Letters in Applied Microbiology ISSN 0266-8254

ORIGINAL ARTICLE

Fermentation of lignocellulosic hydrolysate by the

alternative industrial ethanol yeast Dekkera bruxellensis
J. Blomqvist1, E. South1, L. Tiukova1, M.H. Momeni2, H. Hansson2, J. Stahlberg2, S.J. Horn3,
J. Schnurer1 and V. Passoth1
1 Department of Microbiology, Uppsala Biocenter, Swedish University of Agricultural Sciences, Uppsala, Sweden
2 Department of Molecular Biology, Uppsala Biocenter, Swedish University of Agricultural Sciences, Uppsala, Sweden
3 Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, As, Norway

Keywords
aspen sawdust, Dekkera bruxellensis,
Lignocellulosic hydrolysate, Saccharomyces
cerevisiae.
Correspondence
Johanna Blomqvist, Department of
Microbiology, Uppsala Biocenter, Swedish
University of Agricultural Sciences, Box 7025,
750 07 Uppsala, Sweden.
E-mail: johanna.blomqvist@slu.se
2010 2015: received 9 November 2010,
revised 6 April 2011 and accepted 16 April
2011
doi:10.1111/j.1472-765X.2011.03067.x
Abstract
Aim: Testing the ability of the alternative ethanol production yeast Dekkera
bruxellensis to produce ethanol from lignocellulose hydrolysate and comparing
it to Saccharomyces cerevisiae.
Methods and Results: Industrial isolates of D. bruxellensis and S. cerevisiae were
cultivated in small-scale batch fermentations of enzymatically hydrolysed steam
exploded aspen sawdust. Different dilutions of hydrolysate were tested. None
of the yeasts grew in undiluted or 1 : 2 diluted hydrolysate [final glucose con-
centration always adjusted to 40 g l)1 (022 mol l)1)]. This was most likely due
to the presence of inhibitors such as acetate or furfural. In 1 : 5 hydrolysate,
S. cerevisiae grew, but not D. bruxellensis, and in 1 : 10 hydrolysate, both yeasts
grew. An external vitamin source (e.g. yeast extract) was essential for growth of
D. bruxellensis in this lignocellulosic hydrolysate and strongly stimulated
S. cerevisiae growth and ethanol production. Ethanol yields of 042 001 g etha-
nol (g glucose))1 were observed for both yeasts in 1 : 10 hydrolysate. In small-
scale continuous cultures with cell recirculation, with a gradual increase in the
hydrolysate concentration, D. bruxellensis was able to grow in 1 : 5 hydrolysate.
In bioreactor experiments with cell recirculation, hydrolysate contents were
increased up to 1 : 2 hydrolysate, without significant losses in ethanol yields for
both yeasts and only slight differences in viable cell counts, indicating an ability
of both yeasts to adapt to toxic compounds in the hydrolysate.
Conclusions: Dekkera bruxellensis and S. cerevisiae have a similar potential to
ferment lignocellulose hydrolysate to ethanol and to adapt to fermentation
inhibitors in the hydrolysate.
Significance and Impact of the study: This is the first study investigating the
potential of D. bruxellensis to ferment lignocellulosic hydrolysate. Its high com-
petitiveness in industrial fermentations makes D. bruxellensis an interesting
alternative for ethanol production from those substrates.

environmentally sustainable biofuel feedstock as there is

Introduction
Bioethanol is a liquid fuel with a potential to replace pet-
rol produced from mineral oil. Currently, ethanol is pro-
duced from feedstocks like sugarcane or cereals, which
can also be used as food and animal feed. Lignocellulosic
biomass is the most abundant biomass on earth and
has the potential to become an economically and
less competition with food production (Gnansounou and
Dauriat 2010). Lignocellulosic material consists of cellu-
lose (poly b-1,4-glucose), hemicellulose (mainly poly
b-1,4-xylose in hardwoods) and lignin, which are not
directly fermentable by yeasts. Pretreatment for instance
thermochemical pretreatment like steam explosion
followed by enzymatic degradation of the material is

2011 The Authors

Letters in Applied Microbiology 53, 7378 2011 The Society for Applied Microbiology 73
Lignocellulosic fermentation by Dekkera bruxellensis
required to release the sugar monomers. Thermochemical
pretreatment breaks up the recalcitrant structure of ligno-
cellulose, making the polysaccharides accessible to
enzymes (Alvira et al. 2010). However, the pretreatment
also releases compounds that are toxic to the fermenta-
tion yeasts, for instance furfural, hydroxymethyl furfural
(HMF), acetic acid and phenolic compounds. Acetic acid
originates from the acetyl groups on hemicellulose, furfu-
ral from degradation of pentoses and HMF from glucose
and phenolic compounds from lignin (Alvira et al. 2010).
The classical fermentation yeast Saccharomyces cerevisiae is
usually regarded most appropriate for ethanol production
from lignocellulose, especially when genetically engineered
to ferment pentoses (Bettiga et al. 2009). However, we
have discovered that Dekkera bruxellensis outcompeted
S. cerevisiae in a starch-based industrial fermentation with
cell recirculation (Passoth et al. 2007). The industrial iso-
late was very robust to changes in pH and temperature
and had an equal or higher ethanol yield compared with
industrial S. cerevisiae strains (Blomqvist et al. 2010). This
together with the tolerance of D. bruxellensis to high
numbers of lactic acid bacteria and the ability to ferment
cellobiose (Passoth et al. 2007; Blomqvist et al. 2010)
makes this yeast interesting for the fermentation of ligno-
cellulose, which has not been investigated before. To test
the ability of D. bruxellensis to ferment such a substrate,
ethanol production from steam exploded enzymatically
hydrolysed aspen sawdust was investigated and compared
with S. cerevisiae fermentations.
Materials and methods
Strains and culture media
Yeast strains: D. bruxellensis CBS 11269 and S. cerevisiae
J672 (both isolated from industrial ethanol production
(Blomqvist et al. 2010)). Glucose medium was prepared
according to Blomqvist et al. 2010 with the modifications:
KH2HPO4 689 mmol l)1 and (NH4)2HPO4 201 mmol l)1.
Hydrolysate glucose medium (HGM) contained 1, 1 : 2,
1 : 5 or 1 : 10 volume of the liquid phase of aspen sawdust
hydrolysate (HGM, 05HGM, 02HGM, 01HGM, respec-
tively), (NH4)2SO4, 151 mmol l)1, and yeast extract,
5 g l)1, if not otherwise stated. Glucose was added to the
different dilutions of HGM to reach the same final concen-
tration of 40 g l)1 (022 mol l)1).
Preparation of hydrolysate
Steam exploded aspen sawdust (SEAS) was prepared at
220C, 10 min, in a unit constructed by Cambi A S,
Norway at the Norwegian University of Life Sciences, As,
and stored aseptically at 8C for 24 weeks before use.
74 Letters in Applied Microbiology 53, 7378 2011 The Society for Applied Microbiology
J. Blomqvist et al.
SEAS dry matter (DM) content was estimated to 470%
(w w) using Moisture Analyzer XM 60 (triplicates, at
105C for >30 min; Precisa Instruments AG, Zurich,
Switzerland). Portions of 128 g SEAS (60 g DM) were
pasteurized 1015 min in a boiling water bath.
Accellerase 1000 enzyme solution (Genencor, A
Danisco Division, Palo Alto, CA, USA; 2707 CMC
units g)1, 431 pNPG units g)1 solution) was centrifuged
20 min at 6000 g and filtered through a 02 lm syringe
filter (polyether sulfone; FiltroPur S, Sarstedt AG, Ger-
many). Enzymatic hydrolysis was carried out in two par-
allels at 40C in sodium citrate buffer, pH 50
(33 mmol l)1 final concentration) in 28 l side-baffled
Fernbach flasks (Bellco Glass Inc., Vineland, NJ, USA) at
90 rev min)1, in fed-batch mode: Day 1: One portion of
SEAS (60 g DM, see above) was dissolved in 700 ml dis-
tilled water. The SEAS storage bottle was rinsed with
additionally 50 ml water, which was also added to the
flask. To this hydrolysate suspension, 50 ml 1 mol l)1
sodium citrate buffer, pH 50 and 15 ml enzyme solution
were added. At day 2, one more SEAS portion was added,
including 50 ml water for rinsing the storage bottle. This
procedure was repeated at days 3, 4 and 5. Finally, 300 g
DM SEAS had been added to each flask and the final bio-
mass content was 193% and 199% DM (w w), respec-
tively. Additionally, 15 ml enzyme was added at day 4
(final dosage 01 ml enzyme g)1 DM SEAS). At day 8, the
contents were centrifuged 30 min at 16 300 g and super-
natants were pooled together and sterile filtered (02 lm
Filtropur BT50; Sarstedt AG, Germany). One millilitre
samples taken on days 1, 2, 3, 4, 5, 8 were centrifuged at
13 000 g, filtered (VectaSpin Micro Anopore 02 lm;
Whatman Inc., Piscataway, NJ, USA) and stored at
)20C before carbohydrate analysis by HPAE-PAD.
Batch cultivations with different dilutions of hydrolysate
Precultures were generated as described (Blomqvist et al.
2010). From the precultures, cells were resuspended in
1 ml of the HGM and inocultated to an OD600  10 into
20 ml serum vials (Pharmapack Packaging System, Soder-
talje, Sweden) containing 9 ml of HGM, 05HGM,
02HGM or 01HGM with or without supplementing
yeast extract reaching a final volume of 10 ml. The vials
were closed with a rubber stopper to ensure oxygen limi-
tation. In the case without supplementation of yeast
extract, 65 g l)1 yeast nitrogen base (YNB, Difco; Bec-
ton Dickinson & Co., Franklin Lakes, NJ, USA) was
added at the end of fermentation (see Results). The vials
were incubated on a rotary table (30C, 140 rev min)1).
Growth was monitored twice per day by OD600-measure-
ments and samples were taken out for HPLC analysis at
beginning and at end of experiment.
2011 The Authors
J. Blomqvist et al.
Continuous cultivation with cell recirculation in 6 well
micro-titre plates
Precultured cells were washed with sterile saline (NaCl
solution, 015 mol l)1), and inoculated in 6 well micro-
titre plates (Becton Dickinson & Co.) containing five ml of
glucose medium to an OD600 of approx. 20. The plates
were incubated at slow shaking (50 rev min)1) on a rotary
table at 30C. During the first 2 days 500 ll glucose med-
ium was added to each well four times day (08:00 h,
12:00 h, 16:00 h and 20:00 h). At 12 a.m., two ml of the
cell suspensions was taken out, 02 ml were removed for
OD and viable count (VC) determinations. The remaining
suspension was centrifuged at 8000 g for 1 min. The super-
natant was prepared for HPLC as described (Blomqvist
et al. 2010). The cell pellet was resuspended in 500 ll glu-
cose medium and transferred back to the well. The med-
ium exchange rate corresponded to a total dilution rate of
0017 h)1 (about half of that used in starch-based industrial
processes with cell recirculation, L. Stenmark, Chematur
Engineering, personal communication) calculated by divid-
ing the total added volume during 24 h (2 ml) with the start
volume (5 ml) and time (24 h). From day 2 (12:00 h),
500 ll 01HGM was added instead of glucose medium and
from day 6, at 12:00 h, 02HGM was added to gradually
adapt the cells to the hydrolysate. When necessary, sterile
deionized water was added to the wells to compensate for
volume losses because of evaporation.
Continuous cultivation with cell recirculation in a
bioreactor
Cells were inoculated to an OD600 of approx. 10 to biore-
actors (18 l, Belach, Stockholm, Sweden), containing the
glucose medium with a starting volume of 07 l. Experi-
ments were run at pH 5, 30C, stirring speed
200 rev min)1, dissolved oxygen tension 0% (DO elec-
trode (Broadley James, CA, USA). When the glucose was
consumed (monitored by glucose sticks, Biophan G, Kal-
lies Feinchemie AG, Seibritz, Germany), pumping of
01HGM was commenced at a dilution rate of 0017 h)1.
Once per day, the added volume was pumped out and
samples were taken for OD, HPLC and VC. The remain-
ing cell suspension was centrifuged (3600 g, 15 min); the
pellet was resuspended in sterile deionized water to a vol-
ume of 10 ml and transferred back to the bioreactor. After
6 days, undiluted hydrolysate was pumped in for 2 days.
Analytical methods
Carbohydrates (arabinose, galactose, glucose, mannose,
cellobiose, xylose) were analysed by HPAE-PAD as
described (Dererie et al. 2011).
2011 The Authors
Letters in Applied Microbiology 53, 7378 2011 The Society for Applied Microbiology
Lignocellulosic fermentation by Dekkera bruxellensis
Concentrations of glucose, ethanol, acetate and glycerol
were determined by HPLC as described (Fredlund et al.
2004; Blomqvist et al. 2010). Viability of the yeasts was
determined by VC : Dilution of cell suspensions in saline,
dropping 10 ll of appropriate dilutions onto YPD agar
plates (Blomqvist et al. 2010), incubation at 30C for
24 h (S. cerevisiae) or 72 h (D. bruxellensis). HMF and
furfural were measured by HPLC as described (Almeida
et al. 2008). The hydrolysate was stored for approx.
12 months at +2C before analysis, and some furfural
may have evaporated, thus resulting in a possible under-
estimation of its actual concentration in the hydrolysate
at the time of usage.
Statistical analysis
Students t-test was performed with a significance level of
5% using the software Xlstat (ver. 2010.3.06; Addinsoft,
NY, USA).
Results
Hydrolysate composition
The DM content of the SEAS was 47%. After fed-batch
enzymatic hydrolysis with a final biomass load of 20% DM
(w w), the hydrolysate contained 022 mol l)1 glucose,
120 mmol l)1 xylose, 222 mmol l)1 mannose, 175 mmol
l)1 cellobiose and 017018 mol l)1 acetic acid, and the
pH was 37. The low levels of xylose compared with acetate
strongly indicate that most of the xylan was degraded to
furfural during the SE treatment (Horn and Eijsink 2010).
Indeed, furfural and HMF concentrations were determined
at 159 13 mmol l)1 (15 g l)1) and 131 03 mmol l)1
(165 g l)1), respectively, i.e., above the limit of 1 g l)1
reported to have negative impact on ethanol production
(Wikandari et al. 2010).
Batch cultivations in different dilutions of hydrolysate
We tested growth of D. bruxellensis CBS 11269 and S. ce-
revisiae J672 in different dilutions of hydrolysate in batch
cultures without supplemented yeast extract (HGM,
05HGM, 02HGM, 01HGM). None of the yeasts grew in
HGM and 05HGM. S. cerevisiae grew in 01 and
02HGM, but not D. bruxellensis. However, when 65 g l)1
YNB was added to these cultures at approx. 70 h after
inoculation, D. bruxellensis started growing in 01HGM.
In parallel cultures with yeast extract supplementation,
D. bruxellensis and S. cerevisiae again failed to grow in
HGM and 05HGM. In the 02HGM, only S. cerevisiae
grew, while in 01HGM, both yeasts grew. The maximum
growth rate of S. cerevisiae was ninefold higher in the
75
Lignocellulosic fermentation by Dekkera bruxellensis J. Blomqvist et al.

presence of yeast extract compared with the cultures (a) 10 c e

a d
without addition; the final cell density was about three- 9 b b
fold higher. In the 01HGM, the ethanol yields were
8
042 001 g ethanol (g glucose))1 for both yeast. The
ethanol yield for S. cerevisiae grown in 02HGM was 7

log CFU ml1

049 001 g ethanol (g glucose))1. 6
5
Continuous cultivation with cell recirculation in 6 well 4
micro-titre plates 3

Dekkera bruxellensis has been demonstrated to be very 2

competitive in continuous high cell density fermentations 1
with cell recirculation (Passoth et al. 2007). To test its 0
behaviour in such fermentation with lignocellulose as Glucose 01HGM 02HGM
substrate, we developed a continuous high cell density
cultivation method with recirculation of cells in micro- (b) 035
ab
titre plates to simulate the industrial conditions. To ab
03 ab
define reference values of cell survival and ethanol yield,

Ethanol yield g g1 glucose

cells were first cultivated in glucose medium and then
01HGM was gradually added, as described in Materials
and Methods. In the continuous cultures, the measured
glucose concentrations were 0. When the medium was
completely exchanged to 01HGM no significant differ-
ences in ethanol yield were seen for both yeasts (Fig. 1b),
but the VC was significantly higher in 01HGM (Fig. 1a),
compared with glucose medium for both yeasts. As
D. bruxellensis did not grow in the 02HGM batch cultiva-
tion, we assumed that an inhibitory effect on growth and
ethanol production would be seen when exchanging the
medium to 02HGM. Therefore, 02HGM was gradually
added to determine the minimal inhibition concentration.
Surprisingly, no significant effects on ethanol yield were
seen during medium exchange and the subsequent growth
phase with constant 02 HGM, but the viable counts were
slightly, but significantly lower compared with 01HGM.
The ethanol yield for D. bruxellensis was similar to S. ce-
revisiae, i.e., about 02 g ethanol (g glucose))1. However,
at 02HGM, the yield of D. bruxellensis was slightly, but
significantly lower (P = 0039) compared with S. cerevisiae
(Fig. 1b). The initial pH values in glucose synthetic med-
ium were 50 for both yeasts. When adding the hydroly-
sate medium, pH dropped to 3537 and 3740 for
D. bruxellensis and S. cerevisiae, respectively. In glucose
medium, no differences on growth were observed in a pH
range from 3 to 5 for D. bruxellensis (Blomqvist et al.
2010). However, the impact of pH may be stronger in
hydrolysate environment as the inhibitory effect of weak
acids like acetic acid increases with decreasing pH, thus
the larger pH drop with D. bruxellensis may partially
explain its higher sensitivity. The results imply that it is
possible to adapt D. bruxellensis to higher hydrolysate
concentrations. Ethanol yields in this fermentation were
very low, which may be because of the evaporation of
a a
025 b
02
015
01
005
0
Glucose 01HGM 02HGM
Figure 1 Survival (a) and ethanol yield (b) of D. bruxellensis CBS
11269 (black) and S. cerevisiae J672 (grey) in continuous cultivation
with cell recirculation in 6 well micro-titre plates with increasing con-
centrations of the hydrolysate medium. Columns with at least one
identical superscript letter indicate no significant difference (P < 0.05).
ethanol and or respiratory sugar metabolism instead of
alcoholic fermentation.
Continuous cultivation with cell recirculation in a
bioreactor
To test hydrolysate fermentation under more controlled
conditions, a bioreactor experiment with continuous cul-
tivation and cell recirculation was designed. The yeast
strains were grown in a batch phase in glucose medium
until the glucose was consumed. Then, continuous culti-
vation was initiated by pumping 01HGM to the fermen-
tor. Cell recirculation was achieved as described in
Materials and Methods. After 6 days, pumping of HGM
(undiluted hydrolysate) was commenced, which was con-
tinued for 2 days until the experiment was finished. Dur-
ing that time hydrolysate concentration was increased to

2011 The Authors

76 Letters in Applied Microbiology 53, 7378 2011 The Society for Applied Microbiology
J. Blomqvist et al.
05HGM. As in the micro-titre plate experiments, glucose
concentration was 0. In 01HGM, the VC was
47 108(3 107) CFU ml)1 for D. bruxellensis and
about 13 108(4 107) CFU ml)1 for S. cerevisiae,
which was lower compared with the micro-titre plate
assays. The ethanol yield during growth in 01HGM was
035 001 g ethanol (g glucose))1 for D. bruxellensis and
038 001 g ethanol (g glucose))1 for S. cerevisiae. Sur-
prisingly, when increasing the hydrolysate concentration,
neither ethanol yield nor viable cell count were affected.
Discussion
We have compared for the first time the ethanol produc-
tion potential from lignocellulose of the alternative etha-
nol production yeast D. bruxellensis with that of
S. cerevisiae. For both yeasts, it was necessary to dilute the
material to avoid inhibitory effects. In batch fermenta-
tions, D. bruxellensis failed to grow in 02HGM, in con-
trast to S. cerevisiae, and vitamin addition was essential.
The results indicate high concentrations of inhibitors and
also a lack of growth factors in the hydrolysate, as yeast
extract or YNB substantially improved growth of both
yeasts. D. bruxellensis has an obligate demand for thia-
mine and biotin (van der Walt 1964). The positive effect
on S. cerevisiae indicates a general demand for complex
nutrients when fermenting lignocellulose hydrolysates.
The ethanol yield of D. bruxellensis CBS 11269 was simi-
lar or slightly lower than that of the tested S. cerevisiae
strain. In a recent investigation on glucose medium
(Blomqvist et al. 2010), CBS 11269 had an about 10%
higher ethanol yield than another industrial S. cerevisiae
isolate and almost identical yields compared with S. cere-
visiae J672.
Lower ethanol yields may be because of respiro-
fermentative metabolism in both yeasts. Although the
oxygen tension in the fermentor was 0, there was a
considerable head space in the vessel. As the cells were
glucose limited, respiration was probably not repressed,
enabling utilization of the low oxygen amounts for
respiration. It may also be interesting to note that in the
mitochondrial genome of D. bruxellensis, indications for
the existence of complex I in the respiratory chain were
discovered (Prochazka et al. 2010). This complex could
be responsible for a higher affinity to oxygen, explaining
the slightly lower ethanol yields of D. bruxellensis. In the
batch cultures, respiration was probably repressed by high
initial glucose concentrations. Our investigations were
dedicated to a comparison of both yeasts under similar
conditions and less to an optimization of the fermenta-
tion conditions, but it becomes clear that further investi-
gation is required to identify factors that influence the
ethanol yield.
2011 The Authors
Letters in Applied Microbiology 53, 7378 2011 The Society for Applied Microbiology
Lignocellulosic fermentation by Dekkera bruxellensis
Dekkera bruxellensis had a high ability to adapt to the
harsh conditions in the hydrolysate. When the hydroly-
sate concentration was gradually increased, no decrease in
cell survival and ethanol yield was observed, even at con-
centrations where the cells failed to grow in batch cultiva-
tion. In those fermentations, the performance of
D. bruxellensis was comparable to that of S. cerevisiae.
The ability of rapid adaptation to harsh conditions is an
interesting feature for future applications. A strain CBS
11270, isolated 1 year later than 12269 from the same
industrial ethanol plant, showed improved ethanol pro-
duction parameters (Blomqvist et al. 2010). Thus, our
results suggest that it may be possible to obtain improved
D. bruxellensis strains by laboratory evolution.
S. cerevisiae had in this study slightly better performance
for the production of ethanol, although D. bruxellensis
reached comparable values. In a running investigation, we
found that this yeast can outcompete S. cerevisiae in
co-cultivation (unpublished results). This and its ability to
cope with a high number of lactic acid bacteria in the
fermentation (Passoth et al. 2007) make it an interesting
candidate for industrial ethanol production, as competi-
tiveness against other microbes provides an important
advantage in the largely nonsterile industrial fermentations.
This is probably even more important in lignocellulose
processes, where nonfermentable sugars provide a niche for
eventual contaminants (Schell et al. 2004). Moreover,
understanding the physiological basis of the adaptation to
the higher concentrations of hydrolysate will be highly
relevant for the development of strategies for strain
improvements, both in S. cerevisiaebased and in D. brux-
ellensis-based fermentations. Investigations of stability and
metabolic basis of the adaptations are currently in
progress.
Acknowledgements
This study was financed by the research programme
MicroDrivE (http://microdrive.slu.se). The generous con-
tributions of the MicroDrivE industrial partners Cambi
A S (steam explosion) and Danisco-Genencor (gift of
enzymes) are particularly acknowledged. We are grateful
to Violeta Sanchez Nogue, Applied Microbiology, Lund
University for the help of analysing the inhibitors.
References
Almeida, J.R.M., Modig, T., Roder, A., Liden, G. and Gor-
wa-Grauslund, M.F. (2008) Pichia stipitis xylose reduc-
tase helps detoxifying lignocellulosic hydrolysate by
reducing 5-hydroxymethyl-furfural (HMF). Biotechnol
Biofuels 1, 12.
77
Lignocellulosic fermentation by Dekkera bruxellensis
Alvira, P., Tomas-Pejo, E. and Negro, M.J. (2010) Pretreat-
ment technologies for an efficient bioethanol production
process based on enzymatic hydrolysis. A review. Bioresour
Technol 101, 45814861.
Bettiga, M., Bengtsson, O., Hahn-Hagerdal, B. and Gorwa-
Grauslund, M.F. (2009) Arabinose and xylose fermentation
by recombinant Saccharomyces cerevisiae expressing a fun-
gal pentose utilization pathway. Microb Cell Fact 8, 40.
Blomqvist, J., Eberhard, T., Schnurer, J. and Passoth, V. (2010)
Fermentation characteristics of Dekkera bruxellensis strains.
Appl Microbiol Biotechnol 87, 14871497.
Dererie, D.Y., Trobro, S., Momeni, M.H., Hansson, H.,
Blomqvist, J., Passoth, V., Schnurer, A., Sandgren, M. et al.
(2011) Improved bio-energy yields via sequential ethanol
fermentation and biogas digestion of steam exploded oat
straw. Bioresour Technol 102, 44494455.
Fredlund, E., Blank, L.M., Schnurer, J., Sauer, U. and Passoth,
V. (2004) Oxygen- and glucose-dependent regulation of
central carbon metabolism in Pichia anomala. Appl Environ
Microbiol 70, 59055911.
Gnansounou, E. and Dauriat, A. (2010) Techno-economic
analysis of lignocellulosic ethanol: a review. Bioresour
Technol 101, 49804991.
78 Letters in Applied Microbiology 53, 7378 2011 The Society for Applied Microbiology
J. Blomqvist et al.
Horn, S.J. and Eijsink, V.G.H. (2010) Enzymatic hydrolysis of
steam-exploded hardwood using short processing times.
Biosci Biotechnol Biochem 74, 11571163.
Passoth, V., Blomqvist, J. and Schnurer, J. (2007) Dekkera
bruxellensis and Lactobacillus vini form a stable ethanol-
producing consortium in a commercial alcohol production
process. Appl Environ Microbiol 73, 43544356.
Prochazka, E., Polakova, S., Piskur, J. and Sulo, P. (2010)
Mitochondrial genome from the facultative anaerobe and
petite-positive yeast Dekkera bruxellensis contains the
NADH dehydrogenase subunit genes. FEMS Yeast Res 10,
545557.
Schell, D.J., Riley, C.J., Dowe, N., Farmer, J., Ibsen, K.N.,
Ruth, M.F., Toon, S.T. and Lumpkin, R.E. (2004) A bio-
ethanol process development unit: initial operating experi-
ences and results with a corn fiber feedstock. Bioresour
Technol 91, 179188.
van der Walt, J.P. (1964) Dekkera, a new genus of the Sacchar-
omycetaceae. Antonie Van Leeuwenhoek 30, 273280.
Wikandari, R., Millati, R., Syamsiyah, S., Muriana, R. and
Ayuningsih, Y. (2010) Effect of furfural, hydroxymethyl-
furfural and acetic acid on indigeneous microbial isolate
for bioethanol production. Agric J 2, 105109.
2011 The Authors