194 Release of glycosidic aroma precursors Australian Journal of Grape and Wine Research 21, 194–199, 2015

Evaluation of the inherent capacity of commercial yeast strains

to release glycosidic aroma precursors from Muscat grape must
A. BISOTTO1, A. JULIEN1, P. RIGOU2, R. SCHNEIDER3 and J.M. SALMON2,4
1
Lallemand SAS, Blagnac, France
2
Sciences Pour l’Œnologie, Institut National de la Recherche Agronomique, Montpellier, France
3
Unité Mixte Technologique Qualinnov, Institut Français de la Vigne et du Vin, Gruissan, France
4
Unité Expérimentale de Pech Rouge (UE0999), Unité Mixte Technologique Qualinnov, Institut National de la Recherche
Agronomique, Gruissan, France
Corresponding author: Dr Jean-Michel Salmon, email jmsalmon@supagro.inra.fr

Abstract
Background and Aims: The aim of this work was to establish a general method for evaluating the intrinsic activity
of yeasts towards the release of volatiles terpenes from glycosidic aroma precursors.
Methods and Results: The enzymatic capacity of yeast cells towards glycosidic aroma precursors was assessed on
permeabilised yeast cells. In this way, the pores created into the cell membrane allowed the precursors to freely enter
the cells.
Conclusions: The comparison between the release of terpenes from glycosidic aroma precursors by entire viable cells
during fermentation and by permeabilised cells shows that the permeation of glycosidic aroma precursors through
the yeast cell membrane is the main bottleneck for such release.
Significance of the Study: This work highlights the fact that Saccharomyces species can exhibit β-glycosidase activity
as high as non-Saccharomyces species. This important result points out future prospects for the improvement of
Saccharomyces species by classical genetics.

Keywords: aroma precursor, terpene, wine, yeast

Introduction specific for apioside substrates might be limited in Saccharomyces

During wine fermentation, yeasts are responsible for the trans-
formation of some aroma precursors into aromatic compounds.
It is now well established that certain monoterpenols of grapes,
for example linalool, geraniol, nerol, citronelol, α-terpineol
and linalool oxide, which are linked to diglycosides, such as
6-O-α-L-rhamnopyranosyl-, 6-O-α-L-rhamnofuranosyl- and
6-O-β-D-apiofuranosyl-β-D-glucosides, contribute significantly
to the flavour of wine (Günata et al. 1985, Williams et al.
1989, Schneider et al. 2001). The enzymatic hydrolysis of
these compounds requires a sequential reaction: first, an α-L-
rhamnosidase or a β-D-apiofuranosidase cleaves the (1→6)-
glycosidic linkage, and then, the flavour compounds are
liberated from the monoglucosides by the action of a β-
glucosidase (Günata et al. 1988, 1993). This cleavage can also
occur during wine ageing, under mild acidic conditions. Unlike
acid hydrolysis, enzymatic hydrolysis is efficient and does not
result in modification of the aromatic composition of the bound
fraction (Günata et al. 1990). Yeast glucosidases, however,
exhibit limited activity on monoterpenyl glycosides during
winemaking, and a large fraction of the aromatic precursors
remains unprocessed (Delcroix et al. 1994, Rosi et al. 1994,
Ugliano et al. 2006). Moreover, Gil et al. (2005) have shown
that induced overproduction of an endogenous exoglucanase in
a Saccharomyces cerevisiae strain led to an increase in the release
of glycoside-related volatile compounds in wine, suggesting the
involvement of other enzymatic activities in the hydrolysis of
glycosides by yeast. Furthermore, the work of Ugliano et al.
(2006) indicates that production and/or activity of enzymes
yeast during fermentation, at least in the three strains tested by
these authors.
The terpene profile of Muscat wines fermented by
Saccharomyces species and hybrid yeasts was also investigated
(Gamero et al. 2011). These authors did not find any relation-
ship between β-d-glucosidase activity and the terpene profile
in Muscat wines fermented with Saccharomyces species and
hybrids. The release or formation of aroma compounds from
precursors was found to be strongly linked to the hybrid used,
and, for example, the triple hybrid S. cerevisiae × S. bayanus × S.
kudriavzevii in particular and secondarily the hybrid S.
cerevisiae × S. bayanus were highly efficient in the production of
most varietal terpenols (Gamero et al. 2011). Altogether, these
results suggest that there are two main limiting factors for a
complete release of monoterpenols from grape glycosidic pre-
cursors by yeasts. The first bottleneck deals with the transport of
the precursors into the yeast cells. Aroma glycosidic precursors
are large molecules, which cannot travel through the plasma
membrane by simple diffusion in intact viable cells. Moreover,
Darriet et al. (1988) reported that the β-d-glucosidase activity is
located in the periplasmic space of yeast cell, presumably easily
liberated during yeast autolysis. In addition, it has also been
found that lees from different yeast strains may have a slightly
different ability to release volatile compounds derived from
glycosidic precursors (Loscos et al. 2009).
To our knowledge, however, no scientific data are available
on the specific transport of such molecules in yeasts. The second
point deals with the activity of the enzymes themselves.

doi: 10.1111/ajgw.12127
© 2015 Australian Society of Viticulture and Oenology Inc.
Bisotto et al. Release of glycosidic aroma precursors 195

Previous work has established by working on entire yeast cells ture 245°C; purge on time 0.5-min) onto a DB-Wax column
that only non-Saccharomyces species exhibited β-glycosidase [30 m × 0.25 mm id, 0.25 μm film thickness (Agilent Technolo-
activity (Mendes Ferreira et al. 2001). In contrast, other work gies, Santa Clara, CA, USA)]. Compounds were separated using
has revealed that entire Saccharomyces species cells could exhibit helium carrier gas at 1 mL/min. The temperature program
a low β-glycosidase activity (Delcroix et al. 1994, Zoecklein began with an isotherm at 60°C for 3 min. The temperature of
et al. 1997, Ugliano et al. 2006). Therefore it is possible that the oven was then raised by 3°C/min to 245°C and held for
some yeast strains that exhibit efficient enzymatic β-glycosidase 10-min. The transfer line was held at 250°C, and compounds
activity may remain unable to take up the glycosidic precursors were detected with the source held at 150°C by ionisation by
as substrates. electronic impact generated at 70°C. Full scan mass spectra were
The aim of this work is to establish a general method for recorded between 29 and 350 m/z. Data were acquired and
evaluating the intrinsic activity of yeasts towards the release of treated with the HP 5989 B.05.02 MS Chemstation. The
glycosidic aroma precursors. For this purpose, we use the tech- terpenes identified were semi-quantified using 4-nonanol as an
nique of cell permeabilisation (Salmon 1984) to create pores internal standard. Limits of detection and of quantification of
into the cell membrane allowing the precursors to freely enter the method for the terpenes considered are given in Table 1.
the cells, where the enzymatic release of aglycone terpenes by
yeast enzymatic activity can take place. In this way, the enzy-
matic capacity of the yeast cells can be easily assessed. Yeast strains
All tested yeast strains were commercially available active
dry yeasts (ADY) for winemaking (Lallemand, Montreal, QC,
Material and methods
Canada): five S. cerevisiae strains (Lalvin QA23, Lalvin ICV K1M,
Grape glycosidic precursor purification and analysis Vitilevure DV10, Uvaferm CEG, Lalvin EC-1118); a natural
Preparation of a glycosidic fraction from Muscat must. hybrid of S. cerevisiae and S. uvarum (Lalvin S6U), and two
Juice was extracted by crushing mature Muscat of Frontignan non-Saccharomyces enological strains: Biodiva (Torulaspora
grapes (17°Brix) harvested from the experimental vineyard delbrueckii) and Flavia (Metschnikowia pulcherrima).
of INRA-Pech Rouge (Gruissan, France) with a blender. After
centrifugation (8000 × g), Muscat grape juice (200 mL) was
eluted through a XAD-2 column (Sigma-Aldrich Chimie, Lyon, Growth conditions
France), followed by washing with water (100 mL), then The medium (YEP-20) for yeast cell growth contained yeast
pentane/dichloromethane (2/1 v/v, 100 mL) to remove the free extract 10 g/L, peptone 20 g/L, glucose 20 g/L (pH adjusted to
fraction of terpenes. The bound glycosidic fraction was recov- 3.5). Yeasts were cultured in 100 mL Erlenmeyer flasks contain-
ered by elution with 100 mL methanol (Rosi et al. 1995) and ing 20 mL of growth medium. The flasks were directly inocu-
stored at −18°C. lated with active dry yeasts according to the manufacturer’s
recommendations. Yeast cultures were incubated overnight at
28°C with agitation (180 rpm).
Enzymatic hydrolysis of the Muscat glycosidic frac-
The β-glucosidase activity of entire viable yeast cells was
tion. The glycosidic fraction was dried under vacuum, and the evaluated in a specific synthetic culture medium. This medium
residue was then solubilised in 2 mL of phosphate citrate buffer (MS-50) contained 1.7 g/L yeast nitrogen base (YNB with
(sodium hydrogen phosphate 0.2 mol, citric acid 0.1 mol, pH amino acids, Becton Dickinson and Company, Franklin Lakes,
5.0). An aliquot (200 μL) of an enzymatic preparation (AR2000 NJ, USA), 5 g/L ammonium sulfate, 50 g/L glucose, 3 g/L citric
at 70 mg/mL, DSM Food Specialties, Heerlen, the Netherlands)
in citrate phosphate buffer was added. After mixing, the reac-
tion occurred at 40°C for 16 h. Table 1. Terpene content of the glycosidic fraction from Muscat
must, which was obtained by GC-MS analysis of the volatile fraction
Permeabilised cell assay. Permeabilised yeast cells after hydrolysis by the AR2000 enzymatic preparation.
(4.5 × 108) were added to the glycosidic fraction previously
dried under vacuum and solubilised in 2-mL citrate phosphate Detected compounds 4-Nonanol LOD LOQ
buffer. After mixing, the reaction occurred at 40°C or at room equivalents (μg) (μg)
temperature for 1, 4, 16 or 24 h. A blank experiment was (μg)
realised by replacing the permeabilised cells by the same volume
of imidazole buffer (75 mmol, pH 7.5). Linalool 6.70 0.01 0.04
Hotrienol 2.95 0.04 0.1
Extraction of aglycone terpenes. After enzymatic hydroly- α-Terpineol 1.21 0.03 0.1
sis, or after incubation with permeabilised yeast cells, the vola- Nerol 3.75 0.006 0.02
tile fraction was extracted by five times 2 mL of pentane/ Geraniol 7.10 0.015 0.05
dichloromethane (2/1 v/v) azeotrope. The organic extract was
Linalool hydrate 1.95 0.06 0.2
then dried on anhydrous sodium sulfate. 4-Nonanol (6 µg) was
added as an internal standard. The extract was then concen- Z-8-Hydroxylinalool 2.90 0.04 0.1
trated to about 400 μL by partial rectification at 35°C using a trans-Pyranic linalool oxide 1.28 0.01 0.03
Dufton’s spiral column. The extract was maintained at −20°C 3,6-Terpendiol 1.45 0.02 0.06
until analysis by GC/MS. 3,7 Terpendiol 13.40 0.04 0.1
3,8-Terpendiol 2.15 0.02 0.07
Analysis of the terpenic aglycones by GC/MS. The
aglycone extract was analysed with a Hewlett-Packard (HP) LOD (limits of detection) and LOQ (limits of quantification) of the method were
5890 Series II GC system coupled to a HP 5989 A MS. The calculated using the equations: LOD = 3*Hmax*R and LOQ = 10*Hmax*R,
samples were injected in splitless mode (injector port tempera- where Hmax is the maximum height of the noise and R is the response factor.

© 2015 Australian Society of Viticulture and Oenology Inc.

196 Release of glycosidic aroma precursors Australian Journal of Grape and Wine Research 21, 194–199, 2015

acid and 3 g/L malic acid (pH adjusted to 3.5 with concentrated Table 2. Alcohol dehydrogenase activity measured in permeabilised
KOH). This synthetic medium was supplemented with the solu- cells of eight strains of commercially available active dry yeasts for
tion of glycosidic precursors [dried under vacuum and redis- winemaking.
solved in citrate phosphate buffer (sodium hydrogen phosphate
0.2 mol, citric acid 0.1 mol, pH 5.0)]. Ten millilitres of this Strain ADH activity (nkat/108 cells)
synthetic medium were inoculated with 9 × 108 yeast cells and
incubated in 50 mL-Erlenmeyer flasks for 36 h at 28°C. The Mean Standard deviation
synthetic medium was incubated without yeast inoculation as
EC-1118 10.02 0.47
a blank.
K1M 8.85 0.33
QA23 6.11 0.42
Cell permeabilisation
Cells were permeabilised according to the method described by Flavia 5.08 0.52
Salmon (1984). Ten millilitres of culture were filtered through a CEG 2.02 0.65
Whatman filter GF/C type, and washed with 6 mL of ice-cold S6U 2.05 0.53
water. The cells retained on the filter were then re-suspended DV10 0.31 0.23
in 1 mL of imidazole buffer (75 mmol), KCl 0.1 mol, MgCl2 Biodiva 0.19 0.25
100 mmol (pH 7.5 using HCl). Fifty microlitres reduced
glutathione (0.3 mol), 10 μL Triton X-100 (10%) and 50 μL of Mean and standard deviation of four experiments (nkat/108 cells). ADH, alcohol
toluene/ethanol (1/4 v/v) were then added to the suspension, dehydrogenase; Biodiva, Torulaspora delbrueckii; EC-1118, K1M, QA23, CEG and
which was vigorously shaken by machine for 5 min. After DV10, Saccharomyces cerevisiae; Flavia, Metschnikowia pulcherrima; SU6, natural
mixing, the suspension was filtered, and the cells were washed hybrid of S. cerevisiae.
with 6 mL of ice-cold water. The permeabilised cells were
re-suspended in 3 mL of imidazole buffer. This suspension was Table 3. β-Glucosidase activity measured in the permeabilised cells
kept on ice, and could be used for enzymatic determination of eight strains of commercially available active dry yeasts for
within 6 h (Salmon 1984). winemaking.

Enzyme assays Strain β-Glucosidase activity (pkat/108 cells)

β-Glucosidase activity was determined according to the meth-
odology of Delcroix et al. (1994). One mL of permeabilised Mean Standard deviation
cells was added to 1 mL of ρ-nitrophenyl-β-D-glucopyranoside
Flavia 69.85 0.75
(8 mmol) in sodium acetate buffer (100 mmol, pH 5). After
incubation at 40°C, the reaction was stopped by addition of K1M 21.90 0.45
6 mL of Na2CO3 (1 mol). Absorbance was read at 400 nm, and QA23 22.05 0.39
activity was expressed as picokatals for 108 cells (pkat/108 EC-1118 20.02 0.59
cells). CEG 19.95 0.55
S6U 20.08 0.62
Alcohol dehydrogenase activity. Permeabilised cells (50 to DV10 10.05 0.25
200 μL) were added to 1 mL of glycine/NaOH buffer (75 mmol,
Biodiva 6.98 0.15
pH 9), 100 μL NAD+ (27.7 mmol), 50 μL reduced glutathione
(0.3 mol) and 50 μL pure ethanol. The cells were incubated at
Mean and standard deviation of three experiments (pkat 10−8 cells). Substrate
28°C, and absorbance at 340 nm was recorded continuously for
was ρ-nitrophenyl-β-D-glucopyranoside. Biodiva, Torulaspora delbrueckii;
10 min. Activity was expressed as nanokatals for 108 cells (nkat/ EC-1118, K1M, QA23, CEG and DV10, Saccharomyces cerevisiae; Flavia,
108 cells). Metschnikowia pulcherrima; SU6, natural hybrid of S. cerevisiae.

Analytical methods
Cell enumeration. Yeast culture samples were first diluted
(1000 to 2500 times) with Isoton II (Beckman-Coulter,
Margency, France). After sonication (35 s, 10 W), cells were
counted with a Coulter Z2 electronic counter (Beckman
Coulter, Fullerton, CA, USA), fitted with a 100-μm aperture
probe.
Results and discussion
Efficiency of the cell permeabilisation technique
The efficiency of permeabilisation was assessed by measuring
the alcohol dehydrogenase (ADH) activity of all the yeast strains
tested after permeabilisation. All the permeabilised cells, except
DV10 and Biodiva strains, exhibited a detectable ADH activity
(Table 2) under the conditions of the experiment. The measure-
ment of ADH activity in the permeabilised cells confirmed that
the method used for cell permeabilisation was appropriate,
except for the DV10 and Biodiva strains. Resistance to the
permeabilisation procedure was already observed in previous
work on wine yeast strains, and was mainly attributed to a
specific conformation of the cell membrane (Salmon 1984,
1986). This could be the case in the present study for the
T. delbrueckii Biodiva strain.
β-Glucosidase activity in the permeabilised cells
β-Glucosidase activity was measured with the artificial
substrate, ρ-nitrophenyl-β-D-glucopyranoside (Table 3). The
results obtained are consistent with the previously observed
low level of permeabilisation of the DV10 and Biodiva strains.
The other Saccharomyces strains tested exhibited quite low
β-glucosidase activity of about 20 pkat/108 cells, similar to that
encountered previously (Delcroix et al. 1994). In contrast, the
Flavia strain (M. pulcherrima) exhibited a greater β-glucosidase
activity of about 70 pkat/108 cells. Although previous work
with entire yeast cells showed that only non-Saccharomyces
species exhibited β-glycosidase activity (Mendes Ferreira et al.
2001), we have demonstrated by using permeabilised cells that
Saccharomyces species also exhibit β-glycosidase activity.

© 2015 Australian Society of Viticulture and Oenology Inc.

Bisotto et al. Release of glycosidic aroma precursors 197

Efficiency of permeabilised cells to release terpenes from a
glycosidic Muscat extract
Since some β-glucosidase activity was found in most of the
permeabilised cells tested, the efficiency of permeabilised cells to
release aglycone terpenes from the glycosidic fraction of a
Muscat must was then assayed. The total aromatic potential of
the glycosidic fraction was first evaluated by analysis of the
volatile fraction obtained after hydrolysis by the AR2000 enzy-
matic preparation (Table 1). Second, permeabilised yeast cells
were incubated for 20 h in the presence of the same glycosidic
fraction at either ambient temperature or at 40°C. The increase
in temperature of incubation did not change either the profile or
the quantity of terpenes released but affected only the observed
release rates (data not shown). Only five of the six strains tested
hydrolysed the glycosidic fraction and liberated terpenes
(Table 4). The permeabilised cells of strain S6U, which is a
natural hybrid of S. cerevisiae and S. uvarum, were unable to
release any bound terpenes under the experimental conditions
tested. A great variability was observed in the capacity of the
different permeabilised strains to release specific classes of
bound terpenes during the experiment (Figure 1). Considering
the extent and breadth of terpenes released, the permeabilised
cells of Flavia appeared to possess the greatest glycoside
hydrolase activity among the tested strains (Table 4 and
Figure 1). Globally, the monoterpendiols, as 3,6 and 3,7
monoterpendiols, were the terpenes most released by the tested
strains in their permeabilised state. Interestingly, most of these
odourless molecules can be transformed into other aromatic
molecules during the ageing of wine by many reactions
occurring in acid medium, such as isomerisation, ring closure,

Table 4. Terpenes revealed by permeabilised yeast (after 16 h of incubation at 40°C) or entire viable yeast cells (after 36 h of fermentation
at 28°C) from the glycosidic fraction of a Muscat must.

Detected compounds Yeast strains

QA23 K1M EC-1118 CEG S6U Flavia

PC EC PC EC PC EC PC EC PC EC PC EC

Linalool 4.71 ND ND ND ND ND ND ND ND ND 49.7 ND

Hotrienol 25.0 ND 87.5 23.0 61.3 3.5 36.5 16.7 ND ND 9.1 0.7
α-Terpineol ND ND ND ND ND ND ND ND ND ND 82.9 ND
Nerol 26.3 ND ND ND 8.2 ND 26.6 ND ND ND 40.1 ND
Geraniol 20.4 ND ND ND 8.9 ND 35.0 ND ND ND 39.9 ND
Linalool hydrate 28.0 ND 69.2 ND 48.2 ND 100.0 ND ND ND 25.5 ND
Z-8-Hydroxylinalool 72.3 ND 41.0 ND 48.2 ND 59.0 ND ND ND 94.5 ND
trans-Pyranic linalool oxide ND ND ND ND ND ND ND ND ND ND 100.0 ND
3,6-Terpendiol ND ND 100.0 ND 52.6 ND 100.0 ND ND ND 100.0 ND
3,7-Terpendiol 26.4 ND 53.0 8.7 35.2 7.7 100.0 11.2 ND ND 11.6 1.0
3,8-Terpendiol ND ND 100.0 ND ND ND 93.1 ND ND ND 99.5 ND

EC, entire viable cells; ND, not detectable; PC, permeabilised cells. Results are expressed as a proportion of the total aglycone terpene potential revealed by hydrolysis
with the AR2000 enzymatic preparation (see Table 1).

Figure 1. Monoterpenols
( , ), monoterpenol oxides
( ) and monoterpendiols ( ,
) revealed by permeabilised
yeast cells ( , , ) (16 h
incubation at 40°C) (plain
boxes) and entire yeast cells
( , ) (36 h alcoholic
fermentation at 28°C) (dashed
boxes) from the glycosidic
fraction of Muscat grape juice,
expressed as a proportion of
the total aglycone terpene
potential revealed by hydrolysis
with the AR2000 enzymatic
preparation (see Table 1).

© 2015 Australian Society of Viticulture and Oenology Inc.

198 Release of glycosidic aroma precursors
hydration, dehydration and oxidation (Rapp et al. 1985). The
other major terpenes released were mainly monoterpenols and
monoterpene oxides, with a high efficiency observed with the
Flavia strain. The observed release of monoterpenol oxides was
more surprising. There are two pathways that may explain their
origin: first, some were already present as their glycosylated
forms in the original Muscat extract as shown previously
(Table 1); second, yeast metabolism could oxidise a produced
terpene. As far as is known, however, there are only a few
descriptions of yeast-mediated terpene bio-oxidation processes
(Bicas et al. 2009), mainly by Candida tropicalis, Trichosporum
cutaneum and by ale and lager yeasts. The present study does not
allow a final conclusion to be drawn on the predominant
pathway for their formation.
Comparison between permeabilised cells and entire viable cells
In order to determine whether cell permeabilisation influenced
the release of bound terpenes from the glycosidic fraction of
Muscat must, non-permeabilised yeasts were tested for their
ability to release aglycone terpenes on the same glycosidic
extract as above, during a 36-h alcoholic fermentation. Under
these fermentative conditions, yeast strains were not able to
reveal the same cultivar of compounds as permeabilised cells
(Table 4 and Figure 1). Only hotrienol and 3,7-monoterpendiol
could be detected after fermentation by some strains (K1M, EC
1118, CEG and Flavia) and at a concentration much lower
than that observed with permeabilised cells. This result chal-
lenges the previous findings of Ugliano et al. (2006) who con-
sidered that hotrienol was partially formed through acid-
catalysed hydrolysis of either a glycosidic precursor or aglycons
released by yeast glycosidases. The comparison between the
amount of aglycone terpenes released by whole and
permeabilised cells (Figure 1) clearly shows the benefit of
permeabilisation. Indeed, a considerable increase in the pro-
portion of aglycone terpenes released with permeabilised cells
was observed. These results suggest that the transport of the
glycosidic precursors into the cell is an important limiting
factor for commercial yeast strains to reveal glycosidic aroma
precursors from grape must.
Conclusion
The aim of this work was to study the inherent capacity
of commercial wine yeast strains to reveal glycosidic aroma
precursors from Muscat grape must by using permeabilised
cells. Although a few yeast strains appeared to resist the
permeabilisation treatment, results particularly highlight the
great potential of this technique to improve the actual enzy-
matic capacity of yeast strains to release terpenes from their
glycosidic forms. Surprisingly, some yeast strains appeared to be
able to release the free odourous monoterpenols, while other
release mainly odourless monoterpendiols. The results obtained
confirmed that no concomitant hydrolysis of complex disaccha-
ride glycosides associated with enzymatic activities different
from β-glucosidase (e.g. α-arabinosidase, α-rhamonosidase,
β-apiosidase) occurred in the release of glucosides, as previously
demonstrated by Ugliano et al. (2006).
The results suggest that the transport of glycosidic precursors
is a limiting factor in this aromatic release, which is of specific
physiological interest and could orientate breeding programs for
the construction of new interspecific wine yeasts. Traditional
breeding techniques, mainly based on breeding between S.
cerevisiae or closely related strains, were previously used in the
development of new yeast strains with altered phenotypic char-
acteristics for winemaking (Romano et al. 1985, Chambers et al.
Australian Journal of Grape and Wine Research 21, 194–199, 2015
2009). The generation of an interspecific hybrid, however,
between a commercial S. cerevisiae wine yeast strain and S.
mikatae, a species not previously associated with alcoholic fer-
mentation, was recently described for developing novel yeast
strains that bring greater complexity to wine than strains cur-
rently available to the industry (Bellon et al. 2013). A similar
approach could be used to improve genetically commercial
Saccharomyces wine yeast strains by complementing their inher-
ent glycoside hydrolase enzymatic activity with the ability to
transport actively glycosidic aroma precursors from the grape
must. This will provide additional tools for winemakers to
develop new wine styles.
Acknowledgements
The authors thank Jean-Paul Lepoutre and Nicolas Bouvier for
their technical assistance during aroma analysis. The stay of
Alexandra Bisotto was funded by Lallemand SAS.
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Manuscript received: 3 June 2014
Revised manuscript received: 15 September 2014
Accepted: 24 September 2014