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Staijen, IE, Van Beilen, JB and Witholt, B (2000) Expression, stability and performance of the three-component alkane mono-oxygenase of Pseudomonas oleovorans in Escherichia coli. Eur. J. Biochem. 267:1957-65


We tested the synthesis and in vivo function of the inducible alkane hydroxylase of Pseudomonas oleovorans GPo1 in several Escherichia coli recombinants. The enzyme components (AlkB, AlkG and AlkT) were synthesized at various rates in different E. coli hosts, which after induction produced between twofold and tenfold more of the Alk components than did P. oleovorans. The enzyme components were less stable in recombinant E. coli hosts than in P. oleovorans. In addition, the specific activity of the alkane mono-oxygenase component AlkB was five or six times lower in E. coli than in P. oleovorans. Evidently, optimal functioning of the hydroxylase system requires factors or a molecular environment that are available in Pseudomonas but not in E. coli. These factors are likely to include correct interactions of AlkB with the membrane and incorporation of iron into the AlkG and AlkB apoproteins.




Alkane 1-Monooxygenase; Cytochrome P-450 Enzyme System/biosynthesis; Cytochrome P-450 Enzyme System/genetics; Cytochrome P-450 Enzyme System/metabolism; Enzyme Stability; Escherichia coli/genetics; Mixed Function Oxygenases/biosynthesis; Mixed Function Oxygenases/genetics; Mixed Function Oxygenases/metabolism; Pseudomonas/enzymology; Recombinant Proteins/biosynthesis; Recombinant Proteins/genetics; Recombinant Proteins/metabolism



Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status


Contributes to

GO:0015044: rubredoxin-NAD+ reductase activity



alkT encodes the 3rd component of the alkane hydroxylase complex, the rubredoxin reductase. Transfers electrons from NADH to rubredoxin.

Figure 7 shows the highest specific activity for alkT in P.oleovorans, lower in E.coli HB101 and lowest in E.coli W3110. This concluded that the rubredoxin reductase is present in excess in the E. coli alk+ recombinants

CACAO 4718

See also


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