Reduction of cyclopropene by NifV- and wild-type nitrogenases from Klebsiella pneumoniae.
The nitrogenase from wild-type Klebsiella pneumoniae reduces cyclopropene to cyclopropane and propene in the ratio 1:2 at pH 7.5. We show in this paper that the nitrogenase from a nifV mutant of K. pneumoniae also reduces cyclopropene to cyclopropane and propene, but the ratio of products is now 1:1.4. However, both nitrogenases exhibit the same Km for cyclopropene (2.1 x 10(4) +/- 0.2 x 10(4) Pa), considerably more than the Km for the analogous reaction with Azotobacter vinelandii nitrogenase under the same conditions (5.1 x 10(3) Pa). Analysis of the data shows that the different product ratio arises from the slower production of propene compared with cyclopropane by the mutant nitrogenase. During turnover, both nitrogenases use a large proportion of the electron flux for H2 production. CO inhibits the reduction of cyclopropene by both K. pneumoniae proteins, but the mutant nitrogenase exhibits 50% inhibition at approx. 10 Pa, whereas the corresponding value for the wild-type nitrogenase is approx. 110 Pa. However, H2 evolution by the mutant enzyme is much less affected than is cyclopropene reduction. CO inhibition of cyclopropene reduction by the nitrogenases coincides with a relative increase in H2 evolution, so that in the wild-type (but not the mutant) the electron flux is approximately maintained. The cyclopropane/propene production ratios are little affected by the presence of CO within the pressure ranges studied at least up to 50% inhibition.
Gemoets
J P
JP
Department of Chemistry, University of Southern California, Los Angeles 90089-0744.
Bravo
M
M
McKenna
C E
CE
Leigh
G J
GJ
Smith
B E
BE
D001665:Binding Sites; D002248:Carbon Monoxide / Q000494:pharmacology; D003521:Cyclopropanes / Q000378:metabolism*; D004583:Electrons; D007711:Klebsiella pneumoniae; D009154:Mutation; D009591:Nitrogenase / Q000378:metabolism*; D010084:Oxidation-Reduction
D003521:Cyclopropanes; C021985:cyclopropene; D002248:Carbon Monoxide; C030797:cyclopropane; D009591:Nitrogenase