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A candidate cDNA clone for (−)-limonene-7-hydroxylase from Perilla frutescens
- Source :
-
Phytochemistry . Mar2010, Vol. 71 Issue 4, p373-379. 7p. - Publication Year :
- 2010
-
Abstract
- Abstract: Cytochrome P450 mono-oxygenases from peppermint, spearmint and perilla (all members of the family Lamiaceae) mediate the regiospecific hydroxylation of the parent olefin (−)-limonene to produce essential oil components oxygenated at C3, C6 and C7, respectively. Cloning, expression and mutagenesis of cDNAs encoding the peppermint limonene-3-hydroxylase and the spearmint limonene-6-hydroxylase have allowed the identification of a single amino acid residue which determines the regiospecificity of oxygenation by these two enzymes. A hybridization strategy provided a cytochrome P450 limonene hydroxylase cDNA from perilla with which to further evaluate the structural determinants of regiospecificity for oxygenation of the common substrate (−)-limonene. The perilla cDNA was a partial clone of 1550bp (lacking the N-terminal membrane insertion domain), and shared 66% identity with the peppermint 3-hydroxylase and spearmint 6-hydroxylase at the amino acid level. The perilla cytochrome P450 was expressed in Escherichia coli as a chimeric protein fused with the N-terminal membrane insertion domain of the limonene-3-hydroxylase. The kinetically competent recombinant protein was characterized and shown to produce a mixture of C3-, C6- and C7-hydroxylated limonene derivatives with a distribution of 33%, 14% and 53%, respectively. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 00319422
- Volume :
- 71
- Issue :
- 4
- Database :
- Academic Search Index
- Journal :
- Phytochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 48148319
- Full Text :
- https://doi.org/10.1016/j.phytochem.2009.12.002