Purification and some properties of alcohol oxidase from alkane-grown <i>Candida tropicalis</i>

Dickinson, F M; Wadforth, C

doi:10.1042/bj2820325

Cited by 27 publications

(13 citation statements)

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“…The running buffer and apparatus were chilled on ice before and during the run. The sample was 50 l of 0.2 units of hydroxylapatite-purified C. cloacae material prepared by the method of Dickinson and Wadforth (7). To stain the gel for enzyme activity, buffer, ABTS, and peroxidase at 10 times the concentration used in the standard LC-FAO assay and dodecanol at 2.7 mM were applied to the surface of the gel.…”

Section: Miscellaneous Methodsmentioning

confidence: 99%

“…Purification of Long-chain Fatty-acid Alcohol Oxidase from C. cloacae-We initially adopted a procedure similar to that used for the purification of LC-FAO from C. tropicalis (7). The C. tropicalis procedure did not, however, result in a homogeneous preparation with C. cloacae extracts.…”

Section: Alterations In the Protein Profile Upon Oleic Acid Induction-mentioning

confidence: 99%

“…Flavin-dependent alcohol oxidases have been isolated from a number of different fungal sources (4 -9). In all organisms studied to date, the enzyme is octameric, with the exception of C. tropicalis, where it is dimeric (7). The substrate specificity of the enzyme differs considerably depending on the source of the enzyme.…”

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confidence: 99%

“…The enzymes from Candida maltosa (10), C. tropicalis (7), and C. cloacae (11) all oxidize -hydroxy fatty acids, whereas those from other organisms studied lack the ability to use these substrates. Long-chain alcohol oxidase has been purified from C. tropicalis (7), but to date, there is no gene or amino acid sequence available.…”

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confidence: 99%

“…The substrate specificity of the enzyme differs considerably depending on the source of the enzyme. In C. tropicalis, the enzyme is most active on long-chain fatty alcohols (7), whereas secondary alcohols such as dodecan-2-ol and long-chain -hydroxy fatty acids are also good substrates of the enzyme (7). Alcohol oxidases from Kloeckera sp., Hansenula polymorpha, and Candida boidinii are very active toward methanol and ethanol, but are inactive toward substrates of chain length longer than C 5 and will not oxidize secondary alcohols or -hydroxy fatty acids (4,6).…”

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confidence: 99%

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A Consensus Sequence for Long-chain Fatty-acid Alcohol Oxidases from Candida Identifies a Family of Genes Involved in Lipid ω-Oxidation in Yeast with Homologues in Plants and Bacteria

Vanhanen

West

Kroon

et al. 2000

Journal of Biological Chemistry

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The yeast Candida cloacae is capable of growing on alkanes and fatty acids as sole carbon sources. Transfer of cultures from a glucose medium to one containing oleic acid induced seven proteins of M r 102,000, 73,000, 61,000, 54,000, and 46,000 and two in the region of M r 45,000 and repressed a protein of M r 64,000. The induction of the M r 73,000 protein reached a 7-fold maximum 24 h after induction. The protein was confirmed by its enzyme activity to be a long-chain fatty-acid alcohol oxidase (LC-FAO) and purified to homogeneity from microsomes by a rapid procedure involving hydrophobic chromatography. An internal peptide of 30 amino acids was sequenced. A 1100-base pair cDNA fragment containing the LC-FAO peptide coding sequence was used to isolate a single exon genomic clone containing the full-length coding sequence of an LC-FAO (fao1). The fao1 gene product was expressed in Escherichia coli and was translated as a functional long-chain alcohol oxidase, which was present in the membrane fraction. In addition, full-length coding sequences for a Candida tropicalis LC-FAO (faoT) and a second C. cloacae LC-FAO (fao2) were isolated. The DNA sequences obtained had open reading frames of 2094 (fao1), 2091 (fao2), and 2112 (faoT) base pairs. The derived amino acid sequences of fao2 and faoT showed 89.4 and 76.2% similarities to fao1. The fao1 gene is much more highly induced on alkane than is fao2. Although this study describes the first known DNA sequences encoding LC-FAOs from any source, there are unassigned Arabidopsis sequences and an unassigned Mycobacterium sequence in the GenBank TM Data Bank that show strong homology to the described LC-FAO sequences. The conservation of sequence between yeast, plants, and bacteria suggests that an as yet undescribed family of long-chain fattyacid oxidases exists in both eukaryotes and prokaryotes.

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Section: Miscellaneous Methodsmentioning

confidence: 99%

Section: Alterations In the Protein Profile Upon Oleic Acid Induction-mentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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