Bioresource Technology 225 (2017) 359–366

Contents lists available at ScienceDirect

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

Sustainable biobutanol production from pineapple waste by using

Clostridium acetobutylicum B 527: Drying kinetics study
Manisha A. Khedkar a, Pranhita R. Nimbalkar a, Shashank G. Gaikwad b, Prakash V. Chavan a,
Sandip B. Bankar a,c,⇑
a
Department of Chemical Engineering, College of Engineering, Bharati Vidyapeeth University, Dhankawadi, Pune-Satara Road, Pune 411 043, India
b
Chemical Engineering and Process Development, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune 411 008, India
c
Department of Biotechnology and Chemical Technology, School of Chemical Technology, Aalto University, P.O. Box 16100, FI-00076 Aalto, Finland

h i g h l i g h t s

Characterization of pineapple peel waste.

Drying kinetics of pineapple peel and its mathematical modeling.

Pretreatment and detoxification of wet and dried pineapple peel.

ABE fermentation of detoxified hydrolysates using Clostridium acetobutylicum.

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

Article history: Present investigation explores the use of pineapple peel, a food industry waste, for acetone-butanol-
Received 5 October 2016 ethanol (ABE) production using Clostridium acetobutylicum B 527. Proximate analysis of pineapple peel
Received in revised form 12 November 2016 shows that it contains 35% cellulose, 19% hemicellulose, and 16% lignin on dry basis. Drying experiments
Accepted 14 November 2016
on pineapple peel waste were carried out in the temperature range of 60–120 °C and experimental drying
Available online 18 November 2016
data was modeled using moisture diffusion control model to study its effect on ABE production. The pro-
duction of ABE was further accomplished via acid hydrolysis, detoxification, and fermentation process.
Keywords:
Maximum total sugar release obtained by using acid hydrolysis was 97 g/L with 95–97% and 10–50%
Biobutanol
Drying
removal of phenolics and acetic acid, respectively during detoxification process. The maximum ABE titer
Fermentation obtained was 5.23 g/L with 55.6% substrate consumption when samples dried at 120 °C were used as a
Pineapple peel substrate (after detoxification).
Detoxification Ó 2016 Elsevier Ltd. All rights reserved.

1. Introduction

The rhythm of fossil fuel(s) utilization of current civilization is

clearly unsuitable due to high price of crude oil and adverse envi-
Abbreviations: D0, pre-exponential factor (m2/s); Deff, effective moisture diffu- ronmental impact. Biofuel is a high priority alternative energy
sivity (m2/s); E, activation energy (kJ/mol); k, drying constant (1/min); L, half source because of rapid depletion of fossil fuel and other environ-
thickness of samples (m); M, moisture content for a given time (g/g dry solids); Me, mental issues. This instigated an attention to produce biofuels
equilibrium moisture content (g/g dry solids); Mo, initial moisture content (g/g dry from renewable resources (Ezeji et al., 2013). Currently, a variety
solids); MR, dimensionless moisture ratio; Mt1, moisture contents at time (t1) (g/g
dry solid); Mt2, moisture contents at time (t2) (g/g dry solid); n, positive integer (–);
of alternative fuels such as butanol, ethanol, and methanol have
Nm, mean mass flux of moisture (kg/m2/min); R, universal gas constant (kJ/mol/K); been proposed to replace fossil fuels. Among them, butanol shows
rM, rate of drying (g/g dry solid/min); S, cross sectional area (m2); T, drying air superior chemical properties over ethanol and methanol. It has
temperature (K); t, time (s); t1, t2, drying times (min); Wd, mass bone dry solid (g); higher energy density and low octane number that is comparable
Wt, mass for a given time t (g); qs, density of solid (kg/m3).
⇑ Corresponding author at: Department of Biotechnology and Chemical Technol-
to gasoline. Further, it has low vapor pressure that eases trans-
ogy, School of Chemical Technology, Aalto University, P.O. Box 16100, FI-00076 portation and shows lesser corrosion of contact material (Hu
Aalto, Finland. et al., 2011).
E-mail addresses: [email protected], [email protected] (S.B. Bankar).

http://dx.doi.org/10.1016/j.biortech.2016.11.058
0960-8524/Ó 2016 Elsevier Ltd. All rights reserved.
360 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366

Biobutanol is traditionally produced by acetone-butanol- etha- scrutinized in context of their potential for biochemical conversion
nol (ABE) fermentation with Clostridium spp. A biological route of by Clostridium spp. to biobutanol. Hence, present study is focused
ABE fermentation typically involves four basic steps viz. feedstock on the use of inexpensive and abundantly available lignocellulosic
selection, pretreatment, detoxification, and fermentation. Among biomass to circumvent high-substrate costs. Besides, the develop-
them, selection of feedstock is an important step that contributes ment of a simple, yet economic bioprocess is also desirable to max-
around 60–70% of overall process cost. Therefore, scrutinizing imize the biobutanol production. Therefore, there is an increasing
low cost feedstock such as lignocellulosic biomass is essential. interest in butanol bioconversion, which leads towards significant
Recent progress in agricultural sector is fascinating, and many efforts in commercial production, or retrofitting same existing
researchers are exploring the horizons to improve traditional farm- bioethanol plants for butanol production (Maiti et al., 2016).
ing with advanced irrigation and water management (Valipour, Based on aforementioned discussion, it was thought desirable
2014, 2015; Valipour and Singh, 2016; Yannopoulos et al., 2015). to undertake systematic investigation to produce biobutanol using
With these modern developments, biomass is considered as pineapple peel waste as a feedstock. The present work is catego-
renewable, cheap, and fourth largest source of energy in the world. rized into four parts: (i) characterization of pineapple peel waste,
India alone generates over 400 million tons biomass every year (ii) study of drying kinetics (iii) saccharification and detoxification
directly from plants, agricultural waste, food, and vegetable indus- studies, and (iv) fermentative production of biobutanol from
try waste (Bankar et al., 2013a; Valipour and Singh, 2016). detoxified hydrolysates of pineapple peel waste. This paper essen-
Although, few researchers have made attempts to utilize the agri- tially gives an emphasis on drying study to evaluate its effect on
cultural biomass for biofuel production, the utilization of fruit and saccharification and subsequently on ABE fermentation. The math-
vegetable industry waste is still unattended. India ranks second in ematical model was developed to formulate correlation between
the world in fruit production with an annual output of 48 million experimental and predicated variables during moisture removal.
tons fruits, contributing about 12% of the world’s fruit production. To the best of our knowledge, no attempts have been made on
Pineapple is one of the leading fruits being produced and utilized biobutanol production using pineapple peel waste as a feedstock,
largely in India. Incidentally, India ranks seventh in worldwide till date. The detailed pretreatment and fermentation studies of
pineapple production with annual production of 1.3 million tons pineapple peel biomass will be done in following parts of this pro-
(FAO, 2015). However, the processing and commercial production ject and will be presented in next papers.
of pineapple (juice) generates approximately 20–40% (w/w) waste
in the form of peel and core (Nga and Trang, 2015).
2. Materials and methods
Pineapple peel waste comprises 35–50% cellulose, 20–35%
hemicellulose, and 5–30% lignin (Roda et al., 2014). Hence, the
2.1. Materials
use of pineapple peel waste can be explored in effective bioproduct
generation. Kumbhar et al. (2015) reported higher amount of bac-
Dextrose, dipotassium hydrogen phosphate, ammonium acet-
terial cellulose production (0.3 g/g) from pineapple peel. Besides,
ate, biotin, thiamin, p-aminobenzoic acid, glycerol, sodium potas-
several authors have made an attempts to use pineapple peel as
sium tartarate, sulfuric acid, soluble starch, peptone, L-cysteine
a substrate for production of bromelain, polyphenols (Gautam
hydrochloride, sodium chloride, magnesium sulfate, manganese
et al., 2010), biohydrogen, and biogas (Nga and Trang, 2015). Addi-
sulfate, isopropanol, iron sulfate, sodium hydroxide, butyric acid,
tionally, reports on production of bioethanol from pineapple peel
acetic acid, butanol, acetone, ethanol and 3,5-dinitrosalicylic acid
waste using S. cerevisiae and Enterobacter aerogenes with maximum
were purchased from SRL Ltd, India. Sodium acetate and phenol
production of 9.69 g/L and 1.38 g/L ethanol, respectively are also
were procured from Sigma Aldrich, India. Folin-Ciocalteu reagent,
available (Choonut et al., 2014). Pineapple peel usually contains
sodium carbonate, meat extract and yeast extract were obtained
80–90% (w/w) moisture on wet basis (Namsree et al., 2012), mak-
from Himedia laboratories, India. Hydrochloric acid and sulfuric
ing it compulsory to reduce moisture level (minimum threshold
acid were obtained from Avra, India. All the chemicals used were
value 7% w/w) to circumvent microbial spoilage and deteriora-
of analytical grade.
tion by chemical reaction (Kiranoudis et al., 1995). In this connec-
tion, drying operation substantially reduces the mass and volume
of pineapple peel. With drying operation the storage space, packing 2.2. Organism and revival medium
size and transportation cost can be reduced. Moreover, it also pre-
vents an additional dilution, due to presence of high amount of Bacterial strain, C. acetobutylicum NRRL B-527 was a kind gift
moisture during process operations. from ARS (Agriculture Research Services) culture collection, USA.
Lignocellulosic biomass require crucial step of pretreatment to The lyophilized cells were regenerated by using sterile revival
alter recalcitrant structure. Voluminous reports on pretreatment medium (RM) in 100 mL air tight, anaerobic screw cap glass bottles
of lignocellulosic biomass like grain straw (barley, wheat, sorghum, with working volume of 80 mL. It was grown for 48 h at 37 ± 2 °C
and rice husk), corn stover, and corn fiber (Qureshi et al., 2008) are and subsequently used for the preparation of spore suspension
available in the literature. However, such studies pertaining to and glycerol stock. Revival medium contained (g/L): dextrose 5;
pineapple peel are scarce. Approaches based on pretreatment, peptone 10; beef extract 10; yeast extract 3; sodium chloride 5;
enzymatic saccharification (Roda et al., 2014), physical, and chem- soluble starch 5; sodium acetate 3; L-cysteine HCl 0.5; Resazurin
ical pretreatment methods (Choonut et al., 2014; Rattanapoltee 0.001; water, made up to 1000 mL. Medium pH was adjusted to
and Kaewkannetra, 2014) are known to enhance the sugar release. 6.8 with HCl, if required. The medium was purged with nitrogen
During pretreatment processes, the fermentation inhibitors are to remove dissolved oxygen; sealed and autoclaved at 121 °C for
known to produce, that lead to decrease in solvent productions. 20 min and cooled at room temperature. L-cysteine HCl was added
Thus, detoxification is an essential step to improve solvent yield separately in revival medium using filter sterilization.
and productivity.
Sustainable biobutanol production is still lagged behind due to 2.3. Spore suspension and inoculum preparation
impediments such as high cost of traditional feedstock, low buta-
nol titer due to limited microbial tolerance, and high product Spore suspension of C. acetobutylicum NRRL B-527 was prepared
recovery costs because of low production of butanol and presence by using 60% (w/v) starch solution. Actively growing cells of C. ace-
of other by-products. The pineapple peel waste in India is yet to be tobutylicum NRRL B-527 were inoculated in sterile starch solution
 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366 361

and incubated anaerobically for 6–8 days at 37 °C. Spore suspen- Mt2  Mt1
Drying rate ¼ ð3Þ
sion was stored in a cool and dry place for further studies. A heat t2  t1
shock treatment was carried out to activate the cells in reinforced
where t1 and t2 are drying times in min, Mt1 and Mt2 are moisture
clostridia medium (RCM) as reported by Bankar et al. (2013). Acti-
contents of pineapple peel at time t1 and t2, respectively.
vated spore suspension 2% (v/v) was grown into 100 mL sterile
RCM medium for 18–20 h. After 20 h, 5% (v/v) actively growing
2.5.3. Modeling of drying data
cells (with cell density of OD560-1.2) were inoculated into produc-
An unsteady state shell mass balance approach was used to
tion (P2) medium and in hydrolysates. The P2 medium reported by
model the drying kinetics to study structural changes in
Bankar et al. (2012) was used as control during this study.
pineapple sample during drying operation. Mass balance was
made over a thin shell of finite thickness (Dy) perpendicular to
2.4. Characterization of pineapple peel waste
the direction of moisture transport. The conservation of mass
equation can be expressed by considering a section of sample
Proximate analysis was carried out for pineapple peel waste as
on control volume.
per standardized methods in literature. Moisture and ash content
were determined according to Sluiter et al. (2005). Cellulose Rate of accumulation of moisture
content in biomass was determined by Anthrone method ¼ ðRate of moisture in — Rate of moisture out
(Updegraff, 1969) while hemicellulose estimation was carried
out as per reports of Gao et al. (2014). The methods proposed þ Rate of moisture productionÞ
by Sluiter et al. (2011) were used to determine fat and lignin It can be conveniently assumed that no chemical reaction
content. Analysis of crude fiber, lipid, and total protein content occurs during drying operation, and hence the production term
of the pineapple peel were done as described by Rani and Nand becomes zero. The mass balance equation can be written as
(2004). follows:

2.5. Drying of pineapple peel waste @ðqs MDySÞ

Nm jy S  Nm jyþDy S ¼ ð4Þ
@t
Drying experiments were conducted to study the effect of tem- where Nm is mean mass flux of moisture (kg/m2/min), S is the cross
perature, thickness of biomass, and time of drying operation for sectional area (m2), qs is density of solid (kg/m3) and M is moisture
moisture removal. All the experiments were performed in tripli- content (g/g of dry solid). Using mathematical definition of first
cate and results reported are average of three readings. derivative, following differential equation can be obtained, which
describes mass flux gradient as:
2.5.1. Sample preparation and procedure
Pineapple peel was obtained from local markets of Pune, Maha- @Nm @ qs M
 ¼ ð5Þ
rashtra, India. It was ground by using laboratory grinder (Indica, @y @t
Power 750 W). The wet biomass was squeezed out to remove Moisture transport during the drying operation was assumed to
maximum possible liquid from it. A laboratory hot air tray oven occur solely by molecular diffusion phenomena. Therefore convec-
(Bio-Technic BIT-30, India; 400  380  200 mm) with variable tive moisture transport can be neglected. If the constant moisture
temperature controller was used for drying experiments. Different diffusivity without volume change of drying material is assumed,
pineapple peel samples (10 ± 0.2 g, 67 ± 0.2 g, 160 ± 0.2 g) were then Eq. (5) can be reduced as:
weighed and subjected to drying operation. Drying experiments
were carried out in the range of 60–120 °C and same material @ 2 M @M
was used for saccharification studies. Pineapple peel mass and Deff ¼ ð6Þ
@y2 @t
thickness was measured for all the temperatures, after every
10 min initially, and 60 min during later stage of plateau, till it Moisture transport takes place due to concentration gradient
reaches constant. Moisture content, moisture ratio, and drying rate within solid bed, where concentration is low at surface and high
of samples were calculated with the help of data received. All the at bottom of solid bed. An effective diffusivity (Deff) considers the
experiments were carried out at least in triplicate and results change in volume, shape, and texture as well as change in chemical
reported are average ± standard deviation. composition of drying material. Eq. (6) can be solved analytically if
following assumptions are made: (i) moisture movement is unidi-
2.5.2. Drying calculation mensional, (ii) initial moisture distribution is uniform, (iii) no
The experimental moisture content of pineapple peel is esti- chemical reaction (thermal or chemical) takes place during drying
mated for a given time by following expression: operation, (iv) constant moisture diffusivity, negligible shrinkages
and final equilibrium moisture content is close to zero, and (v) neg-
Wt  Wd ligible external resistances to moisture transfer and isothermal
M¼ ð1Þ
Wd process (Crank, 1975):
  2 
where M is the moisture content for a given time (g/g dry solid), Wt M 8 X
1
1 p Deff t
is a mass for given time t (g), Wd is a mass bone dry solid (g). The MR ¼ ¼ 2 exp ð2n þ 1Þ2 ð7Þ
M0 p n¼0 ð2n þ 1Þ 2
4L2
experimental drying data is analyzed using following non dimen-
sional equation: where Deff is effective moisture diffusivity (m2/s), t is a time (s), L is
M  Me a half thickness of samples (m) and n is a positive integer (–). Eq. (7)
MR ¼ ð2Þ can further be simplified to only first term of series, without affect-
M0  Me
ing accuracy of the prediction (Falade and Solademi, 2010; Kara and
where MR is the dimensionless moisture ratio, M is the moisture Doymaz, 2014):
content for a given time (g/g dry solid), M0 is an initial moisture  
content (g/g dry solid) and Me is equilibrium moisture content M 8 p2 Deff t
MR ¼ ¼ 2 exp  ð8Þ
(g/g dry solid). The experimental drying rate was determined by M0 p 4L2
using following expression:
362 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366

The effective moisture diffusivities can be determined for a 2.8. Fermentative production of biobutanol
given temperature when lnDeff is plotted against time. A slope
(K) of straight line determines the value of effective moisture diffu- Fermentation experiments were carried out in 100 mL air-tight
sivity as: screw cap bottles with 80 mL working volume by using C. aceto-
butylicum B 527. Detoxified hydrolysates obtained from dried and
p2 Deff
K¼ ð9Þ non-dried samples were supplemented with other nutritional
4L2 components, similar to P2 medium. Each hydrolysate, after AC
The temperature dependence of effective diffusivity can be detoxification was supplemented with glucose to reach total sugar
expressed using Arrhenius relationship: concentration of 60 g/L for better comparison in diversified exper-
  iments. A medium pH was adjusted to 6.5 using hydrochloric acid
E if required. Anaerobic conditions were maintained by purging
Deff ¼ D0 exp  ð10Þ
RT nitrogen in bottles containing medium, which were sealed with
butyl rubber stopper fastened with an aluminum crimp and auto-
where D0 is the pre-exponential factor (m2/s), E is an activation
claved at 121 °C for 20 min. All sterile hydrolysate bottles were
energy (kJ/mol), R is universal gas constant (kJ/mol/K), and T is dry-
inoculated with 5% (v/v) 20 h old inoculum and incubated at
ing air temperature (K). The kinetic parameters (E and D0) can be
37 °C for 96 h (Harde et al., 2016). A control experiment with P2
estimated from slope and intercept of a plot of ln Deff verses 1/T.
medium was carried out under same experimental conditions.
Samples were collected and used to analyze the solvents and sugar
2.5.4. Statistical analysis consumption.
The ‘goodness of fit’ of proposed drying model to experimental
data was assessed by means of statistical parameters viz. coeffi-
cient of determination (R2), sum squared error (SSE), and chi- 2.9. Analytical method
square (v2). The highest value of R2 (1.0) and lowest values of
SSE and v2 indicate, best fit of model. The values of statistical Phenol sulfuric acid method was used to calculate total sugar
parameters are determined as follows: (TS) in the hydrolysates (DuBois et al., 1956). Solvents (acetone,
butanol and ethanol) and acids (acetic acid and butyric acid) were
1X N
 2 quantified by using gas chromatography as described by Bankar
SSE ¼ Mexp;j  Mpred;j ð11Þ et al. (2013). Gas chromatography (Agilent Technologies 7890B)
N j¼1
equipped with a flame ionization detector and AB-INNOWAX cap-
PN  2 illary column (30 m  0.32 mm  1 lm) was used. Injector and
j¼1 Mexp;j  M pred;j detector temperature were maintained at 200 °C and 250 °C,
v2
¼ ð12Þ
Nz respectively. The total phenolics were determined with Folin-
Ciocalteu reagent using gallic acid as standard (20–100 lg/mL)
where Mexp, j and Mpred, j are experimental and predicted values of
and absorbance of sample was measured spectrophotometrically
jth moisture content (g/g dry solid), respectively. N is number of
(UV-3000+, Labindia analytical) at 760 nm (Maurya and Singh,
experimental runs, and z is number of constants.
2010).

2.6. Saccharification of pineapple peel waste by acid hydrolysis

3. Result and discussion
The chemical pretreatment method was employed using 2%
(v/v) sulfuric acid, 2% (v/v) hydrochloric acid, and 2% (w/v) sodium 3.1. Feedstock and characterization of pineapple peel
hydroxide in both autoclave (solid to liquid ratio 1:10, g/mL, at
121 °C for 60 min) and water bath (solid to liquid ratio 1:10, Pineapple fruit generally consists of three parts: outer cover
g/mL, at 100 °C for 60 min) to obtain total sugars from sun dried (pineapple peel), crown (top parts), and pulp (inner part). During
pineapple peel waste. Further, pretreatment parameters viz. pineapple fruit processing, a huge amount of pineapple waste
concentration of sulfuric acid, solid to liquid ratio, temperature, (peel, core, stem and leaves) is generated which accounts almost
and time were optimized (unpublished data) to be 1.3% (v/v), 50% (w/w) of total pineapple mass. Among them, peel accounts
1:5 g/mL, 121 °C, and 15 min, respectively. Pineapple peel samples for higher portion of about 30–42% (w/w) (Roda et al., 2014). How-
which were dried earlier at different temperatures (as detailed in ever, due to inconsistency in composition of pineapple biomass, it
Section 2.5.1) were pretreated using optimized condition. Wet is essential to carry out proximate analysis of pineapple peel. Prox-
(non-dried) pineapple peel of 4.4 g (equivalent to 1 g of dried sam- imate analysis experiments showed that pineapple peel contained
ple) was also used as a control, during saccharification experiments 35% cellulose, 19.7% hemicellulose, and 16% lignin. Therefore,
and analyzed for total sugar release. pineapple peel has a huge potential to be considered as promising
feedstock for biobutanol production. Further the total ash, total fat,
2.7. Detoxification of pineapple peel hydrolysates total crude fiber, and total proteins were found to be 4.7%, 0.46%,
23.71%, and 0.33%, respectively. Importantly, pineapple peel also
Detoxification of pineapple peel hydrolysates were carried out showed to have large moisture content in the range of 75–80%
to remove fermentation inhibitors by activated carbon (AC) using on wet basis, which is in close agreement with the reports pre-
the method reported by Hodge et al. (2009). The pH of hydrolysates sented by Rani and Nand (2004).
(pH 2.9) was gradually increased to 10 using sodium hydroxide
solution. Subsequently, 5% (w/v) AC powder was added into hydro- 3.2. Drying of pineapple peel
lysates and kept at constant temperature of 60 °C with continuous
shaking at 200 rpm for 2 h. AC was separated by filtration, and fil- Higher moisture content of pineapple peel (77.2% w/w on wet
trate was used for ABE fermentation. Total phenolics content basis) makes it highly susceptible to microbial spoilage and also
before and after detoxification of dried and non-dried pineapple affects the dilution during saccharification experiments. Therefore,
peel hydrolysates was determined by using Folin-Ciocalteu it is desirable to dry the pineapple peel samples to reduce moisture
reagent. content in order to enhance storability and reduce transportation
 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366 363

cost. Fig. 1 shows the declination of moisture content of pineapple

peel with respect to time for a given temperature. All curves show
an exponential tendency of moisture content (g/g of dry solid)
decreasing rapidly with an increase in drying temperature. It was
also observed that the time require to reduce total moisture con-
tent to fixed values increases, as the temperature decreases. The
drying times required to reach minimal moisture content values
of samples were 200, 120, 80 and 50 min at 60, 80, 100 and
120 °C, respectively. These results are in good agreement with
other reports of drying fruits and vegetables such as potato
(Akpinar, 2006), apple and pumpkin (Karabulut et al., 2007),
wherein the falling rate period was observed to explain drying
operation.
The experimental drying data is converted into moisture ratio
using Eq. (2) and plotted against time to estimate values of effec-
tive diffusivity for a given temperature (Fig. 2). The values of effec-
tive diffusivities are obtained using Eq. (9). An effective diffusivity
value varies in the range of 1.35  108 m2/s to 5.27  108 m2/s
for varied temperatures (60–120 °C). It was observed that effective
diffusivity was increased with an increase in drying temperature. Fig. 2. Kinetics of drying showed by plotting moisture ratio versus time at various
When drying was carried out at higher temperature, a greater pro- temperatures.
portion of heat energy is utilized to increase the movement of
water molecules, leading to higher moisture diffusivity (Xiao
et al., 2010). Table 1 shows that effective diffusivity increased five- since Eq. (8) presumes that kinetics of drying follows first order
fold with an increase in temperature from 60–120 °C. The temper- with respect to moisture content:
ature dependence of effective diffusivity can be illustrated using dM
Eq. (10). A straight-line graph of drying temperature vs effective rM ¼  ¼ kM ð13Þ
dt
diffusivity (not shown in this paper), gave quantitative estimation
of activation energy and pre-exponential factor with slope and where k is a drying constant (1/min) and is equal to p2Deff/4L2. The
intercept, respectively. The values of activation energy and pre- rate of drying can be expressed using values of activation energy
exponential factor were found to be 24.60 kJ/mol and and pre-exponential factor as a function of drying temperature
6.12  103 m2/s, respectively. These results are in good agreement and moisture content.
with reports from Antonio et al. (2012) for olive cake and dry veg-  
2960
etable waste. rM ¼ 151:15  exp  M ð14Þ
T
The moisture content of pineapple peel can now be predicted
when value of effective diffusivity is used in Eq. (8) for a given tem- Fig. 3 shows the fitting of Eq. (14) to experimental data, esti-
perature. Fig. 1 shows that predicted values of moisture content mated using Eq. (3). It can be seen that equation fits well to the
with respect to time match well with experimental data, confirm- experimental data within the operating conditions. Further, it is
ing validity of Eq. (8). The ‘goodness of fit’ of proposed model was also clear that the drying rate decreases exponentially as the mois-
also assessed by estimating statistical parameters (R2, SSE, and v2). ture content reduces close to zero (when moisture is removed
The values of R2, SSE, and v2 are reported in Table 1. Moreover, the completely).
rate of drying can also be predicted using following expression,
3.3. Saccharification of pineapple peel waste by acid hydrolysis

Saccharification is an important step to release total sugar from

complex structure of pineapple peel. An acid hydrolysis conditions
can be deliberated as equilibrium between high hemicellulose and
cellulose alterations for efficient bioconversion. Therefore, various
physical, chemical, and biological pretreatment methods have
been reported in the literature. One such well-known chemical
pretreatment method is dilute acid and/or alkali hydrolysis, which
specifically affects hemicellulose structure. Based on preliminary
reports by different researchers (Amiri and Karimi, 2015; Bankar
et al., 2013a; Karimi et al., 2013, 2015; Rattanapoltee and
Kaewkannetra, 2014), dried biomass was treated with both acids
and alkalis 2% (v/v) to perceive the total sugar release from pineap-
ple peel waste. In current investigation, 2% (v/v) sulfuric acid pre-
treatment of dried biomass at 121 °C for 60 min resulted in highest
total sugar release of 44.25 ± 4.05 g/L. Besides, 2% (v/v) sulfuric acid
pretreatment in water bath (at 100 °C for 60 min) also showed
total sugar release to be 40.09 ± 0.79 g/L. Likewise, 2% (v/v)
hydrochloric acid resulted in total sugar yield of 41.63 ± 2.979 g/L
and 36.87 ± 0.79 g/L in autoclave and water bath, respectively. Fur-
thermore, the treatment with 2% (w/v) sodium hydroxide resulted
in 27.5 ± 0.55 g/L of total sugar in both autoclave and water bath
Fig. 1. Effect of temperature on moisture content versus time during drying. experiments. From aforesaid results, it was concluded that
364 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366

Table 1
Statistical parameters applied to assess the drying model and effective diffusivity.

Temperature, (°C) Statistical parameter Effective diffusivity  108, (m2/s)

R 2
SSE v 2

60 0.992 0.012 0.013 1.35

80 0.993 0.010 0.011 2.43
100 0.996 0.014 0.016 3.72
120 0.992 0.016 0.019 5.27

be 58.53 g/L, which was lower than other dried samples. This
may be due to the dilution effect during pretreatments and favor-
able unpredictable structural changes in pineapple peel samples
during drying operation. Moreover, the amount of acid required
would be higher for non-dried samples to release higher sugars
per unit. Additionally, the non-dried samples were stored for 8–
10 days, before its processing, and hence there is a possibility of
utilization of sugars during microbial spoilage. Based on overall
results, the drying operation has shown significant effect on total
sugar release as compared to non-dried pineapple peel samples
(Table 2). Albanese et al. (2014) and Al-Harahsheh et al. (2009)
reported an importance of drying during tomato pomace process-
ing to prevent microbial spoilage because of high moisture content.
Moreover, some researchers also used dilute sulfuric acid (0.5–4%
v/v) treatment for pineapple peel processing to release maximum
sugars with minimum fermentation inhibitors (Rattanapoltee and
Kaewkannetra, 2014; Huang et al., 2011). Besides, it is worth inves-
tigating the effective pretreatment processes with higher biocon-
version capacity, minimum energy requirements, minimum
fermentation inhibitor formation, and simple methodology. This
Fig. 3. The rate of drying versus time at various temperatures. can be achieved by modification in pretreatment methods or blend
of different pretreatment methods.

treatment at higher temperatures (autoclave 121 °C) resulted in 3.4. Detoxification of pineapple peel hydrolysates
higher total sugar release than treatment at relatively lower tem-
peratures (water bath). In addition, 2% (v/v) sulfuric acid treatment Saccharification methodologies and neutralization are mostly
was selected and further optimized (unpublished data) to get max- accompanied with inhibitor formation that negatively influences
imum total sugars. The optimized parameters include treatment ABE production. The inhibitors released during acid hydrolysis
with 1.3% (v/v) sulfuric acid, solid to liquid (g/mL) ratio of 1:5, include furfural, 5-hydroxymethyl-furfural (HMF), acetic acid,
and treatment time of 15 min. By using optimized conditions, the and phenolic compounds. Moreover, effectual detoxification can
pretreatment experiments were undertaken to understand the be prescribed as medicine to eradicate fermentation inhibitors pre-
effect of drying on total sugar release from pineapple peel waste sent in hydrolysate. The removal of inhibitors has been previously
(dried and non-dried). investigated by different methods such as overliming, vacuum
Total sugar release by previously dried samples (60–120 °C) of evaporation, ion exchange resin, and adsorption using AC (Harde
pineapple peel using optimized pretreatment parameters is sum- et al., 2014). Each method has its own characteristics and a specific
marized in Table 2. Interestingly, highest total sugars of 95.82 g/L mechanism of action for inhibitor removal. The AC detoxification
and 97.25 g/L were obtained from the samples dried at 100 °C method is commonly used due to high adsorption capacity with
and 120 °C, respectively. This could be due to alteration in hemicel- minimum total sugar loss. Hence, in present study, the detoxifica-
lulose components of pineapple peel waste. The further character- tions of five hydrolyzates by AC were carried out. Detoxification
ization is needed to confirm this hypothesis and studies are in treatments lead to removal of 95–97% phenolics (total phenolics
progress. Furthermore, time required for drying at 100 and concentration 0.065–0.11 g/L for all hydrolysates). Similarly, AC
120 °C was much lower than at 60 and 80 °C. This ultimately leads detoxification also removed acetic acid up to 50% when used in dif-
to less energy requirement for the overall process. Concurrently, ferent concentrations with only 10–13% total sugar loss (Table 2).
the total sugar release from non-dried sample was observed to The mechanism of AC detoxification includes adsorptive removal

Table 2
Composition of hydrolyzates before and after AC detoxifications.

Pineapple peel sample Hydrolysates after dilute acid treatment Hydrolysates after AC detoxification
TS (g/L) AA (g/L) TP (g/L) TS (g/L) AA (g/L) TP (g/L)
Non-dried sample 58.53 6.58 2.55 52.23 3.99 0.069
Dried at 60 °C 75.23 5.97 3.48 66.85 3.21 0.065
Dried at 80 °C 86.06 4.45 3.40 74.12 3.25 0.096
Dried at 100 °C 95.82 3.81 3.71 85.28 2.82 0.085
Dried at 120 °C 97.25 3.40 3.84 84.25 3.30 0.111

AC: Activated carbon, TS: Total sugar, AA: Acetic acid, TP: Total phenolics.
 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366 365

Table 3
Effect of pineapple peel drying on total solvents and yield of ABE.

50% hydrolysates plus water after AC IS (g/L) SC (g/L) Total ABE (g/L) TA (g/L) Yields of ABE (g/g) Productivity of ABE (g/L/h)
P2 control 60.67 50.54 10.92 1.55 0.21 0.11
Non-dried sample 59.22 29.62 4.11 3.52 0.12 0.04
Dried at 60 °C 63.00 32.40 4.16 3.62 0.13 0.04
Dried at 80 °C 62.00 32.15 4.67 3.75 0.14 0.04
Dried at 100 °C 60.78 34.34 5.00 3.31 0.14 0.05
Dried at 120 °C 60.66 33.07 5.23 3.73 0.15 0.05

IS: Initial sugar, SC: Sugar consumed, TA: Total acids.

of phenolics and dilute acids (Dina et al., 2012; Tiruneh et al., tion during fermentation was in the range of 50–55% and individ-
2016). Adsorption phenomenon may be driven by physicochemical ual results of sugar consumption for varied dried samples are as
interaction. Physical adsorption is either due to weak forces or shown in Table 3. Hence, drying of pineapple peel waste is efficient
electrostatic interactions, whereas adsorption resulting from at high temperature that does not show any negative effect on total
chemical interaction between adsorbent and adsorbate. Due to sugar release as well as on ABE fermentation. This study apparently
heterogeneity of inhibitory compounds in hydrolysates, the seems contradictory to Oberoi et al. (2007) who reported that the
adsorption mode is expected to be a mixture of physical and chem- cauliflower wastes dried at 50 °C retained the color and resulted in
ical adsorption (Tiruneh et al., 2016). Yamamoto et al. (2014) significant glucoamylases production. However, current study is in
reported the use of activated charcoal to reduce the inhibitor con- line with the reports by Avila-Gaxiola et al. (2016) who presented
centration and further improve ABE fermentation efficiency. positive effect of drying temperature on Agave tequilana leaves
Qureshi et al. (2008) also reported the pre-adjustment to pH 10 and showed improved ethanol fermentation.
decreases inhibitory concentration in dilute acid hydrolysate by As can be seen from Table 3, all the fermentation experiments
using AC detoxification. Similarly, Hodge et al. (2009) reported showed high amount of total acids (acetic and butyric acids) pro-
the use of activated carbon to decrease the inhibitor concentration duction that directly affects ABE production. Ezeji et al. (2007)
of 86–96% of HMF and total phenolics after treatment. The applica- reported that presence of 3 g/L furfural and HMF are stimulatory
tion of AC as a detoxifying agent can be an economic process, if to ABE production, whereas 0.3 g/L p-coumaric and ferulic acids
cheap sources of AC or economic regeneration methods are were inhibitory to C. beijerinckii BA101 and decreased ABE produc-
available. tion significantly. Ezeji et al. (2013) showed another way to
enhance butanol production by integrated fed-batch fermentation
system coupled with gas stripping. Although, the final solvent
3.5. Fermentative production of biobutanol
results out of this study may not seem to be promising, we are in
a process to continuously improve it. Furthermore, the efforts to
Lignocellulosic biomass has been widely explored for ABE fer-
improve ABE solvent titer and yield will be targeted to reduce inhi-
mentation by Clostridia species is evident for biphasic fermentation
bitory components present in the medium. Besides, the large quan-
viz. acidogenic phase and solventogenic phase. In current study,
tity of acids produced during ABE fermentation will also be
the hydrolysates obtained after drying at different temperatures
addressed by genetic engineering aspects as well as by bioprocess
followed by acid hydrolysis and detoxification methods were used
modification. The fed batch and continuous ABE process develop-
for ABE fermentation by using C. acetobutylicum B 527. Addition-
ment are additional target areas to check its commercial feasibility.
ally, Khamaiseh et al. (2014) pointed out the addition of nutrients
other than carbon was essential for the fermentation process to get
optimum ABE solvents. Therefore, all hydrolysates were supple-
mented with nutrient similar to standard P2 medium. In order to 4. Conclusions
maintain total sugar concentration (for effective comparison) to
be 60 g/L, auxiliary glucose was added to the detoxified hydroly- Drying of pineapple peel waste showed significant influence on
sates. Likewise, Harde et al. (2016) studied the fermentation of total solvent production and solvent yield with C. acetobutylicum B
ABE using root hydrolysates and achieved maximum total solvents 527. The dried and non-dried samples were treated at different
after 96 h batch fermentation. Based on this study, all the fermen- temperatures and highest total sugar yield was found to be
tation experiments were carried out at batch level for 96 h and 97 g/L for samples previously dried at 120 °C. The detoxified
samples were analyzed for solvent production and substrate uti- hydrolyzates were subjected to fermentation that resulted in
lization. ABE fermentation titers and yields are shown in Table 3. maximum total solvents and yield to be 5.23 g/L and 0.15 g/g
Standard P2 medium containing glucose as carbon substrate was respectively, with maximum sugar utilization. This process may
used as a control and it produced total solvents and total acids to allow the use of drying at industrial scale and therefore helps to
be 10.92 g/L and 1.55 g/L, respectively. reduce the storability and transportation cost.
For non-dried detoxified hydrolysates, the total solvent produc-
tion was obtained to be 4.11 g/L with yield of 0.12 g/g. While, the
highest total solvent of 5.23 g/L and yield of 0.15 g/g were achieved Acknowledgement
when the samples were dried at 120 °C. It can be inferred that the
total solvent and yield of dried samples were higher when com- The authors gratefully acknowledge, Department of Science and
pared with non-dried sample signifying effectiveness of drying Technology (DST) of Ministry of Science and Technology, Govern-
operation. Further the variations in drying temperatures have ment of India, for providing financial support under the scheme
shown significant effect on total solvent production and yield. of DST INSPIRE faculty award, (IFA 13-ENG-68/July 28, 2014) dur-
For example, the sample dried at 120 °C resulted in total solvent ing the course of this investigation. Authors also would like express
of 5.23 g/L whereas sample dried at 60 °C showed total solvent of their gratitude towards Professor Rekha Singhal from Institute of
4.16 g/L. The reason behind increment in solvent production at Chemical Technology, Mumbai, India for her consistent guidance
higher drying temperature is still unclear. Overall sugar consump- and support.
366 M.A. Khedkar et al. / Bioresource Technology 225 (2017) 359–366

References Karabulut, I., Hayaloglu, A.A., Yildirim, H., 2007. Thin-layer drying characteristics of
kurut, a Turkish dried dairy by-product. Int. J. Food Sci. Technol. 42, 1080–1086.
Karimi, K., Shafiei, M., Kumar, R., 2013. Progress in physical and chemical
Akpinar, E.K., 2006. Determination of suitable thin layer drying curve model for
pretreatment of lignocellulosic biomass. In: Gupta, V.K., Tuohy, M.G. (Eds.),
some vegetables and fruits. J. Food Eng. 73, 75–84.
Biofuel Technol. 53–96.
Albanese, D., Adiletta, G., D’Acunto, M., Cinquanta, L., Matteo, M.D., 2014. Tomato
Karimi, K., Tabatabaei, M., Horvath, I.S., Kumar, R., 2015. Recent trends in acetone,
peel drying and carotenoids stability of the extracts. Int. J. Food Sci. Technol. 49,
butanol, and ethanol (ABE) production. Biofuel Res. J. 8, 301–308.
2458–2463.
Khamaiseh, E.I., Hamid, A.A., Abdeshahian, P., Yusoff, W.M.W., Kalil, M.S., 2014.
Al-Harahsheh, M.A., Al-Muhtaseb, A.H., Magee, T.R.A., 2009. Microwave drying
Enhanced butanol production by Clostridium acetobutylicum NCIMB 13357
kinetics of tomato pomace: effect of osmotic dehydration. Chem. Eng. Process.
grown on date fruit as carbon source in P2 Medium. Sci. World J., 7 395754.
48, 524–531.
Kiranoudis, C.T., Maroulis, Z.B., Marinos-Kouris, D., 1995. Heat and mass transfer
Amiri, H., Karimi, K., 2015. Improvement of acetone, butanol, and ethanol
model building in drying with multiresponse data. Int. J. Heat Mass Transfer 38,
production from woody biomass using organosolv pretreatment. Bioprocess
463–480.
Biosyst. Eng. 38, 1959–1972.
Kumbhar, J.V., Rajwade, J.M., Paknikar, K.M., 2015. Fruit peels support higher yield
Antonio, V.G., Luis, P.D., Roberto, L., Margarita, M., María, J.T., 2012. Mathematical
and superior quality bacterial cellulose production. Appl. Microbiol. Biotechnol.
modeling of thin-layer drying kinetics of cape gooseberry (Physalis peruviana L.).
99, 6677–6691.
J. Food Process. Preserv. 38, 728–736.
Maiti, S., Gallastegui, G., Sarma, S.J., Brar, S.K., Bihan, Y.L., Drogui, P., Buelna, G.,
Avila-Gaxiola, E., Avila-Gaxiola, J., Velarde-Escobar, O., Ramos-Brito, F., Atondo-
Verma, M., 2016. A re-look at the biochemical strategies to enhance butanol
Rubio, G., Yee-Rendon, C., 2016. Effect of drying temperature on Agave tequilana
production. Biomass Bioenergy 94, 187–200.
leaves: a pretreatment for releasing reducing sugars for biofuel production. J.
Maurya, S., Singh, D., 2010. Quantitative analysis of total phenolic content in
Food Proc. Eng. (in press)
adhatoda vasicanees extracts. Int. J. Pharm. Technol. Res. 2, 2403–2406.
Bankar, S.B., Survase, S.A., Ojamo, H., Granstrom, T., 2013. The two stage
Namsree, P., Suvajittanont, W., Puttanlek, C., Uttapap, D., Rungsardthong, V., 2012.
immobilized column reactor with an integrated solvent recovery module for
Anaerobic digestion of pineapple pulp and peel in a plug-flow reactor. J.
enhanced ABE production. Bioresour. Technol. 140, 269–276.
Environ. Manage. 110, 40–47.
Bankar, S.B., Survase, S.A., Singhal, R.S., Granström, T., 2012. Continuous two stage
Nga, N.T., Trang, N.T., 2015. Influence of the fermentation of pineapple wastes with
acetone–butanol–ethanol fermentation with integrated solvent removal using
the use of methanobacterium strains separated in Vietnam on the production of
Clostridium acetobutylicum B 5313. Bioresour. Technol. 106, 110–116.
biogas from them. J. Eng. Phys. Thermophys., 1204–1208
Bankar, S.B., Survase, S.A., Ojamo, H., Granstrom, T., 2013a. Biobutanol: the outlook
Oberoi, H.S., Kalra, K.L., Uppal, D.S., Tyagi, S.K., 2007. Effects of different drying
of an academic and industrialist. RSC Adv. 24734.
methods of cauliflower waste on drying time, colour retention and
Choonut, A., Saejong, M., Sangkharak, K., 2014. The production of ethanol and
glucoamylase production by Aspergillus niger NCIM 1054. Int. J. Food Sci.
hydrogen from pineapple peel by Saccharomyces cerevisiae and
Technol. 42, 228–234.
Enterobacteraerogenes. Energy Process. 52, 242–249.
Qureshi, N., Ezeji, T.C., Ebener, J., Dien, B.S., Cotta, M.A., Blaschek, H.P., 2008. Butanol
Crank, J., 1975. The Mathematics of Diffusion. Clarendon Press, Oxford, London.
production by Clostridium beijerinckii Part I: use of acid and enzyme hydrolyzed
Dina, D.J.D., Ntieche, A.R., Ndi, J.N., Ketcha Mbadcam, J., 2012. Adsorption of acetic
corn fiber. Bioresour. Technol. 99, 5915–5922.
acid onto activated carbons obtained from maize cobs by chemical activation
Rani, D.S., Nand, K., 2004. Ensilage of pineapple processing waste for methane
with zinc chloride (ZnCl2). Res. J. Chem. Sci. 2, 42–49.
generation. Waste Manage. 24, 523–528.
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., Smith, F., 1956. Calorimetric
Rattanapoltee, P., Kaewkannetra, P., 2014. Utilization of agricultural residues of
method for determination of sugars and related substances. Anal. Chem. 28,
pineapple peels and sugarcane bagasse as cost-saving raw materials in
350–356.
Scenedesmus acutus for lipid accumulation and biodiesel production. Appl.
Ezeji, T., Qureshi, N., Blaschek, H.P., 2007. Butanol production from agricultural
Biochem. Biotechnol. 173, 1495–1510.
residues: impact of degradation products on Clostridium beijerinckii growth and
Roda, A., Faveri, D.M., Dordoni, R., Lambri, M., 2014. Vinegar production from
butanol fermentation. Biotechnol. Bioeng. 97, 1460–1469.
pineapple wastes-preliminary saccharification trials. Chem. Eng. Trans. 37,
Ezeji, T., Qureshi, N., Blaschek, H.P., 2013. Microbial production of a biofuel
607–612.
(acetone-butanol-ethanol) in a continuous bioreactor: impact of bleed and
Sluiter, A., Hames, B., Ruiz, R., Scarlata, C., Sluiter, J., Templeton, D., 2005.
simultaneous product removal. Bioprocess. Biosyst. Eng. 36, 109–116.
Determination of ash in biomass. Technical Report, NREL/TP 510-42622.
Falade, K.O., Solademi, O.J., 2010. Modeling of air drying of fresh and blanched sweet
Sluiter, A., Hames, B., Ruiz, R., Scarlata, C., Sluiter, J., Templeton, D., Crocker, D., 2011.
potato slices. Int. J. Food Sci. Technol. 45, 278–288.
Determination of structural carbohydrates and lignin in biomass. Technical
FAO, 2015. Statistical Year Book. Food and Agriculture Organization of the United
Report, NREL/TP 510-42618, Revised July 2011.
Nations, Rome, Italy (www.fao.org). Accessed on September 3, 2016.
Tiruneh, A.T., Fadiran, A.O., Ndlela, W.N., Heikkilä, J., 2016. Assessment of technical
Gao, X., Kumar, R., Wyman, C.E., 2014. Fast hemicellulose quantification via a simple
and economic feasibility of activated charcoal removal of organic matter from
one-step acid hydrolysis. Biotechnol. Bioeng. 111, 1088–1096.
different streams of grey water through study of adsorption isotherms. Am. J.
Gautam, S.S., Mishra, S.K., Dash, V., Goyal, A.K., Rath, G., 2010. Comparative study of
Environ. Prot. 5, 56–64.
extraction, purification and estimation of bromelain from stem and fruit of
Updegraff, D.M., 1969. Semimicro determination of cellulose in biological materials.
pineapple plant. Thai J. Pharm. Sci. 34, 67–76.
Anal. Biochem. 32, 420–424.
Harde, S.M., Bankar, S.B., Ojamo, H., Granstrom, T., Singhal, R.S., Survase, S.A., 2014.
Valipour, M., 2014. Comparative evaluation of radiation-based methods for
Continuous lignocellulosic ethanol production using Coleus forskohlii root
estimation of potential evapotranspiration. J. Hydrol. Eng. 20 (5), 04014068.
hydrolysate. Fuel 126, 77–84.
Valipour, M., 2015. Future of agricultural water management in Africa. Arch. Agron.
Harde, S.M., Jadhav, S.B., Bankar, S.B., Ojamo, H., Granstrom, T., Singhal, R.S., Survase,
Soil Sci. 61, 907–927.
S.A., 2016. Acetone-butanol-ethanol fermentation using the root hydrolysate
Valipour, M., Singh, V.P., 2016. Global experiences on wastewater irrigation:
after extraction of forskolin from Coleus forskohlii. Renewable Energy 86, 594–
challenges and prospects. Maheshwari, B., Singh, V.P., Thoradeniya, B. (Eds.),
601.
balanced urban development: options and strategies for liveable cities, Water
Hodge, D.B., Andersson, C., Berglund, K.A., De, U.R., 2009. Detoxification
Sci. Technol. Lib. 72, 289–327.
requirements for bioconversion of softwood dilute acid hydrolyzates to
Xiao, H.W., Pang, C.L., Wang, L.H., Bai, J.W., Yang, W.X., Gao, Z.J., 2010. Drying
succinic acid. Enzyme Microb. Technol. 44, 309–316.
kinetics and quality of Monukka seedless grapes dried in an air-impingement
Hu, S., Zheng, H., Gu, Y., Zhao, J., Zhang, W., Yang, Y., 2011. Comparative genomic
jet dryer. Biosyst. Eng. 105, 233–240.
and transcriptomic analysis revealed genetic characteristics related to solvent
Yamamoto, M., Iakovlev, M., Bankar, S.B., Tunc, M.S., Van Heiningen, A., 2014.
formation and xylose utilization in Clostridium acetobutylicum EA 2018. BMC
Enzymatic hydrolysis of hardwood and softwood harvest residue fibers released
Genomics 12, 93.
by sulfur dioxide-ethanol-water fractionation. Bioresour. Technol. 167, 530–
Huang, F.C., Jiang, Y.F., Guo, G.L., Hwang, W.S., 2011. Development of a yeast strain
538.
for xylitol production without hydrolysate detoxification as part of the
Yannopoulos, S.I., Lyberatos, G., Theodossiou, N., Li, W., Valipour, M., Tamburrino, A.,
integration of co-product generation within the lignocellulosic ethanol
Angelakis, A.N., 2015. Evolution of water lifting devices (pumps) over the
process. Bioresour. Technol. 102, 3322–3329.
centuries worldwide. Water 7, 5031–5060.
Kara, C., Doymaz, I., 2014. Thin layer drying kinetics of by-products from
pomegranate juice processing. J. Food Process. Preserv. 39, 480–487.