Rubredoxins involved in alkane oxidation.
Rubredoxins (Rds) are essential electron transfer components of bacterial membrane-bound alkane hydroxylase systems. Several Rd genes associated with alkane hydroxylase or Rd reductase genes were cloned from gram-positive and gram-negative organisms able to grow on n-alkanes (Alk-Rds). Complementation tests in an Escherichia coli recombinant containing all Pseudomonas putida GPo1 genes necessary for growth on alkanes except Rd 2 (AlkG) and sequence comparisons showed that the Alk-Rds can be divided in AlkG1- and AlkG2-type Rds. All alkane-degrading strains contain AlkG2-type Rds, which are able to replace the GPo1 Rd 2 in n-octane hydroxylation. Most strains also contain AlkG1-type Rds, which do not complement the deletion mutant but are highly conserved among gram-positive and gram-negative bacteria. Common to most Rds are the two iron-binding CXXCG motifs. All Alk-Rds possess four negatively charged residues that are not conserved in other Rds. The AlkG1-type Rds can be distinguished from the AlkG2-type Rds by the insertion of an arginine downstream of the second CXXCG motif. In addition, the glycines in the two CXXCG motifs are usually replaced by other amino acids. Mutagenesis of residues conserved in either the AlkG1- or the AlkG2-type Rds, but not between both types, shows that AlkG1 is unable to transfer electrons to the alkane hydroxylase mainly due to the insertion of the arginine, whereas the exchange of the glycines in the two CXXCG motifs only has a limited effect.
van Beilen
Jan B
JB
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland. vanbeilen@biotech.biol.ethz.ch
Neuenschwander
Martin
M
Smits
Theo H M
TH
Roth
Christian
C
Balada
Stefanie B
SB
Witholt
Bernard
B
D000473:Alkanes / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D042926:Cytochrome P-450 CYP4A; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000201:enzymology / Q000235:genetics; D007501:Iron / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009795:Octanes; D010084:Oxidation-Reduction; D010802:Phylogeny; D010957:Plasmids; D012416:Rubredoxins / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D016415:Sequence Alignment
D000473:Alkanes; D003470:Culture Media; D009795:Octanes; D012416:Rubredoxins; D003577:Cytochrome P-450 Enzyme System; D007501:Iron; D006899:Mixed Function Oxygenases; D042926:Cytochrome P-450 CYP4A; C026728:octane