Cloning and characterization of the NADPH cytochrome P450 reductase gene (CPR) from Candida bombicola

Abstract Candida bombicola is a yeast with at least two appealing features. The species can grow on alkanes when provided as the sole carbon source, and it produces glycolipids, which have several industrial, cosmetic and pharmaceutical applications. Both metabolic processes require in their pathway...

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Bibliographic Details
Published in:FEMS yeast research 2007-09, Vol.7 (6), p.922-928
Main Authors: Van Bogaert, Inge N.A., Develter, Dirk, Soetaert, Wim, Vandamme, Erick J.
Format: Article
Language:English
Subjects:
Alkanes
Amino Acid Sequence
Base Sequence
Candida - classification
Candida - enzymology
Candida - genetics
Candida bombicola
Carbon sources
Cloning
Cloning, Molecular
COX
CPR
Cytochrome
Cytochrome c
cytochrome c oxidase assembly protein
Cytochrome P450
Cytochrome P450 monooxygenase
Enzymes
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Fungal
Genes, Fungal - genetics
Glycolipids
Homology
Molecular Sequence Data
NADP
NADPH cytochrome P450 reductase
NADPH-ferrihemoprotein reductase
NADPH-Ferrihemoprotein Reductase - chemistry
NADPH-Ferrihemoprotein Reductase - genetics
NADPH-Ferrihemoprotein Reductase - metabolism
phylogenetic tree
Phylogeny
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Summary:Abstract Candida bombicola is a yeast with at least two appealing features. The species can grow on alkanes when provided as the sole carbon source, and it produces glycolipids, which have several industrial, cosmetic and pharmaceutical applications. Both metabolic processes require in their pathway the activity of cytochrome P450 monooxygenase. This enzyme needs and gets reducing equivalents from NADPH cytochrome P450 reductase (CPR). The CPR gene of Candida bombicola was isolated using degenerate PCR and genomic walking. The gene encodes an enzyme of 687 amino acids, which shows homology with known CPRs of other species. The functionality of the gene was proven by heterologous expression in Escherichia coli. The recombinant protein exhibited NADPH-dependent cytochrome c reducing activity. Cloning and characterization of this enzyme is an important step in the study of the cytochrome P450 monooxygenase system of Candida bombicola. The GenBank accession number of the sequence described in this article is EF050789.
ISSN:1567-1356
1567-1364
DOI:10.1111/j.1567-1364.2007.00262.x