24
Additional file 1
Optimization of upstream and downstream process parameters for
cellulase-poor-thermo-solvent-stable xylanase production and extraction by
Aspergillus tubingensis FDHN1
Dharmesh N. Adhyaru1, Nikhil S. Bhatt1*, H.A. Modi2
1
2
P. G. Department of Microbiology, Gujarat Vidyapeeth, Sadra- 382 320, Gujarat, India
Department of Life Sciences, School of Sciences, Gujarat University, Ahmadabad, Gujarat, India
Corresponding author
Dr. Nikhil S. Bhatt*
P. G. Department of Microbiology, Gujarat Vidyapeeth, Sadra- 382 320, Gujarat, India.
Contact No.: (M) +91 98794 83847, (O) 079- 23274274
Fax No.: (079) 232 74 272
E. Mail address: bhattnikhil2114@gmail.com
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Table S1
Effect of metals and modulators on xylanase production from A. tubingensis FDHN1 under SSF
Potential additives
Xylanase activity
(U/g)
Protein
(mg/g)
Specific activity
(U/mg)
Cellulase activity
(U/g)
Control a
Metal compounds
CuCl2
MgCl2
SnCl2
CaCl2
HgCl2
CoCl2
MnCl2
NiCl2
FeCl3
1957 ± 17
27.22 ± 1.36
71.89 ± 2.98
3.12 ± 0.16
2239 ± 20
2446 ± 27
1597 ± 16
2179 ± 17
973.4 ± 14
1316 ± 28
2089 ± 20
1657 ± 24
2047 ± 14
29.07 ± 1.16
25.47 ± 0.99
24.11 ± 1.08
29.95 ± 0.53
26.65 ± 1.22
23.32 ± 0.64
26.81 ± 1.39
24.63 ± 0.75
26.30 ± 1.03
77.02 ± 3.48
96.03 ± 4.21
66.23 ± 3.13
72.75 ± 3.06
36.52 ± 1.44
56.43 ± 2.57
77.91 ± 3.87
67.27 ± 2.41
77.83 ±2.53
1.98 ± 0.10
3.45 ± 0.18
1.80 ± 0.05
2.56 ± 0.17
0.77 ± 0.13
1.54 ± 0.11
4.07 ± 0.14
3.42 ± 0.12
3.11 ± 0.16
2262 ± 19
2449 ± 23
1882 ± 20
2236 ± 9
2071 ± 33
1545 ± 18
1798 ± 30
25.14 ± 0.88
26.87 ± 0.43
15.71 ± 0.67
25.60 ± 1.09
24.15 ± 0.98
25.15 ± 1.12
20.32 ± 0.84
89.97 ± 3.86
91.14 ± 4.33
119.79 ± 4.97
87.34 ± 3.03
85.75 ± 3.12
61.43 ± 2.51
88.48 ± 4.24
3.48 ± 0.15
4.21 ± 0.13
2.70 ± 0.08
4.37 ± 0.17
3.66 ± 0.09
2.91 ± 0.19
2.85 ± 0.11
Modulators
Tween-40
Tween-80
Triton X-100
Glycine
Glycerol
EDTA
SDS
a
SSF was carried using 9 g sorghum straw under optimized physiological conditions (at 5 days incubation, 1:5
(w:v) substrate: moisture ratio, pH 6.0 and 40 °C).
*The control medium used was devoid of any additives.
The data presented are the mean values of three replicates with the standard deviations.
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Figure S1 Effect of sorghum straw concentration (3 to 15 g) on xylanase production. SSF
was carried out by moistening sorghum straw with mineral salt medium pH 5.0 at 35 °C for 6
days.
Figure S2 Effect of xylose concentration (0.1 to 1.1 %) on xylanase production. SSF was
carried using 9 g sorghum straw under optimized physiological conditions (at 5 days
incubation, 1:5 (w:v) substrate: moisture ratio, pH 6.0 and 40 °C).
Figure S3 Effect of organic solvents (10 to 30 %) on the xylanase activity. The crude
xylanase was incubated in the presence of various organic solvents concentration for 30 min
and at the end of reaction residual activities were measured.
Figure S4 Comparison of the un-optimized and optimized extraction conditions on the
xylanase recovery efficiency (%). Optimized extraction conditions were extractant/ solid ratio
12: 1 (v:w), 150 rpm agitation speed and 40 °C. The data presented are the mean values of
three replicates with the standard deviations.
27
Protein
Specific activity
Cellulase activity
1600
80
1400
70
1200
60
1000
50
800
40
600
30
400
20
200
10
0
Protein (mg/g);
Specific activity (U/mg);
Cellulase activity (U/g)
Xylanase activity (U/g)
Xylanase activity
0
3
5
7
9
11
13
Sorghum straw concentration (g)
15
Figure S1
Protein
Specific activity
Cellulase activity
Xylanase activity (U/g)
2500
100
90
80
70
60
50
40
30
20
10
0
2000
1500
1000
500
0
0.1
Figure S2
0.3
0.5
0.7
0.9
Xylose concentration (%)
1.1
Protein (mg/g);
Specific activity (U/mg);
Cellulase activity (U/g)
Xylanase activity
28
Relative activity (%)
100
80
60
10%
40
20%
20
30%
0
Organic solvent
Figure S3
Xylanase recovery efficiency (%)
Unoptimized xylanase recovery
Optimized xylanase recovery
100
90
80
70
60
50
40
30
20
10
0
30
Figure S4
60
90
120
Extraction time (min)
150