Chemosphere 291 (2022) 133003

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Sustainable bioconversion of potato peel wastes into ethanol and biogas

using organosolv pretreatment
Ali Soltaninejad a, Mohammadhadi Jazini a, Keikhosro Karimi a, b, *
a
Department of Chemical Engineering, Isfahan University of Technology, Isfahan 84156-83111, Iran
b
Department of Chemical Engineering, Vrije Universiteit Brussel, 1050, Brussels, Belgium

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Potato peel wastes (PPW) was valorized

to produce ethanol and biogas.
• Organosolv pretreatment significantly
enhanced hydrolysis and fermentation.
• Pretreatment liquid and unhydrolyzed
solids were suitable for biomethane
production.
• From each ton of PPW, 253.5 L gasoline
equivalent energy was obtained.

A R T I C L E I N F O A B S T R A C T

Handling Editor: Konstantinos G Moustakas Potato processing industries generate considerable amounts of residues, i.e., potato peel wastes (PPW). Valori­
zation of PPW for bioethanol and biogas production via a biorefining process was investigated in this study.
Keywords: Organosolv pretreatment was performed on the PPW using 50–75% (v/v) ethanol solution at 120–180 ◦ C with/
Bioethanol without the presence of 1% (w/w) H2SO4 (as a catalyst). After the pretreatment, the solvent, i.e., ethanol, was
Biogas
recovered by distillation. Catalyzed organosolv pretreatment using 50% (v/v) ethanol at 120 ◦ C followed by
Biofuel
enzymatic hydrolysis resulted in a high hydrolysate yield of 539.8 g glucose/kg dry PPW that was successfully
Organosolv pretreatment
Food wastes fermented to 224.2 g ethanol/kg dry PPW. To recover more energy, the liquid fraction of the pretreatment
remained after solvent recovery and the unhydrolyzed solids that remained from the enzymatic hydrolysis were
anaerobically digested. From each kg of dry PPW, the anaerobic digestion produced 57.9 L biomethane. Thus, the
biorefinery comprising ethanolic organosolv pretreatment, solvent recovery, enzymatic hydrolysis, ethanolic
fermentation, and anaerobic digestion of residues was produced 8112 kJ energy per kg of dry PPW.

1. Introduction
The acceleration of industrialization and the growing population
have increased the world energy demand (Zaky et al., 2020). Today, the
large majority (more than 80%) of world energy is supplied by fossil
fuels (Monir et al., 2020). The overexploitation of these fuels has caused
various environmental issues such as climate changes and carbon
emissions (Toor et al., 2020; Haldar and Purkait, 2021). Hence, atten­
tion of researchers has been attached to appropriate alternatives
(Khoshkho et al., 2021). Biofuels (biomass-based fuels), as sustainable

* Corresponding author. Department of Chemical Engineering, Isfahan University of Technology, Isfahan 84156-83111, Iran.
E-mail address: karimi@cc.iut.ac.ir (K. Karimi).

https://doi.org/10.1016/j.chemosphere.2021.133003
Received 13 September 2021; Received in revised form 1 November 2021; Accepted 18 November 2021
Available online 19 November 2021
0045-6535/© 2021 Elsevier Ltd. All rights reserved.
A. Soltaninejad et al.
and renewable energy resources with superior environmental effec­
tiveness, are promising fuel carriers to control energy requirements
(Mohapatra et al., 2017; Moustakas et al., 2020).
Anaerobic digestion (AD), as a microbial process that produces an
energy rich stream of biogas from various organic resources, helps in
reducing the pathogens, odor, and greenhouse gases emissions (Hashemi
et al., 2020). Biogas, one of the most favorable gaseous biofuels, is uti­
lized primarily as a source for electricity and heat production (Feiz et al.,
2020; Lin et al., 2021).
Bioethanol is a green liquid biofuel that can be wildly used in the
transportation industry (Ayodele et al., 2020). The utilization of bio­
ethanol as a substitute for gasoline causes the reduction of carbon
emission up to 80% along with getting rid of acid rain released from
sulfur dioxide (Momayez et al., 2018; Barampouti et al., 2019).
Nowadays, starch-or sugar-based food crops such as sugarcane, sugar
beet, corn, and maize are the mostly-utilized feedstocks for industrial
production of bioethanol (Toor et al., 2020; Shenbagamuthuraman
et al., 2021). The conversion of these edible feedstocks to ethanol re­
quires high water, fertilizers, and arable lands, resulting in food-fuel
conflicts (Xu et al., 2019). Therefore, attention has been attracted to
low value and non-edible alternatives with no compromise regarding
food security, e.g., lignocellulosic materials and agro-industrial wastes
(Cooper et al., 2020; Mohapatra et al., 2020).
Potatoes are the world’s third-largest food crop after rice and wheat.
Every year, about 370 million tons of potatoes are produced worldwide
(www.fao.com/faostat, 2019). Production of potato chips and crisps,
starch, and/or French fries during potato processing causes the accu­
mulation of a large quantity of potato peel wastes (PPW) as the
byproduct. This waste stream, generated by processes of cutting and
peeling, comprises 15–40% of processed potato weight (Liang et al.,
2015; Abedini et al., 2020). Currently, landfilling is the major PPW
management strategy, leading to serious environmental issues. How­
ever, PPW that is mainly composed of starch and lignocelluloses has a
high potential for the production of added-value bioproducts (Atitallah
et al., 2019; Chohan et al., 2020), e.g., biobutanol (Abedini et al., 2020),
biogas (Liang and McDonald, 2015), and bioethanol (Arapoglou et al.,
2010).
Direct bioconversion of PPW sugar polymers to bioethanol is limited
by some compositional and structural obstacles; therefore, extra stages
are necessary for the saccharification of the substrate (Ben Taher et al.,
2017). These stages could be utilized either separately or in combina­
tion. Accordingly, in order to convert sugar polymers into fermentable
sugars, an enzymatic hydrolysis stage has been applied (Zhao et al.,
2009). However, since PPW contains a large portion of lignocellulosic
materials and due to their recalcitrance and complex structure of these
materials, a suitable pretreatment process is essential to enhance cel­
lulose digestibility and, consequently, to improve the yield of ethanol
production (Mahmoodi et al., 2018a; Atitallah et al., 2019; Stoumpou
et al., 2020). Several pretreatment technologies, including biological,
chemical, physical, and physicochemical ones, were proposed to in­
crease the yield of biofuels (Prasad et al., 2019). Amongst these ap­
proaches, organosolv pretreatment, which uses ethanol, was introduced
as a promising chemical technology (Cebreiros et al., 2020). During
organosolv pretreatment of lignocellulosic fraction, hemicellulose and
lignin are solubilized by water-solvent liquor resulting in a remained
solid cellulose-rich phase. In addition, a hemicellulose-rich stream was
obtained after precipitation of lignin by diluting the liquor with water
(Ferreira and Taherzadeh, 2020). Furthermore, in order to improve both
hemicellulose hydrolysis and the degree of delignification, acid catalysts
such as sulfuric acid could be combined with organosolv pretreatment
(Chen et al., 2017; Zhou et al., 2018).
Romaní et al. (2019) used ethanolic organosolv pretreatment on
eucalyptus nitens bark. They reported that the yield of ethanol improved
from 32 to 99% by the pretreatment. In another study, organosolv
pretreatment of sunflower stalks with 50% (v/v) isopropanol and 1%
sulfuric acid at 160 ◦ C for 30 min yielded 278 mL methane/g VS after 45
2
Chemosphere 291 (2022) 133003
days of anaerobic digestion, while only 124 mL/g VS biomethane was
produced from untreated stalk (Hesami et al., 2015). To the best of our
knowledge, organosolv pretreatment of PPW for enhancing the yield of
bioethanol and biogas production has not been studied yet.
The present work was aimed at efficient bioethanol production from
PPW and, subsequently, biomethane production from the process resi­
dues. Ethanol-based organosolv pretreatment was conducted to disrupt
the compact and rigid structure of PPW lignocellulosic fraction. The
solid fraction of pretreatment stage was subjected to cellulose and starch
hydrolysis using three different enzymes followed by ethanolic
fermentation using Saccharomyces cerevisiae. Remained solid of enzy­
matic hydrolysis, together with the ethanol-separated liquid phase of
pretreatment, was anaerobically digested for biomethane production.
2. Materials and methods
2.1. Substrate
PPW was provided from the Isfahan University of Technology
Restaurant, Isfahan, Iran. The potatoes were from the variety of Agria.
They were washed with water, air dried at room temperature, grounded,
and screened to obtain a biomass size of between 0.177 and 0.833 mm.
2.2. Pretreatment
The mixture of 50 or 75% (v/v) ethanol in water with or without 1%
w/w (based on PPW dry weight) H2SO4 as the catalyst at 120, 140, and
180 ◦ C was utilized for organosolv pretreatment of PPW. The solid to
liquid ratio of 1:10 (w/w) was prepared in a 500 mL high-pressure
reactor (SS304, Steel Sanat, Isfahan, Iran). After 1 h treatment at the
desirable temperature, the reactor was quickly put in an ice water
chamber. Afterward, a filter was used to separate liquid and residual
solid from each other. Then, the residual solid was washed with 120 mL
ethanol-water mixture (50% ethanol at 60 ◦ C). The obtained solid
matter was rinsed with distilled water and then air-dried for 2 days. The
weight of the solid matter was measured for material recovery calcula­
tions, and the solid was stored in a sealed bag for further analysis. The
ethanol of pretreatment liquid phase was recovered using a laboratory
distillation. After the evaporation of ethanol, a solid phase that mainly
consisted of lignin was precipitated and separated by centrifugation. The
remained liquid phase (pretreatment liquor) was kept at − 20 ◦ C until
anaerobic digestion.
2.3. Enzymatic hydrolysis
Two-stage enzymatic hydrolysis was applied for fermentable sugar
production from cellulose and starch in the untreated and pretreated
PPW. In the beginning, cellulose hydrolysis was performed by using 20
FPU (Filter paper unit) cellulase per g of dry substrate in 118 mL glass
bottles containing 50 g L-1 (w/w) of dry PPW in 0.05 mol L− 1 citrate
buffer at pH 4.8. Using aluminum caps on rubber stoppers, bottles were
sealed and then put in the shaking incubator at 45 ◦ C and 120 rpm
(Molaverdi et al., 2019). The cellulase enzyme of Cellic® CTec2 that
kindly provided by Novozymes (Copenhagen, Denmark) was used for
cellulose hydrolysis. The cellulase activity of the enzyme was deter­
mined to be 125 FPU/mL according to a standard method (Adney and
Baker, 1996). After 72 h of hydrolysis, the liquid samples were with­
drawn for the analysis of sugar content.
In the later stage of hydrolysis, the starch content was enzymatically
hydrolyzed by α-amylase (Liquezyme, Novozymes A/S, Copenhagen,
Denmark) for the liquefaction step and glucoamylase (Dextrozyme GA,
Novozymes A/S, Copenhagen, Denmark) for the saccharification. To
perform starch hydrolysis, 2 g enzyme/kg starch from each of the en­
zymes (α-amylase and glucoamylase) was added to the 118 mL glass
bottle containing suspension. The pH adjusted to 6, 1 g/l co-factor
(calcium chloride) was added, and liquefaction was carried out for 2 h
A. Soltaninejad et al.
at 90 ◦ C using α-amylase. Afterward, the temperature was decreased to
65 ◦ C, glucoamylase was added, and saccharification was performed for
30 h at 65 ◦ C and pH 4.5 (Mahmoodi et al., 2018b). Then, all samples
were centrifuged (15 min at 4000 rpm) to separate supernatant. To
analyze the sugar content of hydrolysate, 2 mL liquid sample was
withdrawn and the remaining hydrolysate was kept at − 20 ◦ C for
ethanolic fermentation. Furthermore, the air-dried residual solid was
stored at 4 ◦ C for further biomethane production.
The yield of enzymatic hydrolysis was calculated based on g/kg
pretreated or untreated substrate according to Eq. (1):
( )
()
​ Glucose ​ gl
g analysis of lignocellulosic materials (Sluiter and Sluiter, 2008). The
Glucose ​ yield ​ = ( ) × 1000 (1)
kg Untreated ​ or ​ pretreated ​ PPW ​ gl
2.4. Ethanolic fermentation
Saccharomyces cerevisiae (CCUG 53310, obtained from the University
of Gothenburg) was used as the fermenting microorganism. The
fermentation process was conducted in a glass bottle (118 mL) under
anaerobic conditions for ethanol production from hydrolysate of the raw
as well as pretreated PPW. The applied medium for ethanolic fermen­
tation was as follows: 3.5 g L-1 KH2PO4, 1 g L-1 CaCl2.2H2O, 5 g L-1 yeast
extract, 7.5 g L-1 (NH4)2SO4, and 0.75 g L-1 MgSO4.7H2O. The experi­
ments were carried out at 130 rpm and 32 ◦ C for 48 h. The pH was
adjusted to 5 using 2 M NaOH, and the autoclaving of bottles was per­
formed for 20 min at 121 ◦ C. Next, they were cooled to ambient tem­
perature and S. cerevisiae (1 g l-1, based on dry mass) was added to each
medium under sterile conditions (Karimi et al., 2006). Three min in­
jection of nitrogen gas to each medium was performed to provide the
anaerobic conditions. After 48 h of ethanolic fermentation, a sample was
taken to measure the ethanol concentration.
The theoretical yield of ethanol was calculated according to Eq. (2):
( )
g 1.111 × 0.51 × Total ​ glucan ​ (g)
Ethanol ​ theoretical ​ yield ​ = (2)
kg PPW ​ (kg)
where 1.111 is the hydration factor of glucan and 0.51 (g/g) is the
theoretical yield of ethanol from hexoses.
2.5. Biogas production
The ethanol-separated liquid phases of the pretreatment as well as
the solid residues of enzymatic hydrolysis were anaerobically digested
for 45 days in the 118 mL dark glass bottles for biogas production. The
inoculum sludge of the process was provided by a mesophilic anaerobic
digester with 7000-m3 capacity located in the plant of Municipal
Wastewater Treatment in Isfahan city (Iran). According to the method
provided by Hansen et al. (2004), 20 mL of inoculum was mixed with
around 0.25 g of substrate to achieve a double volatile solid ratio of
inoculum to the substrate. Afterward, pure nitrogen gas was purged for
3 min to each glass bottle to establish the anaerobic conditions. Finally,
the 45-day incubation of bottles was performed at 37 ◦ C in an incubator.
Biomethane production from inoculum in the absence of any substrate
was also determined by anaerobic digestion of a mixture containing 20
mL inoculum and 5 mL deionized water and was introduced as a blank
experiment.
The theoretical yield of biomethane was calculated according to Eq.
(3):
( )
L (Carbohydrates ​ (g) × 415.0) + (Protein ​ (g) × 633.6) + (Lipid ​ (g) × 990.4)
Biomethane ​ theoretical ​ yield ​ = (3)
kg
Chemosphere 291 (2022) 133003
where 415.0, 633.6, and 990.4 (mL/g) are the theoretical methane yield
from carbohydrates, protein, and lipid, respectively.
2.6. Analytical methods
To determine starch content of the raw as well as pretreated PPW,
enzymatic hydrolysis of starch was performed by using α-amylase
(Liquezyme, Novozymes A/S, Copenhagen, Denmark) and glucoamylase
(Dextrozyme GA, Novozymes A/S, Copenhagen, Denmark). The starch
percentage of PPW was measured and then the starch was completely
removed through the NREL (TP-510-42624) procedure prior to the
concentration of lignin and structural carbohydrates of PPW were
analyzed using National Renewable Energy Laboratory (NREL) protocol
provided by Sluiter et al. (2008b). The acid soluble lignin content of
PPW was measured using a UV–visible spectrophotometer (UV-1601,
Beijing Rayleigh Co., China) at the wavelength of 320 nm. Moreover, the
absorptivity factor of biomass at the recommended wavelength was 30
L/g cm (Sluiter et al., 2008b)
By using the NREL (TP-510-42621) standard protocol, volatile solids
and total solids content of PPW were also measured (Sluiter et al.,
2008a). Moreover, the protein content of PPW was measured using the
Kjeldahl method (Arapoglou et al., 2010).
The concentration of monomeric sugars and ethanol was analyzed
using a High-performance liquid chromatography (HPLC) system
equipped with RI and UV/Vis detectors (Jasco International Co., Tokyo,
Japan). An Ion-exchange column of Aminex HPX-87P (Bio-Rad labora­
tories, Hercules, USA) operated at 85 ◦ C with the mobile phase of
deionized water (flow rate of 0.6 mL/min) was applied to detect
different sugars (glucose, galactose, arabinose, and xylose). By using a
hydrogen ion-based column of Aminex HPX-87H (Bio-Rad, Hercules,
USA) operated at 60 ◦ C with an eluent of 5 mM sulfuric acid (flow rate of
0.6 mL/min), the concentration of ethanol was measured.
The CH4 and CO2 contents of the produced biogas were measured
using a system of gas chromatograph (GC-2552, Thermal conductivity
detector (TCD), Teif Gostar Faraz Co., Iran) that was equipped with a
stainless-steel packed column (porapak Q, Chrompack, Germany). The
flow rate of carrier gas (nitrogen) was kept at 20 mL/min. The tem­
perature of column, detector, and injector was 40, 150, and 100 ◦ C,
respectively. In all tests, the yields of biomethane production were dis­
played under standard conditions.
2.7. Equivalent gasoline
Since two different biofuels were produced, i.e., ethanol and bio­
methane, and for better comparison between the different pre­
treatments, the total produced energy was calculated based on
equivalent gasoline of biofuels. These comparisons were based on the
utilization of each ton of PPW. The lower heating values of 36.1 MJ/
Nm3, 32 MJ/L, and 22.1 MJ/L were used for methane, gasoline, and
ethanol, respectively (Najafi et al., 2021). Solid recoveries, volatile
solids, and total solids of PPW after the organosolv pretreatment stage
were also used in the calculation.
2.8. Statistical analysis
All experiments and measurements were executed at two replicates,
and the data was demonstrated as mean ± SD (standard deviation). One-
way ANOVA (analysis of variance) and general linear model (GLM) test
PPW ​ (kg) × 1000
3
A. Soltaninejad et al. Chemosphere 291 (2022) 133003

Table 1 from Table 2, increasing the treatment temperature (at constant other
Composition of raw PPW (potato peel wastes). conditions) resulted in a decrease in the amount of material recoveries.
Component Content (% w/w) Through the organosolv treatment with 75% ethanol, 28–62% of starch
content was released into the liquor. Addition of 1% acid and decreasing
Lignocelluloses
Glucan 34.2 ± 0.1 the ethanol concentration to 50% resulted in the highest starch removal
Galactan 3.3 ± 0.2 of 72% at 180 ◦ C. In the absence of acid, when ethanol concentration
Arabinan 4.9 ± 0.2 was decreased to 50%, 31–68% of starch was removed. Organosolv
Acid soluble lignin 0.7 ± 0.1 treatment using 50% ethanol at 120 ◦ C without catalyst led to dissolving
Acid insoluble lignin 11.4 ± 0.3
Starch 32.4 ± 1.6
29.9% of the PPW into the liquor, which consisted of 59.4 g lignin and
Protein 12.5 ± 0.8 99.4 g starch per kg of PPW. The addition of catalyst led to the disso­
Lipid 0.9 ± 0.1 lution of 66.7 g lignin and 122 g starch from each kg of PPW. The
Water and ethanol extractives 27.9 ± 1.9 treatment with 50% ethanol assisted by 1% acid at 180 ◦ C led to the
Total solid 92.1
highest dissolution of PPW. The highest recorded starch recovery was
Volatile solid 85.2
72.4%, obtained from PPW pretreated with 75% ethanol at 120 ◦ C. This
data is in compliance with the starch recovery (65.9%) reported from
were used to compare the means using Minitab 17.1.0 (Minitab Inc, the treatment of municipal solid wastes using 85% ethanol and 1%
USA) software. In all tests, the levels of significances were determined as acetic acid at 120 ◦ C (Ebrahimian et al., 2020). The content of total
0.05 using the Tukey method with 95% confidence. lignin of PPW (12.11%) was considerably affected by organosolv
treatment. The total lignin content in the treated PPW residues was
3. Results and discussion between 3.7 and 9.0% at different conditions. Increasing the treatment
temperature and the addition of catalyst enhanced the total lignin
3.1. PPW composition removal. For instance, increasing the treatment temperature from 120 to
180 ◦ C increased total lignin removal from 46.1 to 76.7% using 75%
The lignocelluloses, starch, and protein contents of PPW were pre­ ethanol. At these conditions, the addition of 1% H2SO4 further increased
sented in Table 1. Based on dry mass, the total solids and volatile solids the corresponding lignin removal values to 53.8 and 84.3%. Interest­
contents of PPW were 92.1 and 85.2%, respectively. Furthermore, this ingly, Jafari et al. (2016) applied acetone for organosolv pretreatment of
waste had remarkable amounts of water and ethanol extractives sweet sorghum bagasse and reported the lignin removal of 75% at op­
(27.9%). PPW was mainly composed of glucan (34.2%) and starch timum conditions (180 ◦ C, 90 min). In another study, 84.5% lignin
(32.4%), which shows its promising potential to be converted into bio­ removal was reported after organosolv pretreatment of municipal solid
ethanol. Based on the glucan to ethanol theoretical conversion yield, waste with ethanol (Farmanbordar et al., 2018). In the present study, the
376.9 g ethanol per kg of PPW could be theoretically produced (Fig. 1). highest total lignin removal was 88.8% achieved after catalyzed treat­
In addition, PPW contained 12.5% protein and 0.9% lipid that, together ment using 50% ethanol at 180 ◦ C.
with carbohydrates, could be subjected to AD for the theoretical As can be inferred from Table 2, organosolv treatment at all applied
methane potential of 377.2 L from each kg of PPW (Fig. 1). conditions was effective in increasing the PPW glucan content. The
recovered materials containing 41.6–55.5% glucan were achieved after
treatments, while the glucan percentage of untreated PPW was 34.1%.
3.2. Pretreatment effects on PPW composition The percentage of glucan was enhanced by the addition of catalyst and
increasing the temperature. Furthermore, increasing the ethanol con­
To enhance the digestibility of lignocellulosic portion of PPW in centration in the treatment had no considerable effect on glucan con­
hydrolysis process, organosolv pretreatment was performed. The pre­ tent. Similar trends were reported for the treatment of wheat straw
treatments were conducted at different ethanol concentrations (50 and (Wildschut et al., 2013).
75% v/v) and temperatures (120, 140, and 180 ◦ C) in the presence of 1% The changes in volatile solids in the pretreatments liquor are re­
w/w or absence of H2SO4 (as a catalyst) for 1 h. The pretreatment ported in Table 3. The amounts of volatile solids in liquors were highly
conditions, the treated PPW composition, and material recoveries were dependent on the treatment temperature. Raising the temperature
given in Table 2. Prior to determining the composition of untreated and enhanced the concentration of volatile solids in liquor. For example,
treated PPW, amylolytic enzymes were added to the solid fractions of increasing the treatment temperature from 120 to 180 ◦ C (in pre­
ethanolic treatment to remove their starch contents. treatments with 50% ethanol) enhanced the concentration of volatile
After performing the organosolv treatment, a considerable amount of solids in the liquor from 10.9 to 20.8 g/l. Furthermore, the addition of
PPW (27.6–63.4%) was solubilized into the liquor. As can be inferred catalysts to the treatment stage led to the increase of volatile solids

Fig. 1. Theoretical yields of biomethane and bioethanol from 1-kg of PPW (potato peel wastes).

4
A. Soltaninejad et al. Chemosphere 291 (2022) 133003

Table 2
Composition of pretreated PPW (potato peel wastes) and the material recovery after each pretreatment.
Pretreatment conditions Material recovery (%) Compositions

T ( C)
◦
Ethanol (%) Acid (%) Glucan (%) Galactan (%) Arabinan (%) Total lignin (%) Starch (%)

120 50 – 70.1 ± 1.3 42.1 ± 3.0 2.6 ± 0.2 6.5 ± 0.2 8.8 ± 0.2 32.0 ± 0.7
140 62.5 ± 1.1 44.9 ± 0.8 2.3 ± 0.2 6.0 ± 0.3 6.2 ± 0.3 29.1 ± 0.2
180 40.1 ± 0.3 53.2 ± 1.2 1.9 ± 0.1 2.2 ± 0.1 5.8 ± 0.3 25.7 ± 1.6
120 75 – 72.4 ± 1.0 41.6 ± 0.5 2.7 ± 0.1 6.5 ± 0.1 9.0 ± 0.6 32.4 ± 1.3
140 66.2 ± 0.4 43.5 ± 0.1 2.4 ± 0.1 5.8 ± 0.2 6.8 ± 0.1 30.9 ± 0.6
180 46.2 ± 0.3 52.7 ± 1.1 2.0 ± 0.1 2.3 ± 0.1 6.1 ± 0.2 26.5 ± 0.7
120 50 1 69.8 ± 1.0 46.1 ± 0.7 2.9 ± 0.1 5.1 ± 0.3 7.8 ± 0.5 28.9 ± 1.4
140 62.0 ± 1.7 48.2 ± 0.4 2.3 ± 0.2 5.0 ± 0.2 5.1 ± 0.4 29.1 ± 1.1
180 36.6 ± 0.6 55.5 ± 0.8 1.8 ± 0.1 1.6 ± 0.1 3.7 ± 0.1 24.8 ± 0.5
120 75 1 71.6 ± 0.9 44.8 ± 0.1 3.2 ± 0.1 5.3 ± 0.2 7.8 ± 0.6 29.1 ± 0.4
140 65.0 ± 0.6 47.7 ± 1.2 2.4 ± 0.1 5.1 ± 0.4 5.5 ± 0.1 28.3 ± 0.9
180 45.1 ± 0.2 55.1 ± 0.3 1.5 ± 0.1 1.5 ± 0.1 4.2 ± 0.1 24.1 ± 0.7

temperature and addition of catalyst could be responsible for these

Table 3
improvements (Zhao et al., 2009; Mirmohamadsadeghi et al., 2014;
Volatile solids content of liquid phase after organosolv pretreatment.
Najafi et al., 2021).
Pretreatment conditions Liquor volatile solids (g/l)

T (◦ C) Ethanol (%) Acid (%)

3.3. Cellulose and starch hydrolysis of organosolv-pretreated PPW
120 50 – 10.9 ± 0.5
140 12.3 ± 0.2
The hydrolysis of cellulose in the solid fractions, obtained from
180 20.8 ± 0.1
120 75 – 10.0 ± 0.2 organosolv pretreatment, was carried out by a commercial cellulase
140 11.1 ± 0.5 enzyme at 45 ◦ C for 72 h at the solid loading of 5% (w/v). Then the
180 15.6 ± 1.1 whole cellulose-degraded samples were exposed to starch hydrolysis by
120 50 1 14.9 ± 0.8 α-amylase for liquefaction stage (90 ◦ C, 2 h) and by glucoamylase for
140 15.2 ± 0.1
180 22.0 ± 0.2
saccharification stage (65 ◦ C, 30 h). Glucose concentrations after cel­
120 75 1 10.9 ± 0.2 lulose and starch hydrolysis as well as glucose yields after the total
140 13.2 ± 0.1 enzymatic hydrolysis process were illustrated in Fig. 2.
180 19.2 ± 0.2 Organosolv treatment, prior to hydrolysis of glucan and starch,
increased the glucose yield in the range of 19.7–39.6%. Subjecting the
organosolv-pretreated PPW to cellulose hydrolysis resulted in hydroly­
content in liquor. The maximum concentration of volatile solids, i.e.,
sates containing 26.4–40.2 g/l glucose. Afterward, starch hydrolysis of
22.0 g/l, was gained by the catalyzed organosolv pretreatment with 50%
the cellulose-degraded samples resulted in the total glucose production
ethanol at 180 ◦ C. Therefore, as can be inferred from Tables 2 and 3,
of 36.4–42.5 g/l. The untreated PPW was also subjected to enzymatic
organosolv pretreatments with lower material recovery had higher
hydrolysis, resulting in 18.35 and 12.06 g/l glucose after cellulose and
volatile solids in the liquor. Higher solubilization of lignin and hemi­
starch hydrolysis, respectively. Therefore, 608.2 g glucose was obtained
cellulose sugars to the liquor by increasing the pretreatment
from each kg of untreated PPW. Hydrolysis of about 82% of glucan

Fig. 2. Glucose concentration obtained from raw and pretreated PPW (potato peel wastes) after 72 h cellulose hydrolysis by cellulase (white) and after starch
hydrolysis by amylases (black), as well as glucose yield (Ο)after both cellulose and starch hydrolysis. The significant differences between the pretreatment conditions
are presented by dissimilar uppercase letters on the columns.

5
A. Soltaninejad et al.
content (consisted of cellulose and starch) resulted in this amount of
glucose.
According to Fig. 2, increasing the temperature of organosolv
treatment enhanced the glucose concentrations obtained from cellulose
hydrolysis, while it clearly decreased the glucose concentration from
starch hydrolysis. For instance, 29.3 and 35.3 g/L glucose was achieved
after cellulose hydrolysis of pretreated PPW at 120 and 180 ◦ C (with
75% ethanol and 1% sulfuric acid), respectively. However, pretreatment
at 180 ◦ C resulted in 39.8% lower glucose concentration after starch
hydrolysis, compared to that of treatment at the same conditions at
120 ◦ C. The glucose concentrations (34.0 and 34.5 g/L) obtained from
cellulose hydrolysis of PPW treated in the absence and presence of 1%
H2SO4, respectively, at 140 ◦ C with 50% ethanol shows the inability of
the acidic catalyst to enhance the glucose amount. Moreover, increasing
the ethanol concentration from 50 to 75% had no considerable effect on
glucose concentration after cellulose hydrolysis. The highest glucose
concentration of 40.18 g/l was achieved from cellulose hydrolysis of
PPW treated with 50% ethanol and 1% H2SO4 at 180 ◦ C. Hence, orga­
nosolv pretreatment had a clear positive impact on the obtained yield of
glucose after cellulose hydrolysis. Lignin removal followed by enhanced
cellulose accessibility to cellulase by the organosolv pretreatment might
be responsible for this improvement (Taherzadeh and Karimi, 2008;
Abedini et al., 2020). Similarly, Abedini et al. (2020) reached the
glucose concentration of 38 g/l applying enzymatic hydrolysis using
cellulase and hemicellulase on pretreated PPW with 75% ethanol and
1% sulfuric acid at 180 ◦ C. In the present study, applying ethanolic
pretreatment at 180 ◦ C with 75% ethanol assisted by 1% sulfuric acid,
the highest glucose yield of 849.2 g per kg of treated PPW was obtained
after cellulose and starch hydrolysis that was 39.62% higher than that of
untreated PPW. The glucose yield (849.2 g per kg treated PPW) obtained
in the present study is in the same order of the glucose yield reported
from municipal solid waste pretreated (817 g per kg of the treated
substrate) by organosolv procedure using 85% ethanol at 120 ◦ C
(Ebrahimian et al., 2020). Also, Ben Taher et al. (2017) reported the
positive effect of hydrothermal, acid, and alkali pretreatments on
glucose concentration from the PPW hydrolysate after enzymatic hy­
drolysis. They reached the maximum hydrolysis yield of 63% from the
hydrothermally pretreated sample, whereas the hydrolysis yield of un­
treated PPW was 20%.
Fig. 3. Concentrations of ethanol and ethanol yields obtained after 48 h fermentation of raw and pretreated PPW (potato peel wastes) at different conditions. The
significant differences between the pretreatment conditions are presented by dissimilar uppercase letters on the columns.
6
Chemosphere 291 (2022) 133003
3.4. Bioethanol production
Anaerobic fermentation of the hydrolysate, obtained from enzymatic
hydrolysis of organosolv-pretreated PPW samples, was performed using
S. cerevisiae at 32 ◦ C for 48 h to produce bioethanol (Fig. 3). The sepa­
rated hydrolysis and fermentation (SHF) of pretreated PPW showed a
48.2% increase in the bioethanol concentration compared to the un­
treated PPW. The maximum bioethanol concentration (18.04 g/L) was
achieved from the PPW pretreated at 180 ◦ C with 75% ethanol and 1%
acid. Notably, this bioethanol yield is in close agreement with the pro­
duced ethanol from acid-pretreated PPW reported previously (17.27 g/
L) (Atitallah et al., 2019).
The SHF of untreated PPW yielded 243.4 g ethanol per kg of biomass
(Fig. 3). The organosolv treatment using 50% ethanol at 120 ◦ C resulted
in an increased bioethanol yield up to 295.2 g/kg treated PPW. Besides
increasing the ethanol concentration from 50 to 75%, the addition of
catalyst led to 9.2% higher bioethanol yield. An increase in the treat­
ment temperature from 120 to 140 ◦ C led to 9.7% increase in bioethanol
production. However, increasing the treatment temperature from 140 to
180 ◦ C had no considerable effect on bioethanol production. As can be
inferred from Figs. 2 and 3, the glucose and ethanol concentrations
achieved after hydrolysis and fermentation processes were completely in
compliance. For example, the maximum glucose concentration (42.5 g/
L) and the maximum ethanol concentrations (18.1 g/L) were obtained
after treatment at the most severe condition (180 ◦ C, 75% ethanol, and
1% catalyst).
3.5. Anaerobic digestion
The liquid fractions of organosolv pretreatment as well as the resi­
dues from the enzymatic hydrolysis stage were used as a substrate for
biomethane production under anaerobic digestion (Fig. 4).
The remained solids from enzymatic hydrolysis of untreated PPW
resulted in 181.3 mL/g VS biomethane. This yield is higher than all the
biomethane yields obtained from the remained solids of pretreated
substrates. The removal of starch and readily biodegradable ingredients
in the hydrolysis and pretreatment stages is the most likely reason for
the lower biomethane production yields from the pretreated PPW
(Ebrahimian et al., 2020).
When pretreated PPW at 120 ◦ C with 50% ethanol subjected to AD,
156.0 mL CH4/g VS and 168.3 mL CH4/g VS were produced from the
A. Soltaninejad et al.
Fig. 4. Produced biomethane after 15 days (black), 30 days (gray), and 45 days (white) anaerobic digestion of pretreatment liquid fraction (A, B) and solid residues
(C, D). The pretreatment was performed with ethanol concentration of 50% (A, C) and 75% (B, D).
liquid fraction of organosolv treatment and the remained solids of hy­
drolysis stage, respectively. Addition of H2SO4 to the pretreatment stage
led to 36.7% increase in the biomethane yield from the liquid fraction of
organosolv treatment. However, the catalyst addition had no significant
influence on the obtained biomethane after AD of the remained solids.
As can be seen in Fig. 4, increasing the temperature from 120 to
140 ◦ C in the treatment processes improved the achieved biomethane
from the pretreatment liquid fraction. However, increasing the ethanol
concentration from 50 to 75% in pretreatment stage had no significant
effect on the biomethane yield from the treatment liquid fraction.
Introducing higher amount of starch and hemicelluloses into the pre­
treatment liquid fraction at a higher temperature is the reason for this
observation. Since, Mahmoodi et al. (2018a) reported that acid pre­
treatment of municipal solid waste (consisted of starchy and lignocel­
lulosic material) using 0.5% sulfuric acid at 160 ◦ C for 1 h yielded a
higher amount of starch, glucose, and fermentable sugars in the treat­
ment liquid phase that yielded higher methane production, compared to
the pretreatment using 0.5% acid at 130 ◦ C for 30 min.
The treatment liquid phase obtained from PPW pretreated with 50%
ethanol and 1% H2SO4 at 140 ◦ C yielded 114.1 mL methane per g VS
after 15 days of AD, and it reached 235.7 mL methane per g VS after 45
7
Chemosphere 291 (2022) 133003
days of AD. In contrast, increasing the treatment temperature to 180 ◦ C
yielded 140.4 mL/g VS methane after 45 days of AD. Overall, increasing
the treatment severity by raising the temperature from 140 to 180 ◦ C
yielded a clear reduction in the methane yield from the treatment liquid
fraction. Formation and realization of inhibitory products to the orga­
nosolv liquid phase at high treatment temperatures could be mentioned
as the main reason for reducing the biomethane yield at higher tem­
peratures (Mirmohamadsadeghi et al., 2014).
3.6. Gasoline equivalent
To compare the pretreatment effects on total energy yield from PPW,
gasoline equivalent calculations were applied, considering ethanol,
biomethane, and solid recoveries in the pretreatments. These calcula­
tions were according to E.q S1 presented in supplementary data. The
calculated bioenergy production after treatment of 1-ton dry PPW at
120 ◦ C with 50% ethanol and 1% acid is presented in Fig. 5. Each ton of
dry PPW can be converted to 539.8 kg glucose. This amount of glucose
can be fermented to 284.1 L ethanol, and 22.1 m3 biomethane can be
produced through the AD of unhydrolyzed remaining solids. Also, 35.8
m3 biomethane was produced from the liquid fraction of pretreatment.
A. Soltaninejad et al.
Fig. 5. The obtained equivalent gasoline from 1-ton of dry PPW (potato peel wastes) at the best conditions.
Therefore, ethanol production, with a bigger order of magnitude in the
yield, played a more important role in the obtained volume of gasoline
equivalent in comparison with biomethane. As shown in Figs. 5 and
188.2 L gasoline equivalent was achieved from ethanol production,
while it was 24.9 and 40.4 L for biomethane production from unhy­
drolyzed solid residues and liquid fraction of pretreatment, respectively.
Chohan et al. (2020) reported the production of 233.2 L ethanol from
each ton of PPW (154.5 L equivalent gasoline) through the simultaneous
saccharification and fermentation process, which is lower than the
present study. Also, the production yields of 22 kg ethanol and 185 kg
butanol were reported from 1 ton of PPW (about 160 L equivalent gas­
oline) (Abedini et al., 2020). In another study, 1 ton of dry potato peel
residues was converted to 100 kg ethanol resulting in about 84 L
equivalent gasoline production (Ben Taher et al., 2017). In this study,
8112.0 KJ energy and 253.5 L equivalent gasoline were achieved from
the conversion of 1-ton of dry PPW to ethanol and biogas production
from residues, indicating notably higher than the mentioned literature.
As can be inferred from Fig. 1, each kg of untreated PPW could theo­
retically result in 10.6 and 13.6 MJ energy from bioethanol and bio­
methane production, respectively. Therefore, the achieved energy
(8112 KJ) in the present study using organosolv pretreatment at opti­
mum condition yielded 80% and 60% of the bioethanol and biomethane
theoretical yields, respectively. Overall, the higher energy and subse­
quently higher equivalent gasoline obtained from the conversion of PPW
to ethanol and biogas through the applied process in this work,
compared to the similar studies.
4. Conclusions
According to the obtained results, it can be inferred that PPW is a
high potential substrate for biofuel production. Catalyzed organosolv
pretreatment was a promising pretreatment for increasing the bio­
ethanol production yield. The pretreatment at optimum condition (with
50% ethanol, 1% acid at 120 ◦ C) resulted in the best glucose yield of
539.8 g/kg dry PPW and the ethanol production of 284.1 mL/kg dry
PPW (59% of theoretical yield). Furthermore, after subjecting the liquid
phase of pretreatment as well as residual solid of hydrolysis step to
anaerobic digestion, a considerable amount of biomethane was obtained
(equivalent gasoline of 65.3 mL/kg dry PPW). On the whole, 8.1 MJ
energy was produced from 1-kg of dry PPW in the proposed process.
8
Chemosphere 291 (2022) 133003
Author’s contribution
Ali Soltaninejad: Investigation, Software, Resources, Formal anal­
ysis, Data curation, Visualization, Writing original draft Mohammad­
Hadi Jazini: Validation, Visualization, Supervision, Writing – review &
editing Keikhosro Karimi: Conceptualization, Supervision, Project
administration, Funding acquisition, Methodology, Writing – review &
editing
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgments
The Institute financially assisted this project for Biotechnology and
Bioengineering, Isfahan University of Technology. The authors are
grateful to Novozymes, Denmark, for providing the enzymes used in this
study.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.chemosphere.2021.133003.
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