Professional Documents
Culture Documents
Production
1
I.
INTRODUCTION
A. Substrates
Sweet sorghum grains were obtained from Indonesian
Bioenergy Foundation and blended into small size of
approximately 20 m to enhance the hydrolysis process.
B. Enzymes
Both -amylase from Bacillus subtilis and glucoamylase
from Aspergillus niger were obtained from enzyme industry
in Riau, Indonesia. The activities of the two enzymes were
identified to be 130,000 U/mL and 25,000 U/mL
respectively.
C. Hydrolysis
The shake flask was filled with 100 ml of distilled water
and heated to 80 C. Then, 25 g of sweet sorghum was
added to the flask (to make 25% (w/v) of substrate). After
that, 0.2% (v/w) of -amylase (from the amount of
sorghum) was added and the mixture was cooked at 80 C
and mixed at 250 rpm for 1 h. After 1 h, the mixture was
cooled down to 60 C and 0.1% (v/w) of glucoamylase was
added and the mixture was left for 4 h with 250 rpm
agitation.
D. Dextrose Equivalent (DE)
Samples dextrose equivalent (DE) determinations were
centrifuged at 5000 rpm for 30 min to remove the substrates.
The supernatant was filtered through a 0.45 m membrane
and analyzed by high performance liquid chromatography
(HPLC) equipped with a refractive index detector. The
column used for separation was a SUPELCOGEL C-610H
column. 10 l of sample was injected into HPLC and
separation was performed at 30 C with 0.1% H3PO4 as the
mobile phase at a flow rate of 0.5 mL/min. Glucose was
used as a standard. DE was calculated as follows:
100
(1)
E. Experimental Design
The 2 Level Factorial Design was used to identify which
factors of hydrolysis process have significant effects on the
response, DE. The factors selected for the experiment were
the amount of substrate (A, % (w/v)), liquefaction
temperature (B, C), liquefaction time (C, h), amount of amylase (D, % (v/w)), saccharification temperature (E, C),
saccharification time (F, h), and amount of glucoamylase
(G, % (v/w)). The factors were examined at two different
levels (low and high) coded (1 and 2, respectively) as shown
in Table I. This design gave an output of eight experimental
runs (combinations) with seven independent variables. All
the experiments were performed in triplicate and the average
of DE was used as the response (dependant variable). Both
the experimental runs and response are shown in Table II.
The 2 Level Factorial Design is based on the first order
model which is as follows:
Symbol
A
Coded levels
Low
High
(1)
(2)
25
35
Liquefaction temperature, C
80
Liquefaction time, h
0.1
0.2
Saccharification temperature, C
50
60
Saccharification time, h
0.1
0.2
III.
Results
0.75029
1.79857
0.93357
144.829
Factors
1
2
3
4
5
6
7
8
TABLE II
THE OBSERVED AND PREDICTED RESULTS FOR DEXTROSE EQUIVALENT VALUE
Run
90
A (% (w/v))
B (C)
C (h)
D (% (v/w))
E (C)
F (h)
G (% (v/w))
25
35
25
35
25
35
25
35
80
80
90
90
80
80
90
90
1
1
1
1
2
2
2
2
0.2
0.1
0.1
0.2
0.2
0.1
0.1
0.2
60
50
60
50
50
60
50
60
4
4
2
2
2
2
4
4
0.1
0.2
0.2
0.1
0.2
0.1
0.1
0.2
Response
Y (% (w/w))
Observed
Predicted
27.70
26.14
51.97
51.25
54.28
53.55
51.26
49.70
61.62
62.35
20.67
22.23
52.04
53.60
41.73
42.45
TABLE III
ANOVA FOR THE ENTIRE MODEL
Source
Sum of
squares
DF
Mean
square
F-value
p-value > F
Model
1389.469
277.8937
46.86677
0.0210
112.5857
112.5857
18.98758
0.0488
174.3111
174.3111
29.39757
0.0324
657.1866
657.1866
110.8345
0.0089
25.87887
25.87887
4.364471
0.1719
419.5063
419.5063
70.74972
0.0138
Residual
11.85888
5.929441
Cor total
1401.328
DISCUSSION
its mean, and the estimated factor effects are real. Also, the
F-value is inversely proportional to p-value > F. Higher Fvalue will result to lower p-value > F.
20.00
14.48
15.00
9.34
10.00
Main Effect
5.00
0.84
0.00
-5.00
-10.00
C
-2.29
F
-3.60
-7.50
-15.00
-20.00
-18.13
CONCLUSION
REFERENCES
[1]
[2]
[3]
[4]
[5]
[6]
[7]
[8]
[9]
[10]
[11]
[12]
[13]
[14]
[15]
[16]
[17]
[18]
[19]