Available online at www.sciencedirect.

com

APCBEE Procedia 2 (2012) 104 109

ICBFS 2012: April 7-8, 2012, Bangkok, Thailand

Role of Lactic Acid Bacteria on Structural and Physicochemical

Properties of Sour Cassava Starch
Widya Dwi Rukmi Putria,*, Haryadib, Djagal Wiseso Marsenob, Muhammad Nur
Cahyanto,b
a

Agricultural Technology Faculty, Brawijaya University, Veteran Street, Malang, Indonesia.

Agricultural Technology Faculty, Gadjah Mada University, Sosio Yustisio, Yogyakarta, Indonesia

Abstract
In this study, the characteristics of cassava starch have been developed through lactic acid fermentation. Structural and
physicochemical properties of native, acidified and fermented cassava starch were analyzed. Carboxyl content of all
samples were increase whereas the carbonyl contents kept unchanged. The starches had similarity on FTIR spectral
pattern although had different absorbance spectra intensities. X-ray diffraction pattern showed the crystallinity value of
native starch was 44.16%. Lactic acid fermentation reduced starch crystallinity ranged from 1.52% to 3.78,%, and the
same effect was also altered the acidified starch with the value decrease by 3,1% in relation to native starch. Fermented
and lactic acidified starches required a longer time to reached their lower peak viscosity than the native starch. The
decrease in crystallinity was attributed to the depolymerization of starch structure, resulting in a weakened granule
organization. Pasting behavior showed the occurrence of starch structure reorganization.

2012
2012 Published
Publishedby
byElsevier
ElsevierB.V.
B.V.
Selection
and/or
peer
review
under
responsibility
of Asia-Pacific

Selection
and/or
peer
review
under
responsibility
of Asia-Pacifi
c
Chemical,
Biological
&
Environmental
Engineering
Society
Chemical, Biological & Environmental Engineering Society Open access under CC BY-NC-ND license.
Keywords: cassava starch, lactic acid fermentation, structural properties

1. Introduction
Fermentation of cassava starch is a common process in Indonesian traditional industries to improve the
textural qualities of the starch. The cassava starch was produced by natural fermentation and then sun dried

* Corresponding author. Tel.: +62-341-569214; fax: +62-341-580106.

E-mail address: widya2putri@ub.ac.id

2212-6708 2012 Published by Elsevier B.V. Selection and/or peer review under responsibility of Asia-Pacific Chemical,
Biological & Environmental Engineering Society Open access under CC BY-NC-ND license.
doi:10.1016/j.apcbee.2012.06.019

Widya Dwi Rukmi Putri et al. / APCBEE Procedia 2 (2012) 104 109

have specific ability to expand during baking or frying. Natural fermentation of cassava starch is characterized
by the presence of a predominantly lactic acid microorganism. The main acid resulted from fermentation,
lactic acid, was oxidized by sun drying and then causing structural changes of cassava starch. Brasil and
Columbia also have been produced sour cassava starch which having baking behavior characteristic, but in
longer time of fermentation (Mestres and Rouau, 1997). Lactic acid bacteria (LAB) play an important role in
sour cassava starch fermentation. LAB fermentation causes the change of starch microstructure because of
their ability to produce lactic acid, specific enzymes and aromatic compound (Camargo et al., 1988; Demiate
et al., 2000; Marcon et al., 2006). Lactic acid produced also induces the amylography and viscosity of starch
(Bertolini et al., 2000). Even, in sour cassava starch (povilho azedo; Brazil), lactic acid fermentation and sun
drying are related to the baking behaviour of this starch (Bertolini et al., 2001; Demiate et al., 2000;
Vatanasuchart et al., 2005). Previous study of the research showed that the amylolytic ability of lactic acid
bacteria used in fermentation influenced the baking properties of the starch (Putri et al., 2011a; Putri et al.,
2011b). Considering the current importance of this fermentation process, this work was conducted to evaluate
the effect of lactic acid fermentation on structural, pasting and physicochemical properties of starches.
2. Material and Methods
2.1. Microorganism
The microorganism used were L. plantarum subsp. plantarum AA11, AA2 and UA3 isolated from soaking
cassava during growol fermentation and L. amylophyllus NBRC 15881 as an amylolytic reference strain
(NITE Biological Resources Center, Chiba, Japan). All of these strains had a different ability to grow in starch
based medium and to degrade raw starch (Putri et al., 2011a) .
2.2. Starch extraction and fermentation
Cassava starch extraction was carried out using a method described by Demiate et al. (2005). Native
cassava starch was made by drying the starch in a cabinet dryer at 60oC for 18 hours after irradiated with
ultraviolet radiation UVA (315 400 nm) for 9 h. Fermentation of raw starch (fermented starch) was carried
out in capped 250 ml Erlenmeyer flasks with 150 ml starch medium (50 g of sterile raw cassava starch
suspended in 150 ml sterile water). For all the strains, the medium was inoculated with 10% (v/v) culture
grown on MRS medium and incubated at 30oC for 48 h. Acidified starch was prepared by soaking raw starch
in 1% lactic acid solution for 30 minutes. Then, starches were washed three times with sterile water before
irradiated with UVA (315 400 nm) for 9 h and dried at 60oC for 18 hours.
2.3. Structural and pasting properties determination
Carboxyl content and pH were analyzed based on Demiate et al. (2000), whereas the carbonyl content and
was measured following the method was done by Kuakpetoon and Wang (2006). The crystallinity pattern of
starch were analyzed using X-ray diffraction (Shimadzu X-ray diffractometer XRD-6000). Pasting
characteristics (10% w/dry weight) was measured using Brabender amilograph (Visco amilograph RV model,
Wingather V2.5, Brookfield Engineering Laboratories).

105

106

Widya Dwi Rukmi Putri et al. / APCBEE Procedia 2 (2012) 104 109

3. Results and Discussion

3.1. Carbonyl and carboxyl contents
As shown in Table 1, there was no differences in carbonyl contents of all cassava starch samples, whereas
carboxyl contents increased as the effect of fermentation and acidification compared to native starch. This
results were in line with the reserach report of Kuakpetoon and Wang (2006), that carboxyl content increased
at a much higher than carbonyl content, which confirmed that hydroxyl groups on starch molecules were
initially oxidized to carbonyl groups and then to carboxyl groups as the primary final product.
Sour cassava starch fermentation by acidication showed the highest value of carboxyl and caused the starch
had the greater ability to expand while baking (Putri et al., 2011b) data not shown in this paper. As Wang and
Wang (2001) reported, the carboxyl group had relationship with molecular fragmentation during oxidation
process resulting molecular structure changes.
Table 1. pH, carbonyl dan carboxyl content of native and sour cassava starches
Sample

Native starch
Acidified starch
1

pH

Carbonyl
content (%)

Carboxyl
content (%)

6,16 0,42 c

0,18 0,96

0,16 0,23 a

4,14 0,66 a

0,18 0,97

0,43 0,60 c

Fermented starch 15881

4,95 0,66 b

0,20 1,02

0,32 0,37 b

Fermented starch AA22

5,01 0,95 b

0,18 0,97

0,24 0,46ab

Fermented starch AA113

4,98 1.03 b

0,19 0,98

0,26 0,33ab

5,01 0,81 b

0,19 0,99

0,24 0,30 ab

Fermented starch UA3

Fermented starch by L. amylophyllus 15881; 2Fermented starch by L. plantarum UA3; 3Fermented starch by L. plantarum AA2;
Fermented starch by L. plantarum AA11. All values are averages of triplicates + standard deviation
Means of each properties followed by same letter are not significantly different p<0,05)

3.2. FTIR spectral pattern

In order to gain more insight into the effect of the treatments on the structural changes of cassava starches,
infrared spectroscopy was used. FTIR spectral pattern of all treatments showed similarity (The pattern were
not shown), although they had different intensity on their absorbance spectra. Cassava starch fermented by L.
amylophylus NBRC 15881 gave low peak in the 1600 cm-1 infrared region that similar with native starch, but
had higher intensity in 1060 cm-1. Demiate et al. (2000) stated, that 1600 cm-1 and 1060 cm-1 spectral region
was necessary for prediction of baking property. The analysis performed on the shorter spectral region gave
bad results, especially on samples, which did not have an important contribution of carboxylate group. It is
possible to observe that the region at 1600 cm-1 is positively correlated to baking property while the other
spectral region around 1060 cm-1 is linked to some non-expansion related structural information.
3.3. X-ray diffraction pattern
X-ray diffraction of native starch, acidified starch and fermented starch presented pattern type A, the same
pattern shown by Atichokudomchai et al. (2002), Cavallini and Franco (2010) and Gomes et al. (2005). All of
diffractograms showed the main peak in 15o, 17o, 18o, and 23o, that correspond to the spacing-d in 5,7, 5,1, 4,8

107

Widya Dwi Rukmi Putri et al. / APCBEE Procedia 2 (2012) 104 109

and 3,8 , respectively. There were no modification in X-ray pattern of the fermented and acidified samples
in relation to native starch, but the diffractograms of the fermented and acidified starches presented decreased
in the diffraction intensity (Figure not shown). The spectrum of native starch sample shows definite
diffraction peaks that presumably reflect crystalline regions in starch The relative crystallinity of native starch
was 44.16%. The treatment samples showed reduction in crystallinities, in the acidified starch was 3,10%
while fermented starch were 1,52% 3,78% in relation to native starch. The fermented starch by the highest
amylolytic of LAB presented the lowest value of relative crystallinity.
Table 2. X-ray crystallinity of native and sour cassava starches
Integrated intensity on diffraction angle (2)

Starch

1
4

15o

17o

18o

Crystallinity index
(%)

23o

Native starch

275

311

275

215

44,16

Acidified starch
Fermented starch
158811
Fermented starch
AA22
Fermented starch
AA113
Fermented starch
UA34

271

306

271

234

41,06

241

278

250

232

40,38

242

277

252

237

41,34

260

302

286

227

42,64

200

236

222

213

40,52

Fermented starch by L. amylophyllus 15881; 2Fermented starch by L. plantarum UA3; 3Fermented starch by L. plantarum AA2;
Fermented starch by L. plantarum AA11. All values are averages of triplicates + standard deviation

3.4. Pasting properties

Native starch reached the peak viscosity in a shorter times while the viscosity value was higher than
another samples (Table 3).. Abera et al. (2003) stated that native starch presented strong connections among
the molecules of starch taking them to get more stable conformation, leading to amylose having a smaller
tendency to leaching out from granules. The acidification and fermentation process reduced the peak viscosity,
paste viscosity , breakdown viscosity and set back viscosity of the starch, while paste temperatures dan peak
times were increased. This results were in line with Gomes et al (2005) which showed the increasing of paste
temperature in annealed povilho azedo (sour cassava starch). The lower viscosity indicated that there were
starch granule disintegration which decreasing the swelling ability. Acid and amylase enzymes easily attacks
the amorphous regions of granules reducing the molecular massa of amylose and amylopectin molecules,
which confirms the high solubility found for the modified starches. The fermented starch by highly amylolytic
LAB (L. amylophylus NBRC 15881) showed the lowest reduction of peak viscosity, which assumed that both
amylose and amylopectin were partially attacked by the starch-degrading enzyme.
Table 3. Pasting properties of native and sour cassava starches

Starches

Native starch

Peak
Viscosity
(cP)

Paste
Viscosity
(cP)

Breakdow
n
(cP)

2169,60

488,00

1681,60

Setback
(cP)

Final viscosity
(cP)

Pasting
Temp.(oC
)

Peak time
(min)

1496,0
0

1984,00

85,10

15,00

108

Widya Dwi Rukmi Putri et al. / APCBEE Procedia 2 (2012) 104 109

Acidified starch
Fermented starch
158811

1216,00

64,00

1152,00

1209,60

166,40

1043,20

1720,00

153,60

1566,40

Fermented starch AA11

1580,80

151,20

1429,60

Fermented starch UA34

1593,60

140,80

1452,80

Fermented starch AA22

3

1
4

1164,8
0
1395,2
0
1587,2
0
1570,4
0
1369,6
0

1228,80

89,70

16,00

1561,60

91,40

17,00

1740,80

92,90

18,00

1721,60

93,80

18,00

1510,40

92,90

18,00

Fermented starch by L. amylophyllus 15881; 2Fermented starch by L. plantarum UA3; 3Fermented starch by L. plantarum AA2;
Fermented starch by L. plantarum AA11. All values are averages of triplicates

4. Conclusion
Results of this study have revealed differences on microstructural and pasting characteristic of starches
with different treatments. The ability of lactic acid bacteria as starter cultures to degrade and produce lactic
acid during fermentation influenced the interrelated parameters of sour cassava starch such as pH, low number
of carboxyl groups formation at 1080,14 cm-1 but had higher number at 1650 cm-1 on FTIR spectroscopy
analysis, reduced crystallinity and modified their pasting properties.
Acknowledgements
This work was funded by Indonesian National Strategic Grant 2009 from Directorate for Higher Education,
National Education Department of Indonesia. I would like to thanks to Clair B. Hicks a professor at
Agriculture Department at the University of Kentucky for their supervision during this work.
References
[1] Abera, S. and Rakshit, S. K. 2003. Comparison of physicochemical and functional properties of cassava starch extracted from
fresh root and dry chip. Starch/Starke 55: 287-296.
[2] Atichokudomchai, N., Shobsngob, S., Chinachoti, P. and Varavinit, S. 2001. A Study of some physicochemical properties of
high-crystalline tapioka starch. Starch/Starke, 53: 577581
[3] Bertolini, A. C., Mestres, C., Raffi, J., Buleon, A., Lerner, D. and Colona, P. 2001, Photodegradation of cassava and corn
starches. Journal of Agricultural and Food Chemistry 49: 675-682.
[4] Camargo, C., Colona, P., Buleon, A. and Molard, D. R. 1988. Functional properties of sour cassava (Manihot utilissima) starch:
polvilho azedo. Journal of the Science of Food and Agriculture 81: 429-435
[5] Cavallini, C. M. and Franco, C. M. L. 2010 Effect of acid-ethanol treatment followed by ball milling on structural and
physicochemical characteristics of cassava starch. Starch/Starke, 62: 236245
[6] Demiate, L. M., Dupuy, N., Huvenne, J. P., Cereda, M. P. and Wosiacki, G. 2000. Relationship between baking behavior of
modified cassava starches and starch chemical structure determined by ftir spectroscopy . Carbohydrate Polymer 42: 149-158
[7] Gomes, A.M.M., Mendes da Silva, C. E., Ricardo, N.M.P.S. 2005. Effects of annealing on the physicochemical properties of
fermented cassava starch (polvilho azedo). Carbohydrate Polymers 60: 16
[8] Kuakpetoon, D. and Wang, Y.J. 2006. Structural characteristics and physicochemical properties of oxidized corn starches
varying in amylose content. Carbohydrate Research 341; 18961915
[9] Marcon, M.J.A., Vieira, M.A. Santos, K. De Simas, K.N., Amboni, R.D.M.C. and Amante, E.R. 2006. The effect of
fermentation on cassava starch microstructure. Journal of Food Process Engineering 29: 362372.

Widya Dwi Rukmi Putri et al. / APCBEE Procedia 2 (2012) 104 109

[10] Mestres, C. and Rouau, X. 1997. Influence of natural fermentation and drying conditions on the physicochemical characteristics
of cassava starch. J Sci Food Agric 74: 147 155.
[11] Putri, W.D.R, Nakagawa, Y., Kawasaki, H. Mika, M. Haryadi, Cahyanto, M.N and Marseno, D.W. 2011a. Differentiation
studies of predominant lactic acid bacteria isolated during growol fermentation by using polyphasic taxonomic characterization. African
Journal of Biotechnology 10 (42): 8194-8204.
[12] Putri, W.D.R, Haryadi, Marseno, D.W and Cahyanto, M.N. 2011b. Effect of biodegradation by lactic acid bacteria on physical
characteristics of cassava starch. International Food research Journal 18(3): 1149-1154.
[13] Vatanasuchart, N., Naivikul, O., Charoenrein, S. and Sriroth, K. 2005. Molecular Properties of Cassava starch Modified by
Different UV Irradiations to Enhance Baking Expansion. Carbohydrate Polymer 61: 80-87.
[14] Wang, L. and Wang, Y. J. 2001. Structures and physicochemical properties of acid-thinned corn, potato, and rice starches.
Starch/Starke, 53: 570576

109