Escolar Documentos
Profissional Documentos
Cultura Documentos
a r t i c l e
i n f o
Article history:
Received 30 August 2013
Received in revised form 5 February 2014
Accepted 8 February 2014
Keywords:
Simultaneous saccharication and
fermentation (SSF)
Very high gravity (VHG)
Ethanol
Cassava our
a b s t r a c t
We developed a simultaneous saccharication and fermentation (SSF) process of cassava our at very
high gravity (VHG). Cassava our (CF) was dissolved in water to reach 315.4 g/l dry matter, and then the
mixture was liqueed at 80 C for 90 min by using alpha-amylase (3532 AAU/kg CF) and beta-glucanase
(2812 U/kg CF). SSF of liqueed mash of cassava was performed at 30 C with the simultaneous addition of
two glucoamylases (Distillase ASP at 540 GAU/kg CF and Amigase Mega L at 0.035% w/w), active dry yeast
(1.5 107 cells/l), urea (12 mM) and KH2 PO4 (4 mM). Under these conditions, the SSF process nished
after 72 h. The ethanol content achieved 17.2% v/v corresponding to 86.1% of the theoretical ethanol yield
at lab scale and decreased to 16.5% v/v corresponding to 83.6% of the theoretical ethanol yield at pilot
scale. Therefore, the SSF of cassava our under VHG condition could have a great potential for the ethanol
industry in Vietnam and South East Asia.
2014 Elsevier B.V. All rights reserved.
1. Introduction
According to the increasing price of oil, bio-ethanol is known
as an ideal candidate to replace the role of fossil fuel. Thus, the
research on this renewable source becomes growingly important
for humans, especially in terms of improving the productivity, the
efciency and decreasing production cost. In Vietnam and in South
East Asia, cassava is considered an attractive raw material for bioethanol production thanks to the following advantages: (i) the ease
of plantation in various soil types and climate conditions; (ii) a
very low input and investment for planting; (iii) all year round
availability of feedstock in the form of fresh roots and dry chips;
(iv) a high starch-containing raw materials and a lower proportion
of bers (Sriroth et al., 2007). Indeed, the Vietnamese Ministry of
Industry and Trade declared that bio-fuel production will achieve
1.8 million tons in 2025, which accounts for 5% of countrys demand
(Ministry of Industry and Trade, 2007b). Moreover, the government
also adapted the policy to improve the beverage ethanol industry
in Vietnam. By the development strategy of beverage ethanol production in Vietnam (Ministry of Industry and Trade, 2007a), ethanol
industry will produce 188 million liters ethanol for food industry
in 2025. Overall, the beverage and bio-ethanol industry has a great
potential in Vietnam in the future.
Corresponding author. Tel.: +84 4 3868 0119; fax: +84 4 3868 2470.
E-mail addresses: son.chuky@hust.edu.vn, kysonchu@gmail.com (S. Chu-Ky).
http://dx.doi.org/10.1016/j.indcrop.2014.02.004
0926-6690/ 2014 Elsevier B.V. All rights reserved.
Besides the conventional process of ethanol production, simultaneous saccharication and fermentation (SSF) process has been
widely used in the world, but only recently introduced to Vietnam
in order to augment ethanol yield and shorten time production.
Indeed, after liquefaction by alpha-amylase, glucoamylase is added
to the slurry, concomitantly with yeasts, and the SSF is conducted
in a single reactor. The presence of yeast along with enzymes minimizes the sugar accumulation in the bioreactor. Moreover, since
the sugar produced during starch or cellulosic breakdown slows
down alpha-amylase action, higher yields and concentrations of
ethanol are possible using SSF (Das Neves, 2006; Klasson et al.,
2013; Molaverdi et al., 2013; Scordia et al., 2013; Wang et al., 2013;
Yingling et al., 2011a,b). The SSF process has been successfully carried out on different substrates such as ax shive (Klasson et al.,
2013), sweet sorghum stalk (Molaverdi et al., 2013), giant reed
(Scordia et al., 2013), sweet sorghum bagasse (Wang et al., 2013),
potato tubers (Srichuwong et al., 2009) and cassava (Chu-Ky et al.,
2009; Yingling et al., 2011b). Therefore, it is of interest to improve
the efciency of the SSF process in the ethanol industry in Vietnam.
Very high gravity (VHG) technology has been introduced to
increase the volumetric productivity and the cost effectiveness
of the SSF process. In VHG technology, mash preparation contains at minimum of 270 g/l dry matter (Bayrock and Ingledew,
2001). This technology has a great deal of advantages in ethanol
production: (i) increasing plant capacity and reduction in capital
costs; (ii) increasing plant efciency; (iii) reducing risk of contaminating bacteria (Thomas et al., 1996; Yingling et al., 2011a,b).
161
Table 1
Characterization of the enzyme products used in this work.
No.
Enzymes products
Nature
Optimal pH
Optimal temperature ( C)
Activity
1
2
3
4
Spezyme Alpha
Optimash TBG
Distillase ASP
Amigase Mega L
Alpha-amylase
Beta-glucanase
Glucoamylase
Glucoamylase
5.75.8
4.56.0
4.04.5
4.04.5
8385
7585
5865
5560
13,775 AAU/ga
5,625 U/gb
580 GAU/gc
a
AAU: Alpha Amylase Unit dened by Dupont (One AAU unit of bacterial alpha-amylase activity is the amount of enzyme required to hydrolyze 10 mg of starch per minute
under specied conditions).
b
U: Unit dened by Dupont (one unit of beta-glucanase activity is dened as the quantity of enzyme which produces reducing sugars equivalent to 1 mol of dextrose
per minute from barley beta-glucan under standard assay conditions).
c
GAU: GlucoAmylase Unit dened by Dupont (One Glucoamylase Unit (GAU) is the amount of enzyme that liberates 1 g of reducing sugars calculated as glucose per hour
from soluble starch substrate under the conditions of the assay).
162
Cassava flour
(CF)
Tap water
Mixture of
suspension
(315.4 g/L)
Spezyme Alpha
(3,532 AAU/kg
CF)
Liquefaction at
800C for 90 min
SSF1
SSF2
Distillase ASP
(540 GAU/kg
CF)
Urea
(12mM)
SSF3
Optimash TBG
(2,812 U/kg
Simultaneous
Saccharification and
Fermentation (SSF)
at 300C for 72 h
Hydrated in
water at 380C
for 20 min
Amigase Mega L
(0.035 % w/w)
Ethanol Red
(1.5x107 cells/ml)
KH2PO4
(4 mM)
Distillation
Byproduct
Ethanol
Fig. 1. Three investigated SSF processes at VHG of cassava our for ethanol production: SSF1 process: Spezyme Alpha + Distillase ASP; SSF2 process: Spezyme Alpha + Distillase
ASP + Optimash TBG; SSF3 process: Spezyme Alpha + Distillase ASP + Optimash TBG + Amigase Mega L.
No.
Components
24 h
36 h
48 h
60 h
72 h
50 rpm
340
270
400
290
1
2
3
4
Maltose (g/l)
Glucose (g/l)
Lactic acid (g/l)
Ethanol (% v/v)
4.3
74.2
0.2
11.2
3.9
17.6
0.5
14.7
4.2
1.8
0.4
16.3
3.5
0.2
0.4
16.5
3.6
0.1
0.5
17.2
350
16
300
250
14
200
12
150
10
100
18
50
0
6
24
Time (hours)
48
Table 3
Evolutions of the concentration of maltose, glucose, lactic acid and ethanol during
SSF3 process.
100 rpm
163
72
Fig. 2. Evolutions of residual sugar and ethanol concentration of the three investigated SSF processes.
17
350
16
300
15
250
14
200
13
150
12
100
11
50
10
164
0
0
24
48
Time (h)
72
88
Acknowledgments
This work was supported by Ministry of Education and Training and Ministry of Science and Technology of Vietnam. We thank
Dupont, DSM and Fermentis for kindly providing us with enzymes
and yeast samples, respectively. We also thank Dr Nguyen TienThanh for his technical assistance in HPLC analysis and Dr Ho
Phu-Ha for her revision of the English text.
Fig. 3. Evolutions of residual sugar, reducing sugar and ethanol concentration of SSF
process at pilot scale.
References
Bayrock, D.P., Ingledew, M.W., 2001. Application of multistage continuous fermentation for production of fuel alcohol by very-high-gravity fermentation
technology. J. Ind. Microbiol. Biotechnol. 27, 8793.
Chu-Ky, S., Nguyen Manh, H., Le Viet, H., Vu Hong, T., Nguyen Thanh, H., Le Thanh, M.,
2009. Optimization for bioethanol fermentation from cassava chips. In: Starch
Update 2009: The Fifth International Conference on Starch Technology, Bangkok,
Thailand, pp. 107112.
Das Neves, M.A., 2006. Bioethanol Production from Wheat Milling By-products.
Graduate School of Life and Environmental Sciences, the University of Tsukuba,
Tsukuba, Japan, pp. 103.
Ingledew, W.M., 2009. Yeast stress in the fermentation process, in: Ingledew W.M.,
Kelsall D.R., Austin G.D., Kluhspies C. (Eds.), The Alcohol Textbook - 5th edition,
pp. 115-126.
Ingledew, W.M., Thomas, K.C., Hynes, S.H., McLeod, J.G., 1999. Viscosity concerns
with rye mashes used for ethanol production. Cereal Chem. 76, 459464.
Klasson, K.T., Dien, B.S., Hector, R.E., 2013. Simultaneous detoxication, saccharication, and ethanol fermentation of weak-acid hydrolyzates. Ind. Crops Prod. 49,
292298.
Miller, G.L., 1959. Use of dinitrosalicylic acid reagent for determination of reducing
sugar. Anal. Chem. 31, 426428.
Ministry of Industry and Trade, 2007a. Development strategy for beverage ethanol
production in Vietnam until 2015 and the vision to 2025. Minister of Trade and
Industrys decision no 2435/QD-BCT dated on May 21, 2009.
Ministry of Industry and Trade, 2007b. Development strategy for beverage ethanol
production in Vietnam from 2007 to 2025. Vietnamese prime ministers decision
no 177/2007/QD-TTg dated on November 20, 2007.
Molaverdi, M., Karimi, K., Khanahmadi, M., Goshadrou, A., 2013. Enhanced sweet
sorghum stalk to ethanol by fungus Mucor indicus using solid state fermentation
followed by simultaneous saccharication and fermentation. Ind. Crops Prod. 49,
580585.
Scordia, D., Cosentino, S.L., Jeffries, T.W., 2013. Enzymatic hydrolysis, simultaneous
saccharication and ethanol fermentation of oxalic acid pretreated giant reed
(Arundo donax L.). Ind. Crops Prod. 49, 392399.
165
Wang, L.J., Luo, Z.L., Shahbazi, A., 2013. Optimization of simultaneous saccharication and fermentation for the production of ethanol from sweet sorghum
(Sorghum bicolor) bagasse using response surface methodology. Ind. Crops Prod.
42, 280291.
Yingling, B., Li, C., Honglin, W., Xiwen, Y., Zongcheng, Y., 2011a. Multiobjective optimization of bioethanol production during cold enzyme starch
hydrolysis in very high gravity cassava mash. Bioresour. Technol. 102,
80778084.
Yingling, B., Zongcheng, Y., Honglin, W., Li, C., 2011b. Optimization of bioethanol
production during simultaneous saccharication and fermentation in very highgravity cassava mash. Antonie van Leeuwenhoek 99, 329339.
Zhang, L., Chen, Q., Jin, Y., Xue, H., Guan, J., Wang, Z., Zhao, H., 2010. Energy-saving
direct ethanol production from viscosity reduction mash of sweet potato at very
high gravity (VHG). Fuel Process. Technol. 91, 18451850.
Zhang, L., Zhao, H., Gan, M., Jin, Y., Gao, X., Chen, Q., Guan, J., Wang, Z., 2011. Application of simultaneous saccharication and fermentation (SSF) from viscosity
reducing of raw sweet potato for bioethanol production at laboratory, pilot and
industrial scales. Bioresour. Technol. 102, 45734579.