April 18, 2015
Supplementary Material
Saccharification of sunflower stalks using lignocellulases from a fungal consortium comprising Pholiota adiposa and Armillaria gemina
Priyadharshini Ramachandran1,+, Tae-Su Kim1,+, Saurabh Sudha Dhiman1, Jinglin Li1, Ji-Hyun Park1, Joon-Ho Choi1, Jae Young Kim2, Dongwook Kim3, Jung-Kul Lee1*
1Department of Chemical Engineering, Konkuk University, Hwayang–Dong, Gwangjin–Gu, Seoul 143-701, Republic of Korea
2School of Civil, Urban and Geosystem Engineering, College of Engineering,
Seoul National University, Gwanak-Ku, Seoul 151-742, Republic of Korea
3Phygen Research Institute, Daejeon, Republic of Korea
+ : These authors equally contributed to this work.
Correspondence: Jung-Kul Lee, Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, Korea
(Tel: +82-2-450-3505; fax: +82-2-458-3504; e-mail: )
Supplementary Fig. S1. Effect of pH on production of different lignocellulases estimated in the crude extract of P. adiposa and A. gemina consortium culture.
Supplementary Fig. S2. Effect of temperature on production of different lignocellulases estimated in the crude extract of P. adiposa and A. gemina consortium culture.
Supplementary Table S1. Effect of carbon sources on EG, CBH, BGL and filter paper activity of consortium of P. adiposa and A. gemina. Each value represents the mean of triplicate measurements and varies from the mean by not more than 10%. Yeast extract (5 g/L) was used as nitrogen sources.
(20 g/L) / EG activity
(U/ml) / CBH activity
(U/ml) / BGL activity
(U/ml) / FPU
(U/ml)
Cellulose / 14.6 ± 1.5 / 9.30 ± 0.92 / 13.4 ± 1.4 / 0.63 ± 0.07
Glucose / 5.15 ± 0.52 / 1.25 ± 0.17 / 1.45 ± 0.15 / 0.25 ± 0.02
Lactose / 4.14 ± 0.49 / 6.14 ± 0.66 / 5.10 ± 0.66 / 0.24 ± 0.02
Maltose / 6.90 ± 0.72 / 4.10 ± 0.42 / 9.60 ± 0.98 / 0.21 ± 0.02
Cellobiose / 4.80 ± 0.52 / 8.41 ± 0.84 / 13.6 ± 1.4 / 0.53 ± 0.05
CMC / 16.7 ± 1.9 / 2.82 ± 0.31 / 6.52 ± 0.66 / 0.23 ± 0.02
Sucrose / 3.70 ± 0.41 / 2.24 ± 0.29 / 6.42 ± 0.62 / 0.22 ± 0.02
Birchwood xylan / 14.3 ± 1.7 / 4.30 ± 0.42 / 7.30 ± 0.69 / 0.16 ± 0.02
Rice straw / 11.6 ± 1.2 / 4.31 ± 0.45 / 13.1 ± 1.4 / 0.53 ± 0.06
Wheat bran / 16.6 ± 0.8 / 6.10 ± 0.34 / 18.0 ± 1.9 / 0.42 ± 0.06
Avicel / 14.5 ± 0.7 / 6.90 ± 0.30 / 16.9 ± 1.7 / 0.51 ± 0.06
Supplementary Table S2. Effect of nitrogen sources on EG, CBH, BGL and filter paper activity of consortium of P. adiposa and A. gemina. Each value represents the mean of triplicate measurements and varies from the mean by not more than 10%. Rice straw (20 g/L) was used as a carbon source.
(20 g/L) / EG activity
(U/ml) / CBH activity
(U/ml) / BGL activity
(U/ml) / FPU
(U/ml)
Yeast Extract / 18.4 ± 1.9 / 13.6 ± 1.4 / 16.6 ± 1.7 / 0.77 ± 0.08
Peptone / 17.3 ± 1.8 / 11.8 ± 1.2 / 12.8 ± 1.3 / 0.47 ± 0.05
Corn steep powder / 22.5 ± 2.3 / 11.5 ± 1.3 / 17.9 ± 1.9 / 0.68 ± 0.07
Urea / 14.5 ± 1.5 / 10.3 ± 1.2 / 15.5 ± 1.7 / 0.33 ± 0.04
(NH)2 SO4 / 9.6 ± 0.9 / 6.1 ± 0.6 / 7.0 ± 0.7 / 0.26 ± 0.03
KNO3 / 16.3 ± 1.7 / 10.3 ± 1.1 / 12.1 ± 1.3 / 0.48 ± 0.05
NaNO3 / 14.6 ± 1.5 / 11.5 ± 1.3 / 11.7 ± 1.3 / 0.44 ± 0.05
Tryptone / 13.6 ± 1.4 / 13.6 ± 1.4 / 16.6 ± 1.7 / 0.72 ± 0.08
Supplementary Table S3. Effect of different inducers, vitamins and amino acids on cellulase production
(0.002 g/L) / EG activity
(U/ml) / CBH activity
(U/ml) / BGL activity
(U/ml) / FPU
(U/ml)
L-ascorbic acid / 16.4 ± 1.8 / 14.7 ± 1.4 / 17.8 ± 1.7 / 0.27 ± 0.08
Folic acid / 19.2 ± 2.1 / 11.9 ± 1.2 / 10.3 ± 1.1 / 0.29 ± 0.03
Alanine / 17.3 ± 1.8 / 11.8 ± 1.2 / 12.8 ± 1.3 / 0.47 ± 0.05
Tryptophan / 22.5 ± 2.3 / 11.5 ± 1.3 / 17.9 ± 1.9 / 0.68 ± 0.07
Thiamine / 24.6 ± 2.6 / 12.4 ± 1.3 / 16.5 ± 1.7 / 0.39 ± 0.04
Vitamin-B1 / 13.4 ± 1.4 / 13.2 ± 1.5 / 15.6 ± 2.1 / 0.42 ± 0.05
Vitamin-B2 / 14.6 ± 1.5 / 13.6 ± 1.4 / 14.8 ± 1.6 / 0.36 ± 0.04
Vitamin-B12 / 19.8 ± 2.1 / 12.8 ± 1.3 / 16.2 ± 1.7 / 0.39 ± 0.05
Gentiobiose / 29.7 ± 3.5 / 16.4 ± 1.7 / 7.7 ± 0.7 / 0.45 ± 0.06
Supplementary Table S4. Enzyme units in the enzyme from the P. adiposa and A. gemina consortium. Each value represents the mean of triplicate measurements and varied from the mean by not more than 10%.
Consortium of A. gemina and P. adiposav (U/mL) / A. gemina SKU2114 (U/mL) [1] / P. adiposa SKU0714 (U/mL) [2]
Filter paper unit (FPU) / 2.57 / 1.72 / 1.25
β-glucosidase / 39 / 15 / 39
Endo-β-1,4 glucanase / 176 / 146 / 26
Exo-β-1,4 glucanase (cellobiohydrolase) / 44 / 34 / 32
Xylanase / 1870 / 1270 / 1680
Laccase / 0.20 / 0.16 / 0.12
[1] Jagtap SS, Dhiman SS, Kim TS, Li J, Lee JK, Kang YC. Enzymatic hydrolysis of aspen biomass into fermentable sugars by using lignocellulases from Armillaria gemina. Bioresource Technology 2013; 133: 307-314.
[2] Jagtap SS, Dhiman SS, Jeya M, Kang YC, Choi JH, Lee JK. Saccharification of poplar biomass by using lignocellulases from Pholiota adiposa. Bioresource Technology 2012; 120: 264-272.
Supplementary Table S5. Experimental design and the results of the central composite design for obtaining the fungal consortium enzyme.
(%) / Predicted Saccharification
(%)
1 / 0 / 0 / 0 / 0 / 77.54 / 77.65
2 / +1 / +1 / -1 / +1 / 84.39 / 84.63
3 / 0 / 0 / 0 / 0 / 68.26 / 77.65
4 / +1 / +1 / -1 / +1 / 46.70 / 51.71
5 / -1 / +1 / -1 / -1 / 34.41 / 31.93
6 / -1 / +1 / +1 / -1 / 44.24 / 33.13
7 / -1 / +1 / -1 / -1 / 60.55 / 59.76
8 / +1 / +1 / +1 / +1 / 67.48 / 63.86
9 / -1 / +1 / -1 / +1 / 83.35 / 79.02
10 / +1 / +1 / +1 / +1 / 9.78 / 20.13
11 / +1 / +1 / -1 / -1 / 44.69 / 37.08
12 / -1 / +1 / +1 / +1 / 10.59 / 10.58
13 / +2 / 0 / 0 / 0 / 83.25 / 78.32
14 / 0 / 0 / -2 / 0 / 63.00 / 68.38
15 / 0 / 0 / +2 / 0 / 19.00 / 10.17
16 / 0 / 0 / 0 / +2 / 58.54 / 43.48
17 / -1 / +1 / -1 / +1 / 47.04 / 45.79
18 / -2 / 0 / 0 / 0 / 48.15 / 51.62
19 / +1 / +1 / +1 / -1 / 15.36 / 13.58
20 / +1 / +1 / +1 / -1 / 41.11 / 51.91
21 / +1 / +1 / -1 / -1 / 81.00 / 77.90
22 / 0 / 0 / 0 / -2 / 6.07 / 17.67
23 / -1 / +1 / +1 / +1 / 27.14 / 44.31
24 / 0 / 0 / 0 / 0 / 80.89 / 77.65
25 / 0 / 0 / 0 / 0 / 81.35 / 77.65
26 / 0 / 0 / 0 / 0 / 80.90 / 77.65
27 / -1 / +1 / +1 / -1 / 5.49 / 4.80
28 / 0 / -2 / 0 / 0 / 8.80 / 9.75
29 / 0 / +2 / 0 / 0 / 81.70 / 81.30
30 / 0 / 0 / 0 / 0 / 73.96 / 77.65