Macrowine 2021
IVES 9 IVES Conference Series 9 Cytochrome P450 CYP71BE5 from grapevine (Vitis vinifera) catalyzes the formation of the spicy aroma compound, (-)-rotundone

Cytochrome P450 CYP71BE5 from grapevine (Vitis vinifera) catalyzes the formation of the spicy aroma compound, (-)-rotundone

Abstract

(-)-Rotundone, an oxygenated sesquiterpene, is a potent odorant molecule with a characteristic spicy aroma existing in various plants including grapes1. It is considered as a significant compound notably in wines and grapes because of its low sensory threshold (16 ng L-1 in red wine, 8 ng L-1 in water) and aroma properties. (-)-Rotundone was first identified in red wine made from the grape cultivar Syrah (regionally called Shiraz) in Australia1, and then it was found in several grape varieties such as Duras, Grüner Veltliner, Schioppettino and Vespolina from Europe2, 3. Several environmental factors affecting the accumulation of (-)-Rotundone during the grape maturation, were reported such as ambient temperature4, soil properties and topography5, soil moisture from irrigation and light exposure in the bunch zone by leaf removal2. However, the biosynthetic pathway of (-)-rotundone in grapevine has been unclarified to date. In this study, we report the identification of VvSTO2 as a α-guaiene 2-oxidase which can transform α-guaiene to (-)-rotundone in the grape cultivar Syrah6. It is a cytochrome P450 (CYP) belonging to the CYP 71BE subfamily, which overlaps with the very large CYP71D family and, to the best of our knowledge, this is the first functional characterization of an enzyme from this family. VvSTO2 generated (-)-rotundone as the major reaction product with α-guaiene, and presumably (2R)-rotundol and (2S)-rotundol as the intermediate compounds to synthesize (-)-rotundone were found at trace levels in vitro enzyme assay. This result suggests that this enzyme can catalyze a one-step oxidation of α-guaiene to (-)-rotundone or a two-step oxidation via a rapid second oxidation from (2R)-rotundol and (2S)-rotundol to (-)-rotundone. VvSTO2 was expressed at a higher level in the Syrah grape exocarp (skin) in accord with the localization of (-)-rotundone accumulation in grape berries. α-Guaiene was also detected in the Syrah grape exocarp at an extremely high concentration. These findings suggest that (-)-rotundone accumulation is regulated by the VvSTO2 expression along with the availability of α-guaiene as a precursor. VvSTO2 expression during grape maturation was considerably higher in Syrah grape exocarp compared to Merlot grape exocarp, consistent with the patterns of α-guaiene and (-)-rotundone accumulation. On the basis of these findings, we propose that VvSTO2 may be a key enzyme in the biosynthesis of (-)-rotundone in grapevines by acting as a α-guaiene 2-oxidase. [References] [1] Wood, C., et al. 2008. J. Agric. Food Chem. 56, 3738-44. [2] Geffroy, O., et al. 2014. Aust. J. Grape Wine Res. 20, 401-408. [3] Mattivi, F., et al. 2011. Rapid Commun. Mass Spectrom. 25, 483-8. [4] Caputi, L., et al. 2011. J. Agric. Food Chem. 59, 5565-71. [5] Scarlett, N.J., et al. 2014. Aust. J. Grape Wine Res. 20, 214-222. [6] Takase, H., et al. 2015. J. Exp. Bot, in press.

Publication date: May 17, 2024

Issue: Macrowine 2016

Type: Poster

Authors

Hideki Takase*, Akira Shinohara, Akira Shinohara, Gen Ikoma, Hideyuki Shinmori, Hironori Kobayashi, Hironori Matsuo, Hiroshi Saito, Kanako Sasaki, Ryoji Takata

*Research Laboratories for Wine

Contact the author

Tags

IVES Conference Series | Macrowine | Macrowine 2016

Citation

Related articles…

Reduction of herbaceous aromas by wine lactic acid bacteria mediated degradation of volatile aldehydes

Consumers typically prefer wines with floral and fruity aromas over those presenting green-pepper, vegetal or herbaceous notes. Pyrazines have been identified as causatives for herbaceous notes in wines, especially Bordeaux reds. However, pyrazines are not universally responsible for herbaceousness, and several other wine volatile compounds are known to produce distinct vegetal/herbaceous aromas in wines. Specifically, volatile aldehydes elicit sensations of herbaceousness or grassiness and have been described in wines well above their perception thresholds.

Impact of industrial-scale serial filtration on macromolecules in red wines

Filtration is a critical step in ensuring the clarity and microbial stability of wine prior to bottling. However the process of filtering potentially reduces red wine quality by removing some of the macromolecules that contribute to the texture of the wine. Commercial red wines, Cabernet Sauvignon (CAS) and Shiraz (SHZ), of two vintages and two grades (premium grade wines from the older vintage: CAS13 and SHZ13; and standard grade wines from a younger vintage: CAS14 and SHZ14) were filtered through industrial-scale commercial filtration units prior to bottling. Samples were taken before and after cross-flow filtration, lenticular filters, 0.65 µm and 0.45 µm pore size nylon membrane filters. The concentration and composition of macromolecules, including tannins and polysaccharides, were measured in all samples as well as particle size distribution and wine colour.

Sensory definition of green aroma concept in red French wines. Evidence for the contribution of novel volatile markers

The aromatic complexity of a wine results from the perception of the association of volatile molecules and each aroma can be categorized into different families. The “green” aromas family in red wines has retained our attention by its close link with the fruity perception. In that study, the “green” olfactory concept of red wines was considered through a strategy combining both sensory analysis and hyphenated chromatographic techniques including HPLC and MDGC (Multidimensional Gas Chromatography). The aromatic space of this concept was specified by lexical generation through a free association task on 22 selected wines by a panel of wine experts. Then, 70 French red wines were scored on the basis of the intensity of their “green” and “fruity” attributes.

Influence of methyl jasmonate foliar application to vineyard on grape volatile composition over three consecutive vintages

An alternative to improve grape quality is the application to the vineyard of elicitors. Although these compounds were first used to increase resistance of plants against pathogens, it has been found that they are also able to induce mechanisms involved in the synthesis of phenolic compounds and some amino acids. However, researches about the influence of elicitors on grape volatile composition are scarcely. Therefore, the aim of this work was to study the influence of methyl jasmonate (MeJ) foliar application on grape aroma composition over three consecutive vintages. MeJ was applied to Tempranillo grapevines at a concentration of 10 mM in 2013, 2014, and 2015 years. Control plants were sprayed with water.

Ageing of sweet wines: oxygen evolution according to bung and barrel type

Barrel ageing is a crucial step in the wine process because it allows many changes to the wine as enrichment, colour stabilization, clarification and also a slow oxygenation. Effects of the oak barrel have to be known to prevent oxidation of the wine. The type of bung used during ageing is also a parameter to consider. Ageing sweet wines in barrel is a real challenge. These wines may need some oxygen at the beginning of ageing but they should be protected at the end of their maturation, to avoid oxidation.