Nitrate reductase-dependent

Pseudomonas aeruginosa can synthesize both types of nitrate reductase, depending upon the environmental conditions, the dissimilatory enzyme being repressed by dioxygen.1049... 

Similar mechanisms operate in the action of nitrate reductase and nitrite reductase. Both of these substances are produced from ammonia by oxidation. Plants and soil bacteria can reduce these compounds to provide ammonia for metabolism. The common agricultural fertilizer ammonium nitrate, NH4NO3, provides reduced nitrogen for plant growth directly, and by providing a substrate for nitrate reduction. NADH or NADPH is the electron donor for nitrate reductase, depending on the organism. 

H. Yamasaki and Y. Sakihama, Simultaneous production of nitric oxide and peroxynitrite by plant nitrate reductase in vitro evidence for the NR-dependent formation of active nitrogen species. FEBS Lett. 468, 89-92 (2000). 

METHOD OF CONTINUOUS VARIATION MOLYBDENUM COFACTOR (MoCo) Molybdenum-dependent reactions, ALDEHYDE OXIDASE MOLYBDOPTERIN NITRATE REDUCTASE NITROGENASE SULFITE OXIDASE XANTHINE DEHYDROGENASE MOLYBDOPTERIN... 

The opposite sequence, reduction of nitrate and nitrite ions, provides a major route of acquisition of ammonia for incorporation into cells by bacteria, fungi, and green plants (Fig. 24-1). Assimilatory (biosynthetic) nitrate reductases catalyze the two-electron reduction of nitrate to nitrite (Eq. 16-61). This is thought to occur at the molybdenum atom of the large 900-residue highly regulated793 molybdopterin-dependent enzyme. In green plants the reductant is... 

The assimilatory nitrate reductase from Chlorella contains the molybdenum cofactor, as evidenced by the ability of the enzyme to donate the cofactor to the nitrate reductase of the mutant nit-1 of N. crassa. Reduction of the enzyme with NADH gives the Mov ESR signal, which is abolished on reoxidation with nitrate. Line shape and g values of the signal show a pH dependence similar to those observed previously for sulfite oxidase. The signal observed at pH 7.0 shows evidence for interaction with a single exchangeable proton.1053... 

Brown, J., Small, I.S. Wray, J.L. (1981). Age-dependent conversion of nitrate reductase to cytochrome c reductase species in barley leaf extracts. Phytochemistry 20, 389-98. 

Although molybdenum and tungsten enzymes carry the name of a single substrate, they are often not as selective as this nomenclature suggests. Many of the enzymes process more than one substrate, both in vivo and in vitro. Several enzymes can function as both oxidases and reductases, for example, xanthine oxidases not only oxidize purines but can deoxygenate amine N-oxides [82]. There are also sets of enzymes that catalyze the same reaction but in opposite directions. These enzymes include aldehyde and formate oxidases/carboxylic acid reductase [31,75] and nitrate reductase/nitrite oxidase [83-87]. These complementary enzymes have considerable sequence homology, and the direction of the preferred catalytic reaction depends on the electrochemical reduction potentials of the redox partners that have evolved to couple the reactions to cellular redox systems and metabolic requirements. 

Plant use of iron depends on the plant s ability to respond chemically to iron stress. This response causes the roots to release H+ and deduct ants, to reduce Fe3+, and to accumulate citrate, making iron available to the plant. Reduction sites are principally in the young lateral roots. Azide, arsenate, zinc, copper, and chelating agents may interfere with use of iron. Chemical reactions induced by iron stress affect nitrate reductase activity, use of iron from Fe3+ phosphate and Fe3+ chelate, and tolerance of plants to heavy metals. The iron stress-response mechanism is adaptive and genetically controlled, making it possible to tailor plants to grow under conditions of iron stress. 

Nitrate Reductase. Nitrate reductase is found widely distributed among plants and microorganisms and catalyzes the reduction of N03 to N02" (16,17,18,19). The physiological role of this enzyme depends... 

All plants depend on nitrate reductase to accomplish the seemingly trivial reaction of nitrate reduction to nitrite, often the first step of nitrogen assimilation into compounds required for growth (5, 22). Many bacteria use molybdenum or tungsten enzymes in anaerobic respiration where the terminal electron acceptor is a reducible molecule other than oxygen, such as nitrate (2, 50), polysulfide (51), trimethylamine oxide (33, 52) or dimethyl sulfoxide (DMSO) (2, 29, 30). 

The postulated catalytic cycles for pterin-containing molybdenum enzymes involve a two-electron change at the molybdenum atom (Mo(VI) Mo(IV)). Microcoulometric titrations of nitrate reductase Chlorella vulgaris) (76), milk xanthine oxidase (77), and sulfite oxidase (78) show that their molybdenum centers are reduced by two electrons. The reduction potentials for the molybdenum center of chicken liver sulfite oxidase are strongly dependent upon pH and upon anion concentration (78). 

Gao, Y., Smith, G. J., and Alberte, R. S. (2000). Temperature dependence of nitrate reductase activity in marine phytoplankton Biochemical analysis and ecological, implications. /. Phycol. 36, 304—313. 

Figure 21. (A) The assembly of an integrated nitrate sensor electrode by the cross-linking of a microperoxidase-11-nitrate reductase (cytochrome-dependent, EC 1.9.6.1) affinity complex on an Au electrode. (B) Cyclic voltammograms of the integrated MP-ll-NR monolayer-modified Au electrode (roughness factor ca. 15). (a) 0.1 M phosphate buffer, pH 7.0 (b) in the presence of KNO3, 20 mM. Potential scan rate, 5 mV s . Inset electrocatalytic cathodic currents [E = —0.6 V vs. SCE) transduced by the modified electrode at different concentrations of KNO3. Measurements were performed under argon.

Recently, de novo-synthesized four-helix polypeptides were applied to mimic functions of cytochrome b and to tailor layered cross-linked electrocatalytic electrodes. A four-helix bundle de novo protein (14728 Da) that includes four histidine units in the respective A -helices was assembled on Au electrodes (Figure 22A). Two units of Fe(III)-protoporphyrin IX were reconstituted into the assembly to yield a vectorial electron-transfer cascade [157]. The de novo-synthesized protein assembly forms affinity complexes with the cytochrome-dependent nitrate reductase (NR) and with Co(II) protoporphyrin IX-reconstituted myoglobin [158]. The resulting layered complex of Fe(III) de novo protein-NR or Fe(lll)-de novo protein-Co(II)-reconstituted myoglobin was cross-linked with glutaric dialdehyde to yield electrically contacted electrocatalytic electrodes. The Fe(lll)-de novo protein-NR electrode assembly was applied for the electrocatalyzed reduction NO3 to NOt" and acted as an amperometric sensor (Figure 22B). The Fe(III)-de novo... 

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