Kinetic studies of Bacillus polymyxa nitrogenase.
Nitrogenase from the facultative anaerobe Bacillus polymxa was separated into its component proteins, which were recombined in the ratio that produced optimal specific activity (125 to 175 nmol of C2H2 reduced/min per mg of total protein). The apparent Michaelis constants (Km)for the magnesium adenosine triphosphate complex, reducible substrates azide, acetylene, and N2 and the nonphysiological electron donor hydrosulfite (S2O42-) were determined to be 0.7, 0.7, 0.2, 0.06, and 0.03 MM, respectively. These apparent Km values are in reasonable agreement with those reported for the nitrogenases of Azotobacter vinelandii and Klebsiella pneumoniae. Either a total lack of cooperativity between binding sites or a single binding site for reducible substrates is indicated by analysis of Hill plots. Hill plot slopes of approximately 1.7 suggest that multiple binding sites exist for both ATP and S2O42-.
D000114:Acetylene / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000693:Anaerobiosis; D001386:Azides / Q000378:metabolism; D001395:Azotobacter / Q000201:enzymology; D001407:Bacillus / Q000201:enzymology*; D001665:Binding Sites; D002474:Cell-Free System; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000201:enzymology; D009584:Nitrogen / Q000378:metabolism; D009591:Nitrogenase / Q000378:metabolism*; D013045:Species Specificity; D013447:Sulfites / Q000378:metabolism
D001386:Azides; D013447:Sulfites; D000255:Adenosine Triphosphate; D009591:Nitrogenase; D009584:Nitrogen; D000114:Acetylene