Assimilatory nitrate reductase from the haloarchaeon Haloferax mediterranei: purification and characterisation.
Haloferax mediterranei can use nitrate as sole nitrogen source during aerobic growth. We report here the purification and biochemical characterisation of the assimilatory nitrate reductase (EC 1.6.6.2) from H. mediterranei. The enzyme, as isolated, was composed of two subunits (105+/-1.3 kDa and 50+/-1.3 kDa) and behaved as a dimer during gel filtration (132+/-6 kDa). A pH of 9 and elevated temperatures up to 80 degrees C (at 3.1 M NaCl) are necessary for optimum activity. The enzyme stability and activity of the enzyme depend upon the salt concentration. Reduced methyl viologen was as effective as the natural electron donor ferredoxin in the catalytic process. In contrast, NADPH and NADH, which are electron donors in nitrate reductases from different non-photosynthetic bacteria, were ineffective.
Martínez-Espinosa
R M
RM
División de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Alicante, Ap. 99, E-03080, Alicante, Spain.
Marhuenda-Egea
F C
FC
Bonete
M J
MJ
D005288:Ferredoxins; D019623:Haloferax mediterranei / Q000201:enzymology* / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D009566:Nitrates / Q000378:metabolism*; D013696:Temperature
D005288:Ferredoxins; D009566:Nitrates; D009565:Nitrate Reductases; D050901:Nitrate Reductase