Amino sugars 5-10% Purine and Pyrimidine Bases trace amounts. Undescribed Lots - non-protein N Crude proteins Lignin - N

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1 N in Soil Note: soil concentrations can be anywhere, depending on vegetation, land use, etc. But a substantial amount indeed most (ca. 99%) soil nitrogen is organic Free amino acids trace amounts Amino sugars 510% Purine and Pyrimidine Bases trace amounts Bound amino acids 2050% of soil N Undescribed Lots nonprotein N Crude proteins Lignin N

2 NITROGEN MINERALIZATION / ASSIMILATION Assimilation incorporation of some substance into biomass also referred to as immobilization Mineralization conversion of organic molecules to some inorganic form e.g., CH 2 O O 2 fi CO 2 H 2 O H 2 NCH 2 COOH fi H 3 CCOOH NH 3 Ammonia (um) is the ONLY inorganic N product of mineralization

3 NITRIFICATION The conversion of ammonia(um) to nitrite then nitrate I. Ammonium oxidation NH 1.5O2 NO H ( H 2O) 3 2 G = 65kcal/mole Carried out by Nitrosomonas II. Nitrite oxidation NO O2 NO3 G = 18kcal/mole Carried out by Nitrobacter

4 Ammonia / Ammonium Equilibrium H 4 NH 3 NH Dissociation constant (K b ) = K B = [ ] NH [ ] [ ] NH H 3 4 K w = K A K B or pk A pk B = 14

5 o f es (mol l o g When ph = pk, the ratio of the acid and base will be 1 E q u i v a l e n t i.e., at pk, [NH 4 ] = [NH 3 ] NH3 NH

6 Rates of nitrification by some heterotrophic and autotrophic nitrifiers Max. product Organism Substrate Product Rate of Formation (µg N/day/ g dry cells) accumulation (µg N / ml) Arthrobacter NH 4 NO (heterotroph) Arthrobacter NH 4 NO (heterotroph) Aspergillus NH 4 NO (heterotroph) Nitrosomonas NH 4 NO million (autotroph) Nitrobacter (autotroph) NO 2 NO million

7 NITRATE REDUCTION a heterotrophic process in 3 forms Assimilatory nitrate reduction: glucose 3NO3 3H 6CO2 3NH3 3H 2O G = ( 429kcal/mole) Not oxygen sensitive used by all organisms that assimilate NO 3 Dissimilatory nitrate reduction: glucose 3NO3 3H 6CO2 3NH3 3H 2O G = ( 429kcal/mole) Oxygen sensitive used by a few facultative anaerobes NH 3 is released to the environment importance? Nitrate is terminal electron acceptor for glucose oxidation.

8 When nitrate and a suitable carbon source are placed together in an anaerobic zone, we see plenty of N 2, N 2 O (occasionally some NO) and possibly some NO 2 NO 3 N 2 N NO 2 N 2 O Denitrification is a heterotrophic process, conditions which favor heterotrophic processes in general will favor denitrification: ph 68, 15 30ºC

9 Denitrification glucose 4.8NO3 4.8H 6CO 2 2.4N2 8.4H2O ( G = 638 kcal/mole) Oxygen sensitive used by some facultative anaerobes to eliminate electrons from the oxidation of glucose 5 NO 3 2e Note: some organisms form NO at this stage NO e N 1 2 O e N 0 2

10 Denitrification is an anaerobic process Occurs in generally anaerobic environments if anaerobic microsites available O 2 O 2 O 2 O 2 O 2 O 2 O 2

11 O 2 acetylene NO 3 NO 2 N 2 O N 2 oxidized reduced e organic CO 2 reduced oxidized By adding a little acetylene to a soil or water sample, the denitrification rate can be determined by the accumulation of N 2 O simple analysis (acetylene block method)

12 N 2 O Produced under varying conditions of soil saturation 450% 350% % NO 3 N Added 100% 75% 50% TIME

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14 Where does the N 2 O (and N 2 ) come from? nitrification denitrification NH 4 NO 2 NO 3? N 2 O N 2

15 Oxygen tolerance for some denitrifiers

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17 HaberBasch Process: N 2 H NH C, 500 Atm of pressure, No O 2, No H 2 O Energy (fossil fuel intensive) Used as source of industrial N and about 65 million metric tons of fertilizer N each year Cost is roughly $ 130 billion / year (global) Remember only about 1/3 of applied fertilizer is used by plants

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19 BIOLOGICAL N FIXATION N 2 H NH 3 Same reaction as the Haber process Occurs at 1 Atm pressure and 1535 C Catalyzed by enzyme NITROGENASE Nitrogenase Anaerobic Process Protection by heterocysts Protection by membrane binding Protection by conversion in presence of oxygen Protection by rapid respiration protection by encapsulation of the organism Active on several other compounds including acetylene

20 Acetylene Reduction Nitrogenase reduces Acetylene to Ethylene Add a little acetylene to system, measure nitrogenase activity as formation of ethylene HC CH H 2 H 2 C=CH 2

21 Estimated Amounts of Nitrogen Fixed during a Year by Wellnodulated Legumes Nitrogen fixed Crop Lb/ac Kg/ha Alfalfa Sweet Clover Red Clover Soybeans Field Beans Field Peas Source: Averaged from several sources

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