Letters in Applied Microbiology ISSN 0266-8254

ORIGINAL ARTICLE

Optimization of process variables for minimization of

byproduct formation during fermentation of blackstrap
molasses to ethanol at industrial scale
M. Arshad1,2, Z.M. Khan1, Khalil-ur-Rehman2, F.A. Shah3 and M.I. Rajoka3
1 Alcohol Division, Shakargnaj Mills Ltd., Jhang, Punjab, Pakistan
2 Department of Chemistry, University of Agriculture, Faisalabad, Pakistan
3 National Institute for Biotechnology and Genetic Engineering, Jhang Road, Faisalabad, Pakistan

Keywords
fermentation, metabolism, production,
sterilization, stress response, test method,
waste water, yeasts.
Correspondence
Muhammad I. Rajoka, Industrial
Biotechnology Division, NIBGE, Faisalabad,
Pakistan. E-mail:
muhammadibrahim_rajoka@yahoo.com
2007 ⁄ 1877: received 21 November 2007,
revised and accepted 1 July 2008
doi:10.1111/j.1472-765X.2008.02446.x
Abstract
Aims: To investigate the effect of molasses concentration, initial pH of molas-
ses medium, and inoculum’s size to maximize ethanol and minimize methanol,
fusel alcohols, acetic acid and aldehydes in the fermentation mash in industrial
fermentors.
Methods and Results: Initial studies to optimize temperature, nitrogen source,
phosphorous source, sulfur supplement and minerals were performed. The essen-
tial nutrients were urea (2 kg in 60 m3), 0Æ5 l each of commercial phosphoric
acid and sulfuric acid (for pH control) added at the inoculum preparation stage
only. Yields of ethanol, methanol, fusel alcohols, total acids and aldehydes per
100-l fermentation broth were monitored. Molasses at 29Brix (degree of dis-
solved sugars in water), initial pH 4Æ5, inoculum size 30% (v ⁄ v) and anaerobic
fermentation supported maximum ethanol (7Æ8%) with YP ⁄ S = 238 l ethanol per
tonne molasses (96Æ5% yield) (8Æ2% increase in yield), and had significantly lower
values of byproducts than those in control experiments.
Conclusions: Optimization of process variables resulted in higher ethanol yield
(8Æ2%) and reduced yield of methanol, fusel alcohols, acids and aldehydes.
Significance and Impact of the Study: More than 5% substrate is converted
into byproducts. Eliminating or reducing their formation can increase ethanol
yield by Saccharomyces cerevisiae, decrease the overall cost of fermentation pro-
cess and improve the quality of ethanol.

ethanol and lower byproduct yields at industrial level

Introduction
The fermentation of sugars to ethanol is a very old and
well-known process, and has great industrial importance.
The production of alcohol and alcohol-related products
make a significant contribution to our national economy
because they are the most heavily taxed products. More-
over, our country mainly exports ethanol and earns huge
foreign exchange to add to its foreign exchange reserves.
The number of jobs provided by alcohol industry makes
it an important source of income for the nations that
produce it. The process is still the subject of much
research and development, with the aim of getting higher
(Taherzadeh 1999).
Saccharomyces cerevisiae produces ethanol with varying
amounts of other substances, namely methanol, alde-
hydes, higher alcohols (fusel alcohols) and organic acids
(acetic acid and lactic acid). These co-products lower the
quality of ethanol and enhance ethanol distillation cost.
Production of ethanol and byproducts from molasses-
based media has been reported on laboratory scale (Eden
et al. 2001; Shen et al. 2003) and in 87Æ5 m3 fermentors
(Abdel-Fattah et al. 2000), but has not been reported at a
level of 300 m3 fermentor. From previous studies (Mun-
ene et al. 2002), it was envisaged that by optimizing

ª 2008 The Authors

410 Journal compilation ª 2008 The Society for Applied Microbiology, Letters in Applied Microbiology 47 (2008) 410–414
M. Arshad et al.
fermentation variables, sugar concentration, pH and yeast
dosage (yeast cell inoculum), yield of product could be
increased in favour of ethanol and byproducts could be
minimized. The present study deals with the optimization
of process variables to lower the synthesis of byproducts
and enhance ethanol formation during fermentation of
molasses by the yeast S. cerevisiae in 300 m3 fermentor
and recovery of ethanol (95%) after distillation in indus-
trial columns.
Materials and methods
Substrate and nutrients
Carbohydrates in the molasses are readily available in
the form of fermentable sugars and need no pretreat-
ment (Murtagh 2003). Sugarcane blackstrap molasses
used was of production season 2004–2005 of Shakarganj
Mills, Jhang, located in the province of Punjab, Paki-
stan. Molasses was diluted by mixing tap water in
60 m3 mild stainless steel tanks to attain desired Brix.
Although molasses generally contain most of the nutri-
ents required for fermentation, ammonium salts and
phosphates are added to supply nitrogen and phospho-
rus (Prescott and Dunn 2002). Commercially available
urea (2 kg) and phosphoric acid (500 ml), previously
optimized as described earlier (Borzani 1996; de Va-
sconcelos et al. 2004), were used as the main nitrogen
and phosphorus sources in inoculum preparation stage.
Sulfuric acid was used to adjust the pH and as a sulfur
source (de Vasconcelos et al. 2004).
Micro-organism, inoculum preparation and fermentation
Compressed S. cerevisiae (Borzani 1996) from Instant
Yeast (France) was used as the fermenting organism. In
inoculum preparation stage, inoculum’s tank (1 m3 ves-
sel) containing molasses (diluted to 8–9Brix), supple-
mented with phosphoric acid and urea (at 0Æ5 l and
2 kg, respectively) as additional nutrients, was sterilized
with live steam at 121C for 30 min, and was cooled
down to 30C with cooling water. One kilogram of
instant yeast (previously optimized after Carvalho et al.
2003) was transferred to 1 m3 vessel (Arshad 2005) and
grown at 30 ± 1C. After 8-h growth, cell count was
above 300 · 106 cells per ml, and was transferred to
next fermentor (10 m3) containing molasses (8–9Brix)
previously sterilized as earlier. This culture was pumped
to next fermentor (60 m3) and grown at 30 ± 1C. The
industrial scale fermentors (3) for three fermenting
experiments at the same time were autoclaved with live
steam for 1 h and cooled to 30C. Inoculum from
60-m3 fermentors (four in number) when cell count
ª 2008 The Authors
Journal compilation ª 2008 The Society for Applied Microbiology, Letters in Applied Microbiology 47 (2008) 410–414
Ethanol production on industrial scale
was 300 · 106 cell per ml was transferred to three
industrial fermentors of working capacity 300 m3 and
fed with diluted molasses of different Brix or of
known Brix at the mash feeding rate of 15 m3 h)1
(level of fermenter rise was 5% h)1). Addition of
molasses ended after 20 h. At this stage (called fermen-
tation stage), no nutrient supplements were added.
Only unused urea and phosphoric acid were sufficient
to give the highest yield of ethanol (Dombek and In-
gram 1987). Fermentation temperature was maintained
at 30–32C by passing cooling water through plate heat
exchangers. After additional 10 h (total time 30 h), fer-
mentation was complete and fermented mash from the
fermentors was transferred to still tank for distillation.
Recovery of fusel alcohols
Fermented mash from the still tank was transferred to the
mash column (C-201) from top. Steam was applied from
bottom maintaining the temperature of bottom at 78–
80C. Vacuum was applied through condensers. Vapours
travelled from this column to condensers (E-205) where
they heated the mash. From evaporator (E-205), vapours
passed to E-206A, condensate passed to column C-203
(called depuration column), where litter water from C-
204 was also mixed. From C-203 mash was fed to column
C-204 (rectification column). Rectification of ethanol was
done in range of 96Æ0–96Æ4%. This column was operated
under pressure and high temperature. From the bottom
of this column, higher alcohols (fusel alcohols) and acids
were removed and quantified as desired.
Analytical methods
Brix was measured with the help of ATAGO densimeter
(model 2312; ATAGO Co. Ltd, Tokyo, Japan). Ethanol in
fermented samples was determined with ebulliometer and
confirmed on HPLC as described previously (Latif and
Rajoka 2001). Analysis of final products (ethanol, acetic
acid, lactic acid) was also done on gas chromatography
(Shimadzu GC-17A version 3Æ0) equipped with flame ioni-
zation detector (FID) as reported earlier (Shen et al. 2003).
Concentration of aldehydes was measured as potassium
permanganate test time (PTT), as described earlier
(ASTM-D-1363). Briefly, 50-ml sample was poured in test
tube and 2 ml of KMnO4 (0Æ02%) was added. Another
test tube was taken as control tube to which 2 ml of
KMnO4 (0Æ02%) was added and made up to 50 ml with
distilled water. The time of change in colour (as com-
pared with control) was noted at the end. Acidity was
measured titrimetrically using phenol red as indicator
(endpoint was pink).
411
Ethanol production on industrial scale
Statistical analysis
The data obtained were subjected to appropriate statistical
analysis (Steel and Torie 1984).
Results
The traditional strategy of optimization, i.e. optimizing
one-at-a-time approach was used to maximize ethanol
production, keeping fermentation temperature (30C)
and pH (4Æ5) constant in control experiments as
described in materials and methods. Molasses were
diluted with tap water to desired sBrix value without fur-
ther treatment and was permitted to flow to the produc-
tion fermentors. Keeping in view the standard operating
conditions in our distilleries, mash feed time was always
kept constant at 15 m3 h)1 (level of fermentor rise was
always 5% h)1). Extensive studies were done to optimize
nitrogen, phosphorous source and other supplements as
described previously (de Vasconcelos et al. 2003). It was
found that urea, phosphoric acid and sulfuric acid (as
described in materials and methods) were sufficient to get
higher productivity of ethanol economically (Arshad
2005). In batch fermentation, the effect of molasses Brix
on per cent ethanol (P £ 0Æ0014) and yield (P £ 0Æ0000)
was found at the highest Brix of 29 used (Table 1). The
optimum (29) Brix supported the lowest methanol
(3Æ34 g per 100 l) (P £ 0Æ0083), fusel (higher) alcohols
(0Æ23 g per 100 l) (P £ 0Æ0000) and aldehydes (14Æ6 min)
(P £ 0Æ0002), and total acidity measured as acetic acid
(0Æ46 g per 100 l) P £ 0Æ0004). Brix 29 was optimized in
these studies and was maintained in all subsequent studies
unless mentioned otherwise.
Dependence of formation of ethanol, methanol, acetic
acid, fusel and aldehydes on pH of the molasses medium
(29Brix) at 30C using 30% inoculum size (Table 2)
indicated that ethanol yield decreased with increasing pH
as were those of acetic acid, fusel alcohol and aldehydes.
Table 1 Effect of molasses medium Brix (initial pH 4Æ5) on production of ethanol, methanol, acetic acid, fusel alcohols and aldehyde in 300 m3
fermentors using 10% inoculum size at 30C
Medium Methanol
Brix Ethanol (%) (g per 100 l)
25 7Æ27 ± 0Æ31d (218d) 3Æ85 ± 0Æ42abc
26 7Æ15 ± 0Æ24d (216e) 4Æ01 ± 0Æ34a
27 7Æ47 ± 0Æ04c (225c) 3Æ89 ± 0Æ07ab
28 7Æ6 ± 2Æ21b (229b) 3Æ52 ± 0Æ047bc
29 7Æ7 ± 0Æ3a (232a) 3Æ34 ± 0Æ38c
F (P £ 0Æ05) 13Æ1 7Æ47
P 0Æ0014 (0Æ0000) 0Æ0083
Each value is a mean of three runs. ±stands for standard deviation among three replicates. Values in parentheses are alcohol yield in l t)1 molas-
ses. Means followed by different letters differ significantly at P £ 0Æ05. F and P values indicated that influence of treatments on all attributes was
highly significant. Aldehydes were determined using potassium permanganate test time (PTT) in minutes.
412 Journal compilation ª 2008 The Society for Applied Microbiology, Letters in Applied Microbiology 47 (2008) 410–414
M. Arshad et al.
Thus, initial pH of 4Æ5 was optimized in these studies and
was maintained constant in the following studies.
Inoculum size-dependent production of ethanol and
by-products following fermentation of molasses medium
(29Brix) of initial pH 4Æ5 at 30C is presented in
Table 3. Ethanol yield increased with increasing inoculum
size and yield of methanol, acetic acid, fusel alcohol or
aldehydes was the lowest at the highest inoculum size
used. Inoculum size (30%) gave the more ethanol and the
least by-product yield (P £ 0Æ0484) (Table 3) and was
optimized for all subsequent experiments.
Discussion
Factors influencing the optimization of molasses for
increase in ethanol production and decrease in by-prod-
uct formation include initial sugar concentration in the
fermentation mash, initial pH of the medium and yeast
cell concentration (dosage) keeping other variables con-
stant (Munene et al. 2002). The sugar concentration opti-
mized at 29Brix (16% total reducing sugars), gave
maximum ethanol yield (232 l t)1, 7Æ7%) and minimum
byproduct formation (Table 1). Ethanol yield was several
fold higher than the values reported by other authors
(73Æ1 kg t)1, Munene et al. 2002). Stehlik-Tomas et al.
(1997) studied initial sugarcane molasses concentrations
at 15, 18, 21 and 24Brix in batch, fed batch and continu-
ous process. They optimized 24Brix as optimal for etha-
nol production and Zn uptake. Other authors also
reported that yeast growth on beet molasses medium was
significantly retarded when the initial sugar concentration
was greater than 120 g l)1 (Wolniewiez et al. 1990). On
the contrary, the amount of all byproducts formed in the
fermentation broth were significantly higher (P £ 0Æ05)
than that supported by molasses at 29Brix.
The pH of the fermentation mash significantly affects
the ionic state of its mineral components and the cellular
exterior surface of the fermenting organism. Significant
Acetic acid Fusel alcohol Aldehydes
(g per 100 l) (g per 100 l) (min)
0Æ56 ± 0Æ06b 0Æ25 ± 0Æ02d 22Æ6 ± 3Æ2a
0Æ57 ± 0Æ5a 0Æ32 ± 0Æ02b 19Æ1 ± 1Æ14b
0Æ58 ± 0Æ01a 0Æ37 ± 0Æ02a 19 ± 0Æ7b
0Æ54 ± 0Æ03c 0Æ29 ± 0Æ02c 18Æ2 ± 1Æ62c
0Æ46 ± 0Æ03d 0Æ23 ± 0Æ02c 14Æ6 ± 0Æ47d
18Æ32 922Æ37 24Æ38
0Æ0004 0Æ0000 0Æ0002
ª 2008 The Authors
M. Arshad et al. Ethanol production on industrial scale

Table 2 Dependence of formation of ethanol, methanol, acetic acid, fusel alcohols and aldehydes on pH of the molasses medium of 29Brix at
30C under nonaerated condition

Medium pH Ethanol (%) Methanol (g per 100 l) Acetic acid (g per 100 l) Fusel alcohol (g per 100 l) Aldehydes (min)
a ab c b
4Æ5 7Æ72 ± 0Æ32 3Æ65 ± 0Æ34 0Æ52 ± 0Æ03 0Æ23 ± 0Æ03 19Æ6 ± 2Æ4b
4Æ7 7Æ31 ± 0Æ32b 3Æ87 ± 0Æ43a 0Æ54 ± 0Æ06b 0Æ32 ± 0Æ04a 23Æ1 ± 3Æ04a
4Æ9 6Æ48 ± 0Æ04d 3Æ54 ± 0Æ33abc 0Æ58 ± 0Æ01a 0Æ34 ± 0Æ03a 19 ± 0Æ7b
5Æ1 6Æ7 ± 0Æ18c 3Æ46 ± 0Æ62bc 0Æ54 ± 0Æ03c 0Æ33 ± 0Æ02a 19Æ0 ± 2Æ23b
5Æ3 6Æ31 ± 0Æ3d 3Æ22 ± 0Æ11c 0Æ58 ± 0Æ01a 0Æ33 ± 0Æ03a 19Æ8 ± 2Æ33b
F (P £ 0Æ05) 75Æ66 5Æ24 5Æ14 76Æ00 12Æ13
P 0Æ0000 0Æ0227 0Æ0238 0Æ0000 0Æ0018

Each value is a mean of three runs. ±stands for standard deviation among three replicates. Means followed by different letters differ significantly
at P £ 0Æ05. F and P values indicated that influence of treatments on all attributes was highly significant. Aldehydes were determined using potas-
sium permanganate test time (PTT) in minutes.

Table 3 Inoculum size-dependent production of ethanol, methanol, acetic acid, fusel alcohols and aldehyde concentrations following fermenta-
tion of molasses medium of 29Brix (initial pH 4Æ5) at 30C under nonaerated condition

Inoculum size (%) Ethanol (%) Methanol (g per 100 l) Acetic acid (g per 100 l) Fusel alcohol (g per 100 l) Aldehydes (min)
a a c b
20 7Æ25 ± 0Æ28 3Æ85 ± 0Æ42 0Æ56 ± 0Æ05 0Æ35 ± 0Æ02 252Æ5 ± 3Æ5a
25 7Æ06 ± 0Æ19c 3Æ77 ± 0Æ39ab 0Æ56 ± 0Æ03a 0Æ42 ± 0Æ01a 18Æ87 ± 1Æ26b
30 7Æ8 ± 0Æ26a 3Æ66 ± 0Æ1b 0Æ53 ± 0Æ03b 0Æ30 ± 0Æ01a 15Æ25 ± 1Æ08c
F (P £ 0Æ05) 198Æ4 8Æ7 6Æ75 141Æ14 21Æ92
P 0Æ0001 0Æ0484 0Æ05228 0Æ0002 0Æ0082

Each value is a mean of three runs. ±stands for standard deviation among three replicates. Means followed by different letters differ significantly
at P £ 0Æ05. Aldehydes were determined using potassium permanganate test time (PTT) in minutes.
F and P at P £ 0Æ05) indicated that the effect of inoculum size on all (except acids) was highly significant.

changes in pH may affect cellular ionic equilibrium and
metabolic enzymes that are particularly sensitive to such
variations (Munene et al. 2002). Under normal fermenta-
tion condition, the optimal pH range for the yeast is 4Æ5–
5Æ5. Variation in these values may influence the metabolic
activities. Fermentation of medium at pH 4Æ5 gave maxi-
mum yield of ethanol at 234 l from 1000-kg molasses
(results not shown) and supported lower values of acids.
Acetic acid accumulation in the medium may change the
pH gradient across the cell membrane down to a pH as
low as 2Æ5. If the extracellular pH is decreased further, the
difference between external and internal pH becomes too
high, which results in acidification of the cytosol (Taher-
zadeh 1999). The undissociated carboxylic acids can dif-
fuse through the cell membrane (Gottschalk 1987;
Verduyn et al. 1990). As the cells usually have a higher
intracellular pH than the medium (Dombek and Ingram
1987), the undissociated acid is partly dissociated into
acetate and hydrogen ions intracellularly. Therefore, the
intracellular pH is decreased and affects the ethanol yield.
It has been reported that acetic acid at a concentration as
low as 0Æ5 g l)1 can inhibit yeast growth (Gottschalk
1987). Variation in pH affected aldehyde content in the
form of elevated pH values.
The higher concentration of byproducts in the cells
may enhance turgor if not effectively exported from the
cell. The decrease of byproduct formation is an example
of a metabolic adjustment by the cells to minimize energy
consumption during metabolism. It is expected that when
the yields of methanol, acids, aldehydes and fusel alcohol
decrease, the metabolism must shift towards ethanol.
Data of Tables 1 and 2 were compared for ethanol
yield (7Æ7 and 7Æ72%, respectively) using molasses of
29Brix at initial pH 4Æ5 but different inoculum size (10%
and 30%, respectively). It revealed that increase in inocu-
lum size although did not significantly increase ethanol
yield, but decreased methanol, acetic acid and fusel alco-
hol in the fermentation mash and corroborated the find-
ings of previous studies (Borzani 2006).
Productivity of ethanol is affected by both inoculum
size and mash feeding size (Carvalho et al. 2003). Carv-
alho et al. (2003) performed experiments in 14-l fermen-
tor and could easily control mash-feeding rate with
peristaltic pump, but our distilleries have a fixed mash-
feeding rate, and this variable was kept constant in these
studies. For known sugar concentration and mash-feeding
size, the ethanol production may attain a maximum value
and formation of byproducts the minimum values when

ª 2008 The Authors

Journal compilation ª 2008 The Society for Applied Microbiology, Letters in Applied Microbiology 47 (2008) 410–414 413
Ethanol production on industrial scale M. Arshad et al.

the yeast viable count in the inocula was 15 · 106 cells Gottschalk, G. (1987) Control of product formation in anaer-
per ml (Munene et al. 2002). Inoculum size (30%) gave obes. Dechema- Monographs 105, 43–53.
the highest ethanol yield but the lowest yield of methanol, Latif, F. and Rajoka, M.I. (2001) Production of ethanol and
acetic acid, fusel alcohols and aldehydes (Table 3), and xylitol from corncobs by yeasts. Bioresour Technol 77, 57–
confirmed the results of previous studies (Remiz et al. 63.
1999). Our current yield testifies that methanol, acids, fu- Munene, C.N., Kampen, W.H. and Njapau, H. (2002) Effects
sel alcohols and aldehyde yields decreased by 33%, 5Æ0%, of altering fermentation parameters on glycerol and bio-
16% and 28% under optimized conditions, respectively, ethanol production from cane molasses. J Science Food Ag-
ric 82, 309–314.
while ethanol yield increased by 8Æ2% at industrial scale
Murtagh, T.E. (2003) Molasses as a Feedstock for Alcohol Pro-
under such conditions.
duction. Wirchester, Virginia: Murtagh and associates.
Prescott, S.C. and Dunn, C.G. (2002) Industrial Microbiology,
References 4th edn. New York: McGraw Hill Book Company Inc.
Remiz, F., Roustan, L., Sablayrolles, J.M., Barre, P. and
Abdel-Fattah, W.R., Fadil, M., Nigam, P. and Banat, I.M.
Dequin, S. (1999) Glycerol overproduction by engineered
(2000) Isolation of thermotolerant ethanologenic yeasts
Saccharomyces cerevisiae wine yeast strains leads to sub-
and use of selected strains in industrial scale fermenta-
stantial changes in by-product formnation and to a stimu-
tion in an Egyptian distillery. Biotechnol Bioeng 68, 531–
lation of fermentation rate in staionary phase. Appl
535.
Environ Microbiol 65, 143–149.
Arshad, M. (2005) Optimization of fermentation conditions
Shen, Y., Moonjai, N., Verstrepen, K.J. and Delvaux, F.R.
for enhanced ethanol production from blackstrap molasses
(2003) Impact of attachment immobilization on yeast
at industrial scale. M Phil thesis, University of Agriculture,
physiology and fermentation performance. J Am Soc Brew
Faisalabad, Pakistan.
Chem 61, 79–87.
Borzani, W. (1996) Ethanol yields during the feeding phase in
Steel, R.G.D. and Torie, J.H. (1984) Principles and Procedures
fed-batch fermentation of sugarcane blackstrap molasses.
of Statistics. New York: McGraw Hill Book Co. Inc.
World J Microbiol Biotechnol 12, 415–416.
Stehlik-Tomas, V., Grba, S. and Runjiperic, V. (1997) Zinc
Borzani, W. (2006) Batch ethanol fermentation: the correlation
uptake of Saccharomyces cerevisiae and its impact on alco-
between the fermentation efficiency and the biomass initial
hol fermentation. Chem Biochem Eng 11, 147–151.
concentration depends on what is considered as produced
Taherzadeh, M.J. (1999) Ethanol from lignocellulose: physio-
ethanol. Braz J Microbiol 37, 87–89.
logical effects of inhibitors and fermentation strategies.
Carvalho, J.C.M., Vitolo, M., Sato, S. and Aquarone, E. (2003)
PhD thesis, Chalmers University of Technology, Goteborg,
Ethanol production by Saccharomyces cerevisiae grown in
Sweden.
sugarcane blackstrap molasses through a fed-batch process.
de Vasconcelos, J., de, N., Lopes, C.E. and de França, F.P.
Appl Biochem Biotechnol 110, 151–164.
(2004) Continuous ethanol production using yeast immo-
Dombek, K.M. and Ingram, L.O. (1987) Ethanol production
bilized on sugarcane stalks. Braz J Chem Eng 21, 357–365.
during fermentation with Saccharomyces cerevisiae: changes
Verduyn, C., Postma, E., Scheffers, W.A. and VanDijken, J.P.
in glycolytic enzymes and internal pH. Appl Environ
(1990) Energetics of Saccharomyces cerevisiae in anaerobic
Microbiol 53, 1286–1291.
glucose-limited chemostat cultures. J Gen Microbiol 136,
Eden, A.L., Nederveld, V., Drukker, M., Benvenisty, N. and
405–412.
Debourg, A. (2001) Involvement of branched chain amino
Wolniewiez, J.J., Ngang, E., Letourneau, F. and Villa, P. (1990)
acid amino-transferases in the production of fusel alcohols
Alcoholic fermentation of beer molasses: study on stillage
during fermentation in yeast. Appl Microbiol Biotechnol 55,
recycle. Biotechnol Lett 12, 73–78.
296–300.

ª 2008 The Authors

414 Journal compilation ª 2008 The Society for Applied Microbiology, Letters in Applied Microbiology 47 (2008) 410–414