title: Olive Recombinant Hydroperoxide Lyase, an Efficient Biocatalyst for Synthesis of Green Leaf Volatiles.
abstract: Volatile C6-aldehydes are the main contributors to the characteristic odor of plants known as "green note" and are widely used by the flavor industry. Biotechnological processes were developed to fulfill the high demand in C6-aldehydes in natural flavorants and odorants. Recombinant hydroperoxide lyases (HPLs) constitute an interesting alternative to overcome drawbacks arising from the use of HPL from plant extracts. Thus, olive recombinant 13-HPL was assayed as biocatalysts to produce C6-aldehydes. Firstly, a cDNA encoding for olive HPL of Leccino variety was isolated and cloned in pQE-30 expression vector. In order to improve the enzyme solubility, its chloroplast transit peptide was deleted. Both enzymes (HPL wild type and HPL deleted) were expressed into Escherichia coli strain M15, purified, characterized, and then used for bioconversion of 13-hydroperoxides of linoleic and linolenic acids. Aldehydes produced were extracted, then identified and quantified using gas chromatography and mass spectrometry. Recombinant HPL wild type (HPLwt) allowed producing 5.61 mM of hexanal and 4.39 mM of 3Z-hexenal, corresponding to high conversion yields of 93.5 and 73 %, respectively. Using HPL deleted (HPLdel) instead of HPLwt failed to obtain greater quantities of hexanal or 3Z-hexenal. No undesirable products were formed, and no isomerization of 3Z-hexenal in 2E-hexenal occurred. The olive recombinant HPLwt appears to be a promising efficient biocatalyst for the production of C6-aldehydes.
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Jacopini
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Mariani
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Virginie Brunini-Bronzini
authors.initials: VB
authors.lastname: de Caraffa
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Gambotti
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Vincenti
authors.affiliation: Laboratoire de Chimie des Produits Naturels, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Desjobert
authors.affiliation: Laboratoire de Chimie des Produits Naturels, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Muselli
authors.affiliation: Laboratoire de Chimie des Produits Naturels, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Costa
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France.
authors.forename: Liliane
authors.initials: L
authors.lastname: Berti
authors.affiliation: Laboratoire de Biochimie et Biologie Moléculaire Végétales, CNRS UMR6134 SPE, Université de Corse, Campus Grimaldi, BP52, 20250, Corte, France. maury@univ-corse.fr.
authors.forename: Jacques
authors.initials: J
authors.lastname: Maury
mesh_terms: D000446:Aldehyde-Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004798:Enzymes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli; D031658:Olea / Q000737:chemistry / Q000235:genetics*; D018515:Plant Leaves / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D055549:Volatile Organic Compounds / Q000737:chemistry / Q000378:metabolism
chemical_list: D000447:Aldehydes; D004798:Enzymes; D011994:Recombinant Proteins; D055549:Volatile Organic Compounds; D003577:Cytochrome P-450 Enzyme System; D000446:Aldehyde-Lyases; C054540:hydroperoxide lyase
keywords: 3Z-Hexenal; Bioconversion; C6-aldehydes; Green note; Hexanal; Hydroperoxide lyase