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

• 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
• ISSN: 1567-1356; 1567-1364
• DOI: 10.1111/j.1567-1364.2007.00262.x

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.