Nitrogenase catalyzes the ATP-dependent reduction of dinitrogen to ammonia,the central process of biological nitrogen *** mechanism of electron transfer from the P-cluster to the iron-molybdenum cofactor(FeMo-co)durin...
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Nitrogenase catalyzes the ATP-dependent reduction of dinitrogen to ammonia,the central process of biological nitrogen *** mechanism of electron transfer from the P-cluster to the iron-molybdenum cofactor(FeMo-co)during the process of substrate reduction remains *** on our previous hypothesis,the dual pathway model,three mutants of Klebsiella oxytoca strain M5al(termed Nb102A,Ka431H and Ia423P)were constructed to map the H?/e-transfer pathway,in which b-102Asn,a-431Lys and a-423Ile of the molybdenum-iron(MoFe)protein of nitrogenases were replaced by alanine,histidine and proline,*** three substitutions affected the diazotrophic growth,especially of the Ia423P strain with very low capability of diazotrophic growth and C2H2reduction *** a 42-L fermenter,the Ia423P strain was induced to the maximal C2H2reduction activity,which was only*17%of that of ***,the transcription level of the nifD gene was approximately four-fold higher than that of the M5al strain,indicating that the low C2H2reduction activity of Ia423P was not because of downregulation of nifD *** maximal C2H2and H?reduction activities of the Ia423P MoFe protein were 13%and 21%of those of the wild-type MoFe protein,*** conclusion,we propose that the changed properties of the Ia423P MoFe protein are related to the disruption of hydrogen bond formation between a-423Ile and homocitrate,and that a-423Ile serves as a gate to the Mo site in FeMo-co during the H?/e-transfer in nitrogenase.
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