Rethinking the cane sugar mill by using selective

fermentation of reducing sugars by Saccharomyces dairenensis,

prior to sugar crystallization
Satoshi Ohara
a
, Yasuhiro Fukushima
b,
*, Akira Sugimoto
c
, Yoshifumi Terajima
c
,
Tetsuya Ishida
a
, Akiyoshi Sakoda
d
a
Research & Development Laboratories for Sustainable Value Creation, Asahi Group Holdings Ltd., 1-1-21 Midori, Moriya,
Ibaraki 302-0106, Japan
b
Department of Environmental Engineering and Sustainable Environment Research Center, National Cheng Kung University,
No. 1 University Road, Tainan City 701, Taiwan
c
Tropical Agriculture Research Front, Japan International Research Center for Agricultural Sciences, 1091-1 Maezato-Kawarabaru, Ishigaki,
Okinawa 907-0002, Japan
d
Institute of Industrial Science, The University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8505, Japan
a r t i c l e i n f o
Article history:
Received 5 January 2012
Received in revised form
27 February 2012
Accepted 27 March 2012
Available online 21 April 2012
Keywords:
Selective fermentation
Invertase-defective yeast
High yield sugarcane
Saccharum spp. hybrids
Reducing sugar
Ethanol
a b s t r a c t
High yield sugarcane is expected to resolve the competitionbetween food and fuel regarding
farmland and biomass resources. However, its higher composition of reducing sugars (i.e.,
glucose and fructose), which inhibit sucrose crystallization, hinders the production of sugar
fromhighyieldsugarcane. Under theconventional integratedsugareethanol manufacturing
system, high biomass yield causes only the increase of ethanol production because of the
increase in unrecovered sugar after extraction, which represents a failure in resolving the
competition. The technology presented here is the worlds rst to solve this problem via
selective ethanol fermentationusing Saccharomyces dairenensis, anunconventional yeast that
ferments only reducing sugars and leaves sucrose untouched. A laboratory-scale test using
sugarcane juices witha highcompositionof reducing sugars (100 g kg
1
) resultedina sucrose
crystal yield increase from a single extraction, from 16.2 to 65.1%, by introducing selective
fermentation. The second extraction, fromthe molasses, which was enabled by the lowered
residual reducing sugar composition, further enhanced the total sugar crystal yield (up to
83.4%). A simulation of the application of this technology in the U.S.A. revealed that both
sugar and ethanol production were enhanced, whereas sugar production declined by the
mere adoption of high yield cultivar, even with the increase in sugarcane yield.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Sugarcane juice contains a variety of carbohydrates, among
which sucrose becomes ne sugar, whereas glucose and fruc-
tose (reducing sugars; RS) are rawmaterials for ethanol. Inraw
sugar factories, not all sucrose is extracted as sugar crystals
because the RS inhibit the crystallization of sucrose [1]. As the
demand for ethanol increases, molasses including residual
sugars, such as RS and unrecoverable sucrose after sugar
crystallization, attracts attention as fermentation substrates
* Corresponding author. Tel.: 886 6 2757575x65838; fax: 886 6 2752790.
E-mail address: fuku@mail.ncku.edu.tw (Y. Fukushima).
Available online at www.sciencedirect.com
ht t p: / / www. el sevi er. com/ l ocat e/ bi ombi oe
b i oma s s a nd b i oe ne r g y 4 2 ( 2 0 1 2 ) 7 8 e8 5
0961-9534/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biombioe.2012.03.024
[2e4]. Most yeast strains produce and release invertase, an
enzyme that splits sucrose into RS, outside the cell [5]. Once
yeast is applied to sugarcane juice or molasses, all the sucrose
content is rapidly degraded into RS, which are eventually
converted to ethanol. Therefore, ethanol production has never
been performed before raw sugar production.
Recently, competition between food and energy produc-
tion from biomass resources has attracted global attention
because of severe shortages caused by population increase [6].
To increase the productivity of both sugar and ethanol,
high yield sugarcanes have been developed through breed
improvement and are considered as raw materials [7e9].
However, these new types of sugarcane are rarely adopted
because of their low sucrose purity (Pur) caused by higher RS
in the crops.
Raw sugar productivity is determined mainly by Pur [10];
therefore, high yield sugarcane can be regarded as effective in
increasing ethanol productionbut inefcient in producing raw
sugar under the existing paradigm: to extract much raw sugar
as the main product rst, and produce ethanol subsequently
from the residue. Ethanol production can be increased easily
by direct fermentation of sugarcane juice without raw sugar
making, as practiced in Brazil [11]. Conversely, it is difcult to
enhance raw sugar production.
Increasing the total productivity and control of produc-
tivity of the desired product at the same time requires not only
high yield sugarcane but also a technology aimed at sepa-
rating sucrose from RS during processing. For example,
a chromatographic separation method can remove 85e90% of
the RS from sugarcane molasses [12]. However, using such
a physical removal method requires an additional pre-
processing of suspended solids in the raw material (i.e., cane
juice) for continuous operation.
Alternatively, a biological removal method in which only
RS are selectively converted to a different material, to enable
its separation from sucrose, is used industrially, e.g., the
production of lactosucrose, in which only the by-product (i.e.,
glucose) is removed, without affecting the raw material (i.e.,
sucrose), by using an invertase-defective yeast [13]. The yeast
used in that study ferments only the RS and leaves sucrose
untouched because of its inability to produce invertase. The
same strategy can be applied to enhance Pur during sugar
production, while producing ethanol from the RS. However,
the application of this strategy to integrated sugareethanol
production has never been reported, because the capability of
yeast to convert sucrose into ethanol is believed to be one of
the most important characteristics; therefore invertase-
defective yeasts have been simply considered useless in the
ethanol industry.
In this paper, an innovative integrated sugareethanol
production process that uses the selective conversion of RS is
proposed. This process is the worlds rst attempt to reverse
the production order of sugar/ethanol and is of value in
turning a mere sugar-inhibitor (e.g., RS) separation into an
output of another main product (e.g., ethanol). The feasibility
and effectiveness of the proposed process are veried using
a laboratory-scale test. First, using sugarcane juices with
varied RS contents as the raw materials, a sugareethanol
production test combined with a selective fermentation (SF)
treatment is executed at the laboratory scale. Next, using the
data from laboratory experiments, simulations of raw sugar
production from juices with various Pur were performed,
to highlight the potential of this strategy to control exibly
the products from productive sugarcanes. Finally, the deploy-
ment of the SF treatment in conventional sugar factories is
discussed.
2. Materials and methods
2.1. Microorganism and culture conditions
The invertase-defective yeast used in this study was the
Saccharomyces dairenensis NBRC0211 strain, which was
obtained from the NITE Biological Resource Center (NBRC).
This strain was isolated from dried persimmon [14]. In a seed
culture, the strain was inoculated into 3 mL of synthetic
medium (YM medium: 10 kg m
3
glucose, 5 kg m
3
peptone,
3 kg m
3
yeast extract, and 3 kg m
3
malt extract) in a 15 mL
test tube, and incubated on a reciprocating shaker for 12 h at
2 Hz. In the preculture, the strain grown in the seed culture
was harvested and placed in 1 L of YM medium in a 2 L ask
with bafes. The culture was incubated on a rotary shaker at
2 Hz for 12 h. The incubation was performed at 28

C.
2.2. SF in the model medium
To investigate the fermentation characteristics of S. dair-
enensis NBRC0211, a fermentation test was conducted using
the model medium (120 kg m
3
sucrose, 30 kg m
3
glucose,
10 kg m
3
yeast extract, 3 kg m
3
malt extract, and 20 kg m
3
peptone), which has a composition similar to that of sugar-
cane juice. The precultured yeast was inoculated into 150 mL
of model medium in a 300 mL ask with a fermentation lock.
The initial cell number was 5.0 10
7
dm
3
. Thereafter,
fermentation was performed on a rotary shaker at 2 Hz for
12 h at 28

C.
The ethanol production rate was calculated by dividing the
total ethanol production per liter of culture at the end of the
fermentation by the fermentation time, whereas the ethanol
yield was calculated relative to the theoretical amount
calculated using chemical stoichiometry, assuming the
complete conversion of the RS in the juice into ethanol.
2.3. Sugar production via SF from sugarcane juices
A schematic of the proposed process using an SF treatment is
shown in Fig. 1A, in contrast to the conventional process
(Fig. 1B). To achieve the same objective, the proposed process
reverses the order of the production of the sugar and ethanol.
2.3.1. Preparation of sugarcane juices
Juice was prepared from Ni15 sugarcane (Saccharum spp.
hybrids), which is the breed used most commonly for sugar
manufacture in Japan [15]. In September of 2004, Ni15 cuttings
were planted at the experimental eld of dark red soil (Shi-
majiri Mahji) on Ie Island (26

43
0
N, 127

47
0
E, Okinawa, Japan).
The inter-row space and the inter-hill space were 1.25 m and
0.30 m, respectively. Chemical fertilizer was applied in the
amounts of N 260 kg ha
1
, P
2
O
5
100 kg ha
1
, and
b i o ma s s a nd b i o e ne r gy 4 2 ( 2 0 1 2 ) 7 8 e8 5 79
K
2
O 100 kg ha
1
. Matured sugarcane (18 months after
planting) was harvested in March of 2006. After sugarcane top
and dead leaf were removed, the stalks of sugarcane were cut
in small pieces about 0.2 m long. Then the feedstock was
immediately crushed with a shredder KS-MS (Matsuo, Japan)
and squeezed with a press machine KS-OP (Matsuo, Japan) to
extract its juice. Thereafter, the juice was frozen quickly and
stored in the freezer at 20

C.
From the juice of Ni15 after thawed at room temperature,
adjustedjuice samples withlower Pur (0.80, 0.70, 0.65, 0.60, and
0.55 g g
1
) were prepared by adding RS (glucose:fructose 1:1).
After adjusting Pur, the juice samples were sterilized in an
autoclave at 120

C for 15 min. The composition of the juice
samples is summarized in Table 1. Pur, dened in equation (1),
is a process control index used widely in sugar manufacture
throughout the world:
Pur
SUC
Bx
(1)
where Pur represents sucrose purity (g g
1
), SUC represents
sucrose content (g kg
1
), and Bx represents soluble solid
content (g kg
1
).
2.3.2. SF using sugarcane juices
The invertase-defective yeast S. dairenensis NBRC0211 was
inoculated into 3 L of each juice in a 5 L ask so that the initial
cell number was 5.0 10
7
dm
3
. Batch fermentation experi-
ments were performed using a rotary shaker at 2 Hz, 28

C,
until the consumption of RS stopped. After the fermentation
was complete, the yeast cells were removed by centrifugal
separation (18,000 m s
2
) and the fermented juice was
collected.
2.3.3. Sugar production after SF
Sugar productionfromw3 L of fermented juice was performed
simultaneously with ethanol extraction using a rotary evap-
orator (NE-2001, Eyela, Japan). First, the fermented juice was
heated under reduced pressure (gauge pressure, 88 kPa), to
evaporate the water and ethanol in the fermented juice, and
syrup (i.e., the highly concentrated juice) with a Bx of
600 g kg
1
was obtained. Here, the heating bath temperature
was 70

C and the rotation speed was 2 rad s
1
. Half of the
syrup was then condensed further to a supersaturated state
(i.e., 110e120% of the saturation solubility of sucrose). At this
point, granulated sugar with a grain size ranging from 250 to
Proposed process
Juice after SF
(Sucrose)
Sugarcane juice
(Sucrose, RS)
Sugar
(Sucrose)
Ethanol
Molasses
(Sucrose)
Selective ethanol
fermentation (SF)
Sugar
production
Ethanol
Conventional
ethanol
production
Sugarcane juice
(Sucrose, RS)
Sugar
production
Sugar
(Sucrose)
Conventional process
Molasses
(Sucrose, RS)
Ethanol
Conventional
ethanol
production
A B
Fig. 1 e Schematic representation of the integrated sugareethanol production from sugarcane. Proposed process (A) and
conventional method (B).
Table 1 e Composition of fresh and Pur-adjusted sugarcane juice samples.
Juice sample Pur
a
[g g
1
] Bx
b
[g kg
1
] Sucrose [g kg
1
] Glucose [g kg
1
] Fructose [g kg
1
]
Fresh juice (Ni 15) 0.833 196 163 9 8
Adjusted juice (1) 0.791 207 164 14 13
Adjusted juice (2) 0.697 223 156 24 22
Adjusted juice (3) 0.669 235 157 31 29
Adjusted juice (4) 0.625 248 155 38 37
Adjusted juice (5) 0.553 268 148 51 49
a Pur, sucrose purity: the ratio of mass of sucrose to the soluble solid content.
b Bx, soluble solid content.
b i oma s s a nd b i oe ne r g y 4 2 ( 2 0 1 2 ) 7 8 e8 5 80
500 mm was added at a ratio of 100 g per kg of syrup, serving as
seed crystals to trigger sucrose crystallization. Throughout the
process, the supersaturation level was kept constant by add-
ing a quantity of syrup that was equivalent to the volume of
water evaporated during heating, thereby achieving sucrose
crystallization in about 3 h. The crystallized sugar was sepa-
rated from molasses using an H-110A centrifugal separator
(Kokusan, Japan) at 7400 m s
2
. Finally, the mass of the sugar
crystals obtained was measured after drying at 60

C for 24 h.
As a control experiment, sugar production tests using each
juice sample were performed under the same conditions, but
in the absence of the SF treatment.
2.4. Analytical methods
Saccharide composition was measured via high-performance
liquid chromatography using an LC-10AD chromatogram
(Shimadzu, Japan) tted with an RID-10A refractive index
detector (Shimadzu, Japan). Saccharides were separated on
a SUGAR SC1011 column (Shodex, Japan) using distilled water
as the mobile phase at a ow rate of 0.8 mL min
1
. Column
temperature was maintained at 80

C. Ethanol concentration
was measured using a BF-5 biosensor (OSI, Japan). The
number of yeast cells was measured using a CYTORECON
automatic cell-imaging counter (ECI, Japan).
2.5. Simulation of sugar yield
To simulate maximum raw sugar yields for the juice samples
with various Pur values, a model was constructed by
combining the results of the experiments. Using the data, the
Pur values of the syrup and molasses and the sugar yield were
modeled using polynomials determined by minimizing the
squared residuals. Sugar extraction can be performed more
than once if the sugar demand is higher; therefore, when the
Pur of molasses is greater than 0.553 g g
1
, sugar from the
consecutive extractionis calculatedupto three times. Intting
the polynomial models, constraints are imposed so that the
values at 0.553 g g
1
equal the value obtained from the exper-
iment. Inthesimulation, theminimumPur for sugar extraction
was determined based on the lower limit of Pur at which sugar
could be extracted in this laboratory experiment. This model
included the material balances of sucrose, RS, and other
components, and no loss of any component was assumed.
3. Results and discussion
3.1. Prole of SF in the model medium
Theprole of fermentationinmodel mediumis showninFig. 2.
Ethanol was generatedselectivelyfromglucose, i.e., nosucrose
in the model medium was consumed by S. dairenensis. This
result conrmed that a systemusing invertase-defective yeast
provides no pathway for sucrose degradation. Glucose was
almost completely consumed. Ethanol yield was 88.8%and the
ethanol production rate was 2.6 kg m
3
h
1
. Yeast propagated
up to three times more than initial cell concentration, regard-
less of the limited availability of carbohydrate. This charac-
teristic is considered favorable to convert only RS into ethanol.
3.2. Effect of SF treatment on the Pur of sugarcane juices
and syrups
The compositions of juice samples after SF treatment are
shown in Table 2. It was conrmed that the Pur of juices after
SF treatment had been improved, as expected. No degradation
or consumption of sucrose caused by SF was noted for any of
the juices. After SF treatment, the juices consisted of
0
30
60
90
120
150
0 2 4 6 8 10 12
Fermentation time [h]
S
u
g
a
r
,

E
t
h
a
n
o
l

[
k
g
m
-
3
]
0
50
100
150
200
250
Y
e
a
s
t

c
e
l
l

n
u
m
b
e
r
[
1
0
6
d
m
-
3
]
Sucrose
Glucose
Ethanol
Yeast
Fig. 2 e Prole of fermentation in the model medium for S.
dairenensis (NBRC0221).
Table 2 e Composition of juice samples after SF treatment and respective RS removal.
Juice sample Pur
a
[g g
1
]
Bx
b
[g kg
1
]
Sucrose
[g kg
1
]
Glucose
[g kg
1
]
Fructose
[g kg
1
]
Ethanol
[g kg
1
]
RS removal
c
[%]
Fresh juice (Ni 15) 0.898 187 168 1 1 5 84.1
Adjusted juice (1) 0.874 189 166 2 1 9 88.9
Adjusted juice (2) 0.850 194 165 2 3 17 91.3
Adjusted juice (3) 0.823 194 160 2 2 24 94.3
Adjusted juice (4) 0.811 196 159 3 2 29 94.0
Adjusted juice (5) 0.775 194 150 2 3 42 95.1
a Pur, sucrose purity: the ratio of mass of sucrose to the soluble solid content.
b Bx, soluble solid content.
c RS removal: the ratio of reducing sugars (RS) in juice removed by SF treatment.
b i o ma s s a nd b i o e ne r gy 4 2 ( 2 0 1 2 ) 7 8 e8 5 81
5e42 g kg
1
of ethanol and an RS removal level of 84.1e95.1%
was achieved. The juice was then condensed, after removal of
yeast by evaporation. Ethanol was simultaneously recovered,
together with evaporated water, in this process. The ethanol
content in syrup was less than 1 g kg
1
, thereby conrming
that ethanol was almost completely recovered.
Fig. 3 contrasts the Pur of syrups with or without SF treat-
ment, demonstrating that the mere removal of impurities and
juice concentration without SF treatment had almost no effect
on Pur. In contrast, a signicant improvement in the Pur of
syrup was noted after SF treatment. In any case, the Pur of
syrup did not reach 1.00 g g
1
even after the SF treatment
because of the presence of other nonfermentable consti-
tuents (minerals, amino acids, etc.), which were present
(42e51 g kg
1
) as the soluble solids in the syrup. Further
improvement is expected if a technology that effectively
removes the nonfermentable constituents is developed.
However, the contribution of such technology would be
limited, as the crystallization inhibition effect of these
constituents is lower than that of the RS [1]. The Pur of syrups
with SF treatment should theoretically be the same across the
cases if the RS are completely removed, because juices in
different cases were prepared by adding only the RS to the
same juices. In this study, the syrup Pur varied within a range
of 0.86e0.91 g g
1
. Residual RS after SF were observed, even in
juices with low content of RS. It is presumed that some RS in
juice turned into nonfermentable material (e.g., melanoidin),
for some reason (e.g., thermal denaturation due to the steril-
ization in an autoclave).
3.3. Effect of SF treatment on sugar production
Table 3 shows the yield and quality of raw sugar, ethanol, and
molasses in the presence or absence of SF treatment in the
laboratory-scale sugar crystallization test.
The sugar yield in the presence of SF was higher than
without SF. A comparison of the raw sugar yields in the
presence or absence of the SF treatment is shown in Fig. 4A.
Without SF treatment, the sugar yield decreased signicantly
as the Pur of the juice decreased. This result is consistent with
existing plant-scale processes [10]. In contrast, with the SF
treatment, a sugar extraction rate of more than 65% was
achieved, regardless of the Pur in the juice samples. In
particular, the improvement in raw sugar yield was larger for
the juice samples with a lower Pur. For example, the rawsugar
yield at a juice Pur of 0.833 g g
1
increased from64.4%to 71.2%,
whereas the raw sugar yield at a juice Pur of 0.553 g g
1
increased from 16.2% to 65.1%.
In addition, the Pur of raw sugar after SF treatment was
constantly above 0.95 g g
1
(0.952e0.973 g g
1
), which was
better than that observed in the control group
(0.818e0.965 g g
1
). After SF treatment, it was easy to separate
molasses from massecuite, i.e., the mixture of sugar crystals
and molasses, via centrifugal separation. The RS, which
formerly caused separation problems due to its high viscosity,
has been removed by SF treatment; therefore, the quality of
the raw sugar improved.
0.5
0.6
0.7
0.8
0.9
1.0
0.5 0.6 0.7 0.8 0.9
Pur of juice [g g
-1
]
P
u
r
o
f

s
y
r
u
p

[
g

g
-
1
]
with SF
without SF (conventional)
(proposed)
Fig. 3 e Improved Pur of syrup after SF treatment. Pur
represents sucrose purity, the ratio of mass of sucrose to
the soluble solid content.
Table 3 e Yield and quality of raw sugar, ethanol, and molasses in the presence or absence of SF treatment using raw
materials with various Pur.
Process Raw material Yield per cubic meter of juice Quality
Raw sugar
a
[kg-dry]
Ethanol
b
[kg]
Molasses
[kg-dry]
Raw sugar
a
Pur
c
[g g
1
]
Ethanol
C
d
[g kg
1
]
Molasses
Pur
c
[g g
1
]
Conventional
(without SF)
Fresh juice (Ni 15) 118.2 e 90.3 0.958 e 0.668
Adjusted juice (1) 105.4 e 106.0 0.965 e 0.618
Adjusted juice (2) 102.6 e 128.1 0.934 e 0.542
Adjusted juice (3) 100.5 e 148.5 0.936 e 0.508
Adjusted juice (4) 67.8 e 209.5 0.818 e 0.532
Adjusted juice (5) 33.3 e 266.5 0.891 e 0.521
Proposed
(with SF)
Fresh juice (Ni 15) 129.2 5.5 66.3 0.952 7 0.753
Adjusted juice (1) 130.1 9.0 65.9 0.963 11 0.747
Adjusted juice (2) 118.2 17.9 75.6 0.969 22 0.719
Adjusted juice (3) 118.3 24.4 79.0 0.969 27 0.698
Adjusted juice (4) 113.4 30.3 88.3 0.973 37 0.660
Adjusted juice (5) 108.9 43.0 83.8 0.960 52 0.675
a Raw sugar: mass of sugar crystal including impurity.
b Ethanol: mass of ethanol content.
c Pur, sucrose purity : the ratio of mass of sucrose to the soluble solid content.
d C, concentration of extracted ethanol.
b i oma s s a nd b i oe ne r g y 4 2 ( 2 0 1 2 ) 7 8 e8 5 82
The Pur of molasses also increased after application of the
SF treatment, as shown in Fig. 4B. When the Pur of molasses is
low, further extraction of sugar becomes difcult. In such
a case, the only option for using the remaining sucrose in the
molasses is the production of ethanol. Increasing the Pur of
molasses allows further extraction of sugar, and molasses can
be used to produce either sugar or ethanol.
3.4. Simulated sugar yield in the presence or absence of
SF treatment
To estimate sugar yields from the juice samples with
various Pur values, a simulation model was constructed by
combining the results of the experiments shown in Fig. 4.
The simulation demonstrated the potential number of sugar
extractions, the sugar yield in each stage, and the maximum
yield, as shown in Fig. 5. The Pur of juices in the simulation
ranged from 0.56 g g
1
to 0.90 g g
1
, based on the results of
the experiments.
Assuming a juice sample with a high Pur (w0.90 g g
1
),
sugar was extracted three times in both cases and the yields
were similarly high, regardless of the use of SF treatment. As
the purity of the raw material juice declined, the difference in
sugar yield became larger. When the juice Pur was less than
w0.70 g g
1
, the effectiveness of the SF treatment became
obvious as, without SF, sugar was extracted only once,
whereas with SF, sugar was extracted twice. For example, at
a low Pur of 0.56 g g
1
, only 19.3% of the sucrose was extracted
using the conventional method. In addition, two or more
sugar extractions were not possible because the Pur of the
molasses obtained was low. Using the SF method, up to 64.8%
of the sucrose was extracted at once from the same juice. This
productivity can be enhanced further to 83.5% by additional
sugar extraction from molasses.
P
u
r
o
f

m
o
l
a
s
s
e
s

[
g

g
-
1
]
R
a
w

s
u
g
a
r

y
i
e
l
d

[
%
]
0
20
40
60
80
100
0.5 0.6 0.7 0.8 0.9
Pur of raw material [g g
-1
]
0.4
0.5
0.6
0.7
0.8
0.5 0.6 0.7 0.8 0.9
Pur of raw material [g g
-1
]
with SF
without SF (conventional)
(proposed)
A B
Fig. 4 e Inuence of raw material Pur on raw sugar yield (A) and Pur of molasses (B). Raw sugar yield indicates the ratio of
mass of sucrose recovered as sugar crystal to the mass of sucrose in syrup. Pur represents sucrose purity, the ratio of mass
of sucrose to the soluble solid content.
0
20
40
60
80
100
0.6 0.7 0.8 0.9
Pur of juice [g g
-1
]
R
a
w

s
u
g
a
r

y
i
e
l
d

[
%
]
without SF
with SF
1
st
sugar
2
nd
sugar
3
rd
sugar
Fig. 5 e Simulated raw sugar yield in the presence or absence of SF treatment, plotted against the Pur of juices. Raw sugar
yield indicates the ratio of mass of sucrose recovered as sugar crystal to the mass of sucrose in syrup. Pur represents
sucrose purity, the ratio of mass of sucrose to the soluble solid content.
b i o ma s s a nd b i o e ne r gy 4 2 ( 2 0 1 2 ) 7 8 e8 5 83
As highlighted by the experiments and simulations, SF
treatment enhances the sucrose fraction in the by-product
molasses, thereby allowing a exible and strategic control of
the production of sugar and ethanol, depending on the market
demands. In this scenario, sugar productivity can be deter-
mined much more exibly and is affected less by the ratio of
RS to total sugar.
Table 4 [9,16e21] shows the yield and Pur of sugarcane
cultivars in the world. High yield sugarcanes tend to have
lower Pur than the conventional types. The simulations per-
formed using the data of the worlds cultivars demonstrate
the enhancement of sugar yield using the proposed tech-
nology (Table 4). For example, in the case of the U.S.A., the
combination of the conventional cultivar (i.e., CP65-357) with
an existing factory produces 13.0 tonnes of raw sugar on the
basis of 1 ha of land. By cultivating L79-1002, high yield
sugarcane, instead of the conventional cultivar, the sugarcane
yield increases 2.4-fold (from 73.3 to 177.2 t ha
1
). Despite the
signicant increase in crop yield, the sugar crystal yield
declines (1.5 t ha
1
) because the recovery rate of sugar
decreases because of its low Pur. The application of the SF
treatment to CP65-357 and L79-1002 allows the enhancement
of raw sugar yield to 13.3 and 14.6 t ha
1
, respectively. Similar
improvements by introducing high yield cultivars and/or SF
treatment are also expected in other areas.
3.5. Deployment of SF treatment technology
Fig. 6 demonstrates a concrete example of introduction of the
SF treatment into an integrated sugareethanol factory.
Despite the fundamental difference in the conceptual design,
the modication of the facility layout required to introduce
the SF treatment is minimal. First, an SF tank is added
between the roll milling and clarication processes of
a conventional sugar factory. Impurities and yeast in the
sugarcane juice after SF can be removed using the contin-
uous vacuum rotating lter used in the conventional clari-
cation and ltration processes. In the subsequent
concentration process to obtain syrup, ethanol is then
vaporized together with water. As shown in Table 3, the
concentration of extracted ethanol in laboratory-scale tests
was 7e52 g kg
1
. The ethanol was almost completely
Table 4 e Yield and Pur of the worlds sugarcane cultivars and simulated production of raw sugar and ethanol.
Country Sugarcane information Simulated production
Conventional process Proposed process
Cultivar Type Cane yield
[t ha
1
]
Pur
a
[g g
1
]
Raw sugar
b
[t ha
1
]
Ethanol
c
[t ha
1
]
Raw sugar
b
[t ha
1
]
Ethanol
c
[t ha
1
]
Japan [16] NiF8 Conventional 75.2 0.850 8.2 1.2 9.1 0.8
KY01-2044 High yield 101.3 0.783 8.4 2.1 9.9 1.4
Brazil [17,18] RB72454 Conventional 89.8 0.833 10.2 1.7 11.4 1.1
RB867515 High yield 103.2 0.735 6.9 2.6 8.8 1.7
Thailand [19] K84-200 Conventional 91.4 0.797 12.5 2.7 14.5 1.9
MPT00-478 High yield 137.4 0.574 3.5 9.7 11.6 5.6
U.S.A. [20] CP65-357 Conventional 73.3 0.927 13.0 0.7 13.3 0.6
L79-1002 High yield 177.2 0.742 11.5 4.1 14.6 2.7
Australia [21] Q117 Conventional 65.0 0.882 9.3 0.9 9.8 0.7
KQ04-6003 High yield 128.0 0.792 13.5 3.1 15.7 2.1
Barbados [9] B77602 Conventional 77.6 0.891 12.1 1.0 12.7 0.8
WI79460 High yield 112.2 0.730 8.0 3.1 10.4 2.1
a Pur, sucrose purity: the ratio of mass of sucrose to the soluble solid content.
b Raw sugar: mass of sucrose as sugar crystal.
c Ethanol: mass of ethanol content.
Fig. 6 e Modication of an integrated sugareethanol production process for the introduction of the SF treatment. The red
lines show the new equipment needed and the remaining equipment used is as in conventional sugareethanol production.
b i oma s s a nd b i oe ne r g y 4 2 ( 2 0 1 2 ) 7 8 e8 5 84
recovered, as stated above. Therefore, new equipment for
ethanol recovery is not necessary. Nevertheless, the
concentration of extracted ethanol will be very low, even in
a factory-scale setting, because the evaporators used in sugar
mills are of the simple distillation type. To further concen-
trate the ethanol extracted, distillation towers and dehydra-
tion equipment for conventional ethanol production can be
used. Only additional piping to send the thin ethanol into
distillation towers is required as the second modication.
The crystallization and centrifugation processes after
concentration are kept the same.
The proposed technology is applicable to unused sugar-
canes. Both productive cultivars and off-season crops tend to
yield a low Pur juice and are thus usually considered unfa-
vorable for sugar extraction. Therefore, productivity is limited
at current levels, and the harvest season has been limited
within a short period to maintain the Pur of juice within
a favorable range (in countries such as Japan and Taiwan).
This results in inefcient, small-scale sugar production using
large-scale equipment for short periods. SF treatment may
resolve these two issues, thereby reducing the production
costs signicantly.
Similarly, SF treatment can be applied to other sugar crops
from which it is difcult to extract sucrose crystals because of
high RS content, e.g., sorghum [22] and maple [23].
A wide range of options to produce either food or biofuel
will be provided by this technology. The use of the untapped
selectivity of microorganisms breaks down stereotypes such
as ethanol vs sugar and will revolutionize the existing
industries. Moreover, this technology offers an innovative
solution to maximize productivity from biomass resources.
4. Conclusion
The laboratory-scale test using actual sugarcane juices
demonstrated that only the RS are selectively converted into
ethanol by invertase-defective yeast, thereby enabling sugar
production with an enhanced yield. Based on experimental
results, the potential of sugar productivity enhancement was
calculated for sugarcane juices with varying Pur. With
a minimal retrot onthe existing sugar factories, the proposed
technology simultaneously allows 1) the enhancement of
overall productivity on farmland and 2) the exible control of
a more desired product (i.e., sugar or ethanol) depending on
economic situation, regardless of RS composition.
Acknowledgments
We thank Dr. Yasunori Kikuchi and Dr. Masahiko Hirao at the
University of Tokyo for discussions.
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