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KMID : 0380020040190040308
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Korean Journal of Biotechnology and Bioengineering
2004 Volume.19 No. 4 p.308 ~ p.314
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Isolation and Nucleotide Sequence Characterization of Novel Cytochrome P450 Hydroxylase Genes from Rare Actinomycetes, Sebekia benihana
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Park Nam-Sil
Park Hyun-Joo
Han Kyu-Boem
Kim Sang-Nyun
Kim Eung-Soo
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Abstract
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A degenerate set of PCR primers based on two conserved regions (heme binding region and oxygen ligand pocket) were designed and successfully applied to amplify DNA fragments of cytochrome P450 hydroxylase (CYP) genes from a rare actinomycetes, S. benihana. The PCR amplified products were employed as a DNA probe to clone the entire CYP genes from S. benihana genomic library. Five different CYP-positive cosmids were isolated by colony hybridization as well as PCR confirmation. The complete nucleotide sequencing of five different CYP genes revealed that each unique CYP showed a significant amino acid homology to previously-known CYP genes involved in streptomycetes secondary metabolism. In addition, four CYP genes (CYP502, CYP503, CYP504, CYP506) were found to be linked to ferredoxin genes in the chromosome, and the CYP503 gene showed the high degree of amino acid similarity to the previously well-characterized CYP105 family in streptomycetes.
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KEYWORD
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Sebekia benihana, cytochrome P450 hydroxylase, actinomycetes
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