Influence of Process Conditions on Digestibility of African Locust Bean (Parkia biglobosa) Starch
This study describes the isolation, digestibility and effect of process conditions on the Parkia biglobosa (African locust bean) starch digestibility. Parkia starch fractions are: Total Starch (TS), Rapidly Digestible Starch (RDS), Slowly Digestible Starch (SDS) and Resistant Starch (RS). The results indicate that processing conditions can be changed to effectively control the relative content of SDS and RS in Parkia starch products. Amylose is the molecular basis of RS while amylopectin is the main constituent of SDS and plays a key role in the structure and digestibility of SDS. This methodology may enable process modifications to influence the functional digestibility properties of prepared Parkia starch products.
May 26, 2012; Accepted: July 09, 2012;
Published: July 24, 2012
Africa locust bean (Parkia biglobosa) is a perennial tree legume which
belongs to the sub-family Mimosoideae and family Leguminosae. It grows in the
savannah region of West Africa (Pelig-Ba, 2009; Ihegwuagu
et al., 2009). Africa locust bean tree is an important food tree
for both man and livestock such as husks and pods and plays a very vital role
in the rural, corresponding author economics of West African countries; virtually
every part of the species is of value as food or fodder (Teklehaimanot,
2004; Tee et al., 2009). Starch is the major
storage carbohydrate in plants. It is produced as granules in most plants cells
and is referred to as native in this state. Native starches from different botanical
sources vary widely in structure and composition but all granules consist of
two major molecular components, amylose (20-30%) and amylopectin (70-80%) (Sankhon
et al., 2012). The physicochemical properties of starch and its use
depend largely on its biological origin and source and the various sources include
cereal, grain, nuts, seeds, leaves, tubers and root. Because starch finds application
in various industries, the researches for new sources of starch, like P.
biglobosa, becomes necessary. Although qualitative determination of the
chemical and nutritional composition of P. biglobosa seeds revealed that
it is rich in starch, lipids, protein, carbohydrates, soluble sugars and ascorbic
acid (Pelig-Ba, 2009; Ihegwuagu
et al., 2009). Studies indicate that the digestibility of starch-based
products are not only affected by food type, degree of maturation, starch structure,
starch content, food ingredients and individual factors (FAO,
1998), processing and storage, pea starch is very susceptible to retrogradation
and thus becomes rich in RS which is naturally indigestible (Skrabanja
et al., 1999). The retrogradation rate and extent are mainly affected
by the inherent starch properties, including molecular and crystalline structure
and by storage conditions, such as temperature, time and water content (Gudmundsson,
1994; Liu and Thompson, 1998). Amylopectin contributes
to the retrogradation occurring in long-term rheological and structural changes,
whereas amylose is usually responsible for the short-term, rapid changes in
food texture (Sathaporn and Jane, 2007). Starch often
changes from an amorphous state to a crystalline state and thus this retrogradation
process includes a recrystallization (Yuan et al., 1993).
In this research, the Parkia starch powder was prepared to apply in
the food industry via. a soaking and cooking process which differed from normal
domestic preparation, therefore, the objectives of this study are to isolate
and investigate how different process conditions affect on the digestibility
of Parkia biglobosa starch and to reduce slow starch digestibility in
favor of resistant starch.
MATERIALS AND METHODS
Africa locust bean (Parkia biglobosa) seeds were purchased from the
local market in Madinah (Conakry, Guinea) in August, 2011 and shipped to Wuxi,
China. Porcine (invertase, pancreatic α-amylase, amyloglucosidase) were
purchased from Sigma Aldrich Co. Ltd. (Shanghai, China). The other chemicals,
potassium hydroxide, sodium hypochlorite, ethanol, 3,5-dinitro salicylic acid
were purchased from Sinopharm Chemical Reagent Co. Ltd. All other reagents used
were of analytical grade.
The different samples ratio material/water (1:2, 1:3, 1:4, 1:5, 1:6 and 1:7
w/v) were prepared in a sterilization equipment chamber pot (YXQ-LS-SII shanghai
Boxun, industry and trade Co. Ltd., medical equipment factory). The following
conditions were applied: time of soaking (10 h) and cooking temperature (90°C)
for 3 h. The samples were cooled at room temperature (28°C) and then placed
in a refrigerator (4°C) for 72 h. Samples were dried in a hot air oven at
45°C until constant weight, after cooling in a desiccator the samples were
ground and sieved using 60 mesh sieve and stored in sealed plastic bags until
analysis for starch digestibility. The different samples ratio were taken one
by one for the fraction of starch digestibility (RDS, SDS and RS). All samples
were treated in triplicate.
This research was conducted in the School of Food Science and Technology Laboratory
and State Key Laboratory of Jiangnan University, Wuxi from August 2011 to May
Starch isolation: The isolation of starch from Parkia biglobosa
seed was performed according to the method of Sira and Amaiz
(2004) with slight modification. Visible dirt and contaminants were removed
from the dark-colored Parkia seed (1 kg) which was then steeped in a
solution of sodium hypochlorite (35 g) and potassium hydroxide (50 g) in water
(2 L) at room temperature (28°C) for 3 h. The pH of the steep solution was
elevated to 9 and the mixture was maintained at 100°C in a thermostat water
bath for 3 h. Then, the solution was drained and the seeds were immersed in
water and left overnight at ambient temperature. Finally, the seeds were thoroughly
washed, manually dehulled and the cotyledon was washed repeatedly until the
wash pH was neutral. The cotyledon was blended with water for 24 h using a domestic
blender. The homogenate was filtered through muslin cloth and the filtrate was
allowed to settle overnight. The supernatant was decanted and the sediment was
centrifuged at 4500 rpm for 10 min using a ZOPR-52D refrigerated centrifuge
(Hitachi Koki Co. Ltd., Tokyo, Japan). The sedimented starch was re-suspended
in water and the process was repeated six times. The resultant starch was dried
at 60°C in a hot air oven, then grounded to powder using a mortar and pestle
and stored in cellophane and wrapped before usage.
In vitro digestibility of Parkia starch: Five hundred
milligrams of starch fractions: Total Starch (TS), Rapidly Digestible Starch
(RDS), Slowly Digestible Starch (SDS) and Resistant Starch (RS) were measured.
Samples were incubated with invertase, pancreatic α-amylase and amyloglucosidase
at 37°C in capped tubes immersed in a water bath shaker according to the
method of Englyst et al. (1992) and the supernatants
were measured at 0, 20 and 120 min for glucose content. The glucose data obtained
was used to calculate the content of various starch types using the following
Total starch (TS) = (Total glucose (TG)-free glucose
Rapid digestible starch (RDS) = (G20-FG)x0.9
Slow digestible starch (SDS) = (G120-G20)x0.9
Resistant starch (RS) = TS-(RDS+SDS)
where, FG, G20 and G120 (mg) represent the amount of glucose in the supernatant
at 0, 20 and 120 min of hydrolysis, respectively. Total Glucose (TG) was measured
with the 3,5-dinitrosalicylic acid according to method of Rose
et al. (1991) after the starch was completely hydrolyzed into glucose
by perchloric acid. The in vitro digestibility of starch was determined
from the calibration curve equation:
y = 0.743x+0.0004 and R2 =0.9958
Processing conditions of starch digestion: To improve the yield of modified
starch from Parkia seeds, different parameters were taken in consideration
according to the digestion method of Englyst et al.
(1992). The sample/water ratio were fixed as (1:2, 1:3, 1:4, 1:5, 1:6 and
1:7) w/v, the soaking time (6, 8, 10, 12 and 24 h) and the cooking temperature
(50,70, 90, 110 and 121°C) for 3 h. Then, the optimal operating conditions
was determined by means of varying one parameter while keeping the others at
a constant level. First the sample/water ratios were varied, after finding the
best ratio; followed by cooking temperatures for 3 h and finally soaking times
were varied to study the effects that affect on the digestibility of Parkia
Differential scanning calorimetry (DSC): Calorimetric measurements (gelatinization
temperature and enthalpy) of the processed Parkia starch product with
conditions of material/water ratio 1:4, soaking for 12 h, 110°C cooking
for 3 h and the samples were cooled at room temperature (28°C) and then
placed in a refrigerator (4°C) for 72 h were analyzed with the Pyris-1 Differential
Scanning Calorimeter (DSC) (PE, USA). Samples (3 mg) were weighed directly into
DSC aluminum pans and deionizer water, after sealing, the pans were left to
equilibrate (24 h) at room temperature and then heated from 30 to 120°C
at 10°C/min. In all measurements an empty pan was used as reference. Onset
(To), peak (Tp) and Endset temperature (Te)
of gelatinization as well as gelatinization enthalpy changes (DH) were determined.
Statistics: The test results were processed by using One-way Analysis
of Variance (ANOVA) test using statistical software (SAS, version 8.1). Differences
at p<0.05 were considered to be significant.
RESULTS AND DISCUSSION
In vitro digestibility of Parkia starch: Based on the
Englyst test, percentages of RDS, SDS and RS in normal Parkia starch
were 10.91, 52.29 and 33.41%, respectively exhibited significant difference
Our results corroborate with the work of Zhang et al.
(2006, 2008) in waxy maize, wheat, rice and maize.
The RDS level (10.91%) in the Parkia starches were generally lower than
those reported for pea (18.2-23.8%), lentil (16.0-16.9%) and cultivars of other
chickpea (21.5-29.9%) starches (Chung et al., 2008a).
The SDS (52.29%) was comparable to those of pea (53.7-59.0%) lentil (58.3-62.2%)
and other chickpea cultivars (45.7-57.7%). Whereas, RS (33.41%) were much higher
than those reported by Chung et al. (2008a, b)
for pea (8.1-12.6%), lentil (13.0-13.2%) and other chickpea cultivars (8.4-18.4%).
SDS content which is considered a desirable form of dietary starch, was 52.29%.
This value are much higher than those reported by Zhang
et al. (2006) for maize (53.0%), waxy maize (47.6%), wheat (50.0%),
rice (43.8%) and potato (15.2%). RS content which is considered a desirable
form of dietary starch, was 33.41%. This value is much higher than those reported
by Pongjanta et al. (2009) of bean varieties
AC Nautica (17.2%), Majesty (21.3%) and red Kanner (21.9%); although measured
using the Englyst method. The RDS, SDS and RS (Fig. 1) levels
of the Parkia starches cannot be compared with those reported for other
legume starches, due to differences in methodology AACC (2000)
vs. Englyst et al. (1992) and to different time
periods of hydrolysis that have been defined for measurement of RDS, SDS and
Generally, digestibility of native starch is influenced by starch source, granule
size, amylose/amylopectin ratio, crystallinity and amylopectin molecular (Chung
et al., 2006; Singh and Ali, 2006). SDS
which leads to a slower entry of glucose into the blood stream and a lower glycemic
response, is digested completely in the small intestine at a lower rate as compared
to RDS, while RS is the starch portion that cannot be digested in the small
intestine but is fermented in the large intestine (Oates,
1997; Amadou et al., 2011).
A moderate postprandial glycemic and insulinemic response of SDS implies that
SDS rich foods may provide wide health benefits in reducing common chronic diseases
such as obesity, diabetes and cardiovascular disease through lessening the stress
on regulatory systems related to glucose homeostasis (Ludwig,
|| Digestibility of Parkia starch by Englyst test
||Effect of the sample cooking (material/water ratio) on in
vitro digestibility of Parkia starch soaked at 10 h, cooked at
90°C for 3 h and after storage at 4°C for 72 h
|Values (Mean±SD) in the same column with different
letters are significantly different at p<0.05, n = 3
Effect of sample/water ratio on the starch digestion: The results exhibited
that ratio 1:4 (Table 1) as the best ratio for the digestion
of Parkia starch. There are significant differences (p<0.05) among
the values 12.09, 39.76 and 44.76%, respectively for RDS, SDS and RS. These
results are comparable to those reported by Yao et al.
(2010) and Zhang et al. (2008). Various studies
have shown that several factors can contribute towards the differences in RS
quantities in foods such as: botanical origin; nature of starch (amylose and
amylopectin content and their ratio); food processing (degree of starch gelatinization
and retrogradation); starch morphology (particle size and cellular structure),
types of starch granules or their crystalline structure (such as A, B and C
or V) and presence of other components (lipids, protein, dietary fiber, antinutrients
and organic acids etc.) (Goni et al., 1997). The
material/water ratio might affect the space available for starch chain extension
in the water/starch suspension. At high concentrations, starch chains may not
be fully extended; and at low concentrations, extended linear and branched starch
molecules have less probability to contact each other thus limiting the formation
of organized molecular arrangements and crystals and subsequently forming high
levels of RS (Yao et al., 2010).
Previous research have demonstrated that in processed food RDS and glycemic
index are highly correlated; while, SDS from the structural perspective show
and prolonged release of glucose of native maize starch. Furthermore, RS is
resistant to digestion and no glucose is available for glycemic response. However,
the fermentation of RS in the large intestine generates short-chain fatty acids
that are beneficial for colonic health. Both SDS and RS either alone or in combination,
contribute to an improved nutritional quality of starch (Englyst
et al., 1999; Seal et al., 2003; Zhang
et al., 2006; Topping and Clifton, 2001).
Effect of cooking temperature on starch digestion: After choosing the
ratio (1:4), the temperatures were varied (50, 70, 90, 110 and 121°C) for
3 h and the soaking time was kept for 10 h and continued the process as in previous
first case (material/water). The result showed in (Fig. 2)
that the ratio (1:4), the temperature of 110°C was the best with RDS, SDS
and RS, 12.97, 37.99 and 45.65%, respectively which are significantly different
(p<0.05). The proportions of RS of cooked temperature (90, 110 and 121°C)
starch samples are good results, exhibiting the best result of 45.65% digestibility.
However, when compared to results following the proportion of SDS and RS fractions
of pea starch (8.5 and 47.7%, respectively) (Yao et al.,
2010) which are slightly lower than our data in the same proportion of SDS
and RS starches used.
||Effect of the cooking temperature on the in vitro digestibility
of Parkia starch, with material/water ratio; 1:4, soaking time; 10
h and cooking temperature; 50, 70, 90, 110 and 121°C for 3 h, after
storage at 4°C for 72 h
Pongjanta et al. (2009) commented on the proportion
of RDS, SDS and RS fractions in legume starches that it is difficult to make
a meaningful comparison of the levels of RDS, SDS and RS proportion of legume
starch since most of these studies used different digestibility methods varying
in time of hydrolysis and enzyme sources. High temperature and high pressure
treatment would lead to full gelatinization of starch granules and complete
migration of amylose molecules. The more, cooking temperature the lower the
viscosity which lead to more free starch. And when lower viscosity, free starch
(such as amylose) molecules have easier access to each other and tend to form
inter-molecular hydrogen bonds which is favorable to full association of amylose
double helix molecules and the formation of RS. Too high a process temperature
will give further decrease of starch molecules to provide low molecular weight
hydrolyzates with limited potential for re-associations (Chinma
and Igyor, 2008; Oates, 1997; Goni
et al., 1997). Chung et al. (2009)
stated that the amylose-amylose interactions which are much stronger than those
of amylose-amylopectin or amylopectin-amylopectin, may have continued to exist
after gelatinization and thereby partly restricting accessibility of starch
chains to the hydrolyzing enzymes.
Effect of soaking time on the digestion of Parkia starch: The
results of RDS, SDS and RS for soaking time were 12.56, 37.82 and 46.22%, respectively.
The best soaking time for the digestion of Parkia starch appear to be
12 h with significant (p<0.05) increase in starch digestibility as it is
shown in Fig. 3. Similar phenomenon was observed in the work
of Siddhuraju and Becker (2001) where soaked and boiled
legumes had significant increase in digestibility.
Soaking affect the starch digestibility, involves entry of water into the legume
kernels, wetting and dissolving soluble nutrients. The observed changes in RS
and SDS level may be explained by the relatively slow cooling after gelatinization,
starch molecules became less energy active and starch molecules of appropriate
sizes rearranged to form orderly crystalline precipitation which made starch
paste retrograde into a gel. Although starch is not generally soluble in cold
water, our study showed that soaking significantly (in most cases) increased
all starch fractions in all sample (Eyaru et al.,
2009). The soaking of seeds in plain or tempered water produces a swelling
of the tissues and water uptake without cell separation; Bishnoi
and Khetarpaul (1993) found that digestibility increased with soaking of
all peas studied.
||Effect of the soaking time on the in vitro digestibility
of Parkia starch with material/water ratio; 1:4, soaking time; 6,
8, 10, 12 and 24 h and cooking temperature; 90°C for 3 h, after storage
at 4°C for 72 h
The observed changes in RS and SDS level may be caused by the relatively slow
cooling after gelatinization, starch molecules became less energy active and
starch molecules of appropriate sizes rearranged to form orderly crystalline
precipitation which made starch paste retrograde into a gel; as soaking time
prolonged, the bundle structure formed between starch chains by hydrogen bonding
may dissociate which is not conducive for the formation of RS but rather favors
the formation of SDS. Conversely, the results of Eyaru et
al. (2009) contributed to the different consequence of soaking and cooking.
They soaked Parkia starch before cooking and this resulted in reduced
starch fractions, possibly due to leaching of soluble fractions.
Differential scanning calorimetry (DSC): DSC is usually a tool used
to investigate the phase transitions of the gelatinization process and it has
been reported earlier that Parkia starch is a less stable one (Ihegwuagu
et al., 2009). The thermal properties of Parkia starches;
onset, peak and endset temperatures (To, Tp and Te,
ΔH, respectively) are summarized in Table 2. This corroborates
our earlier observation that to heat than corn starch (Ihegwuagu
et al., 2009).
In principle, fully gelatinized starch should produce a flat straight line
with no absorption peak in DSC analysis. However, starch molecules rearrange
and retrograde to form many crystal-like structures; breaking these crystal
structures to re-solubilize starch molecules requires external energy. The gelatinization
transition temperatures [onset temperature (To), peak temperature
(Tp), Endset temperature (Te) and the enthalpy of gelatinization
(ΔH) of native and modified starches are presented in Table
2. Cooking starch influenced the gelatinization properties of all the samples.
The gelatinization temperature of native Parkia starch ranged from 83.65-86.23°C
which corroborate with those of other tuber starches, such as potato, cassava
and new cocoyam (Collado et al., 1999; Oladebeye
et al., 2009; Shi and Seib, 1992). The onset
temperature for melting retrograded amylopectin is in the range of 63.74 to
66.30°C. In addition, Table 2 shows that after Parkia
processing, its (ΔH) increased in parallel with its content of RS and SDS
which led to decrease in digestibility of the final Parkia starch product,
thus, this may be attributed to retrograded amylopectin.
|| Differential scanning calorimetry (DSC) of Parkia
native starch and cooked starch
|Values (Mean±SD) in the same column with different
letters are significantly different at p<0.05, n = 3
In conclusion, the study showed that the ratio 1:4; soaking time 12 h at 110°C
for 3 h as the best conditions to produce the high Parkia starch digestion.
However, cooking and processing can be carried out in various ways and under
a range of condition, so that the products can vary widely in the degree of
denaturation, gelatinization and retrogradation they have undergone.
Modifications in processing conditions for Parkia biglobosa starch product
exhibited minimal impact on the content of RDS though had specific effect on
the SDS and RS content. Results deduced that amylose is the molecular basis
of RS and amylopectin plays a key role in the structure of SDS and is the main
constituent of SDS. SDS and RS significantly indicated that various processing
conditions promote the inter-conversion between them. Thus, processing conditions
can be changed to effectively control the relative content of SDS and RS in
Parkia starch products. This methodology may enable process modifications
to influence the functional digestibility properties of prepared Parkia
This article is supported by 111 project-B07029 and PCSIRT062, China. The authors
are also grateful to Fatoumata Sylla manager at ministry of Foreign Affairs
Conakry, Guinea who sent the sample down to Wuxi, China.
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