles biological pump microbial loop viral shunt calcareous ooze siliceous ooze methane cycle atmospheric methane methane clathrate clathrate gun hypothesis arctic methane emissions other cycles aluminum arsenic boron bromine cadmium chlorine chromium copper fluorine gold iodine iron lead lithium manganese mercury ozone–oxygen selenium vanadium zinc related topics biogeochemistry geochemical cycle chemical cycling environmental chemistry biosequestration deep biosphere ocean acidification acid rain biogeochemical planetary boundaries research groups daac geotraces imber nobm solas category atmospheric nitrogen is inaccessible to most organisms, because its triple covalent bond is very strong. most take up fixed nitrogen from various sources. for every 100 atoms of carbon, roughly 2 to 20 atoms of nitrogen are assimilated. the atomic ratio of carbon (c) : nitrogen (n) : phosphorus (p) observed on average in planktonic biomass was originally described by alfred redfield, who determined the stoichiometric relationship between c:n:p atoms, the redfield ratio, to be 106:16:1. nitrogenase the protein complex nitrogenase is responsible for catalyzing the reduction of nitrogen gas (n2) to ammonia (nh3). in cyanobacteria, this enzyme system is housed in a specialized cell called the heterocyst. the production of the nitrogenase complex is genetically regulated, and the activity of the protein complex is dependent on ambient oxygen concentrations, and intra- and extracellular concentrations of ammonia and oxidized nitrogen species (nitrate and nitrite). additionally, the combined concentrations of both ammonium and nitrate are thought to inhibit nfix, specifically when intracellular concentrations of 2-oxoglutarate (2-og) exceed a critical threshold. the specialized heterocyst cell is necessary for the performance of nitrogenase as a result of its sensitivity to ambient oxygen. nitrogenase consist of two proteins, a catalytic iron-dependent protein, commonly referred to as mofe protein and a reducing iron-only protein (fe protein). there are three different iron dependent proteins, molybdenum-dependent, vanadium-dependent, and iron-only, with all three nitrogenase protein variations containing an iron protein component. molybdenum-dependent nitrogenase is the most commonly present nitrogenase. the different types of nitrogenase can be determined by the specific iron protein component. nitrogenase is highly conserved. gene expression through dna sequencing can distinguish which protein complex is present in the microorganism and potentially being expressed. most frequently, the nifh gene is used to identify the presence of molybdenum-dependent nitrogenase, followed by closely related nitrogenase reductases (component ii) vnfh and anfh representing vanadium-dependent and iron-only nitrogenase, respectively. in studying the ecology and evolution of nitrogen-fixing bacteria, the nifh gene is the biomarker most widely used. nifh has two similar genes anfh and vnfh that also encode for the nitrogenase reductase component of the nitrogenase complex. microorganisms main article: diazotroph diazotrophs are widespread within domain bacteria including cyanobacteria (e.g. the highly significant trichodesmium and cyanothece), green sulfur bacteria, azotobacteraceae, rhizobia and frankia. several obligately anaerobic bacteria fix nitrogen including many (but not all) clostridium spp. some archaea such as methanosarcina acetivorans also fix nitrogen,. and several other methanogenic taxa, are significant contributors to nitrogen fixation in oxygen-deficient soils. cyanobacteria, commonly known as blue-green algae, inhabit nearly all illuminated environments on earth and play key roles in the carbon and nitrogen cycle of the biosphere. in general, cyanobacteria can use various inorganic and organic sources of combined nitrogen, such as nitrate, nitrite, ammonium, urea, or some amino acids. several cyanobacteria strains are also capable of diazotrophic growth, an ability that may have been present in their last common ancestor in the archean eon. nitrogen fixation not only naturally occurs in soils but also aquatic systems, including both freshwater and marine. indeed, the amount of nitrogen fixed in the ocean is at least as much as that on land. the colonial marine cyanobacterium trichodesmium is thought to fix nitrogen on such a scale that it accounts for almost half of the nitrogen fixation in marine systems globally. marine surface lichens and non-photosynthetic bacteria belonging in proteobacteria and planctomycetes fixate significant atmospheric nitrogen. species of nitrogen fixing cyanobacteria in fresh waters include: aphanizomenon and dolichospermum (previously anabaena). such species have specialized cells called heterocytes, in which nitrogen fixation occurs via the nitrogenase enzyme. legume family nodules are visible on this broad bean root plants that contribute to nitrogen fixation include those of the legume family—fabaceae— with taxa such as kudzu, clover, soybean, alfalfa, lupin, peanut and rooibos. they contain symbiotic rhizobia bacteria within nodules in their root systems, producing nitrogen compounds that help the plant to grow and compete with other plants. when the plant dies, the fixed nitrogen is released, making it available to other plants; this helps to fertilize the soil. the great majority of legumes have this association, but a few genera (e.g., styphnolobium) do not. in many traditional farming practices, fields are rotated through various types of crops, which usually include one consisting mainly or entirely of clover. fixation efficiency in soil is dependent on many factors, including the legume and air and soil conditions. for example, nitrogen fixation by red clover can range from 50 to 200 lb/acre (56 to 224 kg/ha). non-leguminous a sectioned alder tree root nodule the ability to fix nitrogen in nodules is present in actinorhizal plants such as alder and bayberry, with the help of frankia bacteria. they are found in 25 genera in the orders cucurbitales, fagales and rosales, which together with the fabales form a nitrogen-fixing clade of eurosids. the ability to fix nitrogen is not universally present in these families. for example, of 122 rosaceae genera, only four fix nitrogen. fabales were the first lineage to branch off this nitrogen-fixing clade; thus, the ability to fix nitrogen may be plesiomorphic and subsequently lost in most descendants of the original nitrogen-fixing plant; however, it may be that the basic genetic and physiological requirements were present in an incipient state in the most recent common ancestors of all these plants, but only evolved to full function in some of them. in addition, trema (parasponia), a tropical genus in the family cannabaceae, is unusually able to interact with rhizobia and form nitrogen-fixing nodules. non-legumious nodulating plants family genera species betulaceae alnus (alders) most or all species boraginaceae phacelia phacelia tanacetifolia cannabaceae trema (parasponia) trema orientale trema lamarckiana casuarinaceae allocasuarina casuarina ceuthostoma gymnostoma coriariaceae coriaria coriaria arborea coriaria myrtifolia datiscaceae datisca elaeagnaceae elaeagnus (silverberries) hippophae (sea-buckthorns) shepherdia (buffaloberries) myricaceae comptonia (sweetfern) myrica (babyberries) posidoniaceae posidonia (seagrass) rhamnaceae ceanothus colletia discaria kentrothamnus retanilla talguenea trevoa rosaceae cercocarpus (mountain mahoganies) chamaebatia (mountain miseries) dryas purshia/cowania (bitterbrushes/cliffroses) other plant symbionts some other plants live in association with a cyanobiont (cyanobacteria such as nostoc) which fix nitrogen for them: some lichens such as lobaria and peltigera mosquito fern (azolla species) cycads gunnera blasia (liverwort) hornworts some symbiotic relationships involving agriculturally-important plants are: sugarcane and unclear endophytes foxtail millet and azospirillum brasilense kallar grass and azoarcus sp. strain bh72 rice and herbaspirillum seropedicae wheat and klebsiella pneumoniae maize landrace 'sierra mixe' / 'olotón' and various bacteroidota and pseudomonadota industrial processes historical a method for nitrogen fixation was first described by henry cavendish in 1784 using electric arcs reacting nitrogen and oxygen in air. this method was implemented in the birkeland–eyde process of 1903. the fixation of nitrogen by lightning is a very similar natural occurring process. the possibility that atmospheric nitrogen reacts with certain chemicals was first observed by desfosses in 1828. he observed that mixtures of alkali metal oxides and carbon react with nitrogen at high temperatures. with the use of barium carbonate as starting material, the first commercial process became available in the 1860s, developed by margueritte and sourdeval. the resulting barium cyanide reacts with steam, yielding ammonia. in 1898 frank and caro developed what is known as the frank–caro process to fix nitrogen in the form of calcium cyanamide. the process was eclipsed by the haber process, which was discovered in 1909. haber process equipment for a study of nitrogen fixation by alpha rays (fixed nitrogen research laboratory, 1926) the dominant industrial method for producing ammonia is the haber process also known as the haber-bosch process. fertilizer production is now the largest