Bioresource Technology 251 (2018) 358–363

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Enhancing bioethanol production from water hyacinth by new combined T

pretreatment methods
⁎
Qiuzhuo Zhanga, , Yan Weia,b, Hui Hana, Chen Wenga
a
Shanghai Key Lab for Urban Ecological Processes and Eco-Restoration, School of Ecological and Environmental Sciences, East China Normal University, 200241
Shanghai, China
b
School of Environmental Science and Engineering, Shanghai Jiao Tong University, 200240 Shanghai, China

G RA P H I C A L AB S T R A C T

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

Keywords: This study investigated the possibility of enhancing bioethanol production by combined pretreatment methods
Fossil fuel for water hyacinth. Three different kinds of pretreatment methods, including microbial pretreatment, microbial
Pretreatment combined dilute acid pretreatment, and microbial combined dilute alkaline pretreatment, were investigated for
Water hyacinth water hyacinth degradation. The results showed that microbial combined dilute acid pretreatment is the most
Bioethanol
effective method, resulting in the highest cellulose content (39.4 ± 2.8%) and reducing sugars production
Phanerochaete chrysosporium
(430.66 mg·g−1). Scanning Electron Microscopy and Fourier Transform Infrared Spectrometer analysis indicated
that the basic tissue of water hyacinth was significantly destroyed. Compared to the other previously reported
pretreatment methods for water hyacinth, which did not append additional cellulase and microbes for hydrolysis
process, the microbial combined dilute acid pretreatment of our research could achieve the highest reducing
sugars. Moreover, the production of bioethanol could achieve 1.40 g·L−1 after fermentation, which could pro-
vide an extremely promising way for utilization of water hyacinth.

1. Introduction
Fossil fuels have been excessively exploited and depleted due to
accelerating industrialization and urbanization since the last century.
Renewable energy is recognized as the next great technology that will
one day replace human’s dependency on fossil fuels (Tasnim et al.,
2017). Bioethanol, which could be derived from lignocellulosic bio-
mass, is certainly drawing increasing attention nowadays due to the
advantages of being readily available, low cost and clean to the en-
vironment (Singh et al., 2015; Das et al., 2015).

⁎
Corresponding author.
E-mail address: qzhzhang@des.ecnu.edu.cn (Q. Zhang).

https://doi.org/10.1016/j.biortech.2017.12.085
Received 7 November 2017; Received in revised form 23 December 2017; Accepted 26 December 2017
Available online 27 December 2017
0960-8524/ © 2017 Elsevier Ltd. All rights reserved.
Q. Zhang et al. Bioresource Technology 251 (2018) 358–363

Water hyacinth (Eichhornia crassipes), which was introduced to

China as an ornamental plant in 1930s, has become one of the most
invasive widespread aquatic weeds due to its rapid growth and high
reproducibility in water body (Yan et al., 2015; Ismail et al., 2017; Feng
et al., 2017). It is listed as one of the first batch of nine invasive plant
species by Ministry of Environmental Protection of China (Zhang et al.,
2016a,b). Under proper climatic conditions, the growth rate of water
hyacinth could reach up to 220 kg/ha/day (Bayrakci and Kocar, 2014).
The rapid growth of water hyacinth has occurred a series of environ-
mental problems, such as block river channels, vegetation damage,
water pollution, irrigation problems etc (Yan et al., 2015; Das et al.,
2015; Adanikin et al., 2017). More importantly, there is no effective
method to eradicate them until now (Singh et al., 2015).
However, as a potential lignocellulosic substrate with high hemi-
cellulosic (44.68 ± 0.39%, w·w−1) content, utilization of water hya- Fig. 1. Growth curve of Phanerochaete chrysosporium.
cinth for bioethanol production seems to be a feasible alternative for
managing it and tackling environmental pollution and energy crisis
activate the fungus, it was cultivated in Potato Dextrose Agar (PDA)
(Das et al., 2016; Sindhu et al., 2017). Furthermore, as aquatic biomass,
medium at 28 °C for 7 days. The growth curve of Phanerochaete chry-
water hyacinth offers the advantage over terrestrial lignocellulosic
sosporium was shown in Fig. 1. It was indicated that the lag phase,
biomass because it does not compete with food for land usage (Biswas
logarithmic phase and stationary phase of P. chrysosporium was belong
et al., 2017). Owing to the physical rigidity and chemical recalcitrance
to 0–72 h, 72–108 h and 108–132 h, respectively. Moreover, SEM
of water hyacinth, pretreatment is an indispensable and pivotal step
showed that P. chrysosporium was filamentous fungus (E-supplementary
that should be performed to overcome these restrictions and increase
data for this work can be found in e-version of this paper online).
the enzymatic digestibility (Cheng et al., 2015; Xu et al., 2016). A well-
Yeast Saccharomyces cerevisiae was also procured from China Center
designed pretreatment method needs to be efficient, cost effective and
of Industrial Culture Collection (No. CICC33068). It was cultivated in
environmentally friendly. There are several pretreatment methods de-
nutrient solution at 30 °C, which include 20 g·L−1 glucose, 20 g·L−1
veloped, including mechanical pretreatment, alkali or acid pretreat-
tryptone and 20 g·L−1 yeast extract powder.
ment, steam explosion, irradiation, hot water treatment and super-
critical CO2 treatment (Lin et al., 2015). Unfortunately, these methods
are limited by some drawbacks, such as extreme condition, large energy
2.2. Pretreatment of water hyacinth
consumption, high toxicity, and restricted application (Xu et al., 2016).
Due to the advantages of low energy consumption and high hydrolysis
Three different kinds of pretreatment methods was used for sma-
efficiency, biological pretreatment, especially using white-rot fungi, has
shed water hyacinth, including microbial pretreatment (MB), micro-
attracted much attention nowadays compared to the other pretreatment
bial-dilute acid pretreatment (MB + AC) and microbial-dilute alkaline
methods (Sindhu et al., 2016; Machado and Ferraz, 2017).
pretreatment (MB + AK).
Phanerochaete chrysosporium, which possessed the highest lig-
Phanerochaete chrysosporium was selected in the MB pretreatment.
ninolytic activity among white-rot fungi, has been successfully em-
Nine gram smashed water hyacinth was added into conical flask, which
ployed for lignin degradation and reducing sugars production in several
contained 180 mL inorganic salt medium. The solid-liquid ratio was
studies (Ansari et al., 2016; Harada et al., 2016). It was reported that
maintained for 1:20 in the pretreatment process. The inorganic salt
Phanerochaete chrysosporium could secret extracellular peroxidases in-
medium contains 0.1 g·L−1 MgSO4·7H2O, 1 g·L−1 KH2PO4, 0.4 g·L−1
cluding over ten lignin peroxidases (Lip), five manganese peroxidases
ammonium tartrate, 0.1 g·L−1 CaCl2, 0.05% Tween-80 and 1 mL trace
(Mnp) and some other copper oxidases during secondary metabolism
element. The trace element is mixed with 0.5 g·L−1 MnSO4, 0.02 g·L−1
(Ansari et al., 2016). However, it was sparsely used for water hyacinth
KAl (SO4)2·12H2O, 0.01 g·L−1 H3BO3, 0.1 g·L−1 FeSO4·7H2O, 0.1 g·L−1
biomass degradation. In addition, few publications were available re-
CoCl2·6H2O, 0.1 g·L−1 ZnSO4·7H2O and 0.01 g·L−1 CuSO4·5H2O After
lated to elaborate analysis of monosaccharide in the hydrolysates and
autoclaving at 121 °C for 20 min, 4 loops of Phanerochaete chrysos-
systematic investigation on effective combined pretreatment methods
porium, which was cultured for 4 days (logarithmic phase), were added
to accelerate water hyacinth hydrolysis process.
into the system. The cells were grown under shaking condition
In the present study, Phanerochaete chrysosporium was chosen for
(150 rpm) at 30 °C for 60 h.
microbial pretreatment of water hyacinth. To enhance the reducing
The residue, which was degraded by Phanerochaete chrysosporium
sugars production, dilute acid and alkaline were subsequently added to
after 60 h, was transferred to glass beaker and then dried at 60 °C. Then
microbial pretreatment system, respectively. The efficiency of water
1% H2SO4 and 4% NaOH was added into the pretreated system for 1 h
hyacinth hydrolysis and bioethanol production by the new combined
soaking at 100 °C, which was identified as MB + AC pretreatment and
system was investigated, and mechanism of stimulatory effect of the
MB + AK pretreatment, respectively.
combined pretreatment system was further explored by SEM and FTIR
analyses.
2.3. Production of bioethanol
2. Materials and methods
The residue of dry water hyacinth after MB + AC pretreatment was
2.1. Materials and microorganism
firstly regulated to neutral pH, then it was centrifuged at 5000 rpm for
5 min. 6 g·L−1 Saccharomyces cerevisiae inoculum was added into the
Water hyacinth was obtained from Huangpu River, Shanghai,
liquid supernatant sample for fermentation process. Nitrogen was aer-
China. It was washed after removing its root and dried in oven at 60 °C
ated to exclude air in the system. The fermentation process was carried
till constant weight followed by smashing using grinder below 40 me-
out at 30 °C under shaking condition (120 rpm). After fermentation
shes for further use.
process, the fermented samples were centrifuged at 5000 rpm for 8 min
Phanerochaete chrysosporium, a white-rot fungus, was procured from
and supernatant was used for bioethanol determination.
China Center of Industrial Culture Collection (No. CICC40299). To

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2.4. Analytical methods
2.4.1. Components of water hyacinth
Water hyacinth is mainly composed of cellulose, hemicellulose and
lignin. The components were determined by the method of Goering and
Vansoest (1970).
2.4.2. Composition of reducing sugar
After pretreatment process, the degradation samples of water hya-
cinth were centrifuged at 13,000 rpm for 6 min. The supernatant, which
was filtered by 0.45 µm pinhole filter, was used for determination of
reducing sugars content.
The total reducing sugars were measured by DNS method (Miller,
1959).
The four main monosaccharides, glucose, arabinose, galactose and
xylose, were determined by High Performance Liquid Chromatography
system (HPLC, 1515-2414, Waters, USA). Deionized water was used as
the mobile phase at a flow rate of 0.6 mL·min−1. The oven temperature
was maintained at 80 °C for 30 min. The detector temperature was kept
at 50 °C. It was shown that the appearance time of glucose, arabinose,
galactose and xylose was 12.103 min, 15.751 min, 14.229 min and
13.217 min, respectively.
The bioethanol production after fermentation by Saccharomyces
cerevisiae was measured by headspace sampling Gas Chromatography
(GC, Agilent 7890A) following our previous method (Zhang et al.,
2016a,b).
2.5. SEM and FTIR analysis
The morphology features of water hyacinth after different pre-
treatment methods were investigated by using Scanning Electron
Microscope (SEM, S4800, HITACHI, JPN) and Fourier Transform
Infrared Spectrometer (FTIR, Nicolet Is5, Thermo Fisher Scientific,
USA), respectively.
3. Results and discussion
3.1. Effect of different pretreatment methods on water hyacinth hydrolysis
3.1.1. Constituent of water hyacinth after pretreatment
Three various kinds of pretreatment methods, including MB,
MB + AC and MB + AK, were applied to investigate their effects on
water hyacinth hydrolysis. The constituents of water hyacinth before
and after pretreatment were shown in Table 1.
The original collected sample of water hyacinth constituted of
19.8% cellulose, 49.0% hemicellulose and 4.8% lignin, which was si-
milar to previous reports (Das et al., 2016). Compared to rice straw and
the other plant derived biomass, the relatively higher cellulose and
hemicelluloses content of water hyacinth make it more feasible and
suitable for bioethanol production (Ganguly et al., 2012).
The total dry weight of water hyacinth was all decreased after three
kinds of pretreatment methods. It was decreased by 26.67% after mi-
crobial pretreatment, which was attributed to the contribution of
Phanerochaete chrysosporium that used lignocellulose as carbon source.
The total dry weight of water hyacinth was further decreased after di-
lute acid pretreatment and dilute alkaline pretreatment, which mani-
fested that both dilute acid and dilute alkaline conditions played an
important role for lignocellulosic degradation.
It is obvious that cellulose percentage increased after all kinds of
pretreatment methods, whereas the hemicellulose and lignin percen-
tage decreased. It was reported that although several fungi could in-
crease polysaccharide digestibility, only a small part of species are able
to degrade large amounts of lignin, and preserve cellulose and part of
hemicellulose (Machado and Ferraz, 2017). The single microbial pre-
treatment of our results verified the consequence.
MB + AC made the percentage of cellulose content increased the
highest, which was 1.99 times higher than that in untreated sample.
MB + AK exhibited the most effective lignin removal ability, which
could make the percentage of lignin content decreased by 33.3%. It was
in accordance with the result of Bayrakci and Kocar (2014), in which it
proved that both sodium hydroxide and ammonia could highly disrupt
the lignin content of lignocelluloses. Lai et al. (2017) also indicated that
NaOH could remove lignin more efficiently and reserve the majority of
hemicellulose, which was identified by our results.
3.1.2. Reducing sugars production after pretreatment
The reducing sugars production after three different kinds of pre-
treatment methods were shown in Table 2.
Compared to single MB method, the combined microbial-chemical
method could highly increase the production of reducing sugars in
water hyacinth hydrolysates. The reducing sugars could achieve
430.66 mg·g−1 and 402.10 mg·g−1 after MB + AC and MB + AK pre-
treatment, respectively. Obviously, the combined pretreatment method
was proved to be more efficient for water hyacinth pretreatment.
The four monosaccharides (glucose, arabinose, galactose and xy-
lose), which were recognized as the main constituent of reducing sugars
in water hyacinth hydrolysates, were further detected for their con-
centration by HPLC analysis. It was shown that all of the four mono-
saccharides increased after combined pretreatment compared to single
MB pretreatment, which reconfirmed the favorable efficiency of com-
bined pretreatment method on water hyacinth hydrolysis. After
MB + AC and MB + AK pretreatment, the production of glucose could
achieve 164.11 mg·g−1 and 182.35 mg·g−1, respectively. Dilute acid
was more facilitated for the production of galactose and xylose,
whereas dilute alkaline favored the production of glucose and arabi-
nose. It was reported that hemicellulose disintegrates and xylose gets
release into solution after dilute acid pretreatment, while alkaline
pretreatment preserves part of hemicellulose and mainly removes the
lignin content (Lin et al., 2016; Aswathy et al., 2010), which could il-
luminate our results.
The contents of four monosaccharides in different pretreated sam-
ples could be further explained by the constituent results of water
hyacinth in Table 1. The hexose (glucose) concentration was higher in
MB + AK pretreated hydrolysates than that in MB + AC pretreated
hydrolysates, due to degradation of high cellulose content in MB + AK
process. Pentose, which was mainly consisted of arabinose, galactose
and xylose, was the hydrolysates of hemicellulose. It was shown that
the concentration of pentose was higher in MB + AC pretreated

Table 1
Constituent of water hyacinth.

Samples Cellulose Hemicellulose Lignin Others Total dry weight of biomass

(g) (%) (g) (%) (g) (%) (g) (%) (g)

CK 1.78 ± 0.04 19.80 ± 0.40 4.41 ± 0.23 49.00 ± 2.50 0.43 ± 0.03 4.80 ± 0.30 2.38 ± 0.10 26.40 ± 1.10 9.00 ± 0.00
MB 1.71 ± 0.01 25.90 ± 0.10 1.93 ± 0.14 29.20 ± 2.10 0.28 ± 0.05 4.20 ± 0.70 2.69 ± 0.07 40.70 ± 1.00 6.60 ± 0.30
MB + AC 1.10 ± 0.08 39.40 ± 2.80 0.49 ± 0.03 17.60 ± 1.00 0.13 ± 0.06 4.80 ± 0.20 1.07 ± 0.04 38.20 ± 1.30 2.80 ± 0.60
MB + AK 1.17 ± 0.05 29.90 ± 1.20 0.90 ± 0.03 23.00 ± 0.80 0.12 ± 0.03 3.20 ± 0.80 1.71 ± 0.04 43.90 ± 0.90 3.90 ± 0.50

CK: before pretreatment; MB: microbial pretreatment; MB + AC: microbial-dilute acid pretreatment; MB + AK: microbial-dilute alkaline pretreatment.

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Table 2
Composition of reducing sugars in pretreated water hyacinth hydrolysates.

Samples Reducing sugars Glucose Arabinose Galactose Xylose

(mg·g−1) (mg·g−1) (mg·g−1) (mg·g−1) (mg·g−1)

MB 208.32 ± 0.80 108.26 ± 8.34 94.24 ± 0.49 22.64 ± 5.09 18.40 ± 2.15
MB + AC 430.66 ± 2.32 164.11 ± 5.60 99.45 ± 8.64 119.18 ± 6.67 43.10 ± 3.20
MB + AK 402.10 ± 1.32 182.35 ± 8.67 141.64 ± 7.52 43.98 ± 6.62 27.84 ± 6.70

MB: microbial pretreatment; MB + AC: microbial-dilute acid pretreatment; MB + AK: microbial-dilute alkaline pretreatment.

hydrolysates than that in MB + AK pretreated hydrolysates, which was
coincidence with the results on highly degradation of hemicellulose
after MB + AC pretreatment. Moreover, it was found that the alkaline
environment could facilitate the production of arabinose, whereas the
acid environment facilitated the production of galactose and xylose.
Furthermore, although glucose could be better utilized by yeast for
subsequent fermentation to bioethanol, the utilization of pentose by
genetic engineered microbes became a hot research direction and
proved feasible in recent years (Lopez-Hidalgo et al., 2017; Farias et al.,
2017; Lin et al., 2016). Meanwhile, it was indicated that MB + AC
promoted recovery of hemicelluloses as monomers in the liquid fraction
and high cellulose content in the solid fraction (Dagnino et al., 2013).
Thus, MB + AC was more recommended for water hyacinth pretreat-
ment than MB + AK in consideration of the higher production of re-
ducing sugars and lower cost (Baral and Shah, 2017).
3.1.3. Comparison of different pretreatment methods on water hyacinth
hydrolysis
Several researches are going on the direction for the conversion of
water hyacinth waste to bioethanol production. The comparison of
different pretreatment methods on water hyacinth hydrolysis was listed
in Table 3. Aswathy et al. (2010) used 2% NaOH to react with water
hyacinth samples (10% biomass loading) for 1 h at 95 °C, followed by
hydrolysis of sample by 8 FPUs·g−1 cellulase after adding 0.15% Tween
for 48 h at 45 °C, which could achieve the highest reducing sugars
(731 mg·g−1) in the listed references. However, the expensive cost of
cellulase represents one of the biggest obstacles for competitiveness of
the cellulosic ethanol (Narra et al., 2017). Researchers all over the
world are striving for cutting down the cellulase cost by various
strategies, including screening of higher cellulase producing microbes,
increasing the efficiency of enzymes, adopting cheaper bioprocess
technology and reutilizing the enzymes (Singhania et al., 2015).
Nevertheless, the cost of cellulase could not push down much further,
which perplex the bioethanol production in large scale (Gomes et al.,
2016). In our present study, no additional cellulase was needed in the
process of reducing sugars production, which might be attributed to the
promising effect by Phanerochaete chrysosporium (Li et al., 2017; Han
et al., 2017).
Compared to the studies which did not append additional cellulase
and microbes for hydrolysis process (Awasthi et al., 2013; Thi et al.,
2017), the production of 430.66 mg·g−1 reducing sugars in our present
study were the highest. It should be point out that the production of
reducing sugars in present research was even higher than some of the
studies, which applied hydrolysis process. Meanwhile, as for the con-
stituents of lignocellulose after pretreatment, the cellulose content in
present study was the highest (39.4 ± 2.8%).
Moreover, although 402.93 mg·g−1 reducing sugars were achieved
after sulfuric acid pretreatment and cellulase hydrolysis in our previous
study (Zhang et al., 2016a,b), 750 U cellulase was added for 96 h at
45 °C in the process. This could again verify the advantage of our pre-
sent pretreatment method.
3.2. Structural analysis of water hyacinth after pretreatment
3.2.1. SEM analysis
Since pretreatment could facilitate the water hyacinth hydrolysis
process, it became of interest to examine the morphological changes by
SEM (E-supplementary data for this work can be found in e-version of

Table 3
Comparison of different pretreatment methods on water hyacinth hydrolysis.

References Pretreatment method Hydrolysis condition Reducing sugars Constituents of lignocellulose after pretreatment
(mg·g−1)
Cellulose (%) Hemicellulose (%) Lignin (%)

Sukumaran et al. NaOH T. reesei for 96 h and A. niger for 72 h at 30 °C, then 710 / / /
(2009) hydrolysis at 45 °C
Aswathy et al. NaOH 8 FPUs/g cellulase after adding 0.15% Tween for 48 h 731 / / /
(2010) at 45 °C
Guragain et al. Crude glycerol 210 U cellulase for 2.5 h at 50 °C 451 / / 10.1 ± 0.5
(2011)
Awasthi et al. (2013) H2SO4 No additional hydrolysis 12.63 / / /
Yan et al. (2015) NaOH/H2O2 17.43 FP/mL cellulase for 3 days at 50 °C 223.53 36.33 17.72 6.17
Cheng et al. (2015) CaO2 0.2 mL cellulase and 0.05 mL beta-glucosidase per 325 / / /
gram dry biomass for 72 h at 50 °C
Das et al. (2015) NaOH 49.56 U/g cellulase, 280.33 U/g xylanase and 0.13% 567.2 31.027 17.71 2.834
Tween for 60 h at 50 °C
Zhang et al. H2SO4 750U cellulase for 96 h at 45 °C 402.93
(2016a,b)
Xu et al. (2016) ILM 0.05 mL/g Cellulase and 0.05 mL/g beta-D- 563.7 / / /
Glucosidase for 48 h at 50 °C
Chang et al. (2017) US-IL-SDS 50 FPU/g Cellulose complex enzyme and 320 CBU/ 233.9 38.20 ± 1.02 30.80 ± 1.01 16.20 ± 1.40
mL β-glucosidase enzyme for 48 h at 50 °C
Xiao et al. (2017) Fenton Agent 30 FPU/g cellulase at 50 °C / 35.65 50.92 7.28
Thi et al. (2017) H2SO4/Subcritical No additional hydrolysis 264.4 69.4 / /
water
Present study MB + AC No additional hydrolysis 430.66 39.4 ± 2.8 17.6 ± 1.0 4.8 ± 0.2

ILM: surfactant-free ionic liquid microemulsions; US-IL-SDS: ultrasound-ionic liquid pretreatment assisted by sodium dodecyl sulfate; MB + AC: microbial-dilute acid pretreatment.

361
Q. Zhang et al.
this paper online).
It was indicated that untreated rice straw exhibited a continuous
surface and rigid and highly ordered fibrils, while the fibrils after mi-
crobial pretreatment were distorted. The micro fibrils were separated
from initial connected structure and fully exposed, thus increasing the
external surface area and porosity of water hyacinth, which was in
agreement with our previous study where rice straw was used as sub-
strate (Zhang et al., 2017; Hou et al., 2017). Meanwhile, it was mani-
fested that the secretion of cellulase by Phanerochaete chrysosporium
might play an important role for water hyacinth hydrolysis. The cel-
lulase production by Phanerochaete chrysosporium will be investigated in
future.
After degrading by chemical reagent in both MB + AC and
MB + AK system, basic tissue of water hyacinth was further severely
destroyed; even small holes were breakdown in the water hyacinth
surface. It was reported that NaOH could lead to an increase in internal
surface area, a decrease in the degree of polymerization and crystalline
structure (Bayrakci and Kocar, 2014). Several studies revealed that
H2SO4 was effective in recovering most of the sugars especially from
corn stover as well as other biomasses (Hafid et al., 2017). These above
mentioned researches verified our present study.
3.2.2. FTIR analysis
The characteristic peaks in FTIR spectra were shown in Table 4 (E-
supplementary data for this work can be found in e-version of this paper
online). It was indicated that the main characteristic peaks were basi-
cally the same, which appeared in 3323 cm−1, 2922 cm−1, 1635 cm−1,
1410 cm−1, 1162 cm−1, 1058 cm−1 and 896 cm−1, respectively. There
was no new characteristic peaks appeared after pretreatment. However,
the adsorption strength varied from each other, which manifested that
pretreatment indeed changed the morphological structure of water
hyacinth.
The peaks around 3323 cm−1 stands for stretching vibration and
overlapping of OeH, which is recognized as main infrared sensitive
groups of lignocellulose. The band strengthened after pretreatment,
which illustrated that cellulose structure was fractured after effective
pretreatment method. The peaks around 2922 cm−1, which illustrated
the symmetric or dissymmetric stretching vibration of CeH group, is
one of the characteristic peaks of cellulose (Zhang et al., 2017). The
shape of this peak changed after MB + AK pretreatment, which might
be due to reactions between microbes and the other molecular groups.
The peaks around 1515 cm−1 and 1162 cm−1, which are recognized
as main infrared sensitive groups of lignocellulose, represent benzene
ring vibrations and stretching vibration of CO-OR, respectively. The
band at 1515 cm−1 strengthened tremendously after MB + AK pre-
treatment, suggesting that alkaline played a key role in lignin de-
gradation. The result is consistent with Table 1, which showed that the
lignin content of water hyacinth reduced a lot after pretreatment.
Meanwhile, several previous studies described that phenols were one of
the most abundance compounds in lignocellulosic biomass after alka-
line pretreatment, which is coincidence with our results.
The peak around 1058 cm−1, which stands for CeO stretching of
hemicellulose and cellulose, was tremendously changed after MB + AC
pretreatment. It was reported that MB + AC could promote hydrolysis
Table 4
Characteristic peaks in FTIR spectra of water hyacinth.
Adsorption peak (cm−1) Intensity
3323 Strong
2922 Middle
1635 Relatively strong
1515 Middle
1162 Relatively weak
1058 Strong
896 Weak
362
Bioresource Technology 251 (2018) 358–363
Fig. 2. Bioethanol production after fermentation by Saccharomyces cerevisiae.
of hemicelluloses and part of the amorphous cellulose that results in
high recovery of hemicelluloses, making this kind of pretreatment a
promising method for the production of lignocellulosic bioethanol
(Dagnino et al., 2013).
The peak around 896 cm−1 stands for CeH deformation of skeleton
vibration of saccharides and cellulose, and it is the characteristic peak
of β-glucosidic linkages amid monosaccharide units. The peak was
weaker after pretreatment, which manifested that the fibers of water
hyacinth was exposed and the β-glucoside bond was broken down.
Because the breakage of β-glucoside bond is a rate-limiting step in
lignocellulosic biomass degrading process, the weakened β-glucoside
bond after pretreatment could enormously promote the efficiency of
water hyacinth hydrolysis.
3.3. The production of bioethanol by Saccharomyces cerevisiae
The production of bioethanol by Saccharomyces cerevisiae was
shown in Fig. 2. Because of abundant reducing sugars and mass pro-
pagation of S. cerevisiae, rapid fermentation process occurred at the first
24 h. The production of bioethanol could achieve the maximum value
(1.40 g·L−1) at 24 h. However, bioethanol production was stable after
24 h, which might be due to the inhibition effect of bioethanol on
growth of S. cerevisiae (Xue et al., 2016).
In our previous study, 1.289 g·L−1 bioethanol was produced by
whole plant of water hyacinth at the optimum condition in fermenta-
tion process (Zhang et al., 2016a,b). It was found that 8.61% higher
bioethanol could be achieved in the present research along with no
cellulase was added to the system, which could provide an extremely
promising way for water hyacinth utilization.
4. Conclusion
Using water hyacinth to bioethanol production seems to be a sus-
tainable management of this notorious weed. The results showed that
microbial combined dilute acid pretreatment is a promising pretreat-
ment method for water hyacinth hydrolysis, which could achieve
430.66 mg·g−1 reducing sugars and 1.40 g·L−1 bioethanol without any
additional cellulase. The microbial pretreatment could help in
Affiliation of characteristic peaks
Stretching vibration and overlapping of OeH
Symmetric or dissymmetric stretching vibration of CeH group
Stretching vibration of C]O (lignin)
Characteristic group vibrations of benzene ring (lignin)
Stretching vibration of CO-OR (hemicellulose)
CeO stretching of hemicellulose and cellulose
CeH deformation of skeleton vibration of saccharides and cellulose
Q. Zhang et al.
delignification, and the dilute acid could further break down the
hemicellulose part. Thus, it is highly recommended for water hyacinth
pretreatment in bioethanol production considering both of its extremely
low cost and high production of reducing sugars and bioethanol.
Acknowledgements
This project is funded by National Natural Science Foundation of
China (NSFC, No. 31400513) and Shanghai Science and Technology
Committee (Nos. 17295810603, 17DZ1202804). The authors would
like to thank them for funding this work. The authors are thankful to Dr.
Varenyam Achal for improving the language of the manuscript.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in the
online version, at http://dx.doi.org/10.1016/j.biortech.2017.12.085.
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