The Pseudomonas oleovorans alkane hydroxylase gene: sequence and expression

We have identified and sequenced the Pseudomonas OCT plasmid-encoded alkane hydroxylase gene (alkB) and its promoter. The transcription initiation site of the alkBAC mRNA was determined by nuclease S1 mapping. A putative interaction site with RNA-polymerase was identified based on homology of the al...

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Bibliographic Details
Published in:The Journal of biological chemistry 1989-04, Vol.264 (10), p.5435-5441
Main Authors: KOK, M, OLDENHUIS, R, VAN DER LINDEN, M. P. G, RAATJES, P, KINGMA, J, VAN LELYVELD, P. H, WITHOLT, B
Format: Article
Language:English
Subjects:
alkane 1-monooxygenase
Amino Acid Sequence
arn mensajero
arn messager
Base Sequence
Biological and medical sciences
Cloning, Molecular
Cytochrome P-450 CYP4A
Cytochrome P-450 Enzyme System - genetics
Escherichia coli - genetics
expresion genica
expression des genes
Fundamental and applied biological sciences. Psychology
gene
Gene expression
Genes
Genes, Bacterial
Genotype
hidrocarburos
hydrocarbons
hydrocarbure
messenger rna
Mixed Function Oxygenases - genetics
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
nucleotide sequence
oxidoreductases
oxidorreductasas
oxydoreductase
Phenotype
Plasmids
Promoter Regions, Genetic
Protein Conformation
pseudomonas
Pseudomonas - enzymology
Pseudomonas - genetics
Pseudomonas oleovorans
Restriction Mapping
RNA, Messenger - genetics
secuencia nucleica
Sequence Homology, Nucleic Acid
sequence nucleique
Transcription, Genetic
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Summary:We have identified and sequenced the Pseudomonas OCT plasmid-encoded alkane hydroxylase gene (alkB) and its promoter. The transcription initiation site of the alkBAC mRNA was determined by nuclease S1 mapping. A putative interaction site with RNA-polymerase was identified based on homology of the alk promoter with other Pseudomonas promoters. The alkB gene encodes a 401-amino acid polypeptide which, despite an unusual codon composition, can be expressed at high levels in Escherichia coli and Pseudomonas. The amino-terminal sequence of the purified cytoplasmic membrane alkane hydroxylase was determined and was found to be in agreement with the nucleotide sequence. The translation product of the alkB gene contains nine hydrophobic sequences of which eight are sufficiently long to be membrane-spanning segments. The amino-terminal sequence resembles that of several bacterial integral membrane proteins and is not cleaved off following translation.
ISSN:0021-9258
1083-351X