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A Rose by Any Other Name: Novel Wine Yeast That Impart Floral Aromas

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A Rose by Any Other Name: Novel Wine Yeast That Impart Floral Aromas WINEMAKINGAWRI REPORT A rose by any other name: novel wine yeast that impart oral aromas By Toni Cordente, Mark Solomon, Peter Godden, Chris Curtin and Dan Johnson Managing director The Australian Wine Research Institute. PO Box 197, Glen Osmond, South Australia, Australia Dan Johnson While there is substantial variation amongst existing wine yeast strains in their capacity to influence wine style, some desirable flavour compounds are not produced at concentrations where they significantly impact wine style. As part of ongoing yeast strain development projects at the Australian Wine Research Institute, novel wine yeast that make ‘floral’/’rose’ aroma compounds have been generated. INTRODUCTION AT A GLANCE “A rose by any other name would smell as sweet” (William • Substantial variation is present amongst existing Shakespeare). While varietal ‘floral’ aromas in wine are wine yeast strains in their capacity to influence wine style. typically the product of grape-derived terpenoid flavour compounds, products of yeast metabolism 2-phenylethanol • Some desirable flavour compounds are not present in wine at concentrations where they significantly (2-PE) and 2-phenylethyl acetate (2-PEA) are considered contributed to wine style. important contributors to ‘rose’ aroma (Vilanova et al. 2013). That’s not to say that yeast are what make roses smell • Novel wine yeast that make ‘floral’/’rose’ aroma compounds have been generated. like roses, but that they can enhance the ‘floral’ sensory properties of wine and other fermented foods and beverages • This will enable winemakers to 'dial up' these aromas as desired. (Fukuda et al. 1990, Dueñas-Sánchez et al. 2014). This is but one of several examples where plant and yeast metabolism overlap and make the same or similar compounds. In this case the metabolic pathway involved is named after German biochemist Felix Ehrlich, who noted that stopdust.com.au various flavour-active fusel alcohols had similar chemical Visit structures to amino acids, therefore, he proposed they were derived from them (Hazelwood et al. 2008). During winemaking, flavour compounds formed by yeast via the Ehrlich pathway confer a range of aromas and flavours that contribute to the overall bouquet of wine (Cordente et al. 2012). Generally, in white varietals the concentrations of 2-PE and 2-PEA in finished wines are below their aroma perception thresholds (Vilanova et al. 2013), meaning their contribution to wine style would likely be minimal. A ROSE AMONGST THE THORNS To generate a novel yeast that imparts floral aromas, a Got a dust problem well-known and widely utilised wine yeast strain, AWRI796, was exposed to a chemical selection process previously in your vineyard? applied to saké and baking yeast (Fukuda et al. 1990, Dueñas- Sánchez et al. 2014). A toxic analogue of the amino acid phenylalanine was incorporated into solid agar plates that a large number of AWRI796 cells were spread onto, and You need Road Con a separate set of plates were used for AWRI796 cells that had been previously exposed to a mutagenic agent. Only Polo Citrus Australia would like to take this opportunity to Road Con cells that carry mutations in key phenylalanine biosynthetic introduce you to their exceptional new product. has been designed to suppress dust on unsealed road pathway genes (that enable the cell to make more of its own surfaces for the medium to long term. supply of this essential amino acid) can grow in the presence of this toxic analogue. The mutagenic agent increases the frequency of isolating cells that carry such a mutation. While Ph it may be harsh to refer to AWRI796 as a thorn, cells carrying (03) 9364 9700 www.polocitrus.com.au V31N5 WINE & VITICULTURE JOURNAL SEPTEMBER/OCTOBER 2016 www.winetitles.com.au 39 WINEMAKINGAWRI REPORT ¨ ¥ B ! " # ? CCD E F D CD G @ A A A ^ _ . / 0 1 2 3 4 PR T W KL II H K X M N PR T Y H K X M N Y P T KJ ] Q L I M N § ¦ = > ? @ A 5 6 7 8 9 : ; < Ʃ © \ R PR T J M W [ K X M N ` a b > c d c @ ? F e A § ¥ Z PR T K X N M N V R J M W N N M N © ¦ T PU Q M N T T ] ] IW M N § $ % & ' ( )* ( + , * & -% ¥ Z S T J M [ W Q J M L K M N O P PR S N Q J N M N ¤ £ Ɵ ¢ ¢ £ ¡ ¡ r ` H IJ KL I H M N @ s C A A A CD Figure 2. Odour activity values (OAV) for the ‘rose’ aroma compounds Z S t T J M [ W Q M L K H M N KW W M M N ƟJ 2-phenylethanol (2-PE) and 2-phenylethyl acetate (2-PEA) in u T X M N Z PR X N H M N K ƩLJ Chardonnay wines (20L), produced using f g h i j j l j m n k \ v R S w S T W L Q x W W N M N \ R PR M W [ K X H M N J AWRI 796 (black) and the ‘rose’ strain f o o n j g j p q n Y P R T KJ ] Q L I H M N ^ _ X M z W M M N y AWRI 2940 (blue). Data is expressed PR W KL II [ K X M N as the mean and standard deviations of three fermentation replicates. An Figure 1. Mean ratings for aroma and flavour attributes for Chardonnay wines (20L scale) produced using AWRI 796 and the ‘rose’ strain AWRI 2940 (n = three OAV higher than one (odour threshold) fermentation replicates × 10 judges). ns, not significant; *p < 0.05; ***p < 0.001 indicates a possible contribution of the Aroma and flavour attributes are shown in green, and red, respectively. compound to the final aroma of wine. mutations that allow them to overproduce phenylalanine CONCLUSIONS do indeed smell like roses. A by-product of cells making large amounts of phenylalanine is that more is available By harnessing classical yeast strain development for biosynthesis of the flavour-active compounds 2-PE and techniques, yeast that impart floral aroma during wine 2-PEA. All yeast derived using this and related protocols are fermentation have been isolated and characterised. These not genetically-modified (GM) and are suitable for immediate strains have the potential to provide a tool for winemakers to application by the wine industry. produce more floral wine styles, and tailor new products to The AWRI screened 153 AWRI796 mutants for floral appeal to consumers. aromas, and 17 of those were used to conduct fermentations in the laboratory. The resultant wines were analysed for ACKNOWLEDGEMENTS a suite of volatile aroma compounds. Ten mutants that produced significantly higher concentrations of 2-PE and The authors gratefully acknowledge Petaluma winery 2-PEA had their whole genomes sequenced and, importantly, for the supply of high quality Chardonnay grapes for the six of these were ‘spontaneous mutants’ that had been winemaking trial. This work was financially supported by derived without use of the mutagenic agent, which means Australia’s grapegrowers and winemakers through their they carried a small number of changes ( <10 nucleotides out investment body, Wine Australia, with matching funds from of 12 million) in their genomes. This enabled identification of the Australian Government. The AWRI is part of the Wine two genes in the aromatic amino acid biosynthetic pathway Innovation Cluster. that harboured possible causative mutations. A pilot-scale winemaking trial was performed with one of REFERENCES the ‘spontaneous’ mutants (AWRI2940) using a high quality Cordente, A.G.; Curtin, C.D.; Varela, C. and Pretorius, I.S. (2012) Flavour- Adelaide Hills Chardonnay juice. There was no difference active wine yeasts. Appl, Microbiol. Biotechnol. 96: 601-618. observed between the mutant and parent strain AWRI796 with Dueñas-Sánchez, R.; Pérez, A.G.; Codón, A.C.; Benítez, T. and Rincón, A.M. regard to fermentation kinetics and overall wine chemistry. (2014) Over-production of 2-phenylethanol by industrial yeasts to improve organoleptic properties of bakers’ products. Int. J. Food Microbiol. 180:7–12. Sensory assessment revealed that the wines made with Fukuda, K.; Watanabe, M. and Asano, K. (1990) Altered regulation of AWRI2940 were rated significantly higher in the floral aroma aromatic amino acid biosynthesis in b-Phenylethyl-alcohol-overproducing attribute (Figure 1), and contained 7- and 37-fold higher mutants of Sake yeast Saccharomyces cerevisiae . Agric. Biol. Chem. 54:3151- 3156. concentrations of 2-PE and 2-PEA (Figure 2), respectively. Hazelwood, L.A.; Daran, J.M.; van Maris, A.J.A.; Pronk, J.T. and Dickinson, Less clear are the drivers for perceived differences in palate J.R. (2008) The Ehrlich pathway for fusel alcohol production: a century of terms ‘astringency’ and ‘bitter’, or the apparent slight research on Saccharomyces cerevisiae metabolism. Appl. Environ. Microbiol. increase in ‘yellow colour’; the flow-on impact of altered 74:2259-2266. Vilanova, M.; Genisheva, Z.; Graña, M. and Oliveira, J.M. (2013) amino acid metabolism during wine fermentation is currently Determination of odorants in varietal wines from international grape cultivars being investigated. (Vitis vinifera) grown in NW Spain. S. Afr. J. Enol. Vitic. 34:212-222. WVJ 40 www.winetitles.com.au WINE & VITICULTURE JOURNAL SEPTEMBER/OCTOBER 2016 V31N5.
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