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(2008) 1:239247
DOI 10.1007/s12155-008-9018-6
Introduction
Over the last century, energy consumption has increased
progressively as the result of growing world population and
E. Sasikumar (*) : T. Viruthagiri
Bioprocess Engineering Research Laboratory,
Department of Technology, Annamalai University,
Annamalai Nagar 608 002, India
e-mail: sashikumar_ess@yahoo.co.in
240
241
eter (420820). The sugarcane bagasse sample was analyzed for sugar and Klason lignin content following the
procedures described in NREL Standard Procedure (No.
002). Total reducing sugar concentration was determined by
the dinitrosalicylic acid (DNS) method [26]. Then it was
confirmed by bio spectrophotometer (Elico BL 198) at
510 nm. Ethanol was estimated using NUCON 5765 gas
chromatography (GC) with a flame ionization detector and
CHROMATO-PAK (10%C20 M) column (3.0 m
0.32 cm) using nitrogen as the carrier gas at 40 l/min.
The oven temperature was held at 80C. The injector and
detector temperature was maintained at 200C.
Y b0
k
X
i1
bi Xi
k
X
i1
bij Xi2
k X
k
X
ii <j
bij Xi Xj e
242
Symbols
C
P
T
H
Coded levels
2
10
4
25
24
20
5
30
48
30
6
35
72
40
7
40
96
50
8
45
120
243
Table 2 Four level central composite design and the experimental responses of dependent variable (ethanol yield)
Run order
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
1(20)
1(40)
1(20)
1(40)
1(20)
1(40)
1(20)
1(40)
1(20)
1(40)
1(20)
1(40)
1(20)
1(40)
1(20)
1(40)
2(10)
2(50)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
0(30)
pH
1(5)
1(5)
1(7)
1(7)
1(5)
1(5)
1(7)
1(7)
1(5)
1(5)
1(7)
1(7)
1(5)
1(5)
1(7)
1(7)
0(6)
0(6)
2(4)
2(8)
0(6)
0(6)
0(6)
0(6)
0(6)
0(6)
0(6)
0(6)
0(6)
0(6)
0(6)
Temp. (C)
1(30)
1(30)
1(30)
1(30)
1(40)
1(40)
1(40)
1(40)
1(30)
1(30)
1(30)
1(30)
1(40)
1(40)
1(40)
1(40)
0(35)
0(35)
0(35)
0(35)
2(25)
2(45)
0(35)
0(35)
0(35)
0(35)
0(35)
0(35)
0(35)
0(35)
0(35)
1(48)
1(48)
1(48)
1(48)
1(48)
1(48)
1(48)
1(48)
1(96)
1(96)
1(96)
1(96)
1(96)
1(96)
1(96)
1(96)
0(72)
0(72)
0(72)
0(72)
0(72)
0(72)
2(24)
2(120)
0(72)
0(72)
0(72)
0(72)
0(72)
0(72)
0(72)
Predicted
1.5
2.3
2.1
1.9
1.9
2.6
3.3
2.2
2.2
2.6
1.9
2.9
2.3
2.4
2.5
2.8
2.0
2.8
1.8
2.9
2.1
2.2
2.6
3.8
4.1
4.1
4.1
4.1
4.1
4.1
4.1
1.7
2.4
2.0
2.1
2.1
2.7
3.5
2.3
2.3
2.8
2.1
3.1
2.4
2.5
2.7
2.9
1.8
2.7
1.7
3.1
2.0
2.1
2.8
3.2
4.2
4.2
4.2
4.2
4.2
4.2
4.2
244
Table 3 Results of regression analysis and corresponding t and pvalue of second order polynomial model for optimization of ethanol
production
Term
constant
Regression
coefficient
Std
deviation
tstatistic
Pvalue
Intercept
Ca
Pb
Tc
Hd
CC
PP
TT
HH
CP
CT
CH
PT
PH
TH
4.1000
0.1500
0.1667
0.1167
0.1750
0.4588
0.4713
0.5213
0.2588
0.1250
0.1250
0.1000
0.0875
0.0375
0.1125
0.1179
0.0637
0.0637
0.0637
0.0637
0.0583
0.0583
0.0583
0.0583
0.0780
0.0780
0.0780
0.0780
0.0780
0.0780
34.834
2.354
2.616
1.831
2.746
7.859
8.073
8.929
4.432
1.602
1.602
1.281
1.121
0.480
1.441
<0.001
0.032
0.019
0.086
0.014
<0.001
<0.001
<0.001
<0.001
0.129
0.129
0.218
0.279
0.637
0.169
R2 =0.929
Substrate concentration (g/l)
b
pH
c
Temperature (C)
d
Fermentation time (h)
a
Degrees of
Mean
F-value P-value
freedom (df) square (MS)
Regression 20.4076 14
Linear
2.2683 4
Square
17.1317 4
Interaction 1.0075 6
Residual
1.5592 16
error
Lack-of-fit 1.5592 10
Pure error
0.0000 6
Total
21.9667 30
1.45768
0.56708
4.28293
0.16792
0.09745
14.960
5.820
43.950
1.720
<0.001
0.004
<0.001
0.180
0.15592
0.00000
Estimate
Standard
error
pHtemperature (PT)
Intercept
3.627
0.180
0.319
0.115
P2
0.369
0.115
T2
0.601
0.229
P2 T2
Substrate conc.temperature (CT)
Intercept
3.625
0.183
0.306
0.116
C2
0.369
0.116
T2
0.612
0.233
C2 T2
pHfermentation time (PH)
Intercept
3.443
0.212
0.273
0.135
P2
0.061
0.135
H2
0.771
0.270
P2 H2
Temperaturefermentation time (TH)
Intercept
0.3479
0.204
0.332
0.130
T2
0.070
0.130
H2
0.740
0.260
T2 H2
a
Source
Pvalue
20.151
2.790
3.231
2.630
< 0.001
0.009
0.003
0.014
19.850
2.630
3.170
2.630
< 0.001
0.014
0.004
0.014
16.280
2.030
0.450
2.860
<0.001
0.053
0.656
0.008
17.090
2.560
0.540
2.850
<0.001
0.016
0.595
0.008
245
120
Fermentation time (h)
Temperature (C)
45
40
4
35
3
2
30
96
4
72
3
48
2
25
24
4
6
pH
6
pH
45
Temperature (C)
40
4
35
3
30
2
1
25
1
4
3
2
-1
1
-2
10
20
30
40
Substrate conc. (g/l)
50
-2
-1
Temperature (C)
246
Models
Logistic
X0a
Xmb
0c
md
Kse
f
g
h
i
Avg. error
R2
Logistic
incorporated
leudeking piret
Logistic
incorporated
modified
leudeking piret
1.2 g/l
5.5 g/l
0.20 h1
0.086 h1
0.20 h1
0.52 g/l
0.48
0.018
5.80
0.0542
6.47%
0.942
0.38
0.012
8.80
0.016
4.46%
0.966
0.42
0.014
8.20
0.008
3.02%
0.972
Conclusion
Due to dwindling of fossil fuel microbial production of bio
fuel from biomass (mass of living matter) by products has
acquired significant fuel for the future. Bagasse, the solid
residue after extraction of the sugarcane juice, is mostly
utilized for producing steam and electricity required for the
cane processing plant. The price of bagasse ranges from Rs.
100 to 400 (~$2 to 9) per tonne with an average of about Rs.
100 ($2) per tonne. Because of its high carbohydrate content,
relatively low lignin content and its availability as an
industrial waste product, bagasse is a particularly appropriate
substrate for bioconversion to ethanol. This study examines
the possibility of ethanol production from sugarcane bagasse
using cellulase and K. fragilis. Conventional optimization
studies are time consuming and expensive. To overcome
these problems, a central composite design (CCD) was used
for the optimization of process conditions. From the
experimentation, it is evident that the use of statistical
process condition optimization approach, response surface
methodology has helped to locate the most significant
conditions with minimum effort and time. In addition, it
has also proved to be useful in increasing ethanol concentration. A maximum ethanol concentration of 32.6 g/l was
obtained from 180 g/l in of sugarcane bagasse at the optimized
process conditions of temperature 35C, pH 5.5 and fermentation time 72 h in aerobic batch fermentation. Kinetic models
were evaluated and the model constants were predicted on
ethanol production from pretreated sugarcane bagasse.
Acknowledgements The authors express their sincere thanks to
UGC for the financial support through the Rajiv Gandhi National
Fellowship Scheme (RGNFS) for the fellowship to one of the authors
(E. Sasikumar) and to the Department of Technology, Annamalai
University for providing the necessary facilities for the successful
completion of this research work.
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