Abstract
Several strains that grow on medium-chain-length alkanes and catalyze interesting hydroxylation and epoxidation reactions do not possess integral membrane nonheme iron alkane hydroxylases. Using PCR, we show that most of these strains possess enzymes related to CYP153A1 and CYP153A6, cytochrome P450 enzymes that were characterized as alkane hydroxylases. A vector for the polycistronic coexpression of individual CYP153 genes with a ferredoxin gene and a ferredoxin reductase gene was constructed. Seven of the 11 CYP153 genes tested allowed Pseudomonas putida GPo12 recombinants to grow well on alkanes, providing evidence that the newly cloned P450s are indeed alkane hydroxylases.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alkanes / metabolism*
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Base Composition
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Biodegradation, Environmental
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Cell Membrane / enzymology*
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Cytochrome P-450 CYP4A
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Cytochrome P-450 Enzyme System / genetics
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Cytochrome P-450 Enzyme System / metabolism*
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Gram-Positive Bacteria / enzymology*
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Gram-Positive Bacteria / genetics
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Gram-Positive Bacteria / metabolism
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Mixed Function Oxygenases / genetics
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Mixed Function Oxygenases / metabolism*
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Molecular Sequence Data
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Proteobacteria / enzymology*
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Proteobacteria / genetics
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Proteobacteria / metabolism
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Pseudomonas putida / enzymology
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Pseudomonas putida / genetics
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Analysis, DNA
Substances
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Alkanes
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Recombinant Proteins
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Cytochrome P-450 Enzyme System
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Mixed Function Oxygenases
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cytochrome P-450 CYP153 (Acinetobacter)
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Cytochrome P-450 CYP4A