Two genes encoding proteins with similarities to rubredoxin and rubredoxin reductase are required for conversion of dodecane to lauric acid in Acinetobacter calcoaceticus ADP1.
Mutants of Acinetobacter calcoaceticus ADP1 unable to grow on dodecane, but retaining the ability to grow on lauric acid were isolated after ethylmethanesulphonate (EMS) treatment. This growth deficiency was complemented by a clone from a gene library constructed from chromosomal DNA of the wild-type strain. The complementing DNA mapped in a gene encoding a polypeptide with homology to rubredoxins. The deduced putative rubredoxin amino acid sequence is more similar to related proteins from Gram-positive bacteria than to the Pseudomonas oleovorans rubredoxin involved in alkane oxidation. An adjacent gene encodes a protein with similarity to rubredoxin reductase from Pseudomonas oleovorans and related NAD(P)-dependent reductases. Disruption of the rubredoxin-encoding gene by insertion of a KmR/lacZ cassette rendered the resulting strain unable to grow on dodecane or hexadecane. This demonstrates that these genes are necessary for alkane degradation. Transcriptional fusion of lacZ to the rubredoxin-encoding gene led to low level constitutive beta-galactosidase expression, whereas the fusion oriented in the opposite direction was not expressed.
Haspel
Gerhard
G
Lehrstuhl fr Mikrobiologie, Institut fr Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander Universitt Erlangen-Nrnberg,Staudtstrae 5, 91058 Erlangen, FRG.
Ehrt
Sabine
S
Lehrstuhl fr Mikrobiologie, Institut fr Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander Universitt Erlangen-Nrnberg,Staudtstrae 5, 91058 Erlangen, FRG.
Hillen
Wolfgang
W
Lehrstuhl fr Mikrobiologie, Institut fr Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander Universitt Erlangen-Nrnberg,Staudtstrae 5, 91058 Erlangen, FRG.
D016954:Acinetobacter calcoaceticus / Q000235:genetics* / Q000378:metabolism; D000473:Alkanes / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001673:Biodegradation, Environmental; D016384:Consensus Sequence; D004926:Escherichia coli; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D007850:Lauric Acids / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016254:Mutagenesis, Insertional; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012416:Rubredoxins / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000473:Alkanes; D001426:Bacterial Proteins; D007850:Lauric Acids; D011993:Recombinant Fusion Proteins; D012416:Rubredoxins; C030358:lauric acid; C007548:n-dodecane; C023091:rubredoxin-NAD+ reductase; D009247:NADH, NADPH Oxidoreductases