Membrane-bound alkane hydroxylase

The initial hydroxylation of alkanes is mediated by both membrane-bound and soluble hydroxylases, and the genetics of alkane hydroxylation and alkanol dehydrogenation in pseudomonads is complex involving in Pseudomonas oleovorans the loci alkA, alkB, alkC, alkD, alkE (Fennewald et al. 

To determine the structure of the active site iron center we attempted to over-express various membrane bound enzymes for spectroscopic analysis. While the membrane bound desaturases accumulated to very low levels, the alkane hydroxylase accumulated to very high levels. Membranes containing Fe-enriched alkane hydroxylase were isolated and examined by Mossbauer spectroscopy. Preliminary spectra (Fig. 2) shows the major fraction of the iron in these preparations has Mossbauer parameters similar to those of the soluble castor A desaturase. 

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