Production of a recombinant alkane hydroxylase (AlkB2) from Alcanivoraxborkumensis
Alcanivorax borkumensis is an oil-degrading marine bacterium. Its genome contains genes coding for three cytochrome P450s and two integral membrane alkane hydroxylases (AlkB1 & AlkB2), all assumed to perform hydroxylation of different linear or branched alkanes. Although, the sequence of alkB2 h...
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Veröffentlicht in: | Biotechnology letters 2010, Vol.32 (4), p.497-502 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Alcanivorax borkumensis
is an oil-degrading marine bacterium. Its genome contains genes coding for three cytochrome P450s and two integral membrane alkane hydroxylases (AlkB1 & AlkB2), all assumed to perform hydroxylation of different linear or branched alkanes. Although, the sequence of
alkB2
has been determined, the molecular characterization and the substrate specificity of AlkB2 require more precise investigation. In this study, AlkB2 from
A. borkumensis
SK2 was expressed in
Escherichia coli
to examine the functionality of AlkB2 as a hydroxylating enzyme. Furthermore, the activity of the enzyme in the presence of the accessory proteins, rubredoxin (RubA) and rubredoxin reductase (RubB), produced in
E. coli
BL21(DE3)plysS cells, was determined. Recombinant AlkB2 is produced in an active form and rubredoxin is the intermediate electron donor to AlkB2 and can replace AlkG function, when NADH is the prime electron donor. |
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ISSN: | 0141-5492 1573-6776 |
DOI: | 10.1007/s10529-009-0177-0 |