Letters in Applied Microbiology ISSN 0266-8254

ORIGINAL ARTICLE

Influence of Williopsis saturnus yeasts in combination with

Saccharomyces cerevisiae on wine fermentation
H. Erten and H. Tanguler
Department of Food Engineering, Faculty of Agriculture, Cukurova University, Adana, Turkey

Keywords
flavour production, grape must fermentation,
Saccharomyces cerevisiae, Williopsis saturnus,
wine.
Correspondence
Huseyin Erten, Department of Food
Engineering, Faculty of Agriculture, Cukurova
University, 01330 Adana, Turkey.
E-mail: herten@cu.edu.tr
2009 ⁄ 1732: received 2 October 2009, revised
29 January 2010 and accepted 4 February
2010
doi:10.1111/j.1472-765X.2010.02822.x
Abstract
Aim: To examine the growth and survival of Williopsis saturnus strains along
with wine yeast Saccharomyces cerevisiae in grape must.
Methods and Results: For this study, fermentations were performed in sterilized
grape must at 18C. Inoculum level was 5 · 106 cells per ml for each yeast. The
results showed that W. saturnus yeasts exhibited slight growth and survival
depending on the strain, but they died off by day 5. Saccharomyces cerevisiae,
however, dominated the fermentation, reaching the population of about
8 log CFU ml)1. It was observed that ethanol formation was not affected. The
concentrations of acetic acid, ethyl acetate and isoamyl acetate were found
higher in mixed culture experiments compared to control fermentation. The
results also revealed that higher alcohols production was unaffected in general.
Conclusion: Fermentations did not form undesirable concentrations of flavour
compounds, but production of higher levels of acetic acid in mixed culture
fermentations may unfavour the usage of W. saturnus in wine making.
Significance and Impact of the Study: This study provides information on the
behaviour of W. saturnus together with S. cerevisiae during the alcoholic
fermentation.

formation of volatile acidity; however, it can produce

Introduction
Yeasts, mainly Saccharomyces cerevisiae, are primarily
responsible for the alcoholic fermentation of grape juice
into wine, forming the main metabolic products (ethanol
and CO2) and also several flavour compounds (Fleet
1993; Henschke and Jiranek 1993).
Besides the principal wine yeast S. cerevisiae, the role of
non-Saccharomyces spp. in wine production has been
extensively debated. Non-Saccharomyces yeasts can affect
the chemical and sensory characteristics of wine (Fleet
2003; Jolly et al. 2003a). Kloeckera apiculata (its
telemorph Hanseniaspora uvarum) especially is the pre-
dominant non-Saccharomyces yeast present in grape must
(Fleet and Heard 1993; Pretorius et al. 1999). It usually
forms a high level of volatile acidity, therefore its
potential for a positive contribution in wine making is
low (Jolly et al. 2003b; Mendoza et al. 2009). Candida
pulcherrima can also be isolated in high numbers from
the grape must. It is not normally associated with the
especially relatively high level of esters. Therefore, C. pul-
cherrima can contribute positively to wine with neutral
cultivar characteristics (Jolly et al. 2003a,b). Candida
stellata can also be used to enhance the glycerol content
of wine. However, its low fermentation rate leads to a
limitation in its use in wine making but immobilization
of its cells increased the fermentation rate. Schizosacchar-
omyces pombe can metabolize malic acid but its low
fermentation rate and tendency to form undesirable
flavour compounds limits the possibility of its application
in winemaking (Ciani 1997). Kluyveromyces thermotoler-
ans produces high concentration of l-lactic acid from
glucose and fructose as well as low levels of volatile
acidity and undesirable flavour compounds, therefore low
acidity musts can be improved using Kluyveromyces
thermotolerans (Kapsopoulou et al. 2005, 2007). Torulas-
pora delbrueckii (formerly Saccharomyces rosei) with very
low volatile acidity and acetaldehyde production still has
useful potential in sweet wine making (Bely et al. 2008).

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474 Journal compilation ª 2010 The Society for Applied Microbiology, Letters in Applied Microbiology 50 (2010) 474–479
H. Erten and H. Tanguler
Williopsis (W.) saturnus, formerly known as Hansenula
saturnus (Barnett et al. 1983), synthesizes important levels
of volatile esters, especially isoamyl acetate (banana-like
flavour) and ethyl acetate (Erten and Campbell 2001;
Yilmaztekin et al. 2008, 2009). It is not generally found
from the natural environment of surfaces of grapes and
winery equipments but with the production of desirable
flavour, W. saturnus can potentially enhance the fruity
flavour in wines obtained from neutral cultivar character-
istics. In this article, the behaviour of W. saturnus and
S. cerevisiae was studied in mixed cultures in grape must.
Materials and methods
Yeast cultures
Saccharomyces cerevisiae (Fermiblanc arom) was obtained
from the Gist Brocades Company (France). Williopsis
saturnus HUT 7087, W. saturnus IAM 12217 and
W. saturnus NCYC 22 were obtained from the HUT
(Japan), IAM (Japan) and NCYC (United Kingdom)
culture collections, respectively. Yeasts were maintained
on malt extract agar slants.
Fermentation conditions
White grape must cv. Emir was used for fermentations.
Healthy grapes were obtained from Nevşehir-Urgup
region (ancient Cappadocia) of Turkey. The grapes were
crushed and pressed in a horizontal press, and grape must
was sterilized by autoclaving at 115C for 10 min. The
initial brix and pH of the grape must were 21Æ5 and 3Æ53,
respectively. Mixed culture fermentations of S. cerevisi-
ae ⁄ W. saturnus NCYC 22, S. cerevisiae ⁄ W. saturnus IAM
12217 and S. cerevisiae ⁄ W. saturnus HUT 7087 and single
culture fermentation of S. cerevisiae as control were con-
ducted in 1-l sterile erlenmayer flasks containing 800 ml
of sterile grape must in duplicate. The flasks fitted with
foam bungs were incubated statically at 18C. Fermenta-
tion was followed measuring the specific gravity.
Yeast propagation
A loopful of stock cultures of W. saturnus was plated
onto malt extract agar and incubated for 48 h at 25C. A
single colony was inoculated into 100 ml of sterile grape
juice in a 250-ml sterile conical flask fitted with foam
bung and incubated for 48 h at 25C with orbital shaking
at 160 rev min)1. The yeast cells were centrifuged at
4000 rev min)1 for 10 min at 4C, washed once with cold
sterile water. The pellet was re-suspended in 5 ml of ster-
ile grape must. After counting by haemacytometer, an
initial concentration of 5 · 106 cells per ml for each yeast
ª 2010 The Authors
Journal compilation ª 2010 The Society for Applied Microbiology, Letters in Applied Microbiology 50 (2010) 474–479
Influence of Williopsis saturnus
was added into fermentation medium (Erten and Camp-
bell 2001). Commercial wine yeast S. cerevisiae was sus-
pended in sterile grape juice at 35C for 30 min
according to the producer’s instructions, and an initial
density of 5 · 106 cells per ml was added into fermenta-
tion medium after counting with haemacytometer.
Enumeration of yeasts
Samples were taken under aseptic conditions for counting
yeasts during fermentations. They were diluted in 0Æ25%
saline as necessary, spread inoculated (0Æ1 ml) onto plates
of malt extract agar and l-lysine agar. l-lysine agar was
used to enumerate non-Saccharomyces yeast of W. satur-
nus. The plates were incubated until days 2–4 at 25C
and then examined for the yeast counts (Fleet 1993;
Soden et al. 2000; Jolly et al. 2003a).
Analytic determinations
Density was determined using a density meter. Total acid-
ity was measured by titrating 0Æ1 N NaOH solution using
phenolphthalein as indicator and expressed as g of
tartaric acid l)1. pH was determined using a pH meter
(Inolab WTW, Weilheim, Germany) Ethanol in g l)1 was
calculated from the following formula: ethanol as g
l)1 = ethanol as v ⁄ v · 0Æ789 (Ough and Amerine 1988).
Ethanol, acetic acid, glucose, fructose and sucrose were
analysed by a high performance liquid chromatograph
(Shimadzu, Kyoto, Japan) using an Aminex HPX-87H
column (Bio-Rad, Richmont, CA, USA) at 50C. The
eluent was 5 mmol l)1 H2SO4 in high-purity water at a
flow rate of 0Æ6 ml min)1. Concentrations of ethanol,
glucose, fructose and sucrose were calculated from RI
detector (Shimadzu) and amounts of acetate from UV
detector (Shimadzu) (Erten 1998).
Volatile higher alcohols, esters and acetaldehyde were
measured by a Gas Chromatograph (HP 5890; Hewlett-
Packard, Stockport, UK). Cell-free samples were diluted
to 4% (v ⁄ v) ethanol and 5 ml of diluted sample, 2 g of
NaCl and 50 ll of internal standard (200 mg l)1 of
3-heptanone) were sealed and a 1 ml sample was injected
into a 60 m · 0Æ25 mm i.d. · 0Æ4-lm-thick Chrompac
CP-Wax-57-CB column (Middleburgh, the Netherlands),
temperature-programmed from 40 to 80C. The stream
from the column was split 1 : 1 to flame ionization detec-
tor (Erten and Campbell 2001).
Statistical analysis
Data of ethanol, acetic acid, volatile higher alcohols, esters
and acetaldehyde were analysed for statistical significance
by one-way analysis of variance (anova). Means were
475
Influence of Williopsis saturnus H. Erten and H. Tanguler

9 Table 1 General composition of wine samples*

8 S SxNCYC SxIAM SxHUT Sig.

Yeast count (log cfu ml–1)

7 a b c b
Density (20C) 0Æ9953 0Æ9942 0Æ9929 0Æ9944 ***
6 Ethanol (v ⁄ v) 10Æ15 10Æ20 10Æ19 10Æ22 ns
5 Ethanol (g l)1) 80Æ08 80Æ48 80Æ40 80Æ64 ns
pH 3Æ48 3Æ48 3Æ47 3Æ48 ns
4
Total acidity as tartaric 6Æ59 6Æ14 6Æ22 6Æ36 ns
3 acid (g l)1)
2 Acetic acid (g l)1) 0Æ23c 0Æ71b 0Æ93a 0Æ83a b
***
Glucose (g l)1) 1Æ23 1Æ03 1Æ03 1Æ37 ***
1
Fructose (g l)1) 0Æ91 1Æ02 1Æ02 1Æ01 ***
0 Sucrose (g l)1) 0Æ94 1Æ14 0Æ95 1Æ24 ***
0 1 2 3 4 5 6 7 8
Total residual sugar 3Æ08 3Æ47 2Æ98 3Æ62
Time (days)
(g l)1)
Figure 1 The growth of yeasts during fermentations. Pure culture of Values not sharing the same superscript letter within the horizontal
Saccharomyces cerevisiae (*); mixed culture of S. cerevisiae (D) and line are different according to Duncan test, ns: not significant.
Williopsis saturnus NCYC 22 ( ); Mixed culture of S. cerevisiae (s) ***Displays the significance at 0Æ1% by LSD.
and W. saturnus HUT 7087 (d); Mixed culture of S. cerevisiae (h) and *S: Pure culture of Saccharomyces cerevisiae, SxNCYC: mixed culture
W. saturnus IAM 12217 ( ). of S. cerevisiae and Williopsis saturnus NCYC 22, SxIAM: mixed
culture of S. cerevisiae and W. saturnus IAM 12217, SxHUT: mixed
culture of S. cerevisiae and W. saturnus HUT 7087, Sig.: significance.

compared by Duncan test statistical analysis performed

using the software spss 10.0 for Windows (Ozdamar 1999).
Table 2 Analyses of flavour compounds of wine samples
Results S SxNCYC SxIAM SxHUT Sig.
The growth of S. cerevisiae and W. saturnus in mixed Higher alcohols (mg l )1
)
cultures during the fermentation of grape musts is given n-propanol 21Æ86b 23Æ55a 23Æ15a b
24Æ20a *
in Fig. 1. Isobutanol 36Æ13 37Æ48 36Æ52 37Æ95 ns
The main wine yeast, S. cerevisiae was the dominant 2-methyl butanol 30Æ15 32Æ87 34Æ35 33Æ44 ns
yeast in all mixed fermentations, reaching maximum pop- 3-methyl butanol 120Æ29 130Æ55 135Æ31 132Æ13 ns
Esters (mg l)1)
ulations of 7Æ87–7Æ97 log CFU ml)1 by day 4 and 5. After
Ethyl acetate 39Æ11b 47Æ89a 43Æ36b 38Æ98b *
maximum growth, S. cerevisiae exhibited a stationary
Ethyl butyrate 0Æ22 0Æ25 0Æ24 0Æ22 ns
phase and had the numbers of 7–7Æ64 log CFU ml)1 at Isoamyl acetate 6Æ10 7Æ42 6Æ94 6Æ19 ns
the end of fermentation. With regard to non-Saccharo- Isobutyl acetate 0Æ14b 0Æ16a 0Æ15b 0Æ14b *
myces yeasts, W. saturnus NCYC 22 did not grow Ethyl hexonoate 0Æ44 0Æ52 0Æ52 0Æ41 ns
and died off by day 4. Williopsis saturnus HUT 7087 Ethyl octanoate 0Æ16c 0Æ23ab 0Æ27a 0Æ19bc *
and W. saturnus IAM 12217 exhibited slight growth and Carbonyl compounds (mg l)1)
Acetaldehyde 17Æ31 17Æ89 17Æ14 17Æ13 ns
survival with the maximum numbers of 6Æ86 and
7Æ03 log CFU ml)1 respectively on day 2, but they dis- Values not sharing the same superscript letter within the horizontal
appeared by day 5 of the experiments. line are different according to Duncan test, ns: not significant.
Fermentation was followed by measuring the specific *Displays the significance at 5%, by LSD.
gravity. All fermentations were completed by the 7th day S: Pure culture of Saccharomyces cerevisiae, SxNCYC: mixed culture
of fermentation at 18C (results not shown). Table 1 of S. cerevisiae and Williopsis saturnus NCYC 22, SxIAM: mixed
culture of S. cerevisiae and W. saturnus IAM 12217, SxHUT: mixed
compares the levels of density, ethanol, pH, total acidity,
culture of S. cerevisiae and W. saturnus HUT 7087, Sig.: significance.
acetic acid, glucose, fructose and sucrose in final wines.
Ethanol production (10Æ15–10Æ22% v ⁄ v and 80Æ08–
80Æ64 g l)1) was unaffected (P > 0Æ05) in mixed and mixed culture fermentations with W. saturnus strains
single culture experiments. The concentrations of total produced higher amounts of acetic acid, ranging from
acidity as tartaric acid ranging from 6Æ14–6Æ59 g l)1 0Æ71 to 0Æ93 g l)1 (P < 0Æ001). All sugars in all fermenta-
(P > 0Æ05) and pH levels were not affected (P > 0Æ05). tions were utilized with a density below 1Æ000 and with
Saccharomyces cerevisiae in pure culture experiment <1Æ37 g l)1 levels of glucose, fructose and sucrose
formed the lowest acetic acid level of 0Æ23 g l)1, whereas (P < 0Æ001).

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476 Journal compilation ª 2010 The Society for Applied Microbiology, Letters in Applied Microbiology 50 (2010) 474–479
H. Erten and H. Tanguler
Table 2 gives the concentrations of higher alcohols,
esters and acetaldehyde in wines. Amounts of higher alco-
hols of isobutanol, 2-methyl butanol and 3-methyl buta-
nol were not affected (P > 0Æ05) with the exception of
n-propanol whose concentration slightly increased with
W. saturnus addition from 21Æ86 to 24Æ20 mg l)1. Mixed
cultures of S. cerevisiae ⁄ W. saturnus NCYC 22 and S. cere-
visiae ⁄ W. saturnus IAM 12217 formed relatively higher
amounts of ethyl acetate, 47Æ89 and 43Æ36 mg l)1, respec-
tively in comparison with control S. cerevisiae fermenta-
tion (39Æ11 mg l)1) and mixed culture of S. cerevisiae ⁄
W. saturnus HUT 7087 (38Æ98 mg l)1). The production of
isoamyl acetate was not significantly important
(P > 0Æ05). However, mixed culture of S. cerevisiae ⁄
W. saturnus NCYC 22 produced the highest amount of
7Æ42 mg l)1. Formation of isoamyl acetate in mixed
culture of W. saturnus IAM 12217 ⁄ S. cerevisiae was higher
(6Æ94 mg l)1) than in the single culture fermentation with
S. cerevisiae (6Æ10 mg l)1) and mixed culture of W. saturnus
HUT 7087 ⁄ S. cerevisiae (6Æ19 mg l)1). Combined fermen-
tations with W. saturnus had no effect on concentrations
of ethyl butyrate, isobutyl acetate and ethyl hexonoate
(P > 0Æ05). The amount of ethyl octanoate was signifi-
cantly important (P < 0Æ05).
Discussion
It is believed that non-Saccharomyces spp. such as
K. apiculata ⁄ H. uvarum, C. pulcherrima, C. stellata even-
tually die off with an increase in ethanol level and high
ethanol tolerant S. cerevisiae dominates the fermentation.
Several studies have confirmed that non-Saccharomyces
yeasts, especially K. apiculata and C. pulcherrima, can
survive longer than previously thought in wine fermen-
tations, growing up to 106–107 cells per ml. Such
growth could be significant and can influence the analy-
tical composition and quality of wine (Fleet and Heard
1993; Ciani 1997; Fleet 2003; Jolly et al. 2003b). In this
study, W. saturnus is not able to grow and survive in high
ethanol concentrations in agreement with previous studies
of Erten and Campbell (2001) and Yilmaztekin et al.
(2008).
Ethanol level in all mixed cultures as well as in pure
culture of S. cerevisiae was similar because of the vigorous
fermentative and ethanol-tolerant abilities of the wine
yeast S. cerevisiae in agreement with previous studies of
Pretorius et al. (1999) and Fleet (2003). Residual sugars’
levels showed the complete utilization of glucose, fructose
and sucrose, and a similar behaviour was reported by
Soden et al. (2000), Jolly et al. (2003a) and Kapsopoulou
et al. (2007) who studied the effects of non-Saccharomyces
yeasts on wine fermentation in mixed cultures with
S. cerevisiae.
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Journal compilation ª 2010 The Society for Applied Microbiology, Letters in Applied Microbiology 50 (2010) 474–479
Influence of Williopsis saturnus
Acetic acid, a by-product of yeast metabolism, leads to
wine objectionable near its flavour threshold of 0Æ7–
1Æ1 g l)1 of volatile acidity as acetic acid (Henschke and
Jiranek 1993). Our results are in disagreement with Soden
et al. (2000) and Mendoza et al. (2009) who found the
lower amounts of acetic acid (0Æ3–0Æ69 g l)1) in mixed
cultures of S. cerevisiae and non-Saccharomyces yeasts of
C. stellata and K. apiculata. In this study, mixed cultures
of W. saturnus ⁄ S. cerevisiae produced higher than the
lower threshold value throughout the fermentations
which leads to negative effect on wine quality.
Yeast is the main producer of flavour compounds such
as esters, higher alcohols, organic acids and carbonyl com-
pounds during alcoholic fermentation. A number of
flavour compounds make an important contribution in the
typical odour and taste of wine (Sarris et al. 2009). In other
studies, mixed cultures of S. cerevisiae and non-Saccharo-
myces yeasts of C. stellata, K. apiculata and C. pulcherrima
produced the concentrations of 13Æ50–62Æ9 mg l)1 of
n-propanol, 18Æ05–57Æ6 mg l)1 of iso-butanol and 17Æ8–
57Æ6 mg l)1 of 2-methyl butanol (Ciani 1997; Erten 2002;
Zohre and Erten 2002). n-Propanol has flavour thresholds
of 300–750 mg l)1, isobutanol 75–500 mg l)1 and 2-methyl
butanol 300–330 mg l)1(Etiévant 1991). In this study, the
concentrations of higher alcohols of n-propanol, isobutanol
and 2-methyl butanol were much below than their thresh-
old values given by Etiévant (1991) but within the range of
previous studies reported previously. Although combined
fermentations with W. saturnus did not influence on
3-methyl butanol levels (Table 1), the concentrations
produced in this study was higher than its threshold value
of 14Æ5 mg l)1 given by Etiévant (1991). In other mixed
culture experiments of non-Saccharomyces yeasts and
S. cerevisiae, 73Æ55–166Æ35 mg l)1 of 3-methyl butanol lev-
els were reported (Ciani 1997; Erten 2002; Zohre and Erten
2002).
Esters are important contributors to fruity or floral fla-
vours of alcoholic beverages (Russell 2003). In this study,
mixed cultures formed relatively higher amounts of ethyl
acetate in comparison with control S. cerevisiae fermenta-
tion. Etiévant (1991) states that ethyl acetate is the most
abundant ester and its concentration below 50 mg l)1
does not contribute to the wine flavour, while levels
higher than 200 mg l)1 leads to defects with solvent-like
flavour in wine quality. In this study, the amounts of
ethyl acetate were lower than 50 mg l)1. Similar results
were also obtained by Ciani (1997), Erten (2002) and
Zohre and Erten (2002). Mixed culture fermentations
gave higher amounts of isoamyl acetate compared to
single culture fermentation of S. cerevisiae in this study.
The 1 mg l)1 flavour threshold of isoamyl acetate
reported by Etiévant (1991) was substantially exceeded in
all wines produced by mixed and single culture fermenta-
477
Influence of Williopsis saturnus
tions (Table 1), but up to 16 mg l)1 has been reported in
acceptable wines (Fleet and Heard 1993). Zohre and Erten
(2002) stated 1Æ29 and 1Æ16 mg l)1 of isoamyl acetate in
the mixed culture fermentations of S. cerevisiae ⁄ C. pulch-
errima and S. cerevisiae ⁄ K. apiculata ⁄ C. pulcherrima,
respectively. The concentrations of isobutyl acetate and
ethyl octanoate formed by fermentations did not exceed
their threshold value given by Etiévant (1991). Ethyl
hexonoate with 0Æ1 mg l)1 of flavour threshold gives
apple-like and fruity flavour to wine (Etiévant 1991). In
this study, fermentations produced higher levels of ethyl
hexonoate than its threshold level.
In conclusion, different strains of W. saturnus in mixed
culture experiments inoculated with the main wine yeast
S. cerevisiae survived by 5 days of fermentation, possibly
affecting flavour composition of final wine. Combined
fermentations did not form undesirable levels of flavour
compounds, but their tendency to form high level of
acetic acid may limit their possible usage in wine making.
Acknowledgements
The authors thank G. McKernan and J. MacKinlay of the
ICBD, Heriot-Watt University, Scotland, UK for assis-
tance with the GC analyses.
References
Barnett, J.A., Payne, R.W. and Yarrow, D. (1983) Yeasts: Char-
acteristics and Identification. p. 800. Cambridge, UK: CUP.
Bely, M., Stoeckle, P., Masneuf-Pomarède, I. and Dubourdieu,
D. (2008) Impact of mixed Torulaspora delbrueckii- Sac-
charomyces cerevisiae culture on high sugar fermentation.
Int J Food Microbiol 122, 312–320.
Ciani, M. (1997) Role, enological properties and potential use
of Non-Saccharomyces wine yeasts. Recent Res Dev Micro-
biol 1, 317–331.
Erten, H. (1998) Metabolism of fructose as an electron accep-
tor by Leuconostoc mesenteroides. Process Biochem 33,
735–739.
Erten, H. (2002) Relations between elevated temperatures and
fermentation behaviour of Kloeckera apiculata and
Saccharomyces cerevisiae associated with winemaking in
mixed cultures. World J Microbiol Biotechnol, 18, 373–378.
Erten, H. and Campbell, I. (2001) The production of low-alco-
hol wines by aerobic yeasts. J Inst Brew 107, 207–217.
Etiévant, P.X. (1991) Wine. In Volatile Compounds in Food
and Beverages ed. Maarse, H. pp. 483–546. New York:
Marcel Dekker.
Fleet, G.H. (1993) The microorganisms of winemaking – isola-
tion, enumeration and identification. In Wine Microbiology
and Biotechnology Ed. Fleet, G.M. pp. 1–25 London, UK:
Harwood Academic Publishers.
478 Journal compilation ª 2010 The Society for Applied Microbiology, Letters in Applied Microbiology 50 (2010) 474–479
H. Erten and H. Tanguler
Fleet, G.H. (2003) Yeast interactions and wine flavour. Int J
Food Microbiol 86, 11–22.
Fleet, G.M. and Heard, G.M. (1993) Yeasts – growth during
fermentation. In Wine Microbiology and Biotechnology
ed. Fleet, G.M. pp. 27–54 London, UK: Harwood Academic
Publishers.
Henschke, P.A. and Jiranek, V. (1993) Yeasts – metabolism
of nitrogen compounds. In Wine Microbiology and
Biotechnology ed. Fleet, G.M. pp. 77–164 London, UK:
Harwood Academic Publishers.
Jolly, N.P., Augustyn, O.P.H. and Pretorius, I.S. (2003a) The
effect of non-Saccharomyces yeasts on fermentation and
wine quality. S Afr J Enol Vitic 24, 55–62.
Jolly, N.P., Augustyn, O.P.H. and Pretorius, I.S. (2003b) The
use of Candida pulcherrima in combination with Saccharo-
myces cerevisiae for the production of Chenin Blanc wine.
S Afr J Enol Vitic 24, 63–69.
Kapsopoulou, K., Kapaklis, A. and Spyropoulos, H. (2005)
Growth and fermentation characteristics of a strain of the
wine yeast Kluyveromyces thermotolerans isolated in Greece.
World J Microbiol Biotechnol 21, 1599–1602.
Kapsopoulou, K., Mourtzini, A., Anthoulas, M. and Nerantzis,
E. (2007) Biological acidification during grape must fer-
mentation using mixed cultures of Kluyveromyces thermo-
tolerans and Saccharomyces cerevisiae. World J Microbiol
Biotechnol 23, 735–739.
Mendoza, L.M., Manda de Nadra, M.C., Bru, E. and Farı́es,
M.E. (2009) Influence of wine-related physicochemical fac-
tors on the growth and metabolism of non-Saccharomyces
and Saccharomyces yeasts in mixed cultures. J Ind Microbiol
Biotechnol 36, 229–237.
Ough, C.S. and Amerine, M.A. (1988) Methods for Analysis of
Musts and Wines, 2nd edn. p. 377. New York: John Wiley
and Sons.
Ozdamar, K. (1999) Paket Programlar Ile _ Istatistiksel
_ Veri
Analizi. p. 535. Eskis¸ ehir: Kaan Kitabevi.
Pretorius, I.S., van der Westhuizen, T.J. and Augustyn, O.P.H.
(1999) Yeast biodiversity in vineyards and wineries and its
importance to the South African wine industry. A review.
S Afr J Enol Vitic 20, 61–74.
Russell, I. (2003) Understanding yeast fundamentals. In The
Alcohol Textbook ed. Jacques, K.A., Lyons, T.P. and
Kelsall, D.R. pp. 85–119 Nottingham, UK: Nottingham
University Press.
Sarris, D., Kotseridis, Y., Linga, M., Galiotou-Panayotou, M.
and Papanikolaou, S. (2009) Enhanced ethanol production,
volatile compound biosynthesis and fungicide removal
during growth of a newly isolated Saccharomyces cerevisiae
strain on enriched pasteurized grape musts. Eng Life Sci 9,
29–37.
Soden, A., Francis, I.L., Oakey, H. and Hensche, P.A. (2000)
Effects of co-fermentation with Candida stellata and
Saccharomyces cerevisiae on the aroma and composition
of Chardonnay wine. Aust J Grape Wine Res 6, 21–30.
ª 2010 The Authors
H. Erten and H. Tanguler Influence of Williopsis saturnus

Yilmaztekin, M., Erten, H. and Cabaroglu, T. (2008) Produc-
tion of isoamyl acetate from sugar beet molasses by
Williopsis saturnus var. saturnus. J Inst Brew 114, 34–38.
Yilmaztekin, M., Erten, H. and Cabaroglu, T. (2009) Enhanced
production of isoamyl acetate from beet molasses with
addition of fusel oil by Williopsis saturnus var. saturnus.
Food Chem 112, 290–294.
Zohre, D.E. and Erten, H. (2002) The influence of Kloeckera
apiculata and Candida pulcherrima yeasts on wine fermen-
tation. Process Biochem, 38, 319–324.

ª 2010 The Authors

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