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Universidad de Buenos Aires, Departamento de Industrias, Facultad de Ciencias Exactas y Naturales, Ciudad
Universitaria (1428), Buenos Aires (Argentina)
(Received March 4, 2002; accepted August 26, 2002)
Gelation and melting processes of kappa-carrageenan (k-C) in the presence of beta-lactoglobulin (b-lg), native and denatured soy
protein (NSP and DSP) isolates were investigated by dynamic rheological techniques, apparent viscosity measurements and DSC. In
the presence of proteins, large increases in gelation temperatures and storage moduli of the mixed gels compared to single k-C gel
were observed. Dierences between the proteins in the rst steps of the gelation process were revealed by apparent viscosity
measurements. Although little changes in melting temperatures were observed in the presence of proteins by dynamic rheology, the
DSC analysis showed a large increase in these temperatures. Synergistic eects were also observed on textural properties and water
absorption of the gels. The addition of proteins increased gel hardness, cohesiveness, gumminess, springiness and reduced syneresis.
These results might be due to excluded volume eects that increased the eective concentration of the hydrocolloid and electrostatic
interactions between both biopolymers in solution. The dierent behaviours observed with the three proteins would be related to their
water absorption capacity, molecular size, exibility and supercial charge under the experimental conditions described.
Introduction
Kappa-carrageenan (k-C) is an anionic sulphated
polysaccharide extracted from red algae, which is widely
used in food industry as a thickening, gelling and
stabilizing agent. It forms thermoreversible gels in the
presence of cations (Samant et al., 1993; Ipsen, 1995;
Drohan et al., 1997; Mleko et al., 1997; Lundin and
Hermansson, 1998; Tziboula and Horne, 1998; Ould
Eleya and Turgeon, 2000). By heating aqueous dispersions of k-C at temperatures above 60 1C, the polysaccharide hydrates and adopts a random coil
conformation. Gelation occurs on cooling at a critical
temperature and it has been attributed to a two-stage
reaction involving a coilhelix transition followed by
aggregation of helices (Morris et al., 1980; Morris,
1998). Further cooling enhances the association of
helices into long sti super-strands (Hermansson,
1995). The gelation process is highly inuenced by
To whom correspondence should be addressed.
E-mail: apilosof@di.fcen.uba.ar.
w
Research Fellow, Consejo Nacional de Investigaciones Cient cas y
Tecnicas de la Republica Argentina.
z
Research Fellow, Agencia Nacional de Promocion Cient ca y
Tecnologica, Argentina.
}
Member of Consejo Nacional de Investigaciones Cient cas y
Tecnicas de la Republica Argentina.
0023-6438/02/$35.00
r 2002 Published by Elsevier Science Ltd.
doi:10.1006/fstl.2002.0938
All articles available online at http://www.idealibrary.com on
741
Viscosity measurements
The experimental set-up with two identical vessels,
shown in Fig. 1, was used to acquire temperature and
torque data simultaneously, under identical heat transfer conditions. Solutions (initially at 50 1C) were cooled
in a bath at 19.670.2 1C. Apparent viscosity as a
function of time was recorded for 1 h with a Brookeld
DV-LVT viscometer. The T-spindle (C) was rotated at
0.3 rpm and a helipath stand (pathway 0.4 cm) was used
(the T-spindle performed a helical pathway) to minimize
the eect of shear on gel formation. Concomitantly, the
temperature was recorded. The rate of aggregation
(vagreg) was estimated from the initial slope of the
apparent viscosity increase over time. Maximum viscosity (mmax) and the time at which this value was reached
were also reported.
DSC measurements
A Mettler TA 4000 DSC was used to obtain thermal
transition temperatures on single k-C and mixed
protein/k-C gels. Twenty milligram gel samples were
heated in aluminium pans (40 mL volume) from 5 1C to
70 1C at 10 1C/min. DSC calibration was performed
using ice and indium fusion thermograms, and an empty
aluminium pan was used as a reference. The recorded
thermograms were analysed using the Mettler TA 72
software to obtain the thermal transition temperatures.
The average of two replicates was reported.
742
Table 1 Gelation properties of single k-C (5 g/L) and k-C+protein (5 g/L/20 g/L) systems obtained by dynamic
oscillatory measurements
System
k-C
k-C+b-lg
k-C+NSP
k-C+DSP
tgel
(min)
Tgel
( 1C)
G0 max
(Pa)
19.670.2
14.670.2
16.470.1
14.370.3
11.170.4
20.970.4
17.270.1
21.670.5
45.572.8
154.677.7
129.3710.9
234.2729.2
50
G' (Pa)
40
30
-C
-C + -LG
20
-C + NSP
-C + DSP
10
0
10
20
30
40
T (C)
743
DSP
2000
1500
NSP
1000
500
-lg
-C
0
0
10
20
(a)
30
40
Time (min)
50
60
50-20 C
2000
DSP
-lg
1000
NSP
500
100
80
750
app
60
500
-C
40
G'
250
0
0
(b)
19.5 C
1000
1500
app (cp)
2500
10
15
20
t gel
20
Time (min)
0
0
Fig. 3 Apparent viscosity of single k-C (5 g/L) and mixed kC+protein solutions (5 g/L/20 g/L) during gel formation as a
function of time (a) and initial viscosity increase (b) for gelling
rate determination. (B) k-C, (&) k-C+b-lg, (~) k-C+NSP,
(~) k-C+DSP
G' (Pa)
10
20
30
40
50
Time (min)
Table 2 Aggregation rate (vaggreg), maximum viscosities (mmax) and gelation time obtained from apparent viscosity
measurements on k-C+protein (5 g/L/20 g/L) mixed systems
System
k-C+b-lg
k-CNSP
k-CDSP
vaggreg
(cps/min)
mmax
(cps)
t0 gel
(min)
8274
8772
19676
1057710
1123710
2286715
15.570.5
1671
1670.3
744
Table 3 Melting temperatures obtained by dynamic oscillatory measurements (Tm1) and DSC (Tm2) and thermal
hysteresis (DT) for k-C and k-C+protein systems
DT (Tm1Tgel)
Tm (1C)
System
k-C
k-C+b-lg
k-C+NSP
k-C+DSP
Tm1
3270.5
35.470.9
3370.4
34.570.7
Tm2 (DSC)
(1C)
o35
35.571
4071
4371
20.9
14.5
15.8
12.9
Hardness
Texture properties
60
Adhesiveness
Gumminess
45
30
15
0
-C
Springiness
Cohesiveness
1.0
Resilience
Texture properties
0.9
0.8
0.7
0.6
0.5
-C
+ NSP -C + DSP
-C + -lg -C
Fig. 5 Texture characteristics of k-C (5 g/L) and k-C+protein (5 g/L/20 g/L) mixed gels
745
Synergistic eects have been previously observed between carrageenan and proteins which resulted in much
stronger gels compared to carrageenan alone. Tziboula
and Horne (1998) suggested that in mixed carrageenan
serum proteins, the polysaccharide would interact with
the serum proteins and this interaction would have a
synergistic eect on gel strength. Mleko et al. (1997)
observed higher values of shear stress at fracture for
k-C gels in the presence of whey protein and similar
eects were found by Ipsen (1995) for mixed gels of
k-C with soy or pea protein.
The increased gel strength in the presence of proteins
might be caused by an increased hydrocolloid concentration because of the excluded volume eects. These
results agree with the increased aggregation rate and the
higher gelling and melting temperatures for the mixed
gels. Additionally, electrostatic cross-links between
proteins and k-C might cause a reinforcement of the
network structure.
Conclusions
The data reported in this work demonstrate the eects
of native or denatured proteins on the gelation and
melting of kappa-carrageenan (k-C) solutions and gels.
These eects might be due to thermodynamic incompatibility between k-C and globular proteins that enhanced
the gelling properties of the polysaccharide. The
increased aggregation rate observed during the cooling
process and the shift of peak temperatures under heating
conditions would be due to dierences between the
proteins, related to volume exclusion eects and
electrostatic interactions with k-C molecules.
The use of dynamic oscillatory methodology has proven
to be an excellent tool for characterizing gelation and
melting processes of dierent gel systems. In this work,
the application of complementary methods such as
apparent viscosity measurements and dierential scanning calorimetry gave additional information about the
phenomenon occurring during gelation, involving formation and dissociation of aggregates.
The increased gelation and melting temperatures and the
great improvement in gelation rate and characteristics of
k-C gels in the presence of proteins could be successfully
applied in food technology. The vegetable protein
showed behaviour similar to the animal protein, so the
former could be used as a replacement for the latter in
special foods. Synergistic eects increased with denaturation of soy protein, and this could be applied in the
design of new food formulations.
Acknowledgements
The authors acknowledge the nancial support from
Universidad de Buenos Aires, Consejo Nacional
de Investigaciones Cient cas y Tecnicas y Agencia
Nacional de Promocion Cient ca y Tecnologica de la
Republica Argentina.
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