International Journal of Food Microbiology 144 (2010) 193198

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International Journal of Food Microbiology

j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / i j f o o d m i c r o

Short Communication

Optimization of honey-must preparation and alcoholic fermentation by

Saccharomyces cerevisiae for mead production
A. Mendes-Ferreira a, F. Cosme a, C. Barbosa a, V. Falco b, A. Ins a, A. Mendes-Faia a,
a
Institute for Biotechnology and Bioengineering, Centre of Genomics and Biotechnology, (IBB/CGB-UTAD), Universidade de Trs-os-Montes e Alto Douro,
School of Life Sciences and Environment, Vila Real, Portugal
b
Departamento de Indstrias Alimentares, Universidade de Trs-os-Montes e Alto Douro, Vila Real, Portugal

a r t i c l e i n f o a b s t r a c t

Article history: Mead fermentation is a time-consuming process, often taking several months to complete. Despite of the use of
Received 7 May 2010 starter cultures several problems still persist such as lack of uniformity of the nal products, slow or premature
Received in revised form 2 September 2010 fermentation arrest and the production of off-avors by yeast. Thus the aim of this study was to optimize mead
Accepted 19 September 2010
production through the use of an appropriate honey-must formulation to improve yeast performance alcoholic
fermentation and thereby obtain a high quality product. Honey-must was centrifuged to reduce insoluble solids,
Keywords:
pasteurized at 65 C for 10 min, and then subjected to different conditions: nitrogen supplementation and
Mead
Honey-wine
addition of organic acids. Although the addition of diammonium phosphate (DAP) reduced fermentation length,
Alcoholic fermentation it did not guarantee the completeness of the fermentation process, suggesting that other factors could account for
Saccharomyces cerevisiae the reduced yeast activity in honey-must fermentations. Sixteen yeast-derived aroma compounds which
Assimilable nitrogen contribute to the sensorial quality of mead were identied and quantied. Global analysis of aromatic proles
revealed that the total concentration of aroma compounds in meads was higher in those fermentations where
DAP was added. A positive correlation between nitrogen availability and the levels of ethyl and acetate esters,
associated to the fruity character of fermented beverages, was observed whereas the presence of potassium
tartrate and malic acid decreased, in general, their concentration.
This study provides very useful information that can be used for improving mead quality.
2010 Elsevier B.V. All rights reserved.

1. Introduction
Mead is a traditional honey-derived beverage containing 8 to
18% (v/v) of ethanol, which results from alcoholic fermentation by
yeasts of diluted honey. This beverage has progressively gained eco-
nomic importance (Sroka and Tuszynski, 2007), due to the therapeutic/
nutriceutical properties attributed to honey and by an increasing
demand for gourmet products. Mead fermentation is a time-consuming
process, often taking several months to complete, depending on the type
of honey, yeast strain and honey-must composition (Navrtil et al.,
2001). In most of the mead-producing countries, alcoholic fermentation
is a result of the growth of indigenous microorganisms naturally present
in honeys and always surviving on the substrates and equipment used
(Ashena, 2006; Bahiru et al., 2001). In these cases, alcoholic
fermentation is even more unpredictable and very often at the end of
fermentation, mead is completely spoiled by contaminating yeasts and
bacteria which makes it undrinkable. More recently, selected strains
isolated from honey/honey-wine (Pereira et al., 2009; Teramoto et al.,
2005) or commercial yeasts starter cultures (Sroka and Tuszynski, 2007;
Corresponding author. Tel.: + 351 259350554.
E-mail address: afaia@utad.pt (A. Mendes-Faia).
Ukpabi, 2006; Wintersteen et al., 2005) have been used to reduce the
risks of contamination. However, despite the use of starter cultures for
honey-must inoculation several problems still persist such as lack of
uniformity of the nal products, slow or premature fermentation arrest
and the production of off-avors by yeast (Pereira et al., 2009). These
problems could be due to inappropriate yeast strains that are not
suitable for the specic composition/conditions in honey-musts, or the
result of several stress conditions: high osmotic stress, lack of essential
nutrients such as a deciency in available nitrogen (Maugenet, 1964;
McConnell and Schramm, 1995), low mineral concentration, low pH
(Sroka and Tuszynski, 2007) and low buffer capacity (Maugenet, 1964).
Some of these problems in mead alcoholic fermentation are similar to
those still found in winemaking such as sluggish or stuck fermentations
(Ingledew and Kunkee, 1985; Kunkee, 1991; Salmon, 1989) as well as
the generation of undesirable by-products such as S-off-avors
(Spiropoulos et al., 2000). Previous research suggested that the major
cause of these problems in wines is the limited nitrogen content of some
natural grape-juice (Giudici and Kunkee, 1994; Hallinan et al., 1999;
Spiropoulos et al., 2000; Spiropoulos and Bisson, 2000; Vos and Gray,
1979). There is a lack of scientic information about honey-must
fermentations but it is accepted by mead makers that mead quality
improvement includes the development of the proper additive
formulation and optimization of fermentation conditions. The numerous

0168-1605/$ see front matter 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijfoodmicro.2010.09.016
194 A. Mendes-Ferreira et al. / International Journal of Food Microbiology 144 (2010) 193198
research studies conducted in other fermented beverages may be in
some extent helpful to control honey-must fermentation. Nevertheless,
research is still needed on the physiology and metabolism of
Saccharomyces cerevisiae under the particular harsh honey-musts
environment. Thus, the aim of this study was to optimize mead alcoholic
fermentation, by modulating honey-must composition, in order to
reduce fermentation length and to obtain a consistently higher quality
and microbiologically stable product. The rst task was to improve the
preparation of honey-must before the various supplements have been
tested. The amounts of nitrogen tested were based on previously
published research (Mendes-Ferreira et al., 2004), which are in
agreement with the previous European legislation for grape-must
supplementation (300 mg/L supplied as diammonium phosphate or
ammonium sulphate) and with current legal limit of 1 g/L. Acidity has
extensive and important functions in alcoholic beverages. Organic acids,
such as tartaric and malic acid have important functions for organoleptic
characteristics, and stability of these types of beverages (Boulton et al.,
1996). Moreover, these acids in equilibrium with their salts act as
buffers, maintaining the pH values within an appropriate range. In
synthetic media the addition of tartaric acid helps to prevent sluggish or
stuck fermentation due to its buffering capacity and because it keeps pH
within the optimal for yeast development (Torija et al., 2003).Therefore,
addition of organic acids was considered in amounts approved by
European legislation for wine production, not only for the important
organoleptic functions of acidity but also to act as buffer compounds in
honey-must fermentation. In parallel, the effects of honey-must
composition, and specically the available nitrogen, on the fermentation
performance and formation of aroma compounds were evaluated.
2. Materials and methods
2.1. Yeast strain and culture maintenance conditions
S. cerevisiae UCD522 from the Enology Culture Collection, Department
of Viticulture and Enology, University of California, Davis, USA was used
in this study. The yeast culture was routinely maintained at 4 C on slants
of Yeast Peptone Dextrose agar (YPD), containing per litre: glucose 20 g,
peptone 10 g, yeast extract 5 g and agar 20 g. Before use, the culture was
transferred to a new slant of YPD for 24 h, at 25 C.
2.2. Honey
In this study, we used honey derived from plants of the Ericaceae
(heather) family (Erica spp.) purchased from a local beekeeper from the
north-eastern region of Portugal. The botanical origin of the honey was
conrmed by relative frequency excluding pollen from nectar-less
plants. Counts were done based on a total of at least 500 pollen grains
and pollen-typing according to the methodology previously described
(Valds et al., 1987). Before honey-must preparation, and to conrm
characteristics and satisfactory quality of honeys in accordance to the
legal limits established in the Council Directive (2001/110/EC, 2002),
the following parameters were analyzed: moisture (%), pH, acidity,
hydroxymethylfurfural (HMF), electrical conductivity, ash (%) and
sugars according to validated standard methods (AOAC, 1990).
2.3. Preparation of honey-must for fermentation
To obtain an alcoholic beverage with approximately 11% of ethanol,
honey was diluted with natural spring-water obtained from the market
(37 g:100 mL) and mixed to homogeneity. Any insoluble solids were
removed the mixture by centrifugation (10,000 g for 10 min Sorval
centrifuge) to obtain a claried honey-must. Then the following
parameters were determined: Brix, pH, total acidity and assimilable
nitrogen concentration. Based on the results obtained by these
determinations, adjustments in nitrogen content and titrable acidity
were done in order to optimize yeast performance during honey-must
fermentation for mead production. Consequently, the following
fermentations were carried out: i) with no additionscontrol fermen-
tation; ii) with 5 g/L potassium tartrate and 3 g/L malic acid supple-
mentation (F1); iii) with 300 mg/L of diammonium phosphate (DAP)
supplementation (F2); iv) with 5 g/L of potassium tartrate, 3 g/L of malic
acid plus 300 mg/L of DAP supplementation (F3); and v) with 5 g/L of
potassium tartrate, 3 g/L of malic acid and 1.015 g/L of DAP supple-
mentation (F4). Each one of the honey-musts was pasteurized at 65 C
for 10 min and then immediately cooled. All experiments were
conducted in triplicate.
2.4. Inoculum preparation and fermentation conditions
For all experiments, starter culture was prepared by pre-growing the
yeasts overnight in 100 mL asks, containing 70 mL of Yeast Nitrogen
Base (without amino acids and without ammonium sulphate) with 20%
glucose and 1.26 g/L DAP. Incubation was done at 25 C in an orbital
shaker at 120 rpm. This pre-culture was used to inoculate the honey-
musts with an initial population of 105 colony-forming units (CFU/mL).
The fermentations were done in 250 mL asks lled to 2/3 of their
volume, tted with a side-arm port sealed with a rubber septum for
anaerobic sampling and maintained at 22 C without shaking. Fermenta-
tions were daily monitored by weight loss as an estimate of CO2
production. Aseptic sampling for assessing fermentation and growth
parameters was done using a syringe-type system as previously described
(Mendes-Ferreira et al., 2009). At the end of alcoholic fermentation
samples were taken from all fermented media to determine the
production of aroma compounds by solid phase micro-extraction
(SPME) coupled to gas chromatography-mass spectrometry (GC-MS).
Yeast cell population and culture dry weight as well as residual nitrogen
were also determined.
2.5. Determination of different growth parameters during fermentation
Yeast growth was followed by periodic measurement of the optical
density (660 nm) of appropriately diluted culture samples in a
Schimadzu UV-2101 spectrophotometer (Schimadzu, Kyoto, Japan)
and by counting the CFUs in solid YPD plates after incubation at 30 C for
48 h. Culture dry weight was determined in samples of 3 50 mL,
centrifuged in pre-weighed tubes for 10 min at 2300 g, washed twice
with sterile deionised water, dried for 24 h at 100 C, and stored in a
desiccator before weighing. Maximum fermentation rate was deter-
mined from the slope of the linear dependence of the steepest decline in
weight (g) at the corresponding time points (h).
2.6. Oenological parameters
Total and free SO2, pH, titrable acidity, volatile acidity and ethanol
content were determined according to validated standard methods
(Organisation International de la Vigne et du Vin, 2006). Yeast
assimilable nitrogen (YAN) was determined according to a method-
ology previously described (Filipe-Ribeiro and Mendes-Faia, 2007).
All physicochemical analyses were performed prior to and at the end
of alcoholic fermentations.
2.7. Solid-phase micro-extraction (SPME) procedure and GC-MS analysis
Chromatographic analyses were performed using an Agilent
6890 N gas chromatograph equipped with a 5973 N mass spectrom-
eter. The target analytes were separated using an Innowax capillary
column, 30 m 0.25 mm with 0.5 m lm thickness (Agilent, Santa
Clara, CA, USA). The column was maintained at 40 C for 5 min after
desorption, ramped at 4 C/min up to 200 C, and then ramped at
10 C/min up to 240 C, where it was held for 15 min. Helium was
used as the carrier gas at 34 cm/s average linear velocity. A 0.75-mm
liner was used and analysis was performed in the splitless mode. All
 A. Mendes-Ferreira et al. / International Journal of Food Microbiology 144 (2010) 193198 195

mass spectra were acquired in electron impact (EI) mode at 70 eV,

using full scan with a scan range of 26250 atomic mass units, at a rate
of 6.12 scans/s. Identication of all compounds was conrmed by
comparing mass spectra and retention indices with those of authentic
standards and quantication was performed in the selected ion
monitoring (SIM) mode, by selecting for each compound their most
characteristic ions (Mendes-Ferreira et al., 2009). To eliminate
variations in extraction efciency caused by small differences in the
sample matrix (particularly ethanol content), internal standardization
using 2-octanol as the internal standard was applied to quantify the
analytes. SPME procedure ber calibration and extraction were
performed according to previously described methods (Mendes-
Ferreira et al., 2009). All experiments were repeated at least twice and
all reported data are mean values.

2.8. Statistical analysis

The data are presented as mean values with their standard deviation.
One-way analysis of variance (ANOVA) and comparison of treatment
means (LSD, 5% level) of the physicochemical parameters of the meads
produced from honey-musts with different formulation were per-
formed using Statistica 6.1 software (StatSoft, Tulsa, OK, USA).

3. Results and discussion

In the course of mead production the delay and/or premature

cessation of the alcoholic fermentation constitutes a major problem,
as seen in other alcoholic beverage industries. In this study the
popular wine yeast strain S. cerevisiae UCD522 was chosen due to its
recognized high fermentative performance and average nitrogen
demands. Honey-must composition was modied by manipulating
yeast assimilable nitrogen and titrable acidity through the addition of
DAP and/or potassium tartrate and malic acid. The effects of such
treatments on yeast growth and fermentative activity as well as on the
production of aroma volatile compounds were evaluated.

3.1. Yeast growth and fermentation behavior in honey-musts

A global view of the fermentation and growth proles is presented
in Fig. 1A, B.
The results showed that the yeast specic growth rate appears not to
be affected by the different supplementations (Fig. 1A). In control and F1
fermentations an earlier slowdown of cell growth was observed as
compared to F2, F3 and particularly to F4 fermentations, underlying the
effect of DAP supplementation on yeast growth (Bely et al., 1990;
Mendes-Ferreira et al., 2004; Varela et al., 2004). Accordingly, a more
relevant increase in yeast cell biomass was only observed in the media
with the highest initial nitrogen concentration (F4), which could explain
a faster alcoholic fermentation in this media. However, the initial
nitrogen concentration had no effect on the extent of nitrogen
consumption by the yeast strain as shown in Table 1; almost 30 mg/L
of available nitrogen remained in the media at the end of fermentations.
Moreover, maximum cell populations in all fermentations were similar
and lower than those observed in other alcoholic fermentations using
different growth media (Bely et al., 1990; Mendes-Ferreira et al., 2004;
Varela et al., 2004). These observations suggest that nitrogen is not the
only growth-limiting factor. To address the question if the yeast strain
could be inadequate to such a harsh environment several new
experiments using different yeast strains, different honey varieties
and yeast culture adaptation prior to inoculation were evaluated.
Neither yeast pre-adaptation nor honey botanical and geographic origin
led to a modication in maximum yeast cell population (results not
shown). Therefore, the inhibitory effect of honey-must on yeast cells
needs to be claried.
No effect of organic acid (potassium tartrate and malic acid) addition
was detected on either on yeast growth or fermentative activity, as it can
Fig. 1. Growth (A) and fermentation (B) proles of S. cerevisiae UCD522 grown under
different conditions. Controlwithout additions; F15 g/L potassium tartrate, 3 g/L
malic acid; F2300 mg/L DAP; F35 g/L potassium tartrate, 3 g/L malic acid and
300 mg/L DAP; F45 g/L potassium tartrate, 3 g/L malic acid, 1.015 g/L of DAP.
be seen by comparing fermentations, F1 with control and F3 with F2
(Table 2 and Fig. 1). Conversely, the initial nitrogen concentration in the
honey-must had a strong impact on yeast fermentative activity. It is
pertinent to point out that some residual sugars, other than glucose,
remained in all meads after alcoholic fermentations (results not shown).
Moreover, the duration of fermentation was reduced from 25 days at
control fermentation to 11 days at F4 (1.015 g/L of DAP) and 14 days at
F2 and F3 (300 mg/L of DAP).
3.2. Evolution of pH throughout honey-musts fermentation
Low pH, among other factors, has been pointed out as one cause of
sluggish or premature fermentation arrest in alcoholic beverages
(Bisson, 1999). Considering the low pH (Sroka and Tuszynski, 2007)
and low buffer capacity (Maugenet, 1964) of honey-musts the
monitoring of pH during alcoholic fermentation was performed in all
experiments to assess whether or not incomplete sugar break down
could be accounted for low pH in this type of beverage (Fig. 2).
Accordingly, in the media in which higher amounts of DAP were added,
the pronounced fall of pH indicates ammonium uptake by yeast cells
through an ammonium/hydrogen ion antiport mechanism (Colombi
et al., 2007; Roos and Luckner, 1984). Nevertheless, it was in the control
fermentation that a more accentuated fall of pH was observed, probably
196 A. Mendes-Ferreira et al. / International Journal of Food Microbiology 144 (2010) 193198

Table 1
Physicochemical characteristics of honey-musts and respective meads obtained after fermentation by yeast strain UCD522. Results are shown as mean values with their standard
deviation.

Control F1 F2 F3 F4

Honey-musts
Brix 22.2 0.2 22.3 0.2 22.0 0.0 22.2 0.1 22.3 0.2
pH 4.47 0.10 3.50 0.10 4.72 0.10 3.54 0.10 3.68 0.10
Titrable acidity (g/L tartaric acid) 0.5 0.1 5.7 0.1 0.5 0.0 5.7 0.0 5.7 0.0
Initial nitrogen (mg/L YAN) 35.0 4.9 35.0 4.9 98.0 4.9 98.0 4.9 259.0 14.8

Meads
pH 3.67 0.13a 3.44 0.07b 3.64 0.08a 3.42 0.06b 3.27 0.16c
Final nitrogen (mg/L YAN) 24.5 4.9a 24.5 4.9a 29.8 7.4a 29.8 2.5a 36.8 7.4a
Alcohol (%) 10.8 0.7a 10.7 0.8a 11.3 0.0a 11.4 0.1a 11.4 0.2a
Titrable acidity (g /L tartaric acid) 3.0 0.1a 7.1 0.0bc 3.1 0.1a 7.5 0.3c 7.7 0.6c
Volatile acidity (g/L acetic acid) 0.63 0.04a 0.84 0.00b 0.57 0.04 ac 0.65 0.06a 0.51 0.04c
Total SO2 (mg/L) 30.7 0.4a 40.8 2.4b 32.1 1.0a 36.2 4.1ab 29.9 4.9a
Free SO2 (mg/L) 0.0 0.0a 1.0 0.4b 0.0 0.0a 1.0 0.3b 0.0 0.0a

Controlwithout additions; F15 g/L potassium tartrate, 3 g/L malic acid; F2300 mg/L DAP; F35 g/L potassium tartrate, 3 g/L malic acid and 300 mg/L DAP; F45 g/L potassium
tartrate, 3 g/L malic acid, 1.015 g/L of DAP. YANyeast assimilable nitrogen. Values in the same line shown with different letters differ signicantly from one another (p b 0.05).

Table 2
Maximum fermentation rate, yeast cell biomass and CFU/mL of Saccharomyces cerevisiae UCD522, in the meads obtained from different supplemented honey-must.

Control F1 F2 F3 F4

Maximum fermentation rate (g/h) 0.069 0.068 0.133 0.134 0.183

Final biomass (g/L dry weight) 1.478 0.076 1.513 0.008 1.709 0.073 1.667 0.067 2.407 0.025
Final CFU/mL 1.5E + 07 1.6E + 07 2.3E + 07 2.0E + 07 2.8E + 07

Controlwithout additions; F15 g/L potassium tartrate, 3 g/L malic acid; F2300 mg/L DAP; F35 g/L potassium tartrate, 3 g/L malic acid and 300 mg/L DAP; F45 g/L potassium
tartrate, 3 g/L malic acid, 1.015 g/L of DAP.

due to lower buffer capacity of this honey-must, but incomplete sugar
consumption could not be accounted for by such pH level.
3.3. Analysis of mead composition
At the end of the alcoholic fermentation, samples were taken to
evaluate mead nal composition (Table 1). The amounts of ethanol
produced by yeast in all of the honey-must fermentations were not
signicantly different and ranged from 10.7 to 11.4% (v/v). Sulphur
dioxide was produced by the yeast in similar quantities in all
experiments. Titrable acidity increased after alcoholic fermentation
which indicates high production of acids by yeast (Boulton et al., 1996).
In other meads the dominant acids responsible for the increase in
titrable acidity are reported to be succinic and acetic acid (Sroka and
Tuszynski, 2007). However, meads obtained in this study showed a
volatile acidity (0.510.84 g/L acetic acid) below the legal limits
specied for this and other alcoholic beverages. The decrease in volatile
acidity observed in this study in response to increasing nitrogen is
inconsistent with the results obtained by other authors using grape-
juice (Barbosa et al., 2009; Bely et al., 2003). From a practical point of
view, this result is very interesting for mead production considering that
volatile acidy, mainly acetic acid, should be as low as possible to avoid
the vinegar off-character.

3.4. Mead aroma composition

To assess how mead aroma composition could be affected by the

Fig. 2. Evolution of pH throughout alcoholic fermentation of different honey-musts.
Controlwithout additions; F15 g/L potassium tartrate, 3 g/L malic acid; F2300 mg/L
DAP; F35 g/L potassium tartrate, 3 g/L malic acid and 300 mg/L DAP; F45 g/L potassium
tartrate, 3 g/L malic acid, 1.015 g/L of DAP.
fermentation conditions evaluated in this study, all meads were
analyzed by GC-MS at the end of alcoholic fermentation. Sixteen
yeast-derived aroma compounds which contribute to the sensorial
quality of mead have been identied and quantied (Fig. 3). Global
analysis of aromatic proles revealed quantitative differences between
the different meads conrming the contribution of both yeast
metabolism and honey-must composition to the diversity of bouquet.
The results obtained herein are in line with those obtained in previous
studies conducted with the same (Carrau et al., 2008; Mendes-Ferreira
et al., 2009) or with other wine strains (Carrau et al., 2008; Hernandez-
Orte et al., 2006; Mendes-Ferreira et al., 2009; Miller et al., 2007;
Vilanova et al., 2007), showing that manipulation of nitrogen concen-
tration of natural or synthetic grape-must modulates the sensory prole
of the nal product. Also, in this work the total concentration of aroma
compounds in meads was higher in those fermentations where DAP was
added. In particular, medium chain fatty acids (MCFA) concentration
were four to vefold higher in F2, F3 and F4 than in control fermentation.
 A. Mendes-Ferreira et al. / International Journal of Food Microbiology 144 (2010) 193198 197

Fig. 3. Concentration of aromatic compounds in the different honey-musts fermented with UCD522 strain. HMhoney-must no fermented. Controlwithout additions; F15 g/L
potassium tartrate, 3 g/L malic acid; F2300 mg/L DAP; F35 g/L potassium tartrate, 3 g/L malic acid and 300 mg/L DAP; F45 g/L potassium tartrate, 3 g/L malic acid, 1.015 g/L of
DAP.

The fresh fruity aroma of fermented beverages derives, to a great extent,
from the mixture of esters produced by the yeast during fermentation
(Saerens et al., 2008). In the present study, the levels of the MCFA ethyl
esters, mirrored the behavior of their respective acids yet, no differences
were found between F3 and control fermentations. This result suggests
that the conversion of MCFA into their corresponding ethyl esters was
strongly affected by the presence of organic acids (potassium tartrate
and malic acid). Also, a positive correlation between nitrogen
availability and ethyl and isoamyl acetates levels was observed,
corroborating the fact that acetate esters production increases with
higher carbon or nitrogen concentrations (Saerens et al., 2008). Ethyl
acetate, a compound with a solvent-like odor, considered an off-avor,
was, quantitatively, the most important ester found, but at a
concentration below its detection threshold (Guth, 1997).
The levels of 2-phenylethanol and isoamyl alcohol were higher in F2
and F3 fermentations. It is widely accepted (Hernandez-Orte et al.,
2006; Mendes-Ferreira et al., 2009; Rapp and Versini, 1991; Vilanova
et al., 2007) that there is an inverse correlation between higher alcohols
and nitrogen levels. Nevertheless, in this study this was only observed in
the range of 100 to 300 mg of YAN/L.
To our knowledge, there are only three reports investigating
some aroma compounds of meads (Sroka and Tuszynski, 2007;
Vidrih and Hribar, 2007; Wintersteen et al., 2005). In these studies it
has been shown that the higher alcohols and esters analyzed varied
198 A. Mendes-Ferreira et al. / International Journal of Food Microbiology 144 (2010) 193198
considerably depending upon honey oral source (Vidrih and Hribar,
2007; Wintersteen et al., 2005), and heat treatment of honey-must
(Wintersteen et al., 2005). The levels of MCFA determined by Sroka and
Tuszynski (2007) were markedly higher than those obtained in this
study. It should be underlined that the comparison of our results with
those previously reported is somehow restricted by the differences in
honey oral source, honey-must preparation, yeast strain and fermen-
tation conditions.
In summary, scientic studies on mead are very rare. Nitrogen
supplementation is a widely accepted practice in the mead production
community yet, there are no previous reports into its effect on yeast
growth and fermentation activity as well as on the sensorial quality of
this fermented alcoholic beverage. In this study we showed that,
addition of organic acids (potassium tartrate and malic acid) had no
effect on growth and fermentation prole. Conversely, addition of DAP
reduced fermentation length, improved the breakdown of sugars and led
to the generation of more interesting aroma compounds by the yeast
strain. Although more studies are needed, our results suggest that the
reduced yeast fermentative ability, and thereby increased risk of delayed
or arrested fermentations, is due to factors other than nitrogen limitation
in honey-musts. Scale-up studies under these optimized conditions are
still needed, as well as the associated organoleptic evaluations of the new
meads.
Acknowledgements
This work was carried out in the framework of project PTDC/AGR-
ALI/68240/2006. C.B. has a fellowship from FCT (SFRH/BD/61881/2009).
The authors thank Miguel Maia for the pollen analysis of the honey used
in this study and Dr. R. N. Bennett for the English revision of the
manuscript.
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