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KMID : 0545120060160121995
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Journal of Microbiology and Biotechnology
2006 Volume.16 No. 12 p.1995 ~ p.1999
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Cloning of p-Hydroxybenzoate Degradation Genes and the Overexpression of Protocatechuate 4,5-Dioxygenase from Pseudomonas sp. K82
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Yun Young-Ho
Park Soon-Ho
Leem Sun-Hee
Kim Seung-Il
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Abstract
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Pseudomonas sp. K82 cultured in p-hydroxybenzoate induces protocatechuate 4,5-dioxygenase (PCD 4,5) for p-hydroxybenzoate degradation. In this study, a 6.0-kbp EcoR1 fragment containing p-hydroxybenzoate degradation genes was cloned from the genome of Pseudomonas sp. K82. Sequence analysis identified four genes, namely, pcaD, pcaA, pcaB, and pcaC genes known to be involved in p-hydroxybenzoate degradation. Two putative 4-hydroxyphenylpyruvate dioxygenases and one putative oxidoreductase were closely located by the p-hydroxybenzoate degradation genes. The gene arrangement and sequences of these p-hydroxybenzoate degradation genes were similar to those of Comamonas testosteroni and Pseudomonas ochraceae. PcaAB (PCD4,5) was overexpressed in the expression vector pGEX-4T-3, purified using a GST column, and confirmed to have protocatechuate 4,5-dioxygenase activity. The N-terminal amino acid sequences of overexpressed PCD4,5 were identical with those of purified PCD4,5 from Pseudomonas sp. K82.
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KEYWORD
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p-Hydroxybenzoate, protocatechuate 4,5-dioxygenase, pcaA and pcaB, Pseudomonas sp. K82
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