Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive
Hydroxytyrosol derivatives are the most important phenolic components in virgin olive oil due to their well-demonstrated biological activities. In this regard, two phenyl acetaldehyde reductase genes, <i>OePAR1.1</i> and <i>OePAR1.2,</i> involved in hydroxytyrosol synthesis,...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-06-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/10/7/1268 |
| _version_ | 1827689128431452160 |
|---|---|
| author | Rosario Sánchez Cristina Bahamonde Carlos Sanz Ana G. Pérez |
| author_facet | Rosario Sánchez Cristina Bahamonde Carlos Sanz Ana G. Pérez |
| author_sort | Rosario Sánchez |
| collection | DOAJ |
| description | Hydroxytyrosol derivatives are the most important phenolic components in virgin olive oil due to their well-demonstrated biological activities. In this regard, two phenyl acetaldehyde reductase genes, <i>OePAR1.1</i> and <i>OePAR1.2,</i> involved in hydroxytyrosol synthesis, have been identified from an olive transcriptome. Both genes were synthesized and expressed in <i>Escherichia coli</i>, and their encoded proteins were purified. The recombinant enzymes display high substrate specificity for 2,4-dihydroxyphenylacetaldehyde (3,4-DHPAA) to form hydroxytyrosol. The reaction catalyzed by OePAR constitutes the second, and last, biochemical step in the formation of hydroxytyrosol from the amino acid L-3,4-dihydroxyphenylalanine (L-DOPA) in olive. OePAR1.1 and OePAR1.2 enzymes exhibit high thermal stability, similar pH optima (pH 6.5), and high affinity for 3,4-DHPAA (apparent <i>K</i>m 0.6 and 0.8 µmol min<sup>−1</sup> mg<sup>−1</sup>, respectively). However, OePAR1.2 exhibited higher specific activity and higher expression levels in all the olive cultivars under study. The expression analyses indicate that both <i>OePAR1.1</i> and <i>OePAR1.2</i> genes are temporally regulated in a cultivar-dependent manner. The information provided here could be of interest for olive breeding programs searching for new olive genotypes with the capacity to produce oils with higher levels of hydroxytyrosol derivatives. |
| first_indexed | 2024-03-10T10:10:28Z |
| format | Article |
| id | doaj.art-2cd8ebfcc39d4565a9962f0ece73408f |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T10:10:28Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-2cd8ebfcc39d4565a9962f0ece73408f2023-11-22T01:10:38ZengMDPI AGPlants2223-77472021-06-01107126810.3390/plants10071268Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in OliveRosario Sánchez0Cristina Bahamonde1Carlos Sanz2Ana G. Pérez3Department of Biochemistry and Molecular Biology of Plant Products, Instituto de la Grasa (CSIC), 41013 Sevilla, SpainDepartment of Biochemistry and Molecular Biology of Plant Products, Instituto de la Grasa (CSIC), 41013 Sevilla, SpainDepartment of Biochemistry and Molecular Biology of Plant Products, Instituto de la Grasa (CSIC), 41013 Sevilla, SpainDepartment of Biochemistry and Molecular Biology of Plant Products, Instituto de la Grasa (CSIC), 41013 Sevilla, SpainHydroxytyrosol derivatives are the most important phenolic components in virgin olive oil due to their well-demonstrated biological activities. In this regard, two phenyl acetaldehyde reductase genes, <i>OePAR1.1</i> and <i>OePAR1.2,</i> involved in hydroxytyrosol synthesis, have been identified from an olive transcriptome. Both genes were synthesized and expressed in <i>Escherichia coli</i>, and their encoded proteins were purified. The recombinant enzymes display high substrate specificity for 2,4-dihydroxyphenylacetaldehyde (3,4-DHPAA) to form hydroxytyrosol. The reaction catalyzed by OePAR constitutes the second, and last, biochemical step in the formation of hydroxytyrosol from the amino acid L-3,4-dihydroxyphenylalanine (L-DOPA) in olive. OePAR1.1 and OePAR1.2 enzymes exhibit high thermal stability, similar pH optima (pH 6.5), and high affinity for 3,4-DHPAA (apparent <i>K</i>m 0.6 and 0.8 µmol min<sup>−1</sup> mg<sup>−1</sup>, respectively). However, OePAR1.2 exhibited higher specific activity and higher expression levels in all the olive cultivars under study. The expression analyses indicate that both <i>OePAR1.1</i> and <i>OePAR1.2</i> genes are temporally regulated in a cultivar-dependent manner. The information provided here could be of interest for olive breeding programs searching for new olive genotypes with the capacity to produce oils with higher levels of hydroxytyrosol derivatives.https://www.mdpi.com/2223-7747/10/7/1268hydroxytyrosolphenyl acetaldehyde reductaseolive fruitphenolic compoundsvirgin olive oil<i>Olea europaea</i> |
| spellingShingle | Rosario Sánchez Cristina Bahamonde Carlos Sanz Ana G. Pérez Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive Plants hydroxytyrosol phenyl acetaldehyde reductase olive fruit phenolic compounds virgin olive oil <i>Olea europaea</i> |
| title | Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive |
| title_full | Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive |
| title_fullStr | Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive |
| title_full_unstemmed | Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive |
| title_short | Identification and Functional Characterization of Genes Encoding Phenylacetaldehyde Reductases That Catalyze the Last Step in the Biosynthesis of Hydroxytyrosol in Olive |
| title_sort | identification and functional characterization of genes encoding phenylacetaldehyde reductases that catalyze the last step in the biosynthesis of hydroxytyrosol in olive |
| topic | hydroxytyrosol phenyl acetaldehyde reductase olive fruit phenolic compounds virgin olive oil <i>Olea europaea</i> |
| url | https://www.mdpi.com/2223-7747/10/7/1268 |
| work_keys_str_mv | AT rosariosanchez identificationandfunctionalcharacterizationofgenesencodingphenylacetaldehydereductasesthatcatalyzethelaststepinthebiosynthesisofhydroxytyrosolinolive AT cristinabahamonde identificationandfunctionalcharacterizationofgenesencodingphenylacetaldehydereductasesthatcatalyzethelaststepinthebiosynthesisofhydroxytyrosolinolive AT carlossanz identificationandfunctionalcharacterizationofgenesencodingphenylacetaldehydereductasesthatcatalyzethelaststepinthebiosynthesisofhydroxytyrosolinolive AT anagperez identificationandfunctionalcharacterizationofgenesencodingphenylacetaldehydereductasesthatcatalyzethelaststepinthebiosynthesisofhydroxytyrosolinolive |