Waste Management xxx (2017) xxx–xxx

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Waste Management
journal homepage: www.elsevier.com/locate/wasman

Over production of fermentable sugar for bioethanol production from

carbohydrate-rich Malaysian food waste via sequential acid-enzymatic
hydrolysis pretreatment
Halimatun Saadiah Hafid a, Nor ’Aini Abdul Rahman a,⇑, Mohd Noriznan Mokhtar b, Ahmad Tarmezee Talib b,
Azhari Samsu Baharuddin b, Umi Kalsom Md Shah a
a
Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia
b
Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia

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

Article history: In Malaysia, the amount of food waste produced is estimated at approximately 70% of total municipal
Received 30 November 2016 solid waste generated and characterised by high amount of carbohydrate polymers such as starch, cellu-
Revised 8 May 2017 lose, and sugars. Considering the beneficial organic fraction contained, its utilization as an alternative
Accepted 11 May 2017
substrate specifically for bioethanol production has receiving more attention. However, the sustainable
Available online xxxx
production of bioethanol from food waste is linked to the efficient pretreatment needed for higher pro-
duction of fermentable sugar prior to fermentation. In this work, a modified sequential acid-enzymatic
Keywords:
hydrolysis process has been developed to produce high concentration of fermentable sugars; glucose,
Food waste
Acid-pretreated
sucrose, fructose and maltose. The process started with hydrothermal and dilute acid pretreatment by
Enzymatic hydrolysis hydrochloric acid (HCl) and sulphuric acid (H2SO4) which aim to degrade larger molecules of polysaccha-
Fermentation ride before accessible for further steps of enzymatic hydrolysis by glucoamylase. A kinetic model is pro-
Saccharomyces cerevisiae posed to perform an optimal hydrolysis for obtaining high fermentable sugars. The results suggested that
Bioethanol a significant increase in fermentable sugar production (2.04-folds) with conversion efficiency of 86.8%
was observed via sequential acid-enzymatic pretreatment as compared to dilute acid pretreatment
(
42.4% conversion efficiency). The bioethanol production by Saccharomyces cerevisiae utilizing fer-
mentable sugar obtained shows ethanol yield of 0.42 g/g with conversion efficiency of 85.38% based on
the theoretical yield was achieved. The finding indicates that food waste can be considered as a promising
substrate for bioethanol production.
Ó 2017 Published by Elsevier Ltd.

1. Introduction to monosaccharide sugars and other high value products through

In Malaysia, it is estimated 33,000 tonnes of waste produced
daily in 2012 and the amount of waste generated are keep increas-
ing with rapid urbanisation, population growth and industrialisa-
tion especially in the urban areas (News Strait Times, 2013).
Organic waste particularly food waste are mainly discharged from
households, restaurant and leftovers from food industries (Hafid
et al., 2011). Food waste indeed not only contains high moisture
and humidity, but it is mainly composed of high carbohydrate
polymers such as starch (more than 40%), proteins (4–5%), cellu-
lose (2–3%), and total sugars (more than 50%) can be converted
⇑ Corresponding author.
E-mail addresses: hs_hafid@yahoo.com (H.S. Hafid), nor_aini@upm.edu.my
(A.R. Nor ’Aini), noriznan@upm.edu.my (M.N. Mokhtar), mezee.t@gmail.com
(A.T. Talib), azharis@upm.edu.my (A.S. Baharuddin), umikalsom@upm.edu.my
(M.S. Umi Kalsom).
environmental-friendly approaches (Kiran et al., 2014; Uncu and
Cekmecelioglu, 2011). Interestingly, recent studies shows that
the carbohydrate compound of food waste can be potentially used
for fuel bioethanol production in which is perceived will not com-
pete with global food supply (Uncu and Cekmecelioglu, 2011; Ma
et al., 2008).
Hydrolysis of carbohydrate polymers in food waste through
pretreatment and enzymatic hydrolysis resulting in breakage of
glycosidic bonds with releasing monosaccharides sugars such as
glucose, mannose, sucrose and fructose, which are more amenable
to fermentation for production of high value materials such as bio-
fuels including methane, bioethanol, hydrogen and biodiesel (Kiran
et al., 2014). However, due to the complexity composition of food
waste, the hydrolysis of food waste is not straightforward. In addi-
tion, the effective pretreatment method in the hydrolysis step is
usually considered as the rate limiting steps in overall process.

http://dx.doi.org/10.1016/j.wasman.2017.05.017
0956-053X/Ó 2017 Published by Elsevier Ltd.

Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
2 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx
Therefore, numerous pretreatments by thermal, chemical and bio-
logical means is aimed to disrupt some structural characteristic of
food waste and increase the accessibility of enzyme to hydrolyse
polysaccharide into sugars monomers.
In the fermentable sugar and bioethanol production from food
waste, the cost of traditional hydrolytic enzyme such as amylase
is a major economic barrier due to the cost of the amylase and long
incubation period. Therefore, an alternative pretreatment method
in replacing the use of enzyme are of interest in the conversion
of food waste into fermentable sugar and bioethanol. Of the many
pretreatment technologies, dilute acid pretreatment stands out
having been already observed in a large number of studies as well
as appearing more economically feasible at larger scale than other
pretreatment technologies (Redding et al., 2011). Dilute acid pre-
treatment has proven to be effective in recovering most of the sug-
ars especially from corn stover as well as other biomasses (Shen
and Wyman, 2011). Furthermore, hydrolysis of carbohydrate poly-
mers by dilute acid has various advantages such as short reaction
time and much simpler pretreatment (Guerra-Rodriguez et al.,
2012). A lot of studies has been done on the conversion of lignocel-
lulosic materials to fermentable sugars with different reactants
such as sulphuric acid and hydrochloric acid (Shen and Wyman,
2011; Zhao et al., 2014; Yu et al., 2013). The hydrolysis of food
waste by dilute acid pretreatment is postulated to be highly depen-
dent on an increase in temperature, acid concentration, reaction
time, substrate concentration and composition that will directly
speed up the hydrolysis. Hydrolysis of polysaccharide by
hydrothermal and dilute acid pretreatment is assume as a
pseudo-homogeneous irreversible reaction in order to overcome
the problem of the heterogeneity and complexity of the biomass
(Jin et al., 2011). The kinetic model was developed based on the
hypothesis of that the whole biomass could be hydrolysed into
monosaccharides, therefore, the overall scheme of the reaction
was generalised to;
Polymer ! monomers ! decomposition product
The main drawbacks of hydrothermal and dilute acid pretreatment
is generation of degradation products due to the extreme condition
applied such as high temperature and low pH during hydrolysis
process (Khawla et al., 2014). The formation of degradation prod-
ucts such as furfural, 5-hydroxymethyl-2-furaldehyde (HMF) and
acetic acid continuously may reduce the concentration of fer-
mentable sugar particularly glucose in the hydrolysate. The hydrol-
ysis strategies that can lead to high productions sugars and
simultaneously control the formation of degradation products in
food waste is very limited documented in the literature as com-
pared to other lignocellulosic biomass. Therefore, this study pro-
posed a new sequential hydrothermal and dilute acid pretreament
aiming at sustaining high concentration of fermentable sugar pro-
duced for longer incubation time before degraded. The proposed
strategies take into account the interdependent of both hydrolysis
method where enzymatic hydrolysis needs pretreatment either by
hydrothermal or dilute acid to gelatinized a carbohydrate compo-
nents of food waste that’s allows the enzymes to penetrate easily
into carbohydrate linkages contributing to a more efficient reaction.
Meanwhile, additional of enzymes in the hydrolysate needs supple-
mentary steps for cooling and neutralize the acidic medium, thus,
controlling the lethal concentration of degradation products. This
work deals with the kinetic study of hydrothermal and dilute acid
pretreatment in order to provide an in-depth understanding of
the kinetics behaviour of polysaccharide hydrolysis during pretreat-
ment for further optimization processes. Food waste was subjected
to hydrothermal, sulphuric acid and hydrochloric acid–catalysed
systems in various reaction temperatures (80–100 °C) and concen-
trations (0.5–2%). Experimental data were obtained by measuring
the fermentable sugars released from the substrate and the intro-
Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
duced model was applied to fit the experimental data for estimating
unknown parameters. The effects of temperatures, acid reactants
and concentrations to the production of fermentable sugars were
evaluated. A sequential dilute acid-enzymatic pretreatment was
carried out in laboratory-scale hydrolysis reactor to examine the
efficacy of the pretreatment in dealing with high solid content,
heterogeneity and inconsistent composition of the food waste,
notwithstanding to ensure complete degradation occurs and
enhanced the sugar formation. Despite of the many pretreatment
method tested in the literature, there is still a need to establish
more defined pretreatment based approach for food waste conver-
sion and evaluate the processing conditions for an effective fer-
mentable sugar production. Subsequently, the recovered
fermentable sugar of the hydrolysate will be tested for its fer-
mentability for bioethanol production by locally isolated yeast Sac-
charomyces cerevisiae in separate hydrolysis and fermentation.
2. Materials and methods
2.1. Raw material
The food waste used in this study was collected from food
courts in Universiti Putra Malaysia, Serdang, Selangor. Plastics,
glasses, tissues and other inorganic materials were separated out.
In order to standardize the composition of food waste throughout
the study, the remaining organic fraction; rice, meats and vegeta-
bles were set at 3:1:1 ratio and ground with water at 1:1 vol ratio
using a laboratory Waring blender to ensure liquidity of the sam-
ple. The samples were stored at 20 °C (Hafid et al., 2011). The
overview of the experimental design is illustrated in Fig. 1.
2.2. Hydrothermal and dilute acid pretreatment
Samples were taken before and after each treatment for evalu-
ð1Þ ation of the physicochemical properties of the food waste. Blended
food waste was subjected to treatments with hot distilled water as
a reactant, and a dilute acid pretreatment using HCl and H2SO4 at
various concentrations (0.5%, 1.0%, 1.5%, 2.0% v/v). Both hot water
and dilute acid pretreatments were conducted at several of tem-
perature values (80 °C, 90 °C, and 100 °C) and incubation times
(0, 30, 60, 90, 120, 180, 240, 300 min). All treatment methods were
carried out in 250 mL shake flask containing 100 mL of blended
food waste. The food waste was incubated in a water bath shaker
at 100 rpm and treated with respect to incubation time, tempera-
ture and different chemicals and concentrations as stated before.
After the treatment, both raw and treated food waste were anal-
ysed for organic compound contents. Three replicates were done
for each treatment.
2.3. Mathematical modelling
The kinetic modelling of dilute acid hydrolysis of food waste
polysaccharides is described by Saeman’s model (Zhao et al.,
2014) and represented in the following equation;
r r¼1
PolysaccharideðFood wasteÞ mi¼1 ! Monosaccharide ðaqÞ
r r¼2
 ðFermentable sugarsÞmi¼2 ! Degraded material mi¼3 ð2Þ
where mi represents the concentration of components such as food
waste (i = 1), sugars (i = 2) and degraded material (i = 3) (g/L) and
rr=1 is the reaction rate of monosaccharides or fermentable sugar
formation (g/Lh) and rr=2 is the reaction rate of fermentable sugar
decomposition (g/L h); Based on the reaction model, the mass bal-
ance for food waste hydrolysis is;
mi ¼ V:C i ð3Þ
 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx 3

Food waste preparation

Collected kitchen waste
Restaurant waste/ cafeterias/
residential areas

Recycling Centre

Rice Meats Vegetables In-organic

Ground kitchen waste

Thermometer
Sequential acid-enzyme
hydrolysis system

pH Liquid sampling
Enzyme meter valve
reservoir
Acid tank Hydrolysate
reservoir

Water bath

Inoculum Hydrolysate
Solid-liquid
separation
S. cerevisiae
Bioethanol production

Preculture
Liquid
sampling

Fermentation
broth
Bioethanol production

Bioethanol Solid-liquid separation Solid

Fig. 1. Experimental setup for sequential acid-enzymatic hydrolysis pretreatment for fermentable sugar and bioethanol production.

2 3
where V is the volume of reactant solution (L) and C is the concen- 1 0
6 7
tration of component. The overall component balance equation for lir ¼ 41 1 5 ð7Þ
food waste was assumed as;
0 1

C i¼1 ¼ C i0¼1  C i¼2  C i¼3 ð4Þ

where for the r1, the hydrolysis reaction started with the degrada-
tion of food waste [1] to fermentable sugar [0] whereas for the
where Ci0=1 is initial concentration of the food waste, 117 g/L. r2, the negative number of fermentable sugar [1] indicated the
The production of total sugars (Cp) is introduced as; degradation of sugar to degraded material [1].
C p ¼ C i¼2  C i0¼2 ð5Þ The following equation was used to calculate the rate of sugar
formation (rr=1) and degradation (rr=2) during hydrothermal with-
where Ci0=2 is initial concentration of total sugars. The mass rate or out and with dilute acid hydrolysis as follow:
kinetic of mass is;
!
X2 C 2i
dmi r r ¼ kr  ð8Þ
¼ V: ðlir  r r Þ ð6Þ C 2i þ K 2Sr
dt r¼2

The stoichiometric coefficient of the reaction can be represented in where kr is the hydrolysis rate coefficient (min1); KSr is the sub-
matrix form as: strate limitation constant (g/L). The hydrolysis rate coefficient (kr)

Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
4 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx
for dilute acid reaction is assumed to have temperature and acid
concentration dependence as:
 br  mr  (Hafid et al., 2016). The enzymatic hydrolysis was performed in
T CH
kr ¼ kr0   þ1
373 K Hr
kr0 is a constant rate of hydrolysis (min1), T is the temperature (K);
br and mr are constants for temperature and acid concentration,
respectively; CH is the acid concentration (%) and KHr is the limita-
tion constant for acid concentration (%). The values of mr and KHr
were interrelated in which at high value of KHr will bring down
the overall reaction rate. The approach of these models take into
account the hydrolysis rate without acid addition, and therefore,
for the hydrothermal reaction, the hydrolysis rate coefficient, kr,
was assumed to be independence of the acid concentration factor
which is defined as follows:
 br
T
kr ¼ kr0 
373
During the hydrolysis of food waste, the presence of the dilute acid
in the substrate affected considerable amounts of free H+ that pro-
mote the protonation reaction with the nitrogen lone pair of the
amino group from the food waste (Yu et al., 2013). Therefore, this
model also takes into consideration the consumption of acid. The
rate of acid consumption can be described as:
dC H X trate was kept at 20 °C and was subsequently used as a fermen-
¼ ðY Hr  rr Þ
dt
where YHr is the yield of acid consumption (% acid per g/L substrate).
This model proposes that fermentable sugar is released by the
hydrolysis of food waste polysaccharide of food waste and further
decomposed to degraded materials under certain conditions. Both
of the release and decomposition are generally considered to be
occurred at temperature less than 100 °C with acid reactant of
below than 2% concentration.
2.4. Statistical analysis and parameter estimation
The determination of means and standard deviation of samples
were analysed using IBM SPSS Statistics Version 20.0 (IBM Corp,
Armonk, NY). Difference means of samples were assessed using
one way analysis of variance (ANOVA) and post-hoc analysis using
Duncan Multiplle Range Test with significance level p < 0.05. The
parameter estimation and dynamic simulation were conducted
using gPROMSÒ 3.6.0 (Process Systems Enterprise Ltd, UK). The
experimental data used for data analysis of model parameters were
the average of duplicates experimental results. The kr=1 and kr=2
can be obtained from estimated parameters of the model because
the food waste and fermentable sugar concentration as a function
of time was known.
2.5. Sequential acid-enzymatic hydrolysis
One L of ground food waste (1720 g wet basis) was subjected to
pretreatment by dilute hydrochloric acid of 1.5% (v/v) at 90 °C for
3 h incubation time in 2 L of hydrolysis reactor constructed with
a heating shell (Fig. 1). The temperature was kept constant by set-
ting the temperature of the thermal fluid circulation through the
outer heating shell of the reactor (Li and Jin, 2015). Agitation was
applied at 100 rpm and the samples were withdrawn for every
1 h. The pH value was monitored at certain interval time. After
dilute acid pretreatment, the food waste hydrolysate was then kept
cooling to 60 °C with the adjusted initial pH of 5.0 using 1 M HCl
and 5 M NaOH. The pretreated food waste was subjected to enzy-
matic hydrolysis using glucoamylase enzyme (extracted from
Aspergillus niger) provided by Sukanan (Weifang) Biotechnology
Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
Co. Ltd with an initial enzyme loading of 85 U/mL. The activity of
glucoamylase was determined according to Standard Method
the same reactor at 100 rpm for another 6 h (Hafid et al., 2015).
ð9Þ
The hydrolysate obtained was analysed for sugar concentration.
The apparent total sugar production efficiency (ATPE) and
degree of food waste decomposition (DoD) were defined as;
DC s
ATPE ¼  100 ð12Þ
C ITCHO
C cTCHO
DoD ¼ 1   100 ð13Þ
C ITCHO
where DCs is different in sugar concentration (g/L); CITCHO and CCTCHO
are initial concentration of total carbohydrate and current concen-
tration of total carbohydrate, g/L, respectively.
ð10Þ 2.6. Recovery of sugar hydrolysate
Once the physical-chemical hydrolysis and enzymatic sacchari-
fication was completed, the hydrolysate was then centrifuged at
10,000 rpm for 15 min and filtered on 0.8 lm, cellulose acetate fil-
ter paper by using vacuum pump filter. The filtrate was evaporated
using rotary evaporator (Buchi Rotavapor R-200, Swiss) until
desired concentration sugar achieved (Omar et al., 2009). The fil-
ð11Þ tation feedstock in the bioethanol fermentation.
2.7. Bioethanol production from food waste hydrolysate
2.7.1. Microorganisms and inoculums preparation
Saccharomyces cerevisiae used in this study was isolated from
Bakers yeast and grown on yeast peptone dextrose (YPD) agar
plates containing yeast extract 10 g/L; peptone 20 g/L; D-glucose
20 g/L; and bacto agar 20 g/L. The yeasts were incubated at 30 °C
for 48 h. Then, a loopful of yeasts were transferred in 100 mL
growth media containing 10 g/L of yeast extract; 20 g/L of peptone
and 20 g/L of glucose and incubated at 30 °C for 12–20 h for pre-
culture (Natarajan et al., 2012).
2.7.2. Batch ethanol fermentation
Ethanol fermentation was carried out in 250 mL of shake flasks
containing 100 mL of sterile fermentation mixture composed of the
following substances per L distilled water; 5 g yeast extract; 5 g/L
of peptone; 20 g glucose; 1 g KH2PO4; and 1 g MgSO47H2O The fer-
mentable sugars in food waste hydrolysate were set at 20 g/L and
commercial glucose was used as a control. Sulphuric acid (5 M) and
sodium hydroxide (5 M) were used to adjust the initial pH at 6.5–
7.0. The yeasts inoculums were added at a ratio of 10% (v/v) to the
fermentation media under aseptic conditions. The flasks were
incubated at temperature and agitation speed of 30 °C and
120 rpm, respectively. Samples were taken every six hours inter-
vals prior analysis of dry cell weight, ethanol, and reducing sugars.
The fermentation period was kept for 120 h.
2.8. Analytical methods
2.8.1. Physicochemical analysis of food waste hydrolysate
The ground food waste was initially analysed for physicochem-
ical characteristics including determination of pH, total solid (TS),
total volatile solid (TVS), total suspended solid (TSS), and volatile
suspended solid (VSS). The analyses were based on the Standard
Method (APHA, 1985). Total chemical oxygen demand (TCOD)
and soluble chemical oxygen demand (SCOD) were measured by
HACH COD (USA) analyser in accordance with HACH method. The
measurement of nitrogen content in the food waste was according
 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx
to the Kjeldahl method. Crude fiber was determined using acid
base digestion and solubilisation whereby crude fat was measured
by the Soxhlet extraction method using petroleum ether (APHA,
1985). Moisture and ash content were determined based on the
Standard Method (APHA, 1985). The remaining percentage based
on dry weight basis was assumed to be the total carbohydrate con-
tent (Uncu and Cekmecelioglu, 2011). For the determination of
monosugars and organic acids before and after each pretreatments,
the food waste hydrolysate was centrifuged at 4000 rpm for
20 min to separate liquid and solid particles before filtered through
0.45 mm Nylon Whatman filter. Monosugars produced from food
waste hydrolysis were further analysed by using high performance
liquid chromatography (JASCO, Japan) with LiChroCart @
250 mm  4.6 mm, Purospher @ STAR NH2 (5 mm) column
equipped with Jasco RI-1530 Intelligence RI detector at a flow rate
of 2 mL/min. Organic acids derived from pretreatment were anal-
ysed by HPLC- SPDA-10A, UV–VIS Detector; LC-10 AS Liquid Chro-
matography Shimadzu, Japan column packed with ion exclusion
HPLC organic acids analysis column (AminexÒHPX-87H)
300 mm  7.8 mm, Biorad Laboratories at a flow rate of 0.6 mL/
min and detected with UV detector at 215 nm (Hafid et al., 2011).
2.8.2. Bioethanol analysis
For estimation of dry cell weight of yeast, ethanol and sugar
concentration, fermentation broth was centrifuged at 10,000 rpm
for 10 min to separate cell and supernatant. The dry cell weight
of the yeasts was quantified based on different grams of cell before
and after drying the cells. The supernatant was filtered using cellu-
lose nylon filter paper (0.45 mm) for analysis. The ethanol was
detected using a gas chromatograph (Model GC-17A, Shimadzu,
Japan) equipped with a flame ionization detector (FID) and capil-
lary column packed with polar BP 20. The ethanol concentration
was quantified based on peak areas of the samples over the peak
area of the standards solution. The ethanol yield (Yp/s) and conver-
sion efficiency (%) were calculated using the following equation;

½Ethanol gL
Yp=s ¼ g
½Sugar L
Yp=s
Conversionefficiency ð%Þ ¼  100
0:051
where Yp/s is ethanol yield (g/g). The conversion efficiency (%) was
obtained based on the theoretical yield of calculated bioethanol.
0.51 is the maximum ethanol yield per unit of glucose.
3. Results and discussion
3.1. Characterisation of food waste
The fermentable sugar production was highly affected by the
components of food waste. The major compositions of Malaysian’s
food waste were analysed as 60.78 ± 2.02% of carbohydrate,
20.53 ± 1.77% of protein, 13.65 ± 1.74% of fat and 1.67 ± 0.05% of
fiber from the main constituents of rice, meat and vegetables
(Table 1). The results of nutritional components such as carbohy-
drate and protein are slightly higher than that reported by other
researcher (Tian et al., 2015; Wang et al., 2014; Shen et al.,
2013). The high carbohydrate content (>60%) at a high proportion
making Malaysian food waste an attractive feedstock for sacchari-
fication and fermentation process (Grohman and Bothast, 1997). A
large fraction of TSS/TS (
0.60 ± 0.09) made the pretreatment nec-
essary to release fermentable sugars where the high ratio of VSS/
TSS (approx. 0.9 ± 0.08) indicated high content of organic matter
in the food waste. This value was in agreement with Kuo and Lai
(2010), who stated that the ratio of VSS/TSS of approximately 0.9
Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
5
suggested the biodegradability properties of the substrate. The
VSS comprised of biodegradable volatile solid, which included
starch, organic acids, lipids and protein (Qiao et al., 2011). Maltose
(10.53 ± 0.15 g/L), glucose (3.45 ± 0.05 g/L), fructose (0.43 ± 0.23 g/
L) and sucrose (0.14 ± 0.12 g/L) were the most abundant fer-
mentable sugar contain in raw food waste. It is known that maltose
is a disaccharide produced after the starch being breakdown by
amylase. In the raw food waste, high concentration of maltose
leads to the utilization of glucoamylase enzyme only (in which
the use of amylase can be neglected) to further breakdown the gly-
cosidic linkage of food waste after pretreated with dilute acid for
fermentable sugar. Very small amounts of the weak organic acids;
lactic acid (1.32 ± 0.02 g/L), acetic acid (5.89 ± 0.12 g/L), and propi-
onic acid (0.79 ± 0.07 g/L) were noticed in the food waste. Accord-
ing to Feng et al. (2012), a weak acid such as acetic acid does not
exert distinct inhibitory effect on hydrolysis of substrate to sugars.
In addition, the presence of volatile organic acids associated with
carbohydrate indicated the low activity of indigenous microbial
inhibitors due to the freshly prepared samples.
3.2. Optimization of food waste pretreatment
The hydrothermal and dilute acid pretreatment of food waste
was aimed to enhance as much as possible the fermentable sugars
produced before enzymatic treatment was applied to the substrate.
Fig. 2 shows the effects of temperature on fermentable sugar pro-
duction from food waste by hydrothermal and dilute HCl and
H2SO4 at 0.5% concentration. Both hydrothermal and dilute acid
pretreatment shows an increase in fermentable sugar production
as temperature increase from 80 to 100 °C. Hydrothermal pretreat-
ment shows an increase of 27.59 g/L of fermentable sugar when
the temperature changes to 90 °C whereas no significant fer-
mentable sugar increment observed when the temperature shift
to 100 °C (p > 0.05). This result was due to the increase of ioniza-
tion constant of water at high temperature, where, the concentra-
tion of H+ and OH increased thus facilitate the hydrolysis of
carbohydrate into monosaccharides (Lin et al., 2015).
ð14Þ
Meanwhile, a similar trends of fermentable sugar was observed
for dilute acid pretreatment in which both HCl and H2SO4 pretreat-
ment showed an approximately 33.47 g/L and 40.45 g/L increase of
ð15Þ production at 90 °C (p < 0.05), however, the total fermentable
sugar concentration were nearly equal and shows not significantly
different at 100 °C (p > 0.05) . High temperature could promote
multi-phase reaction between substrate and hot water or acid
and reduce the Maillard browning reaction, thus enhance the
hydrolysis of carbohydrate to monosaccharides sugar (Yu et al.,
2013; Vavouraki et al., 2012). The hydrothermal reaction of the
food waste took place at the pH ranged from 4.7 to 5.2 whereas
the pH of food waste treated with acid ranged from pH 1 to 3. At
lower pH resulting in high H+ concentration stimulate the protona-
tion of the reactive amino group with the hydrogen ion from the
acid resulting from the ionization process that makes it unreactive.
On contrary, heated solution in hydrothermal pretreatment with
low H+ concentration promote interactions between the amino
groups of amino acids and the carbonyl groups from the glucose
and fructose that lead to high sugar decomposition and production
of undesirable by-products and inhibitors (Borne et al., 2012).
At the same reaction temperature (90 °C), the pretreatment effi-
ciency is distinctly affected by the type and concentration of acid
catalyst used more than an increase in reaction time. In dilute
HCl pretreatment, an increase of 15.93 g/L of fermentable sugar
was observed when 1.5% of acid concentration used. In particular,
dilute H2SO4 pretreatment shows highest increment of fer-
mentable sugar (approximately 10.87 g/L) at 1% of acid concentra-
tion but not significantly different with statistical results (p > 0.5)
(Fig. 3). This was probably due to the higher acid strength of
6 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx

Table 1
Characteristic of kitchen waste used in this study.

Composition This study Tian et al. (2015) Wang et al. (2014) Shen et al. (2013)
pH 5.20 ± 0.32 nd 5.08 nd
Total solids (g/L) 117.47 ± 0.78 188.1 ± 0.12 221.7 ± 1.57 226.1
Total Suspended solids (g/L) 82.50 ± 2.09 nd nd nd
Volatile suspended solid (g/L) 74.25 ± 0.35 172.4 ± 0.08 178.7 ± 1.28 179
Moisture (%) 70.06 ± 2.33 nd 77.83 77.39
Carbohydrate (%) 60.78 ± 2.02 46.43 ± 0.32 nd 33.28
Crude Protein (%) 20.53 ± 1.77 14.97 ± 0.35 16.88 ± 1.24 14.71
Fat (%) 13.65 ± 1.74 30.27 ± 0.21 33.82 ± 5.04 28.85
Fiber (%) 1.67 ± 0.05 17.64 ± 0.52 nd 8.61
Ash (%) 1.97 ± 0.10 nd nd nd

nd – not determined.

by hydrolysis of polysaccharides into monosaccharides. However,

Fig. 2. Effect of hydrothermal and dilute acid pretreatment on fermentable sugar
concentration at different temperatures (the fermentable sugar concentration was
taken at highest production at 180 min of incubation time).
60
Changes in fermentable sugar
this model is only applicable for pure substrate hydrolysis but defi-
cient for complex biomass food waste hydrolysis. Many research-
ers have proven that Saeman’s model must be modified to more
accurately describe the kinetic behaviour of polysaccharide hydrol-
ysis in the biomass and in the formation of decomposition products
(Zhao et al., 2012). Food waste hydrolysis catalysed by dilute acid
is usually recognised as a series of complex pseudo-homogeneous
irreversible reactions. The use of pseudo-homogeneous reaction
makes it possible to overcome the heterogeneity of the reaction
medium (Jin et al., 2011).
Food waste was pretreated with hot distilled water and differ-
ent concentrations of HCl and H2SO4 at different temperatures.
Fig. 4 shows a time course of experimental and predicted values
of fermentable sugars produced during hydrolysis of food waste.
All values were first transformed by subtracting the initial sugar
concentration produced. The results of the kinetic model were con-
structed to represent fermentable sugars at temperature of 90 °C
for all pretreatment. The dilute acid hydrolysis system assumes
that chemical equilibrium was rapidly reached to the larger mole-
cules of polysaccharides in the food waste when concentration of
1.0% of HCl and 1.5% of H2SO4 used which then further degraded
to monomers such as glucose. For the hydrothermal and dilute

50 acid-catalysed system, model showed a good fit between the

experimental data and the predicted data with R2 of all treatment
production (g/L)

40 condition > 0.9 indicating that the model was reliable.

30
20
10
0 2012). Tentatively, the estimated mr values in a dilute acid pre-
0.5 1 1.5
Acid Concentration (%)
Fig. 3. Effect of different dilute acid catalyst and concentration on fermentable
sugar production at 90 °C and 180 min of incubation time. Dilute acid catalyst (D:
HCl and Ο: H2SO4).
H2SO4 that could dissolve and hydrolyse the original carbohydrate
in the food waste to fermentable sugars more effective as com-
pared to low acid strength of HCl. It can be observed that an addi-
tion of 1.5% and 2.0% H2SO4 only produced access of H+ in the
reaction system thus does not give significant influence on the rate
of hydrolysis, therefore, it can be neglected.
3.3. Determination of kinetic constants, modelling and simulation
Saeman’s model is the simplest model which assumes that the
process of polysaccharide dissolution involves a direct conversion
Referring to the estimated unknown parameter shown in
Table 2, the estimated value of br, the dependence of reaction rate
on temperature was clearly observed in kr=1. The rate constant of
kr=2 shows that the temperature is not affect the rate of fer-
mentable sugar decomposition in all pretreatments as the sugar
degradation in mild acid hydrolysis most apparently occurred at
the reaction temperature of more than 100 °C (Vavouraki et al.,
2
treatment were higher in kr=2 than kr=1 for both HCl and H2SO4
reaction suggested that the fermentable sugar degradation is
highly dependence to acid concentration. However, it is worthy
to note that the values of mr and KHr were interrelated. A high val-
ues of KHr will actually bring down the overall reaction of kr=2 indi-
cated that acid concentration shows a negatively effect on
fermentable sugar decomposition. This was supported by the val-
ues of acid consumption, YHr which were very small.
An inverse effect was found for kr=1 where the value of mr and
KHr were estimated very small. A small value of KHr constant irre-
versibly increased the overall reaction. Based on the experimental
values, the kr=1 increased with the acid concentration. In this
model, the estimated parameters indicated that kr=1 values was
higher than kr=2 at all temperatures and acid concentration tested
shows that the sugar formation was more favoured than its degra-
dation. This was in agreement with Jin et al. (2011). The kr=1 for
dilute acid pretreatment was two times higher than for the
hydrothermal reaction at 90 °C. By comparison, based on the value

Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx 7

60

Changes of fermentable sugar concentration,

50

1.0% H2SO4, 90 C
40
1.5% HCl, 90 C

Ci=2 (g/L)
30

20
Hydrothermal, 90 C

10

0
0 50 100 150 200 250 300
Time (min)
Fig. 4. Experimental data and predicted model (line) of fermentable sugar production from kitchen waste by hydrothermal, dilute HCl and H2SO4 pretreament.

Table 2
Estimated parameters of hydrothermal and dilute-acid catalysed system of kitchen waste hydrolysis.

hot dH2O HCl H2SO4

Parameter r=1 r=2 r=1 r=2 r=1 r=2
br 16.35 0.00 16.35 0.00 16.35 0.00
kr0 1.07 0.15 1.07 0.15 1.07 0.15
KHr Nil Nil 1.21 2.04 0.49 1.99
KSr 122.88 14.24 122.88 14.24 122.88 14.24
mr Nil Nil 1.38 50.00 0.78 12.64
YHr Nil Nil 0.000 0.002 0.004 0.003

of kr=1, the formation of fermentable sugars occurred more exten- future changes that can be used in the similar process. The reaction
sively in dilute H2SO4 (2.32 g/L/min) as compared to dilute HCl temperature significantly affected the fermentable sugar produc-
(1.61 g/L/min) and hydrothermal (0.686 g/L/min) pretreatment at tion. It was expected by the model that higher temperature could
the same temperature applied. This suggesting that the acid addi- increase the rate of food waste hydrolysis thus indirectly increasing
tion resulted in the rate-controlling steps conversion of food waste the rate of fermentable sugar production (Lin et al., 2015). The fer-
polysaccharides to fermentable sugar compared to hydrothermal mentable sugar production increased from 70 °C to 150 °C, implying
pretreatment, which explain the reason of higher yield of dilute that fermentable sugar production were affected at changes of tem-
acid pretreatment than hydrothermal pretreament from reaction peratures within the specified time. When the temperature increase
mechanism. It was supported by Shen and Wyman (2011) who to 110 °C and 150 °C, the reaction rate, kr=1 is increasing based on
reported the rate constant for bioconversion of xylan to xylose the calculated curves of fermentable sugar presented faster peak
monomers in dilute acid pretreatment is 4.9 times higher than tendency over time at approximately 75 min and 25 min, respec-
hydrothermal pretreatment. Thus, the reaction rate coefficient for tively. However, it could be seen that, at higher temperature applied
total sugars production can be written as in Eqs. (16)–(18). could drastically decrease the fermentable sugar production that
For hydrothermal reaction; was attributed to the degradation of fermentable sugar to degrada-
 16:35 tion products (Lin et al., 2015). According to Timung et al. (2016), a
T
kr¼1 ¼ 1:07  ð16Þ
373
150°C
For dilute HCl-catalyst system;
 16:35 " 1:38 #
T CH 110°C
kr¼1 ¼ 1:07   þ1 ð17Þ
373 1:21
90°C
For dilute H2SO4-catalyst system;
 16:35 " 0:78 # 70°C
T CH
kr¼1 ¼ 1:07   þ1 ð18Þ
373 0:49

Fig. 5 depicts the result of the simulation of predicted fermentable

sugar production by dilute 1.5% of HCl at different temperatures. It
is aimed to sustain high concentration of fermentable sugar pro-
duced for a longer incubation time before degraded. Testing of Fig. 5. Simulation of different temperatures on fermentable sugar production at
many different temperature provide a prediction and potential 1.5% HCl.

Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
8 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx
drop in fermentable sugar was attributed to the formation of the
inhibitory products in the hydrolysate such as organic acids, fur-
fural and HMF. High rate of sugar degradation into inhibitory prod-
ucts subsequently affect further recovery process in which
detoxification step need to be included in the process to remove
the fermentative inhibitor in the bioethanol production (Raghavi
et al., 2016). On the contrary, the fermentable sugar production at
70 °C and 90 °C were remained constant with low rate of degrada-
tion to by-products. It is expected that the concentration of
degraded product or inhibitor in the hydrolysate was low thus the
detoxification step can be omitted. Therefore, temperature of
90 °C is optimal for getting better sugar production and sustained
at longer incubation time compared to higher temperature before
being degraded. The estimation of fermentable sugar produced
was successfully explained by the model.
3.4. Sequential acid-enzyme pretreatment in hydrolysis reactor
Food waste which was subjected to acid pretreatment by 1.5%
of HCl was then served as a substrate for batch enzymatic hydrol-
ysis by 85 U/mL of glucoamylase enzyme loading (Hafid et al.,
2016) to see the effect of acid pretreatment and enzyme addition
on hydrolysis of food waste and formation of fermentable sugar.
Initially, a separate experiment was conducted using individual
1.5% of HCl and glucoamylase hydrolysis, a yield of 50.5 g/L of total
sugar with a conversion of carbohydrate to sugar is 0.424 g sugar/g
carbohydrate was obtained in acid hydrolysis, whereas, a slightly
higher of total sugar (60.32 g/L) with conversion of 0.506 g sugar/
g carbohydrate was obtained through enzymatic hydrolysis
(Table 3).
Noticeably, the acid hydrolysis or glucoamylase hydrolysis
alone was insufficient for complete hydrolysis of food waste for
sugar production. Khawla et al. (2014) reported that the acid
hydrolysis stage alone was inadequate and a supplementary sac-
charification stage by enzyme was needed. Therefore, for this rea-
son, the use of combination of 1.5% HCl and glucoamylase was
necessary for an effective hydrolysis of food waste. Fig. 6 shows
the virtual fermentable sugar released after combination of 1.5%
of HCl hydrolysis incubated for 3 h and subjected to enzymatic
hydrolysis by glucoamylase under non-control pH. Noticeably,
the pH value was almost constant at pH 1.8 throughout acid
hydrolysis steps and maintained at pH 5 during enzymatic hydrol-
ysis. This indicates that the positive charge of ion H+ is not being
consumed during the hydrolysis reaction but bound with the bio-
mass to form a cyclic carbonium ion to cleave the glycoside bond
and catalyse the hydrolysis reaction (Zhao et al., 2014).
The highest sugar concentration (103.4 g/L) was observed par-
ticularly after glucoamylase enzyme addition (85 U/mL), which
increased by 2.04 and 1.71 folds comparing to acid and enzymatic
Table 3
Summary of fermentable sugar production at different pretreatment.
Treatment
Parameter Acid
hydrolysis
Reagent 1.5% of HCl
Temperature (°C) 90
Neutralization no
Production of total sugar (g/L) 50.5
Conversion of total CHO to sugar 0.424
(g sugar/g CHO)
Conversion efficiency (%) 42.4
Hydrolysis rate (g/L/min) 0.608
Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
hydrolysis with the maximum hydrolysis rate of 0.344 g/L/min. It
is speculated that acid hydrolysis accelerated the process of food
waste hydrolysis in which enhanced the accessibility of glucoamy-
lase to total carbohydrate, hence, marginally increased the fer-
mentable sugar production. The decreased in the content of total
carbohydrate by 66% would result in the decrease in viscosity
which benefited the hydrolysis efficiency that can be seen from
the glucoamylase enzyme action simultaneously hydrolysed the
remaining carbohydrate releasing more sugar in the hydrolysate
(Gao et al., 2014). The end product inhibition by high concentration
of glucose may decrease the activity of glucoamylase, therefore,
the fermentable sugar concentration on prolonged incubation
remained constant (Kuhad et al., 2010).
The apparent total sugar production efficiency (ATPE) and
degree of food waste decomposition (DoD) after sequential pre-
treatments are represented in Fig. 7. As can be seen, ATPE values
ranging from 30% to 44% were obtained in the first 3 h of acid
hydrolysis experiment with a maximum food waste decomposition
of 34%. Li et al. (2017) stated that dilute acid was aimed to remove
unnecessary component of food waste that covering the starch
component and simultaneously degrade the resistant starch of
food waste that hindered the efficiency of enzyme attack. The ATPE
values were enhanced obviously mainly after glucoamylase addi-
tion that shows higher total sugar approximately 75–90% of pro-
duction efficiency. The important increase in the ATPE during the
sequential acid-enzyme pretreatment does agree with degree of
food waste decomposition in which a dramatically decrease of car-
bohydrate content were observed in the hydrolysate. Under such
conditions, about 43–80% of food waste has been degraded and sol-
ubilized into fermentable sugar leaving untreated carbohydrate
still retains
20% before partially degraded to other products.
The integrated combination pretreatment based on acid and enzy-
matic hydrolysis is undeniable improve the bioconversion of food
waste, liberates a higher amount of fermentable sugar.
3.5. Mass balance of hydrolysis process for biosugar production
Fig. 8 summarizes mass balances of the overall process under
the combined 1.5% HCl-glucoamylase pretreatment condition from
1000 mL of ground food waste containing 117 g of carbohydrate.
The food waste hydrolysate contained high total sugar concentra-
tion (103.4 ± 0.04 g/L) consist of 98.13 g/L glucose, 1.74 g/L fruc-
tose, 0.69 g/L sucrose and 2.84 g/L maltose with overall sugar
released was over 80% confirming that pretreatment conditions
can preserve most of the sugars. The overall sugar released is
almost the same as reported by Wei et al. (2012) using combina-
tion of dilute acid and enzymatic hydrolysis of biomass eucalyptus.
The unhydrolysed solid removed from the liquid phase through
solid-liquid separation step whereby to eliminate the problem of
Enzymatic Sequential
hydrolysis acid-enzyme
hydrolysis
85 U/mL of glucoamylase 1.5% of HCl followed by 85
U/mL of glucoamylase
55–60 90 °C followed by cooling stage
and adjusted temperature to 55–60 °C
5 M of NaOH to pH 5 5 M of NaOH to pH 5
60.32 103.4
0.506 0.868
50.6 86.8
0.1 0.344
 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx 9

120 Acid Cooling phase Enzymatic 100

Changes in fermentable sugar and total

hydrolysis hydrolysis 90

carbohydrate concentration (g/L)

100
80

pH, Temperature (C)

70
80
60
60 50
40
40
30
20
20
10
0 0
0 1 2 3 4 5 6 7 8 9 10
Time (hour)
Fig. 6. Profile of virtual fermentable sugar production and total carbohydrate degradation in sequential acid-enzyme hydrolysis system under non-control pH with (*)
temperature; (+) pH.

Acid Cooling Enzymatic

hydrolysis phase hydrolysis
100
Sugar production and food waste

DoD
80
ATPE
degradation (%)

60

40

20

0
1 2 3 4 5 6 7 8 9 10
Time (hour)
Fig. 7. Profile of apparent total sugar production efficiency (ATPE) and degree of food waste degradation (DoD) during sequential acid-enzyme hydrolysis pretreatment
system.

hydrolysate viscosity that may decrease the yield of further fer- months storage in 4 °C at pH 4.8. The concentration of fermentable
mentation process. The increased in viscosity due to solid content sugar, minerals, and elemental composition exhibit very little
may create mixing problem in the reactor that may disrupt the changes and can be neglected. In addition, the fermentation studies
sugar uptakes of the ethanol fermenting microorganisms and yeast cell growth assays of Escherichia coli and Saccharomyces
(Palmqvist and Lidén, 2014). In addition, solid-liquid separation cerevisiae showed insignificant different of its fermentability utiliz-
is needed to remove oil compound in the food waste hydrolysate ing fresh hydrolysate and stored hydrolysate. This indicates that
that typically generates problems of flotation, clogging and mass the recovered sugar from this study can be stored and stable at
transfer in the reactor thus hinder the efficiency of the hydrolysis low temperature (4 °C) for further application. The remaining
and fermentation process (Yasin et al., 2013). In order to obtain sugar in the acid-enzyme hydrolysis residue has not been detected
very viscous saccharified liquid containing high sugar, vacuum as it implies that there was a nearly complete hydrolysis of food
rotary evaporator system has been adopted to remove residual waste to fermentable sugar during previous hydrolysis.
water present in the saccharified liquid by boiling at 60 °C. The
vapour will be condensed with cooling or chilling water. The mate-
3.6. Fermentation of food waste hydrolysate for bioethanol production
rials balance indicated loss of some mass during pretreatment,
solid-liquid separation and evaporation steps maybe due to either
Food waste hydrolysate contains approximately 80.04% of glu-
loss as solid and water residue, gaseous compounds or conversion
cose, 10.12% of maltose, 6.14% of fructose, and 3.72% of sucrose.
to other compound that not be determined in this study. The over-
Food waste hydrolysate was directly used as substrate to produce
all sugar recovery was increased to 150.5 ± 0.11 g/L after evapora-
bioethanol by Saccharomyces cerevisiae in a shake flask fermenta-
tion steps. About 128.4 g/L glucose, 6.24 g/L fructose, 5.59 g/L
tion. Due to the different in pretreatment condition between
sucrose, and 10.18 g/L maltose can be recovered in the concen-
sequential acid-enzyme method and bioethanol production, the
trated liquid sugar. Jin et al. (2013) reported that the concentration
separated hydrolysis and fermentation (SHF) process is the best
of liquid sugar hydrolysate is not significantly changed upon three
option. The fermentation process in the first 24 h incubation time

Please cite this article in press as: Hafid, H.S., et al. Over production of fermentable sugar for bioethanol production from carbohydrate-rich Malaysian food
waste via sequential acid-enzymatic hydrolysis pretreatment. Waste Management (2017), http://dx.doi.org/10.1016/j.wasman.2017.05.017
10 H.S. Hafid et al. / Waste Management xxx (2017) xxx–xxx

Kitchen waste (1000 mL, 1720 g) that, S. cerevisiae is able to convert the sugar compound present
15 mL of 1.5% HCl in the food waste into bioethanol. Obviously, the increased in the
yeast cell density after the fermentation will bring additional rev-
Acid hydrolysis
enue from food waste hydrolysate fermentation into bioethanol.
8.5 mL of glucoamylase 10 mL of 5M NaOH
Inhibitor analysis of the hydrolysate revealed that it was devoid
of major fermentation inhibitors like furfural and 5-
Enzymatic hydrolysis hydroxymethyl furfural. The presence of organic acids like lactic
acid, acetic acid, propionic acid, and butyric acid shows no inhibi-
tory effect of the pretreated food waste to fermentation medium.
Centrifugation and filtration Solid residue (122 ± 0.1 g) Hence the hydrolyzate can be used for fermentation without any
elimination of inhibitory material and the process conversion of
Saccharified liquid sugar (750 mL, 800 ± 0.1 g) food waste to bioethanol are more technically practical and eco-
Total Sugar: 103.4 ± 0.04 g/L nomically feasible.
Glucose: 98.135 ± 0.01 g/L
Fructose: 1.747 ± 0.02 g/L
Sucrose: 0.694 ± 0.02 g/L 4. Conclusions
Maltose: 2.824 ± 0.03 g/L
Evaporation Vapor and water condensed out In this study, food waste showed as potential substrate for
(275 mL, 275 ± 0.3 g) bioethanol production after appropriate hydrolysis. Initially, the
reaction kinetic model to predict the fermentable sugar production
Concentrated kitchen waste hydrolysate containing sugar (475 mL, 520 ± 0.2 g)
at a defined temperature and acid concentration was proposed for
Total Sugar: 150.5 ± 0.11 g/L food waste hydrolysis. A good fit was observed between the exper-
Glucose: 128.472 ± 0.1 g/L imental data and the kinetic model in all pretreatment conditions.
Fructose: 6.247 ± 0.13 g/L
The model was found to be reliable in describing the kinetic beha-
Sucrose: 5.598 ± 0.1 g/L
Maltose: 10.183 ± 0.2 g/L viour of food waste hydrolysis at lower and higher temperatures
under the hydrothermal and acid-catalysed system. The fer-
Fig. 8. Mass balance for the recovery of fermentable sugar from kitchen waste mentable sugar produced upon food waste hydrolysis via sequen-
hydrolysate. tial HCl and glucoamylase pretreatment was 103.4 g/L, which was
respectively 2.04 and 1.71 folds higher compared to individual acid
Table 4
and enzymatic hydrolysis. In addition, highly concentrated sachar-
Summary of kinetic data of bioethanol production utilizing food waste hydrolysate by ified sugar of 150.5 g/L were obtained through further recovery
S. cerevisiae at 24 h and 48 h incubation time. process by solid-liquid separation and evaporation. The sacchari-
Parameter 24 h 48 h
fied liquid sugar was successfully converted to bioethanol by S.
cerevisiae at 85.38% conversion efficiency with no effect of inhibi-
Initial cell (Xo, g/L) 0.44 0.44
Maximum cell (Xmax, g/L) 4.73 5.76
tors indicated that food waste is highly promising substrate for
m max (h1) 0.36 0.36 bioethanol production.
Cell yield (Yx/s) 0.19 0.23
Initial sugar (So, g/L) 25.00 25.00
Final sugar (Sf, g/L) 2.83 1.63 Acknowledgement
Sugar consumption (%) 88.68 93.48
Initial ethanol (Po, g/L) 1.52 1.52 The authors are thankful to the Ministry of Higher Education,
Maximum ethanol (Pmax, g/L) 10.92 8.32
Malaysia for supporting of this work under research grant No.
Ethanol Yield (Yp/s) 0.42 0.29
Conversion efficiency (%) 85.38 65.05
02-03-11-1008FR.
Ethanol volumetric productivity (gL1 h1) 0.46 0.17

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