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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

Utilization of Sago Solid Waste for Bioethanol Production With

Variation of Sago Solid Waste-Solvent Ratio and Pre-treatment
Time

Chairul1*, M H Ridho2, Zultiniar1, Alfino Hendra1

1)
Pulp Making Laboratory Chemical Engineering Department Riau University,Binawidya
Campus H.R. Soebrantas Street Km 12.5 Simpang Baru Panam,Pekanbaru Indonesia
28293.
2)
Chemical Engineering Department Hacettepe University. Hacettepe Beytepe Kampüsü
No:35, 06800 Çankaya/Ankara. Turkiye.
*Corresponding e-mail: E-mail: chairul@lecturer.unri.ac.id

Abstract. Sago solid waste is a lignocellulosic material that can be used as a raw material for making
bioethanol. The lignin component of lignocellulose can slow saccharification by preventing cellulase
enzymes from accessing cellulose. The pre-treatment stage is the first stage of the lignocellulose-to-
bioethanol conversion process, and it is used to lower lignin levels in lignocellulose. Saccharification
and fermentation are the next processes in turning lignocellulosic material to bioethanol. The pre-
treatment stage was carried out in this study using a 10% NaOH solution at a 4 bar pressure reactor
and a temperature of 140 oC, while the saccharification and fermentation stages were carried out
simultaneously (SSF) using cellulase enzymes and yeast Saccharomyces cerevisiae at 30 oC and pH
5. The ratio of sago solid waste to NaOH solution and the period of the pre-treatment process were
varied in the study. The sago solid waste to solvent ratios were 1:5, 1:6, and 1:7, with processing
times beginning at 20 minutes, 30 minutes, and 40 minutes. The maximum lignin reduction results
(60.11%) were obtained at a ratio of 1:7 and a pre-treatment time of 40 minutes, while the highest
yield of sago solid waste mass (35.14%) was obtained at a ratio of 1:5 and a pre-treatment process
duration of 20 minutes. The results of the initial treatment stage were then continued with SSF for 0
hours, 24 hours, 48 hours, and 72 hours with the highest bioethanol content of 12% at 72 hours.

1. Introduction
The decreasing potential of fossil energy, especially oil and gas, encourages the government to make
Renewable Energy (RE) a top priority to maintain energy security and energy self-sufficiency. Renewable
energy, such as geothermal, wave, ocean currents, biofuels, and OTEC (Ocean Thermal Energy
Conversion), has become the renewable energy of preference [1]. Biofuels are all fuels derived from
vegetable oils. Therefore, biofuels can be in the form of biodiesel, bioethanol, and bio-oil [2]. Bioethanol is
an alcohol compound obtained through a fermentation process using the yeast strain Saccharomyces
cerevisiae, which can convert reducing sugars such as glucose into ethanol [3]. One of the biomasses that

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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

has the potential to be used as bioethanol is sago solid waste. Sago solid waste is a lignocellulosic material
composed of lignin, cellulose, hemicellulose, extractive substances, and starch.
Riau Province, especially the Meranti Islands District, is the largest sago-producing area in Indonesia
which in 2016 produced 326.755 tons of sago or around 85.1% of Indonesia's total sago production [4]. In
this area, there are sago processing plants that produce by-products in the form of sago solid waste. Sago
solid waste is one of the second-generation bioethanol raw materials in the form of biomass waste containing
lignocellulose. With a large amount of sago solid waste, it has the potential to be processed into bioethanol.
Bioethanol production technology from lignocellulosic materials generally includes the stages of
pretreatment, hydrolysis, fermentation, and product separation/distillation. The main obstacle in the
technological process of making bioethanol from lignocellulosic materials is the presence of lignin which
can inhibit the rate of hydrolysis by blocking the access of cellulase to cellulose, so it is necessary to choose
the right pretreatment stage for bioethanol production from lignocellulosic materials. This stage determines
the quantity and quality of bioethanol produced later [5]. Alkaline pretreatment is mostly done using NaOH.
Treatment with NaOH was chosen because this solution is quite effective in improving the hydrolysis results
and is relatively cheaper than other chemical reagents [6]. Chemical pretreatment with NaOH generally
occurs at low temperatures but has the disadvantage of a long process time that is above 1 hour, even almost
24 hours [7].
Hydrolysis and fermentation processes to produce bioethanol can be done separately or through Separate
Hydrolysis and Fermentation (SHF). However, the process is still less effective because it is carried out in
two reactors and is not carried out continuously or simultaneously without going through a long grace period
[8]. Another method of hydrolysis and fermentation is Simultaneous Saccharification and Fermentation
(SSF). In SSF, hydrolysis and fermentation are carried out in a single reactor, and enzymes and yeast are
brought together so glucose is quickly converted to ethanol [9]. The SSF process allows for maximum
hydrolysis, especially in processes that use enzymes in the hydrolysis process. This can occur because the
results of hydrolysis are directly fermented by microorganisms so that enzyme inhibitors in the form of
cellobiose and glucose can be minimized [10].
This paper will discuss the variation of chemical pretreatment process conditions using NaOH on sago
solid waste and determine the effect of sago solid waste-solvent ratio and pretreatment process time as the
initial stage for the bioethanol production process from lignocellulose.

2. Research Methodology

2.1. Material
The raw material in this research is sago solid waste obtained from Meranti Islands District. The materials
used include commercial cellulase enzyme with NOVOzymes brand, Saccharomyces Cerevisiae yeast,
NaOH, KH2PO4, MgSO4, (NH4)2SO4, H2SO4, buffer pH 5, glucose, Nelson's reagent, arsennomolybdate
solution, and distilled water.

2.2. Research Procedure

The stages of making bioethanol from sago solid waste through the process of saccharification and
simultaneous fermentation go through several stages, namely: preparation of raw materials, delignification,
saccharification process, and simultaneous fermentation (SSF). Lignocellulose characteristics of raw
materials and after pre-treatment using the Chesson method, while bioethanol content is measured using an
alcoholmeter.

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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

2.2.1. Raw Material Preparation

The raw material used in this research is sago solid waste obtained from Meranti Islands District. Before
use, sago solid waste was washed thoroughly and dried in an oven at 105 oC, after which it was mashed and
then sieved to a size of 30-40 mesh.
The delignification or pretreatment process uses the PARR Reactor using a 10% NaOH solution. The
ratio between the mass of sago solid waste and NaOH solution varied from 1:5; to 1:6; to 1:7. The pressure
was set at 4 bar at the beginning of the delignification process. The temperature used in this study was 140
o
C, while the processing time was 20, 30, and 40 minutes. After the process was completed, the mixture
consisting of lignocellulose and NaOH was filtered to separate the solids of sago solid waste and black
liquor. The filtered sago pulp was washed until the pH of the washing liquid was neutral, then dried in an
oven at 105 oC until the weight remained constant, and then used in the simultaneous saccharification and
fermentation (SSF) process.

2.2.2. Simultaneous Saccharification and Fermentation (SSF)

This SSF process combines enzyme hydrolysis and fermentation reaction simultaneously in one reactor.
The enzyme used is cellulase enzyme, and the yeast used is Saccharomyces cerevisiae. The medium for SSF
is 2000 mL consisting of a sago solid waste sample, nutrient medium, acetate buffer (pH = 5), cellulase
enzyme 1% of the total fermentation medium, yeast inoculum, and distilled water. The sago solid waste
used was a mixture of various pre-treatment results, and its lignocellulose content was recalculated. Nutrients
medium consisted of (NH4)2PO4 1.0 g/L, MgSO4 .7H2 O 0.05 g/L and yeast extract 2 g/L. All materials and
apparatus, except enzyme and inoculum, were sterilized for 15 minutes at 121 o C using an autoclave.
Enzymes and inoculum were added after the sterile media cooled. Then, it was shaken at 150 rpm for 0, 24,
48, and 72 hours. Cultivation was taken and put into a microcentrifuge tube. Separate the clean liquid and
sediment in the centrifuge tube, and then the liquid is separated between ethanol-water and nutrient salts
using a rotary evaporator at a temperature of 80-85 oC. The ethanol concentration was measured using an
alcohol meter.

3. Results and discussion

3.1. Characteristics of Sago Solid Waste

The characteristics of sago pulp before pre-treatment can be seen in Table 1. According to Table 1, sago
solid waste has a fairly high cellulose content of 33.04%. The value of the lignocellulose content of sago
solid waste in this study is not much different from that of the previous study [11]. This high cellulose
content of sago solid waste has the potential to be used as raw material for making bioethanol. However,
the lignin content in sago solid waste, which is 12.46%, needs to be reduced in order to improve the enzyme
performance during hydrolysis or saccharification of sago solid waste.

3.2. Effect of Pretreatment on Lignocellulose Composition

Pre-treatment/delignification is carried out to reduce lignin content, acetyl group compounds and dissolving
a little hemicellulose. In the pre-treatment process, in addition to producing sago solid waste, it will also
produce waste in the form of black liquor. This black liquor contains lignin, hemicellulose, and residual
NaOH from the pre-treatment process. Black liquor smells bad because it contains chemical compounds such

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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

as metal mercaptan, dimethyl sulfide ((CH3)2S), and dimethyl disulfide (CH3-S-S-CH3). These gases are
formed by the reaction of breaking the methyl aryl ether bond in one of the constituent units of lignin [12].

Table 1. Lignocellulose Composition of Sago Solid Waste

Composition (%)
Component Ahmad et al., This research
(2020)
Lignin 9.75 12.46
Cellulose 35.6 33.02
Hemicellulose 22.43 18.87
Ash - 4.52
HWS - 31.13

3.3. Effect of Pretreatment on Lignin Content

In the pretreatment process of sago solid waste, it is expected that the cellulose content is high and lignin is
low. The effect of cooking time and pretreatment solvent sago solid waste ratio on lignin content is shown
in Figure 1.

14
12
lignin content (%)

10
8
1: 5
6
1: 6
4
2 1: 7

0
0 20 30 40
time (min)

Figure 1 Effect of sago solid waste-solvent ratio and pretreatment time on lignin content

The results showed that the lowest lignin content was in the treatment of 40 minutes pretreatment time
and 1:7 solvent sago solid waste ratio with a lignin content of 4.97% or reduced by 60.11%, the highest
lignin content was in the treatment of 20 minutes pretreatment time and 1:5 solvent sago solid waste ratio
with a lignin content of 11.17% or reduced only by 10.35%. The results showed that with the increase in
the ratio of solvent sago solid waste, lignin content will decrease, likewise, the increase in pretreatment time
will decrease lignin content. This is in accordance with the research of [13] who delignified the TKKS with
the best cooking time at 40 minutes resulting in degraded lignin reaching 86.92%.

3.4 Effect of Pretreatment on Hemicellulose Content

The effect of cooking time and pretreatment solvent sago solid waste ratio on hemicellulose composition
levels is shown in Figure 2 with hemicellulose levels before pretreatment of 12.46%. In pretreatment with

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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

a time of 40 minutes with a ratio of 1:5; 1:6; and 1:7, the hemicellulose content was 14.42%, 13.33%, and
12.75%, respectively. Based on these data, it can be concluded that the more NaOH used, the smaller the
hemicellulose at the end of pretreatment.
The longer the pretreatment time, the smaller the hemicellulose content tends to be. This can be seen in
the ratio of 1:7 solvent sago solid waste with pretreatment times of 20, 30, and 40 minutes, respectively, the
content becomes 14.29%, 14.00%, and 12.75%. Hemicellulose is a carbohydrate substance that is insoluble
in boiling water, but soluble in dilute alkali and dilute acid [14].

20
Hemiselulosa content (%)

15

10 1: 5
1: 6
5
1: 7

0
0 20 30 40
time (min)

Figure 2 Effect of sago solid waste-solvent ratio and pretreatment time on hemicelluloses content

3.5 Effect of Pretreatment on cellulose Content

The effect of cooking time and pretreatment sago solid waste-solvent ratio on cellulose composition content
is shown in Figure 3, with cellulose content before pretreatment/delignification at 33.02%. The results
showed that the highest cellulose content was in the treatment of 40 minutes pretreatment time and 1:7
solvent sago pulp ratio with 70.49% cellulose content, while the lowest cellulose content was in the
treatment of 20 minutes pretreatment time and 1:5 solvent sago pulp ratio with 64.08% cellulose content.
The results show that the greater the ratio between sago solid waste and NaOH solvent and the longer the
pretreatment time, the cellulose content will increase.

80
70
60
Selulose content (%)

50
40 1: 5
30 1: 6
20
1: 7
10
0
0 20 30 40
time (min)

Figure 3. Effect of Sago Solid Waste-solvent ratio and pretreatment time on cellulose content.

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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

Based on the results of the study, sago solid waste that has been pretreated with NaOH will increase the
cellulose content from 33.02% to reach a cellulose content of64.08-70.59%. The increase in cellulose
content is due to the large loss of lignin and hemicellulose as shown in Figures 1 and Figure 2. Hemicellulose
molecules that dissolve in alkali are short chains and branch chains and acetylation of substituent groups on
hemicellulose occurs, while intra-molecular glycosidic bonds of hemicellulose are difficult to hydrolyze.
NaOH can remove some of the lignin and hemicellulose that protects the cellulose molecules of sago pulp
powder, as well as being able to break hydrogen bonds, especially inter-molecular cellulose bonds so that
cellulose is in an unbound state. This situation causes cellulose to become loose both against bonds with
non-cellulosic components and on its cellulose so that the cellulase enzyme can more easily contact with
cellulose which ultimately hydrolysis of cellulose into simple sugars can run more perfectly [15].

3.6 Effect of Pretreatment on Mass Yield

Sago solid waste that has undergone a pretreatment process will experience changes in the composition of
lignin, hemicellulose, and cellulose. The pretreatment process using NaOH can also cause a reduction in the
mass of sago solid waste produced. The amount of pulp mass gain after pretreatment is shown in Figure 4.

40
35
Percentage of

30
1:5
mass Yield

25 1:6
20
1:7
15
10
5
0

10 20 30 40
.
Time (min)

Figure 4. Sago solid waste yield after pretreatment

In general, sago solid waste that has been pretreated will lose more than half of its initial mass. The
pretreatment process time affects the mass yield of sago solid waste, the longer the processing time, the
lesser the mass yield of sago solid waste. Likewise with the treatment between the ratio of sago solid waste
and the solvent, the greater the ratio of sago solid waste than the pretreatment solvent will cause the
acquisition of lesser sago solid waste mass. The highest mass yield occurred at a time of 20 minutes with a
ratio of 1:5, which is around 35.14%. This is due to the length of pretreatment time and the large ratio of
sago solid waste, the more lignin and hemicellulose dissolved in black leachate. These results are in line
with the results of research conducted by [16], [17], and [13] who conducted chemical pretreatment with
NaOH on Miscanthus TKKS with the results of biomass acquisition of 32.0% to 72.7%

3.7 Effect of Saccharification Time and Concurrent Fermentation on Bioethanol

The simultaneous saccharification and fermentation process of sago pulp that has undergone pre-treatment
is carried out under anaerobic conditions with the help of cellulase enzymes and Saccharomyces cerevisiae
yeast. The sago solid waste used in the SSF process is a mixture of several pre-treatment results with various
conditions and the lignocellulose content is recalculated, the lignocellulose content of the sago pulp used in the
SSF process is 66.04% cellulose, 15.0% hemicellulose, 9.27% lignin, and 2.92% ash.

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SAFE-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

The effect of saccharification time and simultaneous fermentation on glucose content can be seen in
Figure 5. The concentration of bioethanol at 0 hours, 24 hours, 48 hours, and 72 hours sampling increased
by 1%, 5%, 11%, and 12%, respectively. This shows that the longer the fermentation will increase the
amount of alcohol produced, but the length of the fermentation process has a maximum limit [18].
Bioethanol concentration in this study at 72 hours has not seen the peak condition of bioethanol production
and the possibility if the SSF time is extended will produce higher bioethanol concentration.

14
Bioethanol Concentration (%)

12
10
8
6
4
2
0
0 24 48 72
Time (hours)

Figure 5. Effect of SSF process time on bioethanol concentration

The results of the acquisition of bioethanol concentration in this study can be said to be better when
compared with the research of [19] who converted sago solid waste into bioethanol with SSF process
without pretreatment resulting in a bioethanol concentration of 8% with 72 hours fermentation time. This
proves that the pretreatment process is very effective to increase the conversion of sago solid waste into
bioethanol. With the pretreatment process, the lignin content in sago solid waste that can hinder the
performance of cellulase enzymes in the process of converting cellulose into glucose can be reduced.

4. Conclusion
Sago solid waste can be used as raw material for making bioethanol because it has a high cellulose content
of 33.02%. The best pre-treatment process conditions were obtained at a pre-treatment condition of 1:7 sago
solidwaste-solvent ratio and 40 minutes with the acquisition of 4.97% lignin, 12.75% hemicellulose, and
70.59% cellulose. The highest recovery of sago solid waste after pre-treatment was obtained under the process
conditions of a sago solid waste-solvent ratio of 1:5 and a time of 20 minutes with a sago solid waste
recovery of 35.14%. The best time for simultaneous saccharification and fermentation process was obtained
at 72 hours with a 12% bioethanol concentration.

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IOP Conf. Series: Earth and Environmental Science 1228 (2023) 012034 doi:10.1088/1755-1315/1228/1/012034

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