Accepted Manuscript

Title: Characteristics of glucomannan isolated from fresh

tuber of Porang (Amorphophallus muelleri Blume)
Author: Anny Yanuriati Djagal Wiseso Marseno Rochmadi
Eni Harmayani
PII:
DOI:
Reference:

S0144-8617(16)31032-3
http://dx.doi.org/doi:10.1016/j.carbpol.2016.08.080
CARP 11505

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Received date:
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Accepted date:

24-3-2016
15-8-2016
25-8-2016

Please cite this article as: Yanuriati, Anny., Marseno, Djagal Wiseso., Rochmadi,
., & Harmayani, Eni., Characteristics of glucomannan isolated from fresh
tuber of Porang (Amorphophallus muelleri Blume).Carbohydrate Polymers
http://dx.doi.org/10.1016/j.carbpol.2016.08.080
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Characteristics of glucomannan isolated from fresh tuber of Porang (Amorphophallus muelleri

Blume)
Anny Yanuriatia,b,*, Djagal Wiseso Marsenob, Rochmadi,c Eni Harmayanib
a

Department of Agricultural Technology, Sriwijaya University, Jl. Palembang Prabumulih Km. 32,
Inderalaya, Ogan Ilir 30662, South Sumatra, Indonesia
b
Faculty of Agricultural Technology, Gadjah Mada University, Jl. Flora No.1, Yogyakarta 55281,
Indonesia
c
Chemical Engineering Department, Faculty of Engineering, Gadjah Mada University, Jl. Grafika No.
2,Yogyakarta 55281, Indonesia

*Corresponding author:
Anny Yanuriati, Department of Agricultural Technology, Sriwijaya University, Jl. PalembangPrabumulih Km. 32, Indralaya, Ogan Ilir 30662, South Sumatra, Indonesia. Tel. +6281367740860; Fax:
+62711580276.
E-mail address: annyyanuriati@fp.unsri.ac.id

Highlights

Glucomannan could easily and quickly be isolated from fresh porang tubers.

Isolation was performed by repeated fresh tuber milling in ethanol and filtration.

Isolation of glucomannan from fresh porang tubers required no further purification.

Impurities of glucomannan isolated from fresh tubers could be removed completely.

Glucomannan isolated from fresh tuber had high purity, viscosity and transparency.

Abstract
Porang is a potential source of glucomannan. This research objective was to find a direct glucomannan
isolation method from fresh porang corm to produce high purity glucomannan. Two isolation methods
were performed. In first method, sample was water dissolved using Al2(SO4)3 as flocculant for 15 (AA15)
or 30 (AA30) minutes with purification. In second method, sample was repeatedly milled using ethanol
as solvent and filtered for 5 (EtOH5) or 7 (EtOH7) times without purification. The characteristics of
obtained glucomannan were compared to those of commercial porang flour (CPF) and purified konjac
glucomannan (PKG). High purity (90.98%), viscosity (27,940 cps) and transparency (57.74 %) of
amorphous glucomannan were isolated by EtOH7. Ash and protein level significantly reduced to 0.57%
and 0.31%, respectively, with no starch content. Water holding capacity (WHC) of EtOH7 glucomannan
significantly enhanced, whereas its solubility was lower than those of PKG due to its ungrounded native
granular form.
Keywords: porang, Amorphophallus muelleri Blume, glucomannan, characteristics, isolation, fresh tuber.

1. Introduction
Amorphophallus muelleri Blume, locally called porang or iles kuning, is one of glucomannan potential
source in Indonesia due to its high level of glucomannan content. The tubers are not consumed and
unpalatable as they contain high levels of calcium oxalate. As highly demanded export commodity, the
tubers are sliced, dried to chips, milled to flour and exported without further processed into glucomannan,
despite its wide application, due to the complicated long process. Glucomannan has diverse function in
food, pharmaceutical (Alonso-Sande, Teijeiro-Osorio, Remun-Lpez, & Alonso, 2009; Tester & AlGhazzewi, 2013; C. Zhang, Chen, & Yang, 2014), cosmetics and chemical industries (Zhang, Xie, & Gan,
2005).
Available commercial glucomannan are extracted from dried tuber, especially konjac. Konjac
tuber parenchyma cortex comprises ordinary cells and large cells (idioblasts). The glucomannan granules
are located in egg-shaped idioblasts within parenchyma as single cells (Chua, Hocking, Chan, & Baldwin,
2013), encapsulated by scale like cell walls and dispersed all over the tuber (Takigami, Takiguchi, &
Phillips, 1997). Starch, cellulose and nitrogen containing materials in ordinary cells around idioblasts are
small (approximately 0.004 mm), while idioblasts contain very hard glucomannan granules with oval or
round shape, translucent, large with diameter between 0.25 0.75 mm and sized more than 5 10 times
than those of ordinary cells. The starch as agglomerated granules is easily broken into very fine particles
and insoluble in cold water whereas glucomannan granules are very difficult to be disintegrated for its
very hard form. The particles are bigger than the impurities (Zhao, Zhang, Srzednicki, Kanlayanarat, &
Borompichaichartkul, 2010) and soluble in water (Enomoto-rogers, Ohmomo, Takemura, & Iwata, 2014;
Luo, He, & Lin, 2013), but insoluble in ethanol (Li et al., 2014). Based on these characteristics, it is
possible to isolate glucomannan granules directly from fresh tuber by water dissolving with flocculants to
remove impurities followed by purification or only by repeated milling using ethanol and filtration
without further purification.

Konjac glucomannan, a non-ionic polysaccharide with molecular weight > 1 x 106 Da (Chao et
al., 2012; Lin et al., 2010), consists of mannose and glucose residues linked by -(1-4) with ratio of 1.6:1
(Koroskenyi & Mccarthy, 2001; Ratcliffe, Williams, Viebke, & Meadows, 2005) and the acetylation
degree around 5-10% (Gao & Nishinari, 2004). The ratio of glucose and mannose, the acetylation degree,
and the molecular weight could be different depends on the source of glucomannan (Gao & Nishinari,
2004; Koroskenyi & Mccarthy, 2001).
Basically, previous commercial glucomannan granules are extracted by slicing, drying, milling to
remove impurities which adhere to the glucomannan granules, followed by pulverizing to flour, sifting
and air clarifying (Ohashi, Shelso, Moirano, & Drinkwater, 2000; Zhao et al., 2010). Some remaining
impurities attached on the surface of glucomannan granules reduce the glucomannan concentration,
viscosity and transparency of konjac flour sol.

The impurities encapsulated on the surface of the

glucomannan granules are not easy to be removed (Zhao et al., 2010).

They can only be detached partially as drying makes them attach to the granule surfaces more
firmly. Purification process is required to remove them for they turn the gel or sol to have higher
turbidity, with milky-white or cloudy appearance (Ohashi et al., 2000). Some methods were developed to
remove them by stirring in ethanol 50% (Chua et al., 2012) and simple centrifugation after flocculation
using Al2(SO4)3 (Tatirat & Charoenrein, 2011). The 2 methods required further long time purification
using high amount of high concentrated ethanol, while the others were by one-step procedure of
azeotropy-assisted acidic ethanol using citric acid (W. Xu, Wang, Jin, et al., 2014), and dimethyl
sulfoxide addition (Ye et al., 2014). All the methods used konjac flour from chips as material and still
need further ethanol washing.
Up to now, reports on direct glucomannan isolation from fresh tuber without drying are very rare.
The impurity removal will be more easily if the granules are isolated directly from fresh tuber without
tuber slices drying. The impurities surrounding the granules in fresh tuber could be detached more easily
since more firmly attaching impurities on the surface of glucomannan granules from drying could be
hindered.

As a result, the obtained glucomannan will have higher yield, purity, viscosity, and
5

transparency even without further purification. This research objective was to find a direct glucomannan
isolation method from fresh porang corm to produce high purity glucomannan.

Hypotheses
The purity and the yield of porang glucomannan could be enhanced significantly as well as
viscosity and transparency of the sol by a direct isolation from fresh tuber since more firmly attaching
impurities on the surface of glucomannan granules from drying could be avoided, which resulted in easier
removal of the impurities.

2. Materials and methods

2.1. Materials
The materials were 2 year old porang tubers with average weight about 2.250 250 g obtained
from Klangon Village, Madiun, Jawa Timur, Indonesia, ethanol 96%, and deionized water for
glucomannan isolation. All other chemicals were analytical grade from Merck Co.

2.2. Isolation and purification of glucomannan from Amorphophallus muelleri fresh tubers
Glucomannan were directly isolated from fresh tubers of porang using 2 methods, by water
dissolving using Al2(SO4)3 as a flocculant, followed by purification using ethanol and by repeated milling
using ethanol and glucomannan granules filtration to separate them from impurities without further
purification. In the first method, peeled tubers were cut and shredded. The shredded tubers were put in
sodium metabisulphite solution 200 ppm (w/v) and agitated for 4 hours. The glucomannan extract
dissolved in water (sol) was filtered and separated from the solid. The sol then was heated up to 40oC
with agitation, added with 10% Al2(SO4)3 for 15 or 30 minutes. Then the sol was centrifuged at 4000 rpm
for 20 minutes.

The supernatant was then centrifuged at 4000 rpm for another 20 minutes and

precipitated using ethanol 96%. The precipitate was filtered, the residue of ethanol on the flakes was
evaporated, and the flakes were subsequently dried, ground, and sifted through a 60 mesh sieve.
6

In second method, the clean tubers were sliced and milled in 50% ethanol around 12.000 rpm for
5 minutes, then filtered and pressed to get crude glucomannan. These milling, filtering and pressing
processes were repeated for 5 or 7 times and the obtained granules were dried at 40oC. The obtained
glucomannan characteristics were compared to CPF and PKG.

2.3. Glucomannan content

Glucomannan content was analyzed using 3,5-DNS colorimetric assay (Chua et al., 2012).
Glucomannan (0.2 g) was added to formic acid-sodium hydroxide buffer (0.1 mol/L; 50 ml) and
magnetically stirred for 4 hours at room temperature. The mixture was then diluted with formic acidsodium hydroxide buffer to 100 ml in a volumetric flask, followed by centrifugation (4000 rpm, 40 min,
25oC) to get supernatant as the glucomannan extract.
The glucomannan extract (5 ml) was put into a 25 ml volumetric flask and added with 3 M
sulphuric acid (2.5 ml). The resultant solution was mixed using a vortex, subsequently hydrolyzed for 90
min in a boiling water bath and allowed to cool to room temperature before the addition of 6 M sodium
hydroxide (2.5 ml). The solution was then made up to 25 ml with deionized (DI) water to form the
glucomannan hydrolysate. Both the glucomannan extract and hydrolysate were subjected to colorimetric
reactions and deionized water was used as a blank.
D-glucose standard solution (1 mg/ml) was diluted to 0.20%, 0.40%, 0.80%, 1.20%, and 1.60%
using deionized water (DI). About 1.5 ml of 1% 3,5 DNS solution was added to 2 ml of the sugar
standards. Each mixture was heated for 5 min in a boiling water bath and cooled to room temperature
before being diluted to 25 ml with DI water in a volumetric flask. Absorbance was then measured at 550
nm and a plot of the measured absorbance against the glucose content (mg) was constructed. A Dmannose standard curve was built up by the procedure as described for glucose.
The glucomannan content (db) was calculated by following equation:
GM content (%) =

Where f = correction factor, T = glucose content of glucomannan hydrolysate (mg), To = glucose content
of glucomannan extract (mg), m = mass of glucomannan (200 mg) and w = water content of glucomannan

2.4. Chemical Compositions

The content of moisture, protein and ash were determined according to AOAC methods (Tatirat
& Charoenrein, 2011). The starch were analyzed qualitatively on glucomannan granules by staining
using I2-KI (Zhao et al., 2010). The presence of dark blue colour after staining indicated high starch
content of glucomannan.

2.5. Yield
The glucomannan yield (db) was calculated by following formula:

Where m1 = mass of dried glucomannan, m2 = mass of wet peeled porang tuber, w1 = water content of
dried glucomannan, and w2 = water content of wet peeled porang tuber.

2.7. Colour
Lightness (L*) of glucomannan powder was analyzed with a Minolta spectrophotometer. Dried
samples were put in a quartz silica cylinder and the lightness values were measured (Tatirat &
Charoenrein, 2011).

2.8. Morphology
Morphology of glucomannan granules obtained from repeated milling using ethanol and filtration
for 5 or 7 times was analyzed using Scanning Electron Microscope (SEM) (Tatirat & Charoenrein, 2011).
Dried glucomannan was placed on a stub and coated with thin layer gold sputters at 55 nm thickness onto

the samples and observed with a SEM-EDS Merck FEI type S50, EDAX AMETEK. The magnification
and accelerating voltage were displayed on the micrograph.

2.9. Transparency
One gram of glucomannan and 99 g or deionized water was dissolved until completely hydrated.
Transparency of the glucomannan sol was measured by UV-VIS Spectrometer at 550 nm (Ye et al.,
2014).

2.11. Viscosity apparent

Sol glucomannan (1%) was agitated at 150 rpm until completely hydrated. The measurement was
taken using a Brookfield Viscometer Model LVTDV-II at 25oC with spindle no.64 at 20 rpm.

2.12. Solubility
Glucomannan (0.1 g) was dispersed to 24.9 g deionized water and stirred for 1 hour. The mixture
was centrifuged for 20 minutes at 4000 rpm. About 10 g of the supernatant was dried to constant weight
at 105oC. The solubility was calculated by following equation (Du, Li, Chen, & Li, 2012):

Where m is the weight of soluble component in 10 g upper solution, W is the total weight of
glucomannan.

2.13. Water Absorbency

Glucomannan (0.1 g) was placed in previously weighed falcon and added with 30 g of deionized
water. The mixture was kept stay for 1 hour. The mixture was then centrifuged at 4000 rpm for 30
minutes. The supernatant was removed and the remained substance was weighed. The weight difference

between the two measurements was taken as the weight of the absorbed water (Koroskenyi & Mccarthy,
2001).

2.14. X-Ray Diffraction (XRD)

XRD patterns were determined using a Lab X XRD-6000 Shimadzu (Japan) equipped with a Cu
K target 40kV and 30 mA with a scan rate of 4oC/min. The diffraction angle ranged from 2 = 5 to 2
= 60o. Crystallinity percentage (%) = area under the peaks/total curve area x 100 (Wang, Zhou, Wang, &
Li, 2015).

3. Results and discussion

3.1. Isolation of glucomannan from fresh tubers
The purity and the yield of glucomannan isolated by EtOH7 were enhanced significantly to
90.98% with low percentage of ash (0.57%) and protein (0.61%) without starch content (Table 1).

Table 1
Ash, starch, protein and yield of porang glucomannan compared to those of PKG and CPF
Treatment
Glucomannan
Ash
Starch
Protein
Yield
(%)z
(%)
(I2-KI test)
(%)
(%)z
a
b
bc
AA15
76.83
2.07
+++++
3.39
59.02b
b
a
b
AA30
85.72
1.40
+++++
2.52
50.02a
bc
a
a
EtOH5
88.67
0.65
0.99
65.23c
EtOH7
90.98c
0.57a
0.61a
61.05b
d
a
a
PKG
94.42
0.60
+
0.31
Na
CPF
79.91a
5.51c
+++
1.86b
Na
Different superscripts among mean values in each column indicated significant difference (p 0.05).
z
data were based on dry basis
na = not available
- = no color, + = very light blue, ++ = light blue, +++ = blue, ++++ = dark blue, +++++= very dark blue

Compared to PKG, EtOH7-isolated glucomannan had lower purity, however, remained considered as
purified glucomannan based on the standard by The Ministry of the Peoples Republic of China (2002).
The level of ash and protein on EtOH7 isolated glucomannan by was not significantly different from that
on PKG.

10

The glucomannan granules of porang were similar to those of konjac. During milling, the softer
starch around the glucomannan and fiber cells from peripheral cell layers of the corms were first
disintegrated to tiny ash particles (Zhao et al., 2010). However, the glucomannan granules could not be
broken to fine particles as they were very hard. In addition, the size of glucomannan granules was bigger
than that of the impurities. These differences resulted in an easy separation of starch particles and other
impurities from glucomannan granules during milling and further filtration. The impurities leached out by
ethanol during filtration. The higher purities of glucomannan granules could be produced by longer
milling time than those by EtOH7. However, the low yield probably due to the partial small granules
leaching during filtration resulted in the decreasing of the EtOH7 isolated granules.
During milling in ethanol, the impurities (starch, protein, and ash) previously present interstitially
among the glucomannan granules in fresh tuber could be more easily detached from glucomannan
granules than those in flour derived from chips. Drying process hardened porang slices and stuck
impurities covering glucomannan granules more firmly. Smirnova, Mestechkina & Shcerbukhin (2001)
removed pigments and low molecular substances from glucomannan by boiling in 70% aqueous ethanol
solution for 1 hour. Xu, Wang, Ye, et al. (2014) also found that konjac flour was effectively purified by
using ethanol with controlling its temperature at 68oC. In this research, the increase in ethanol solution
temperature to 45-50oC during milling resulted in significant higher removal of impurities.
Milling and polishing of chips or konjac flour were only able to remove partial impurities while
some others were still attached to the glucomannan granules. Consequently, the impurity level of the
glucomannan granules was still high. Further purification by water dissolving and precipitation using
higher volume of high concentrated ethanol was required, even after stirring the konjac flour in 50%
ethanol for 90 minutes (Chua et al., 2012) or after simple centrifugation of water dissolved glucomannan
with flocculating agent (Tatirat & Charoenrein, 2011). Other methods, after impurities removal using
68oC citric acid ethanol (W. Xu, Wang, Jin, et al., 2014) and DMSO (Ye et al., 2014), the obtained
glucomannan need to be washed using higher amount of high concentrated ethanol to get high purity
glucomannan. On the other hand, the impurities covering glucomannan in fresh corm could be removed
11

more easily during milling in ethanol than those in chips or konjac flour, thus the EtOH7 glucomannan
had high purity without purification. The EtOH7 isolated glucomannan already contained a low level of
ash although slightly higher than those obtained by Tatirat & Charoenrein (2011) and by Xu, Wang, Jin,
et al. (2014) methods.
The ash content including calcium oxalate could be totally removed at longer duration of tuber
milling. This method would be easier and faster than the previous methods as drying of tuber, or further
washing or purification using higher amount of high concentrated ethanol as well as a flocculating agent
were no longer required.

3.1. Characteristics of porang glucomannan

3.1.1. Lightness values
Glucomannan lightness value isolated by both EtOH7 and by AA15 was not significantly
different from PKG (Fig.1).

Fig 1. Lightness values of CPF, PKG and glucomannan isolated by AA15, AA30, EtOH5 or EtOH7.
Each lightness value is shown beside of each picture label. Different superscripts among mean values
indicate significant difference (p 0.05).

The impurities covering glucomannan granules were more easily broken up to very fine particles
while the glucomannan granules were very hard to be ground. During milling in ethanol, the impurities in

12

the fresh tuber of porang can be gradually removed from surface of glucomannan granule. Longer milling
caused significantly higher impurities detachment which could be separated and ditched from
glucomannan granules during filtration.

The higher removal level of impurities resulted in whiter

glucomannan granules. In addition, the native carotenoid on the tuber (Wootton, Luker-Brown, Westcott,
& Cheetham, 1993) could be removed as the pigment could be dissolved by ethanol and discarded during
filtration as well as browning inactivation due to ethanol temperature increase to 45-50oC during milling .

3.1.2. Morphology of glucomannan

SEM images of the glucomannan morphology are shown in Fig. 2.

Fig 2. SEM photographs of PKG (A), glucomannan granules isolated by EtOH5 (B) or EtOH7 (C) with
70X (1). 200x (2) and 750X magnification
13

Shape of native porang glucomannan granules isolated by EtOH7 and EtOH5 was round or oval with
rough, scale like-pattern while PKG was irregular as the glucomannan had already been ground. Surface
of CPF was smooth with blur scale-like pattern inside as the surface of the glucomannan granules was
still covered by impurities (Yanuriati, Haryadi, Rochmadi & Harmayani, 2013). The granules were
dispersed in the corm as single cells covered with scale-like cell walls (Takigami et al., 1997) and the
scale like pattern became more clearly appear on the surface of purified glucomannan granules
(Takigami, 2000). Li et al. (2009) found that native structure of konjac glucomannan was primarily
composed of lamella structure units. It appeared that scale like pattern on first lamella structure units of
glucomannan granules isolated by EtOH7 thinned and began to crack (Fig 2.C.). The layer could be
released by longer milling duration.

3.1.3. X-ray diffraction

Spectrums of X-ray diffraction describe the correlation graph between the diffraction of intensity
and angle. The sharp diffraction peak with light baseline is illustrated at a crystalline state, whereas the
widened peak diffraction appears solid and amorphous state. The X-ray curves of porang glucomannan
were shown in Fig 3.

Fig 3. XRD pattern of PKG, glucomannan isolated by EtOH7, AA15, and CPF

14

Patterns of all porang glucomannan at 2 = 5 - 50o exhibited a very broad band with high peaks
around 19o-20o and small peaks around 35o. Native glucomannan had low degree of crystallinity around
or less than 5.43%, which indicated that all of them almost fully amorphous (Table 3). The same
patterns were also found in native konjac glucomannan (S. Wang et al., 2015; Yao et al., 2011) and
Amorphophallus corrugates glucomannan (An, Thien, Dong, Dung, & Du, 2011).

The degree of

crystallization of native konjac glucomannan was higher than that of native porang glucomannan, about
16% (Pan et al., 2003 in Li et al., 2009). Figure 3 showed that an increase of crystallinity levels occurred
during glucomannan isolation. The highest increase of crystallinity degree was detected on EtOH7
isolated glucomannan (5.43%), followed by CPF (2.78%), AA15 (0.033%), and PKG (0.026%),
respectively.
The higher level of crystallinity on EtOH7 isolated glucomannan was stimulated by the use of
50% ethanol and ethanol temperature increase during milling. The crystallinity of glucomannan was
enhanced by ethanol (Wang, Zhong, Chen, Li, & Lv, 2011) since dehydration of glucomannan granules
strengthened the intermolecular and intramolecular hydrogen bonds and increased the crystallinity. In
addition, the exothermic ethanol during milling between 45oC-50oC also contributed to the increase in
crystallinity of the glucomannan granules. The amorphous parts of glucomannan granules were removed
by milling in ethanol. Li et al. (2009) found that 50% ethanol containing water activity around 0.25 had
only few number of activated water molecules which broke the hydrogen bonds between lamellar
structure units of konjac glucomannan granules. Thus, the crystallinity was still strongly maintained by
the inter and intramolecular bonds.
On the other hand, AA15 isolated glucomannan had lower crystallinity than those isolated by
EtOH7 and CPF which probably caused by purification process. Purification was performed by dissolving
in water, clarification using centrifugation and precipitation using ethanol. Prawitwong, Takigami, &
Phillips (2007) described that water was initially sorbed in the amorphous regions with less hydrogen
bonds. From these points, it could be hypothesized that the glucomannan chain could expand and the
water molecule had capability to break down the inter and intramolecular hydrogen bonds of
15

glucomannan. The weakness of the hydrogen bonds allowed water to form hydrogen bonds with the
molecules and increase of amorphous space region.

3.1.4. Transparency of porang glucomannan sol

Transparency of porang glucomannan was shown on Table 2. The most transparent sol of
glucomannan was produced by EtOH7 followed by EtOH5, which appeared more transparent compared
to PKG. In contrast, the transparency of glucomannan sols isolated by AA15 or AA30 was far lower
(Table 3) than those by EtOH7 or EtOH5. The presence of yellowish color in AA15 and AA30 isolated
glucomannan sols indicated that glucomannan still contained high level of impurities, such as protein,
starch (Table 1) including the native carotenoid.
Table 2
Physical characteristics of porang glucomannan
Treatments
Viscosity (cps)
Solubility (%)

WHC (g water/g GMP)

Transparency
(%)
AA15
102a
83.16b
24.21a
0.44a
a
a
a
AA30
134
74.16
24.91
2.15b
b
a
c
EtOH5
24,650
71.38
50.91
51.81e
b
a
c
EtOH7
27,940
74.59
52.11
57.74f
PKG
27,300b
98.43c
58.12d
47.83d
a
a
b
CPF
1,700
73.74
36.99
19.47c
Different superscripts among mean values in each column indicated significant difference (p 0.05).
In addition to the presence of a low level of starch content (Table 1), the less transparent of PKG
sol compared to the glucomannan sol from EtOH7 and EtOH5 could also be affected by the milling
process. The PKG had more further processes, such as purification, precipitation, drying and milling to
powder. During purification and drying, the intermolecular hydrogen bonds of glucomannan molecules
became closer and stronger (Alonso-Sande et al., 2009) to produce the very hard glucomannan, thus the
dried glucomannan precipitate was not easy to be milled. The precipitate powdering required a high
speed grinder resulted in high temperature frictions. The high temperature oxidized the glucomannan
granules which brought about decolourization and reduction on transparency and viscosity (Ohashi et al.,
2000).

16

3.1.5. Viscosity
Viscosity of glucomannan sol can be observed in Table 2. The highest viscosity level was found
in EtOH7 isolated glucomannan which was not significantly different from PKG and EtOH5 isolated
glucomannan. However, the sol viscosity of CPF, AA15 and AA30 isolated glucomannan were
significantly very low.
Glucomannan has poor solubility despite its hydrophilic property (Pan et al., 2013). The low
viscosity of the 2 last glucomannan was accounted for the low solubility of glucomannan. Not all
glucomannan granules had been dissolved during 4 hours extraction using water. As a result, some high
molecular granules might lose during centrifugation which lowered their glucomannan content.
Viscosity level was affected by molecular weight and purity. It was proportional to the molecular
weight (Luo, Yao, Zhang, Lin, & Han, 2012). Glucomannan had high molecular weight and very high
viscosity (Ojima et al., 2009). The viscosity lowered as the molecular weight reduced (Ojima et al., 2009;
Tatirat, Charoenrein, & Kerr, 2012). In this study, the molecular weight analysis by gel permeation
chromatography was conducted on EtOH7 isolated glucomannan whose Mn (number average molecular
weight), Mw (weight average molecular weight) and Mw/Mn were 6.47 x 105, 1.27 x 106 and 1.96
respectively due to its highest purity, viscosity and transparency. The AA15 and AA30 isolated
glucomannan still had higher impurities (ash, protein and starch) than EtOH7 and EtOH5 isolated
glucomannan (Table 1). The impurities had lower viscosity than glucomannan, resulted in reduction of
viscosity.
3.1.6. Solubility
Solubility of EtOH5, EtOH7, or AA30 isolated glucomannan was not significantly different,
except AA15 isolated glucomannan had significant higher solubility than the others. However, its
solubility remained significantly lower than PKG.
Solubility of glucomannan was affected by molecular weight and by the material morphology.
Lower molecular weight was less compact and had more porous particle (Luo et al., 2012) leading to
higher solubility. The solubility of glucomannan was also related to the hydroxyl and O-acetyl group
17

(Luo et al., 2013). In addition to high purity, the highest level of solubility of the PKG could be the
results of purification and grinding. Purification was performed by dissolution of glucomannan in water
and precipitation of the glucomannan sol in ethanol. During dissolving in water, the molecular chain
expanded (Luo et al., 2013) and the acetyl groups became be more exposed. The acetyl groups enhanced
the solubility and dispersion of glucomannan as the groups prevented the formation of intra and
intermolecular hydrogen (Alonso-Sande et al., 2009; Chen, Li, & Li, 2011) which became closer and
tighter during drying (Xu, Willfr, & Holmbom, 2008). The changes of the molecular orientation did not
revert totally after drying and enhanced the solubility.
EtOH5 and EtOH7 isolated glucomannan were still in native granule form. Fig. 2B and 2C
showed the glucomannan granules were covered by first lamella structure which might also hinder the
solubility. Li et al. (2009) found that the native konjac glucomannan consisted of lamella structure units
containing both crystalline and amorphous regions. The connection zones between lamellar structures
contained booth loose and tight aggregation regions. These lamella structure units contributed the slow
hydration due to the gradual solubility. The solubility was higher in the amorphous than in the crystalline
regions.

3.1.7. Water holding capacity (WHC)

WHC of EtOH5 and EtOH7 isolated glucomannan rose significantly. On the other hand, WHC
of AA15 and AA30 isolated glucomannan was reduced significantly. Nonetheless, WHC of EtOH5 or
EtOH7 isolated glucomannan remained significantly lower than the PKG (Table 2). Strong structural
hydrogen bonds stimulated the low aqueous solubility (Kohyama, Sano, & Nishinari, 1996). However,
the formation of strong hydrogen bond between hydroxyl groups and water stimulated the high water
absorbency. The high water absorbency of native konjac glucomannan could reach 105.4 g water/g KGM
(Koroskenyi & Mccarthy, 2001). The cracks on some parts of first lamella unit on the EtOH7 isolated
glucomannan (Fig. 2C) contributed to the rate of water dispersion through the amorphous regions in the
lamellar structural units. The existence of water molecules in sufficient amount weakened and broke the
18

hydrogen bonding which resulted in the occurrence of bonds between water molecules and hydroxyl
group. However, the presence of some impurities covering glucomannan granule restrained the adsorption
of water which resulted in the lower WHC in AA15 and AA30 isolated glucomannan.
Conclusions
Repeated milling of sliced fresh porang tuber in ethanol followed by filtration without further
purification could be developed as an easier and faster novel method to isolate glucomannan, with
significantly high purity (90.98%), viscosity (27,940 cps) and transparency (57.74 %), since the level of
ash and protein content was reduced significantly to 0.57%, 0.31% with no starch content. The yield of
glucomannan was also enhanced by EtOH7. Besides no corm drying process was required for this
method, some other common steps could be eliminated, such as addition of antibrowning as well as
further washing or purification process using flocculants and higher volume of high concentration of
ethanol. The native glucomannan granules were still amorphous. The WHC was enhanced significantly
while the solubility was significantly lower than commercial PKG. In addition to high molecular weight
of native granular form, the lower solubility of the glucomannan could be contributed the gradual
solubility due to the lamellar structure units of glucomannan.

Acknowledgements
This research was partly supported by Post Graduate Grant in 2013 no. LPPMUGM/1396/LIT/2013 and Doctoral Dissertation Grant in 2015 no. 115/UN9.3.1/LT/2015 from Ministry
of Research, Technology and Higher Education, Republic of Indonesia. The authors also expressed
greatly thank to Dr. Anni Faridah for commercial PKG support.

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