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A candidate cDNA clone for (−)-limonene-7-hydroxylase from Perilla frutescens

Authors :
Mau, Christopher J.D.
Karp, Frank
Ito, Michiho
Honda, Gisho
Croteau, Rodney B.
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