Production of a recombinant alkane hydroxylase (AlkB2) from Alcanivorax borkumensis.
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.
Miri
Mandana
M
Science and Research Branch of Tehran, Islamic Azad University, Tehran 1477893855, Iran.
Bambai
Bijan
B
Tabandeh
Fatemeh
F
Sadeghizadeh
Majid
M
Kamali
Nasrin
N
D053521:Alcanivoraceae / Q000201:enzymology* / Q000235:genetics; D015153:Blotting, Western; D042926:Cytochrome P-450 CYP4A / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012416:Rubredoxins / Q000378:metabolism
D011994:Recombinant Proteins; D012416:Rubredoxins; D042926:Cytochrome P-450 CYP4A