Research Article
Effect of pH and temperature on vinasse decolorization
by lactic acid bacteria in batch processes
Marta Wilk,1 Małgorzata Krzywonos,1 Przemysław Seruga,1 Ewa Walaszczyk2
1
Department of Bioprocess
Engineering, Wroclaw University of
Economics, Wrocław, Poland
2
Department of Biotechnology and Food
Analysis, Wroclaw University of Economics,
Wrocław, Poland
Received 10 July 2018; Revised 9 October
2018; Accepted 30 October 2018
National Science Centre (Poland), Grant/
Award Number: N N312 421940
Correspondence to: Marta Wilk,
Department of Bioprocess Engineering,
Wroclaw University of Economics,
Wrocław, Poland.
Email: marta.wilk@ue.wroc.pl
Published online 28 February 2019 in Wiley
Online Library (wileyonlinelibrary.com)
DOI: 10.1002/wer.1065
© 2019 Water Environment Federation
Water Environment Research • 91: 573–580, 2019
• Abstract
The waste-­free policy is part of the process of sugar production from beets in which the
resulting molasses are used for ethanol production. However, during this process an-
other byproduct, namely vinasse, is created. Therefore, there is a problem with the
utilization of wastewater, which cannot be disposed to the environment without being
treated. Melanoidins, caramels, and hexoses alkaline degradation products contained
in the vinasse give it a dark brown color. The aim of the study was to investigate the
effect of the pH and the temperature on the decolorization of vinasse by lactic acid
bacteria (Lactobacillus plantarum, L. casei, and Pediococcus parvulus). Experiments
were performed in batch mode in a BioStatB bioreactor for 72 hrs. The medium con-
sisted of 25% v/v sugar beet molasses vinasse, 77.34 gdm−3 of glucose, and 2.24 gdm−3
of yeast extract. The maximum decolorization was 25.14% and was achieved at non-
controlled pH 6.5 and at 30°C.   © 2019 Water Environment Federation
• Practitioner points
• Lactobacillus plantarum, L. casei and Pediococcus parvulus showed potential for de-
colorization of sugar beet molasses vinasse.
• Controlled pH has a negative effect on sugar beet molasses vinasse decolorization.
• Toxic substances, i.e. acrylamide, 4-methylimidazole , 5-hydroxymethylfurfural and
furfural after decolorization was not detected.
• Bacteria showed high degradation potential of 2-acetyl-4-(1,2,3,4)-tetrahydroxy-
butylimidazole.
• Key words
decolorization; lactic acid bacteria; vinasse; wastewater
Introduction
The biofuel produced in the world in the largest quantity is ethanol. Brazil and the United
States supply most of the ethanol on the market, and their production is mainly based on
sugar cane and maize, respectively. According to OECD (2017) overview, the world etha-
nol production will grow by 19∙109 L in the next 10 years, so it is approximately 73% less
than between 2004 to 2006 and 2014 to 2016. The slowdown of ethanol growth is because
of the stagnating mandated ethanol use in the United States, but it is forecast that demand
will be partially compensated by developing countries where ethanol is produced from
molasses. In the European Union, ethanol is mostly produced from sugar beet, maize,
and wheat (OECD, 2017, 2018). The sugar industry promotes no waste process ideas and
claims that ethanol production could be more efficient if distilleries were to use interme-
diates and byproducts like molasses besides sugar beets (Gumienna et al., 2016). It is eco-
nomically justified because 100 to 120 L of ethanol can be obtained from 1 ton of sugar
beets; whereas 300 L of ethanol can be obtained from 1 ton of beet molasses (Alexiades,
Kendall, Winans, & Kaffka, 2018; Golisz & Wójcik, 2013). The problem is that distilleries,
in addition to ethanol, obtain waste water called sugar beet molasses vinasse (BMV) in
the amount of 8 to 15 L per liter of alcohol (Naik, Jagadeesh, & Alagawadi, 2008).
Sugar beet molasses vinasse is characterized by a high load of pollutants, an
acidic pH, and a dark brown color (Krzywonos, Chałupniak, & Zabochnicka-­Świątek,
Research Article
2017; Wilk, Krzywonos, & Seruga, 2017). Colored com-
pounds, which are formed during sugar beet and then
molasses processes, are called the following: melanoidins—
from Maillard reaction; caramels—from decomposition of
sucrose; and hexoses alkaline degradation products (HADP)
(Schlumbach, Pautov, & Flöter, 2017; Wilk et al., 2017). In
vinasse, one can also find toxic or carcinogenic substances,
like furfural, 5-­hydroxymethylfurfural (5-­HMF), acrylamide,
4-­methylimidazole (4-­MeI), and 2-­acetyl-­4-­(1,2,3,4)-­tetrahy
droxy-­butylimidazole (THI), which appear during very com-
plex and a plurality of successive and parallel chemical reac-
tions (Coca, García, González, Peña, & García, 2004; Lee,
Jang, & Shibamoto, 2013). Chemical oxygen demand (COD)
and biochemical oxygen demand (BOD) value (80 to 100 g
O2 dm−3 and 40 to 50 g O2 dm−3, respectively) and color make
the BMV difficult to degrade (Krzywonos & Seruga, 2012).
It is possible to use vinasse as a fertilizer, but only in a low
concentration. Research shows that long-­term fertilization of
the fields with the vinasse causes soil saturation and contam-
ination of nearby bodies of water as well as contamination of
the aquifer (España-­Gamboa et al., 2017; González & Mejía,
2015). Moreover, vinasse causes the inhibition of seed germi-
nation (Ale, Jha, & Belbase, 2008). Disposal of such a dark and
polluted byproduct into the water is also dangerous because
it could endanger the existing flora and fauna (Christofoletti,
Escher, Correia, Marinho, & Fontanetti, 2013). It is therefore
necessary to treat the vinasse before it is discharged into the
environment. There are known physicochemical and micro-
biological methods of COD and BOD reduction (Fuess, de
Araújo Júnior, Garcia, & Zaiat, 2017; Lutosławski, Ryznar-­
Luty, Cibis, Krzywonos, & Miśkiewicz, 2011; Petta, De Gisi,
Casella, Farina, & Notarnicola, 2017; Rodrigues et al., 2017;
Santos, Ricci, França Neta, & Amaral, 2017), but the sec-
ond stage of vinasse treatment should be the removal of the
color. The physicochemical treatment of vinasse is not eco-
nomically viable (Campos, Mesquita, Silva, & Schwan, 2014).
This is why many researchers focus on microbial decoloriza-
tion (Limkhuansuwan & Chaiprasert, 2010; Rajasundari &
Murugesan, 2011; Ravikumar, Vasanthi, & Saravanan, 2013).
In these bioprocesses, temperature and pH value have a very
important role (Kumar et al., 1997; Santal, Singh, & Saharan,
2016; Tiwari, Gaur, & Singh, 2012; Zuraida, Nurhaslina, & Ku,
2013). The aim of this work is investigation of the influence of
pH and temperature on the decolorization of BMV by a con-
sortium of lactic acid bacteria.
Methodology
Materials
Distillery wastewater. The BMV was collected from CHECO
Manufacturing Plant, Ltd., Włocławek, Poland, and was stored
in a sealed container, at −20°C. Vinasse characterization and
data reported in the literature are shown in Table 1.
Microorganisms
Lactobacillus plantarum MiLAB393 and Pediococcus parvulus
MiLAB099 were obtained from the Department of Microbiology
574
of the Swedish University of Agricultural Sciences in Uppsala.
Lactobacillus casei 0848 was obtained from the Department
of Food Chemistry of the Polish University of Technology in
Łódź. The strains were stored in the MRS medium (de Man,
Rogosa and Sharpe; Biocorp, Poland) with 10% v/v glycerol, at
65°C.
Process conditions
The experiments were carried out in duplicate for 72 hrs in a
5 dm3 working volume stirred-­tank bioreactors (Biostat B, B.
Braun Biotech International) with a stirrer speed of 200 rota-
tions per minute, no aeration, at 30 or 37°C, pH 6.0 or 6.5.
These conditions and medium composition have been chosen
according to the authors’ previous experiments and literature
(Adikane, Dange, & Selvakumari, 2006; Kaushik & Thakur,
2009; Limkhuansuwan & Chaiprasert, 2010; Zuraida et al.,
2013). Moreover, the processes were carried out with both con-
trolled (pH) (parameter value was maintained automatically
by using 2 M H2SO4 and 33% NaOH throughout the entire
process) and noncontrolled (pH0) pH levels. The authors’
previous experiments were conducted in the shake-­flask and
the medium composition was optimized using experimental
design of experiments (manuscript in preparation). On this
basis, the authors have formulated the medium consisting of
BMV (25% v/v), 77.34 g dm−3 of glucose (Chempur, Poland)
(added separately after medium sterilization at 121°C for
15 min), and 2.24 g dm−3 of yeast extract (Biocorp, Poland).
After sterilization, the culture medium was inoculated with
20 cm3 of a bacteria suspension in MRS medium, which cor-
responded to 1.5 g dm−3 of bacteria dry weight.
Methods
The samples were centrifuged at 9000 g (Sigma® 4K15) for
15 min. The decolorization effectiveness and biomass growth
were evaluated spectrophotometrically (475 nm and 620 nm,
respectively).
Melanoidins, caramels, and HADP content were deter-
mined using the Ivanov-­Sapronov method (details were pro-
vided in Krzywonos, Seruga, Wilk, Borowiak, & Stelmach, 2016).
Melanoidins glucose-­ glycine (Glu-­Gly), acrylamide (Sigma,
Poland), 4-­methylimidazole (4-­MeI) (Alfa Aesar, Germany), 2-­
acetyl-­4-­(1,2,3,4)-­tetrahydroxy-­butylimidazole (THI) (Sigma,
Poland), 5-­hydroxymethylfurfural (5-­HMF) (Aldrich, Poland),
furfural (Acros Organics, Poland), glucose and lactic acid
content were measured by HPLC (Knauer). Chemical oxygen
demand was established spectrophotometrically using Hach-­
Lange cuvette tests. Details of all used methods were provided
previously (Wilk, Krzywonos, Borowiak, & Seruga, 2019).
The decolorization effectiveness and reduction of com-
ponents and indicators of vinasse were calculated from the
formula:
(A0 − At )
%A = ⋅ 100% (1)
A0
where:
A0 is the initial value, and
At is value in the time t.
Wilk et al.
 Research Article

Table 1. Vinasse characterization

a
VALUE
CIBIS, RYZNAR-­LUTY,
KRZYWONOS, LUTOSŁAWSKI, RYZNAR-­LUTY
PARAMETER WILK ET AL. (2019) AND MIŚKIEWICZ (2011) ET AL. (2008)
pH 5.0 4.97 ± 0.01 5.4
COD 89.3 ± 4.8 57.39 ± 0.35 126.7
BOD5 256 ± 10.7 36.40 ± 2.14 nd
Glycerol 3.9 ± 0.19 3.333 ± 0.117 3.74
Glucose 1.42 ± 0.07 nd nd
Total nitrogen 5.075 ± 0.205 4.004 ± 0.165 3.63
Total phosphorus 0.1 ± 0.02 0.056 ± 0.0021 0.0057
Lactic acid 20.4 ± 1.02 2.913 ± 0.148 20.76
Acetic acid 2.22 ± 0.11 1.509 ± 0.082 12.11
Pyroglutamic acid 8.51 ± 0.43 5.568 ± 0.178 nd
Succinic acid 11.65 ± 0.58 nd nd
Isobutyric acid 21.07 ± 1.05 nd 1.65
Tartaric acid 1.04 ± 0.05 nd nd
Gluconic acid 2.17 ± 0.11 0.871 ± 0.032 nd
Invert degradation products of 20.07 ± 1 16.45 ± 1.41 nd
alkaline hydrolysis
Caramels 1.75 ± 0.09 2.401 ± 0.208 nd
Melanoidins 2.91 ± 0.15 0.879 ± 0.115 nd
Acrylamide 0.011 ± 0.001 nd nd
4-­methylimidazole 0.602 ± 0.031 nd nd
2-­acetyl4-­(1,2,3,4)-­tetrahydroxy-­ 0.062 ± 0.003 nd nd
butylimidazole
furfural 0.083 ± 0.004 nd nd
5-­HMF 0.016 ± 0.001 nd nd
Notes. nd, no data.
a
All values except pH are expressed in g dm−3.

Results and discussion
It was observed that control of the pH is critical for color
removal. In experiments at noncontrolled pH, decolorization
was two times higher than at a controlled pH. The highest color
removal, 25.14%, was achieved in experiment at initial pH 6.5,
noncontrolled. The percentages of decolorization of BMV by
using lactic acid bacteria are shown in Figure 1. During the first
12 hrs, a decrease of pH from 6.5 to 4.6 and almost 2/3 of total
color reduction was noted. During the next 5 hrs, pH decreased
averagely approximately 0.05 units per hour. After this time,
significant changes in pH were not observed and at the termi-
nation of fermentation the pH was at 3.6 (Figure 2). Different
microorganisms have been used for decolorization (Table 2).
In the literature, there are references where better results have
been achieved for decolorization with mixed culture than
individual microorganism (Kumar & Chandra, 2006; Sharma
& Mittal, 2014; Tiwari, Gaur, & Singh, 2014). Kumar and
Chandra (2006) explained this phenomena as the enhanced
effect of coordinated metabolic interactions on decolorization.
Moreover, Ryznar-­Luty, Krzywonos, Cibis, and Miśkiewicz
(2008) reported that, compared to monocultures, nutritional
requirements of mixed cultures are lower and their activity is
higher. Regarding the process conditions, it is assumed that
favorable temperature for decolorization depend on the strains
and their genetic variety (Tiwari et al., 2014). Adikane et al.
(2006) showed a critical role of the pH decrease in decoloriza-
tion, based on the result of experiments in which less decol-
orization or increase in color was achieved when pH increased.
Seyis and Subasioglu (2009) also found a relationship between
the molasses decolorization and acidic conditions after incuba-
tion. They observed that color removal by fungi was achieved
with the pH value lower than 4.9. España-­Gamboa et al. (2015,
2017) recognized the increasing pH inside the air-­pulsed bio-
reactor during decolorization by Trametes versicolor, as the
reason for no significant color removal. Kaushik and Thakur
(2013) reported that the relationship between pH and decol-
orization was connected with the bioadsorption potential of
vinasse on the cell walls of microorganism. They explained
that complex polymeric organic compounds with different
aromatic rings and functional groups in vinasse have a wide
range of ionization potentials at different pH values. The ini-
tial pH value also has a critical influence on decolorization
(Jiranuntipon, Chareonpornwattana, Damronglerd, Albasi, &
Delia, 2008). Tiwari et al. (2014) studied the effect of initial pH

Water Environment Research • 91: 573–580, 2019 575

Research Article
(A)
(B)
Figure 1. Effect of process conditions (T = 30°C or 37°C and pH = 6.0 or pH = 6.5) on decolorization under (a) without pH control (pH0) and
(b) controlled pH (pH).
Figure 2. pH value ( ) during the process at T = 30°C and
pH0 = 6.5 and its effect on decolorization ( ).
in the range from 4.0 to 7.0, at intervals of 0.5, on distillery
spentwash decolorization by consortium of Pediococcus acidi-
lactici and Candida tropicalis. They noted the highest efficiency
of color removal in an experiment with pH 6.0; moreover,
beyond this value there was a decrease in decolorization.
Ravikumar et al. (2013) optimized the parameters of the
decolorization by Cladosporium cladoporioides using response
surface methodology. They found that carbon concentration
576
and pH are highly significant factors, because in an optimum
pH of vinasse the organisms might acquire a sufficient carbon
source. They achieved the highest color removal at pH 6.0. In
this study during the pH controlled process (pH = 6.5), bacteria
assimilated 81.36% of glucose, whereas during noncontrolled
pH experiment (pH0 = 6.5), concentration of carbon source
decreased by only 23.54% (Figure 3). Interestingly, despite the
low level of glucose reduction, the increase in biomass and
decolorization were 2.7 and 2 times higher, respectively, com-
pared to the values obtained in controlled pH processes. It can,
therefore, be assumed that the bacteria, under noncontrolled
pH conditions, assimilated the original carbon source found
in the vinasse, for example, from colored compounds. Under
controlled pH, the bacteria used glucose for the production of
lactic acid, which was noted to be five times higher than in non-
controlled pH.
In controlled pH experiments, an increase in surface area
characteristic of Glu-­Gly melanoidins was observed (Figure 4).
Under noncontrolled pH conditions, the highest removal of
melanoidins Glu-­Gly was reported for experiments at 37°C. In
the experiment in which vinasse decolorization was the high-
est (pH0 = 6.5, T = 30°C) (Figure 1a), the removal of melanoi-
dins Glu-­Gly was 28.64%, and it was the lowest result among
experiments conducted under noncontrolled pH (Figure 4). It
Wilk et al.
 Research Article

Table 2. Microorganism and process conditions used in decolorization of distillery effluent

COLOR INITIAL TEMPERATURE
NAME REMOVAL (%) PH (°C) REFERENCES
soil sample 69 6.0 30.0 Adikane et al. (2006)
Trichoderma viride 53.5 <4.9 30.0 Seyis and Subasioglu (2009)
Trametes versicolor 18 4.5 25.0 España-­Gamboa et al. (2015)
Klebsiella oxytoca, Serratia marcescens, and 19.3 4.0 30.0 Jiranuntipon et al. (2008)
Citrobacter sp.
Pediococcus acidilactici and Candida 82 6.0 45.0 Tiwari et al. (2014)
tropicalis
Cladosporium cladoporioides 62.5 6.0 37.0 Ravikumar et al. (2013)
Lactobacillus plantarum MiLAB393 26 6.5 35.8 Wilk et al. (2017)
Lactobacillus plantarum MiLAB93 67 6.25 37.0 Seruga and Krzywonos (2015)
Lactobacillus plantarum SF5.6 60.91 6.0 30.0 Limkhuansuwan and
Chaiprasert (2010)
Paracoccus pantotrophus 81.2 7.0 37.0 Santal et al. (2016)
Bacillus megaterium ATCC 14581 38 7.0 37.0 Krzywonos et al. (2017)
Lactobacillus plantarum MiLAB393, L. casei 25.14 6.5 30.0 This work
0848, and Pediococcus parvulus MiLAB099

(A) 90 800
80 700
Lactic acid and glucose

70
concentration [g/dm3]

Biomass growth [%]

600
60
500
50
400
40
300
30
20 200

10 100
0 0
0 12 24 36 48 60 72
t [hr]
Lactic acid Glucose Biomass

(B) 90 800
80 700
Lactic acid and glucose

70
concentration [g/dm3]

Biomass growth [%]

600
60
500
50
400
40
300
30
20 200

10 100
0 0
0 12 24 36 48 60 72
t [hr]
Lactic acid Glucose Biomass

Figure 3. Biomass growth and concentration of lactic acid and glucose during decolorization under noncontrolled pH0 = 6.5 (a) and con-
trolled pH = 6.5 (b) at 30°C.

can be assumed that under these conditions there was a higher an increase in the content of caramels in the medium. Hexoses
reduction of other color compounds than in experiments alkaline degradation products and melanoidins contents were
with pH0 = 6.0 at T = 30°C or T = 37°C and with pH0 = 6.5 at reduced, while under controlled pH conditions the melanoidins
T = 37°C. It was found that under noncontrolled pH, there was content increased, and the HADP and caramels concentration

Water Environment Research • 91: 573–580, 2019 577

Research Article

T = 30°C T = 37°C
pH
pH

pH = 6.0
pH

pH = 6.0
pH T = 37°C
T = 30°C
pH = 6.5

–105 –85 – 65 – 45 –25 –5 15 35 55 pH = 6.5

Reduction of melanoidin Glu-Gly content [%]
Figure 4. Reduction of melanoidins glucose-­ glycine (Glu-­
Gly)
content depending on the process conditions under controlled pH
(pH) and without pH control (pH0) at 30°C and 37°C.
decreased (Table 3). The reduction of the melanoidins content
in the experiments in which the pH decreased over time resulted
from the fact that these compounds are easier to remove in an
acidic medium (Rajasundari & Murugesan, 2011).
The authors of this study tested at temperatures of 30°C
and 37°C because they are the best for lactic acid bacte-
ria growth. The highest color removal was achieved at 30°C
(Figure 1). For Lactobacillus plantarum MiLAB393 used in
the previous study (Wilk et al., 2017) and for L. plantarum
MiLab93 used in the study of Seruga and Krzywonos (2015),
optimum temperatures were 35.8°C and 37°C, respectively.
Optimum temperatures were higher than for the bacteria con-
sortium. Limkhuansuwan and Chaiprasert (2010) also tested
the ability of lactic acid bacteria to effect decolorization, but
in a wider range of temperatures: 20, 25, 30, 37, 43, and 50°C.
They reported that the degree of decolorization increased as the
temperature increased to 30°C. Above this value, there was a
decrease in color removal. Santal et al. (2016) found the same
correlation but decolorization increased to the level of tempera-
ture of 37°C. Zuraida et al. (2013) used Lactobacillus delbruckii
in the decolorization of batik wastewater. They monitored the
process at different temperatures (25, 30, 37, and 42°C) and
initial pH (4, 5, 6, 7, and 8). It was interesting that the opti-
mum temperature for bacteria growth and decolorization was
37°C, but the optimum initial pH was different: 7.0 and 6.0,
respectively. A range of pH from 6.0 to 8.0 was used by Yadav
–5 0 5 10 15
Reduction of COD [%]
Figure 5. Chemical oxygen demand (COD) reduction depending
on the process condition under pH with control (pH) and without
control (pH0) at 30°C and 37°C.
and Chandra (2012) to examine the effect of pH on the degra-
dation of extracted molasses melanoidins by bacterial consor-
tium. The best results were recorded at pH 7.0; pH higher or
lower than this value adversely influenced the bacterial growth
and degradation capability. They also noted that temperature
affects the microbiological decolorization effectiveness. On the
other hand, the authors of this study have recorded that effect
of temperature on decolorization is not significant like pH con-
trolling (Figure 1). Tiwari et al. (2014) noted that consortium
of Pediococcus acidilactici and Candida tropicalis was active in
all tested temperatures (it exhibited about 80% decolorization
in all experiments) so mix culture of microorganism was not
affected by temperature.
For the experiment that was carried out at 37°C with con-
trolled pH of 6.0, the highest COD reduction of 14.5% (Figure 5)
and the lowest degree of decolorization (8.87%) was recorded
(Figure 1). To maintain the pH at level 6.0, NaOH was added
to the medium throughout the experiment. In the process with
the highest degree of decolorization, the value of COD during
the process decreased by 1.6%. In this process, bacterial acid-
ification took place and the pH decreased to 3.6. Krzywonos
et al. (2017) have noted that during BMV decolorization by
Bacillus megaterium ATCC 14581, the acidification of the
medium influenced the increasing of the COD value. They also

Table 3. Hexoses alkaline degradation products (HADP), melanoidins, and caramels reduction depending on the process condition
PROCESS CONDITIONSa REDUCTION OF COLORED COMPOUNDS (%)
T (°C) HADP MELANOIDINS CARAMELS
pH0 = 6.5 30 14.97 2.50 −35.57
pH0 = 6.5 37 11.65 9.51 −32.42
pH0 = 6.0 30 15.85 7.99 −27.63
pH0 = 6.0 37 6.69 18.99 −37.72
pH = 6.5 30 25.01 −34.61 42.33
pH = 6.5 37 13.26 −14.50 32.63
pH = 6.0 30 15.56 −55.54 26.16
pH = 6.0 37 6.89 −4.61 24.96
Notes. Minus before the value means the increase in the content of a colored compound.
a
pH0 = 6.0 and pH0 = 6.5 means noncontrolled pH; pH = 6.0 and pH = 6.5 means controlled pH.

578 Wilk et al.


reported that BMV concentration affected COD removal. The
best results they obtained were in the run with 5% v/v addition
of vinasse. Higher vinasse concentrations resulted in a smaller
(or no) COD reduction. Considering that the authors of this
study used 25% v/v of the vinasse in the medium, the report
by Krzywonos et al. (2017) may confirm the low, or no, reduc-
tion of COD in the present study. Moreover, when the authors
herein used single strain of lactic acid bacteria in the previous
study (Wilk et al., 2017), the COD reduction was not greater
than 14%. Hence, it can be concluded that the use of a lactic
acid bacteria consortium did not improve the COD reduction.
In an experiment conducted at 30°C with a pH controlled
at 6.5, complete removal of THI was noted. In the processes
with pH = 6.5 at T = 37°C, pH = 6.0 at T = 37°C, and T = 30°C,
reduction of the content of this compound was approximately
80%. In the process in which decolorization efficiency was at
the highest level (pH0 = 6.5, T = 30°C), the THI decrease was
about 62%, and in the experiments with pH0 = 6.5 at T = 37°C,
pH0 = 6.0 at T = 37°C, and T = 30°C, this value was in the range
of 26% to 31%. In the case of other toxic substances (i.e., acryl-
amide, 4-­MeI, furfural, and 5-­HMF), their content in the decol-
orized medium was not recorded (72 hrs).
Conclusions
Controlled pH has a negative effect on sugar beet molasses
vinasse decolorization. It was observed that decrease of pH dur-
ing the process influences positively on color removal. Optimal
initial pH and temperature for decolorization are 6.5 and 30°C,
respectively. Due to the low use of an external carbon source
by bacteria (after completion of the experiment, approximately
76% of glucose remained in the medium), in the next stage of
the study it is necessary to investigate how lowering the dose
of glucose will affect the effectiveness of BMV decolorization.
However, it can be stated that the bacterial consortium used
can be applied to one of the vinasse treatment stages, which is
decolorization.
Acknowledgment
This study was financed by the National Science Centre
(Poland) under Project No. N N312 421940.
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