Renewable Energy 102 (2017) 9e14

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Renewable Energy
journal homepage: www.elsevier.com/locate/renene

Ethanol production by solid-state saccharication and fermentation in

a packed-bed bioreactor

udia Alessio a, Edson L. Foletto a, Raquel C. Kuhn a,
Nicholas I. Canabarro a, Cla
Wagner L. Priamo b, Marcio A. Mazutti a, *
a
Department of Chemical Engineering, Federal University of Santa Maria, Av. Roraima, 1000, Santa Maria, 97105-900, Brazil
b
Department of Food Technology, IFRS - Campus Serta~o, Serta
~o, RS, 88040-900, Brazil

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

Article history: In this work was presented a sequential strategy to optimize and scale-up the production of ethanol by
Received 21 November 2015 solid-state saccharication and fermentation using rice bran as substrate. In the rst step, fermentation
Received in revised form was carried out in Erlenmeyers to study the inuence of eight variables by means of a Plackett Burman
17 August 2016
design. After the choice of most signicant ones, a central composite rotational design (CCRD) for three
Accepted 13 October 2016
Available online 14 October 2016
independent variables (rice bran concentration, moisture content and inoculum size) was conceived to
optimize the ethanol production in a packed-bed bioreactor. From Erlenmeyers to packed bed bioreactor
the process was scaled-up 10 times. Maximum ethanol production in the packed-bed bioreactor was
Keywords:
Rice processing residue
135 10.8 g kg
1 at inoculum size, rice bran concentration and moisture content of 10% v/v, 62.5% w/w
Solid-state saccharication and and 65% w/w, respectively. The ethanol yield obtained in the packed-bed bioreactor was similar to that in
fermentation the erlenmeyers asks (138.7 g kg
1), validating the strategy adopted for optimization and scale-up.
Second generation ethanol 2016 Elsevier Ltd. All rights reserved.

1. Introduction technologies [9e11].

Overall energy consumption is increasing with the growing
world population and rapid industrial growth, reducing fossil fuels
reservoirs and increasing environment problems caused by
greenhouse gases accumulation [1e3]. For this reason, countries
worldwide have paid much attention to the non-fossil energy,
especially, the cleaning renewable biomass fuels [3e5]. Lignocel-
lulosic biomass has been considered as the ideal feedstock for
biofuel production as it does not compete with food resources and
can potentially reduce carbon dioxide emission by up to 75%
compared to fossil fuels [1,6e8].
Second generation biofuels from lignocellulosic biomass or
agricultural wastes have advantages, for instance, most second
generation biofuels are considered to be able to deliver substantial
greenhouse gases emissions reductions when compared with pe-
troleum. Development of second generation biofuels has been
challenging, because it is need ensure low-cost and stable feed-
stocks, minimizing land use and changes caused by demand for
biomass feedstocks and optimizing bioethanol production
* Corresponding author.
E-mail address: mazutti@ufsm.br (M.A. Mazutti).
The effective conversion of agricultural residue to sugar and its
recovery can be regarded as the key to the success of this tech-
nology. Enzymatic hydrolysis is reported as the most promising
technique for converting lignocellulosic compounds into ferment-
able sugars such as glucose, which can be used as a cheap carbon
source for ethanol production [12]. Enzymatic hydrolysis of agri-
cultural residues is mainly accomplished with simultaneous
saccharication and fermentation processes in liquid media [13].
Therefore, more energy to concentrate the ethanol produced due to
low solid content (substrate) and a large quantity of waste water is
required [14].
Solid-state saccharication and fermentation is dened as the
simultaneous hydrolysis and fermentation in absence (or near
absence) of free water, it can overcome the above problems because
of the many benets such as high substrate concentration and
product yield, simple and controllable operation, less efuent
wastewater, and less energy consumption. The main advantage of
this procedure is the fact that the process is conducted in the
absence of free aqueous phase, resulting in minimum water con-
sumption and thus a low efuent production. As the amount the
water can be adjusted to a minimum during the extraction of
ethanol produced, the resulting solution is more concentrated that
traditional liquid fermentation [12,15]. Some studies had

http://dx.doi.org/10.1016/j.renene.2016.10.026
0960-1481/ 2016 Elsevier Ltd. All rights reserved.
10 N.I. Canabarro et al. / Renewable Energy 102 (2017) 9e14
demonstrated that solid-state saccharication and fermentation of
cellulosic materials can effectively reduce the production cost,
improving the commercial application of bioethanol [13,15].
However, it is fundamental to dene a strategy to optimize and
scale-up of solid-state saccharication and fermentation process.
Solid-state fermentation, unlike submerged fermentation, has
many challenges to be solved before the scale-up. One of these
difculties is the choice of bioreactor, since it provides the envi-
ronment for growth and activity for the microorganisms, which
cause the biological reaction. The most used bioreactors for solid-
state fermentation are tray, packed bed and rotary drum [16]. The
packed-bed reactor seems to be a promising bioreactor congura-
tion, since they are capable of supporting microbial growth for long
culture periods under low shear conditions, due to the immobili-
sation of cells within macroporous matrices.
Based on these aspects, the main objective of this work was to
dene a strategy to optimize and scale-up the ethanol production
using rice processing residue as substrate by solid-state sacchari-
cation and fermentation. In the rst step, fermentations in
erlenmeyers asks were carried out select the most important
variables. In the second one, the optimization and scale-up were
accomplished in a packed-bed bioreactor.
2. Material and methods
2.1. Materials
The rice husk and bran were obtained in a local rice milling
(Santa Maria-RS, Brazil), which were maintained under refrigera-
tion (4  C) until the experiments. The composition (%wt) of rice
bran was determined using the methodology described by Van
Soest et al. [17]: Lignin (5.4), hemicelluloses (22.9), cellulose (9.2),
fat (16.5), starch (30.8) and protein (15.2). Corn steep liquor (CSL)
was obtained from Ingredion (Mogi Guau, Brazil). Soybean bran
was obtained in a local market. The enzymes used in this study
were the cellulolytic complex from Trichoderma reesei NS50013
(Novozymes Latin American) and the amylolytic complex Spi-
rizyme Fuel (Novozymes Latin American).
2.2. Microorganism and inoculum
A commercial strain of Saccharomyces cerevisae (Fleischmann)
was used [18]. Cell production for pre-inoculum was carried out in
Erlenmeyers with 10 mL of medium containing (g.L
1): sucrose
(20.0), yeast extract (5.0), K2HPO4 (5.0), NH4Cl (1.5), KCl (1.15) and
MgSO4$7H2O (0.65). The medium was inoculated with 1 g of
dehydrated yeast and incubated in orbital shaker (INNOVA 44R,
New Brunswick Scientic), at 30  C, 150 rpm for 24 h.
2.3. Experimental procedure for solid-state fermentations
The experiments for optimization of ethanol production were
carried out in two steps. In the rst one, the fermentations were
performed in Erlenmeyers containing 100 g of dry solid substrate,
where the effects of eight variables were studied. The second step
consisted in the selection of most signicant effects of rst one to
optimize the process in a packed-bed bioreactor containing 1 kg of
dry substrate (scale-up of 10 times).
For the rst step, 100 g of dry solid substrate, which was
composed by different proportions of rice bran and rice husk (rice
husk was used only as support) were charged into 500 mL Erlen-
meyer asks. Afterwards, the solid substrate was supplemented
(CSL and soybean bran) and the moisture content adjusted at
specied level. Each Erlenmeyer was covered with hydrophobic
cotton and autoclaved at 121  C for 20 min. Preliminary
experiments showed that no changes in moisture content of the
substrate after autoclaving were detected. After cooling, the en-
zymes (amylase and cellulase) and microbial cells were inoculated
and incubated at specied temperature for 24 h in a chamber with
temperature and humidity control (POL-EKO, model KK 350). The
variables investigated in this step were: temperature (30e40  C),
rice bran concentration used in the solid substrate (50e75% w/w),
inoculum concentration (5e15% v/v), moisture content (60e80% w/
w), CSL concentration (5e15% w/w), soybean bran concentration
(5e15% w/w), amylase concentration (0.2e0.6% v/w) and cellulase
concentration (0.2e0.6%v/w). All concentrations were dened for a
dry mass of 100 g. The effect of variables was investigated by a
Plackett-Burman design (PB16).
In the second step was employed a packed-bed bioreactor that
consists of a cylindrical stainless bed connected to a thermostatic
bath for temperature control. It was lled with 1 kg of dry solid
substrate and supplemented with soybean bran and CSL. The
moisture content was corrected to desirable level and autoclaved at
121  C for 20 min. After cooling, the enzymes (amylase and cellu-
lase) and microbial cells were inoculated and incubated at 36  C for
24 h. A central composite rotational design (CCRD) for three inde-
pendent variables was conceived to investigate the inuence of rice
bran concentration in the solid substrate (41.5e83.5% w/w), mois-
ture content (39.8e90.2% w/w) and inoculum (1.6e18.4% v/v).
2.4. Fermentable sugar and ethanol determination
After the fermentations, ethanol and sugar were extracted from
the solid material following methodology described by Canabarro
et al. [19]. After the extraction, 1 mL of supernatant was used to
determine the amount of fermentable sugars by 3.5-dinitrosalicylic
acid method [20]. A liquot of 5 mL of supernatant was used for
determination of ethanol content using an alcolyzer at 25  C
(Alcolyzer Wine M/WE e Wine Analysis System - Anton Paar). The
results were expressed in g of ethanol per kg of dry substrate.
2.5. Statistical analysis
All the results were analyzed using the software Statistica 7.0
(Statsoft Inc., Tulsa, OK, USA), considering a signicance level of 90%
for Plackett-Burman design and 95% for the CCRD.
3. Results and discussion
3.1. Selection of variables in erlenmeyers
Table 1 present the results of the PB16 to select the most sig-
nicant variables on ethanol production by solid-state sacchari-
cation and fermentation in Erlenmeyers. The ethanol
concentrations ranged from 149.2 (run 8) to 17.7 g kg
1 (run 2),
with great variation among the runs. Central points of PB16 pre-
sented low experimental variation, showing a good reproducibility
of data. From data of Table 1 it was evident that the studied process
variables affect the production. Aiming to select the most signi-
cant variables, data of Table 1 were used to determine the effects of
independent variables on the response, which are presented in
Table 2.
The media supplements (CSL and soybean bran) did not inu-
ence the ethanol production in the evaluated range. Concerning the
enzymes used to convert substrate (starch or cellulose) into
fermentable sugars, amylase showed a positive effect, whereas
cellulase was not signicant. This can be due to fact that the main
sugar source in the solid material is starch (cellulose content is
about three times lower) and, by this reason, the contribution of
amylase in comparison with cellulase activity to provide sugar for
 N.I. Canabarro et al. / Renewable Energy 102 (2017) 9e14 11

Table 1
Variables reals and coded (in parentheses), results of PB16 for screening of signicant variables for ethanol production.

Run x1 x2 x3 x4 x5 x6 x7 x8 EtOH (g.kg
1)

1 40 (1) 50 (
1) 5 (
1) 60 (
1) 15 (1) 5 (
1) 0.2 (
1) 0.6 (1) 47.4
2 40 (1) 75 (1) 5 (
1) 60 (
1) 5 (
1) 15 (1) 0.2 (
1) 0.2 (
1) 17.7
3 40 (1) 75 (1) 15 (1) 60 (
1) 5 (
1) 5 (
1) 0.6 (1) 0.2 (
1) 29.5
4 40 (1) 75 (1) 15 (1) 80 (1) 5 (
1) 5 (
1) 0.2 (
1) 0.6 (1) 26.8
5 30 (
1) 75 (1) 15 (1) 80 (1) 15 (1) 5 (
1) 0.2 (
1) 0.2 (
1) 32.3
6 40 (1) 50 (
1) 15 (1) 80 (1) 15 (1) 15 (1) 0.2 (
1) 0.2 (
1) 44.3
7 30 (
1) 75 (1) 5 (
1) 80 (1) 15 (1) 15 (1) 0.6 (1) 0.2 (
1) 31.8
8 40 (1) 50 (
1) 15 (1) 60 (
1) 15 (1) 15 (1) 0.6 (1) 0.6 (1) 147.2
9 40 (1) 75 (1) 5 (
1) 80 (1) 5 (
1) 15 (1) 0.6 (1) 0.6 (1) 14.7
10 30 (
1) 75 (1) 15 (1) 60 (
1) 15 (1) 5 (
1) 0.6 (1) 0.6 (1) 93.4
11 30 (
1) 50 (
1) 15 (1) 80 (1) 5 (
1) 15 (1) 0.2 (
1) 0.6 (1) 35.7
12 40 (1) 50 (
1) 5 (
1) 80 (1) 15 (1) 5 (
1) 0.6 (1) 0.2 (
1) 21.7
13 30 (
1) 75 (1) 5 (
1) 60 (
1) 15 (1) 15 (1) 0.2 (
1) 0.6 (1) 43.4
14 30 (
1) 50 (
1) 15 (1) 60 (
1) 5 (
1) 15 (1) 0.6 (1) 0.2 (
1) 99.7
15 30 (
1) 50 (
1) 5 (
1) 80 (1) 5 (
1) 5 (
1) 0.6 (1) 0.6 (1) 110.1
16 30 (
1) 50 (
1) 5 (
1) 60 (
1) 5 (
1) 5 (
1) 0.2 (
1) 0.2 (
1) 47.2
17 35 (0) 62.5 (0) 10 (0) 70 (0) 10 (0) 10 (0) 0.4 (0) 0.4 (0) 94.9
18 35 (0) 62.5 (0) 10 (0) 70 (0) 10 (0) 10 (0) 0.4 (0) 0.4 (0) 96.7
19 35 (0) 62.5 (0) 10 (0) 70 (0) 10 (0) 10 (0) 0.4 (0) 0.4 (0) 89.0

Additional experiments to evaluate the amylase concentration

20 35 62.5 10 70 5 5 0.0 0.2 35.3

21 35 62.5 10 70 5 5 0.5 0.2 109.9
22 35 62.5 10 70 5 5 1.0 0.2 125.6
23 35 62.5 10 70 5 5 1.5 0.2 136.1
24 35 62.5 10 70 5 5 2.0 0.2 138.7

X1 Temperature ( C), X2 rice bran concentration (% w/w), X3 inoculum size (%, v/v), X4 moisture content (% w/w), X5 soybean concentration (% w/w), X6 Corn steep liquor
concentration (w% w/w), X7 Spirizyme Fuel concentration (% v/w), X8 cellulase concentration (% v/w). Experimental error: 5%.

Table 2 was xed at 1.0 wt% for the CCRD.

Effects of independent variables in the ethanol production for runs of the PB16.

Variables Effect Standard error t (10) p-value 3.2. Optimization in packed-bed bioreactor
Mean/Interc. 0.43 0.04 9.61 <0.0001
Temperature
0.04 0.10
0.44 0.6723 The rice bran concentration, inoculum size and moisture con-
Rice bran
0.27 0.10
2.76 0.0202
tent were optimized in a packed-bed bioreactor by means of a CCRD
Inoculum 0.18 0.10 1.86 0.0920
Moisture content
0.26 0.10
2.66 0.0241
for three independent variables. The results obtained in the CCRD
Soybean 0.08 0.10 0.87 0.4062 are presented in Table 3, where it is seen that ethanol concentration
CSL 0.10 0.10 1.07 0.3091 ranged from 86.6 (run 2) to 129.5 g kg
1 (run 16). The highest
Amylase 0.27 0.10 2.76 0.0202 concentration obtained in the packed-bed bioreactor (~125 g kg
1)
Cellulase 0.16 0.10 1.53 0.1560
was similar to the values obtained in the fermentations in
Erlenmeyers.
To better understand the inuence of process variables on the
fermentation is much higher. The increase of inoculum size showed ethanol production, data of Table 3 were used to estimate the co-
a positive effect on ethanol production, whereas moisture content efcients of a quadratic model (Table 4), where the coefcients are
and rice bran concentration had negative effects. The inoculum size directly related to the effect of each variable on the response. Linear
for solid-state fermentation should be higher than for submerged and quadratic terms for rice bran concentration, inoculum size and
one, because the growth in the solid material is non-homogeneous. moisture content were statistically signicant (p < 0.05). Rice bran
The negative effect of rice bran concentration would be correlated concentration presented positive effect on the ethanol production,
with inhibition of amylase activity due to high concentration of indicating that the increase from level
1 to 1 of the CCRD would
glucose in the media. However, this hypothesis was not proven lead to higher concentration. However, the analysis of quadratic
since the sugar content was not determined. Moisture content is an term for this variable is indicating the presence of a maximum
important factor and inuences the enzymatic hydrolysis and point in the studied region, since its value was negative. For other
fermentation performance. The water content of material should be variables, the increase from level
1 to 1 did not inuence sta-
adjusted to adequate range to increase the conversion efciency. tistically the production of ethanol, but the negative sign of
The most signicant variables in the ethanol production in quadratic terms indicate the presence of a maximum point.
Erlenmeyers were moisture content, amylase concentration, rice Based on the statistical analysis of coefcients presented in
bran concentration and inoculum size. Before conceiving a new Table 4, the coded model to predict the ethanol concentration from
experimental design with a great number of variables selected in another condition than those experimentally evaluated in the CCRD
the PB16, some additional experiments were carried out to inves- is:
tigate the effect of amylase concentration on ethanol production in
Erlenmeyers (runs 20e24 of Table 1). Increasing the amylase con- EtOH 128:28 12:68,RB
8:32,RB2
7:16,I 2
8:46,M 2
centration from 0 to 2.0 wt% was veried an increase in ethanol
(1)
production from 35.3 to 138.7 g kg
1. However, for amylase con-
centration higher than 1.0 wt% there is no statistical difference where EtOH is the ethanol concentration (g.kg
1), RB, I and M are
among the results (p < 0.05). For this reason, amylase concentration the coded values for rice bran concentration, inoculum size and
12 N.I. Canabarro et al. / Renewable Energy 102 (2017) 9e14

Table 3
Matrix of the results obtained in the CCRD for optimization of ethanol production in the packed-bed bioreactor.

Run Moisture (% w/w) Inoculum (% v/v) Rice bran concentration (% w/w) EtOH (g.kg
1)

1 50 (
1) 5 (
1) 50.0 (
1) 88.6

2 80 (1) 5 (
1) 50.0 (
1) 86.6
3 50 (
1) 15 (1) 50.0 (
1) 91.9
4 80 (1) 15 (1) 50.0 (
1) 89.9
5 50 (
1) 5 (
1) 75 (1) 119.4
6 80 (1) 5 (
1) 75 (1) 111.4
7 50 (
1) 15 (1) 75 (1) 118.1
8 80 (1) 15 (1) 75 (1) 123.5
9 39.8 (
1.68) 10 (0) 62.5 (0) 99.3
10 90.2 (1.68) 10 (0) 62.5 (0) 111.4
11 65 (0) 1.6 (
1.68) 62.5 (0) 110.0
12 65 (0) 18.4 (1.68) 62.5 (0) 108.0
13 65 (0) 10 (0) 41.5 (
1.68) 88.6
14 65 (0) 10 (0) 83.5 (1.68) 122.9
15 65 (0) 10 (0) 62.5 (0) 128.2
16 65 (0) 10 (0) 62.5 (0) 129.5
17 65 (0) 10 (0) 62.5 (0) 126.8

Fixed conditions: Amylase concentration (1.0% v/w); Temperature (35  C); soybean bran concentration (5% w/w), CSL concentration (5% w/w), cellulase concentration (0.2% v/
w).

Table 4 this time an accentuated decreasing was veried, probably due to

Estimated coefcients for quadratic model from data of CCRD and signicant anal- evaporation or adsorption into solid structure.
ysis of parameters. In the simultaneous hydrolysis and fermentation processes, the
Coefcient Std. Err. of Coeff. p-value process should be operated at temperatures and times which meet
Mean/Interc. 128.28 2.95 <0.0001
both active microorganism and enzyme complexes. In this work,
(1)Moisture(L) 1.01 1.39 0.4921 the temperature was ideal for microbial growth, but for enzymes
Moisture(Q)
8.46 1.53 0.0009 the optima is around 50  C [20]. For this reason, there is a fast
(2)Inoculum(L) 1.03 1.39 0.4827 consumption of sugar and ethanol production until 24 h. The en-
Inoculum(Q)
7.16 1.53 0.0022
zymes continue acting in the process as can seen for the period
(3)Rice bran(L) 12.68 1.39 <0.0001
Rice bran(Q)
8.32 1.53 0.0010 between 32 and 40 h, where the sugar concentration increased
1L by 2L 1.68 1.81 0.3863 again. Due to fact that temperature is not the optima for enzyme
1L by 3L 0.18 1.81 0.9258 hydrolysis, the time to reach maximum conversion is higher than
2L by 3L 0.53 1.81 0.7805
for a separated hydrolysis and fermentation.
It is important emphasize that there is no difference between
ethanol production in erlenmeyers asks (138.7 g kg
1) and in the
moisture content, respectively. This model was validated by anal- packed-bed bioreactor (135 g kg
1). This is a desirable result,
ysis of variance and presented a determination coefcient (R2) of because the production in scale-up was maintained at the same
0.9515. The validated model was applied to drawn contour plots. level of bench scale. The strategy to optimize and scale-up the
The ethanol production in function of inoculum size and mois- process showed satisfactory results.
ture content, rice bran concentration and moisture content and rice The ethanol yield in this work was 135 8.2 g kg
1, which is in
bran concentration and inoculum size are depicted in Fig. 1(aec), good agreement with other studies reported in literature using rice
respectively. Analyzing Fig. 1(aec) it is seen that maximum ethanol bran as substrate. Tiwari et al. [21] studied solid simultaneous
production was obtained for rice bran concentration in the range of saccharication and fermentation of rice straw to bioethanol using
62e78 wt%, moisture content in the range of 60e70% wt% and novel bacteria (Bacillus cereus strain McR-3) reported in ethanol
inoculum size in the range of 8e13 wt%. Based on this, the opti- production. The authors set parameters like temperature and pH
mized condition for ethanol production was obtained at inoculum reaching 9.36 0.44% ethanol yield. In this work was achieved a
size, rice bran concentration and moisture content of 10 wt%, total amount of ethanol by SSF process (135 8.2 g kg
1) using a
62.5 wt% and 65 wt%, respectively. commercial yeast, making the fermentation process more practical
An additional fermentation was carried out to validate experi- than others process found in literature. Michel et al. [18] produced
mentally the optimized condition and also to investigate the ki- 175 5.8 g kg
1 of ethanol by simultaneous saccharication and
netics of sugar and ethanol concentration. The results are presented fermentation of rice bran in submerged fermentation with high-
in Fig. 2, where ethanol was 135 8.2 g kg
1 (Fig. 2) which is in solid loading at 35  C during 48 h. Arokiasamy et al. [22] ob-
good agreement with the value predicted by the model at specied tained the maximum ethanol yield of 0.2 g kg
1 using rice bran as
condition (128.3 g kg
1). Ethanol concentration increased linearly substrate by simultaneous saccharication and fermentation pro-
until 24 h of fermentation, decreasing after this period. This cess. Wang et al. [12] studied the ethanol production by solid
reduction in the concentration would be related to evaporation due simultaneous saccharication and fermentation using rice straw,
to metabolic heat generated by the microorganism or by adsorption reaching a total amount of 4.079 g g
1.
into solid material. Initial sugar concentration was about 10 g kg
1, Many authors have described ethanol production from
increasing to 70 g kg
1 until 8 h of fermentation due to action of numerous raw materials. Cuevas et al. [23] produced 133 g kg
1 of
amylase and cellulase. After this period, an accentuate decreasing ethanol by simultaneous saccharication and ethanol fermentation
was veried until to reached around 10 g kg
1 after 20 h due to of pretreated olive stones. Yu et al. [24] obtained the maximum
consumption for growth and ethanol production. Maximum ethanol yield of 79 g kg
1 sweet sorghum fresh stalks by solid state
ethanol production was 135 g kg
1 at 24 h of fermentation. After fermentation using thermotolerant yeast. Kwon et al. [25] reached
 N.I. Canabarro et al. / Renewable Energy 102 (2017) 9e14
Fig. 1. Contour plots expressing the inuence of process variables on the ethanol
production: a) inoculum and moisture content; b) rice bran concentration and mois-
ture content and c) inoculum and rice bran concentration. Third variable was main-
tained at the central point.
the highest ethanol yield of 250 g kg
1 fresh stalks of sweet sor-
ghum using a deep-bed bioreactor. Mohanty et al. [16] reached
13
160
Ethanol
140 Glucose
120
Concentration (g/kg)
100
80
60
40
20
0
0 10 20 30 40 50
Fermentation time (h)
Fig. 2. Prole of sugar and ethanol at optimized condition of packed-bed bioreactor.
maximum ethanol productivity around 225 4.0 g kg
1 by solid
state fermentation from mahula owers (Madhuca latifolia L.) and
Huang et al. [26] performed a simultaneous saccharication and
fermentation (SSF) from tuber of Canna edulis ker, reaching a total
ethanol production of 70 g kg
1. On the other hand, some re-
searchers have conducted studies on ethanol production at high
solid concentrations, obtaining better results than in solid-state
fermentation. Pietrzak and Kawa-Rygielska [27] studied simulta-
neous saccharication and ethanol fermentation at high solids
loading from waste wheat-rye bread, yielding about 425 g kg
1 of
dry matter. Martins el al. [28] produced ethanol from 100.9 to
135.4 g kg
1 of dry matter when was investigated the production by
SSF process at high solids loadings of sugarcane bagasse.
4. Conclusions
This study evaluated the ethanol production by solid-state
saccharication and fermentation in a packed-bed bioreactor.
From fermentations in the Erlenmeyers the variables most impor-
tant were inoculum size, rice bran concentration and moisture
content. These variables were optimized in a packed-bed bioreactor
and maximum ethanol production was 135 10.8 g kg
1 obtained
at inoculum size, rice bran concentration and moisture content of
10 wt%, 62.5 wt%1 and 65 wt%, respectively. The highest ethanol
produced on fermentation in Erlenmeyers was similar to results
obtained in the packed-bed bioreactor. The results demonstrated
that is possible to obtain comparable ethanol yield by solid-state
fermentation with liquid fermentation with high-solid loading.
Acknowledgements
The authors would like to thank CAPES and CNPq (454645/
2012-0) for the nancial support.
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