Food Hydrocolloids 19 (2005) 549–556

www.elsevier.com/locate/foodhyd

Gelation of globular protein in presence of low methoxyl pectin: effect

of NaC and/or Ca2C ions on rheology and microstructure of the systems
L. Donato, C. Garnier*, B. Novales, J.-L. Doublier
Unité de Physicochimie des Macromolécules, INRA, Rue de la Géraudière, BP 71642, 44316 Nantes Cedex 03, France

Abstract
The present investigation aimed at understanding how the type of cations (NaC and/or Ca2C) influences heat-induced gelation of globular
protein/LM pectin mixtures in comparison to the globular protein only. Two globular proteins (b-lactoglobulin or bovine serum albumin)
were compared. On the basis of viscoelastic and microscopic observations (CLSM), it has been shown that the two cations play a major role
in gelation of the individual biopolymers and, therefore, influence strongly the properties of the mixture. These differences are related to a
competition between gelation of the protein (in absence of Ca2C) or of both biopolymers (if Ca2C is added) and phase separation processes
taking place before the mixed system is entirely gelled.
q 2004 Elsevier Ltd. All rights reserved.

Keywords: Globular protein; LM pectin; Phase separation; Gelation; Rheology; Salts; Confocal laser scanning microscopy

1. Introduction the main protein in whey with a molar mass 18.6 kg molK1
and the same isoelectric point as BSA (Dufour, 2004).
Globular proteins and polysaccharides are two gelling Under heat treatment, globular proteins undergo denatura-
biopolymers used in food industry for their wide range of tion and, depending on medium conditions, a gel can be
textural properties (Dickinson & Mc Clements, 1995). formed (Clark & Lee-Tuffnell, 1986; Gosal & Ross-
When mixed together, a phase separation process often Murphy, 2000). Hydrogen bonding, electrostatic and
occurs because of thermodynamic incompatibility or hydrophobic interactions have been suggested as the
depletion–flocculation mechanisms (Doublier, Garnier, major forces involved in gelation (Kinsella & Whitehead,
Renard, & Sanchez, 2000). If one or both biopolymers 1989). It has also been shown that formation of disulfide
form a gel, mixed gels are formed and a kinetic competition linkages and sufhydryl-disulfide interchange are involved in
between gelation and phase separation takes place. The heat-induced gelation of globular proteins. In addition, salts
control and understanding of the relative rates of these present in the system play a major role in the rate of gelation
processes by way of external factors such as medium and in textural properties of the protein gels (Matsudomi,
conditions can lead to a wide range of microstructures and a Rector, & Kinsella, 1991; Mulvihill & Kinsella, 1988;
large variety of textures (Tolstoguzov, 2003). The present Haque & Aryana, 2002; Jeyarajah & Allen, 1994; Yasuda,
study aimed at evaluating the properties of a model system Nakamura, & Hayakawa, 1986). Depending on the nature of
composed of a globular protein, bovine serum albumin globular protein and on the concentration and valence of
(BSA) or b-lactoglobulin (b-Lg), and an anionic poly- salts added in the medium specific interactions (protein–
saccharide, a low methoxyl pectin (LM pectin). BSA is the ions cross-linkage) or electrostatic interactions (salting in)
most abundant protein in plasma that contributes to colloid can take place and enhance the aggregation process. Above
osmotic blood pressure. Its molar mass is 68 kg molK1 and a critical concentration of added salts, a salting out process
its isoelectric point is around 5.2 (Peters, 1975). b-Lg is can also take place (Relkin, 1996).
Pectins are complex polysaccharides that belong to cell
* Corresponding author. Tel.: C33 240 67 50 45; fax: C33 240 67 50 43.
walls of plant materials. The main chain of pectins is a
E-mail address: [email protected] (C. Garnier). polygalacturonic acid, partially esterified by methoxyl

0268-005X/$ - see front matter q 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodhyd.2004.10.019
550 L. Donato et al. / Food Hydrocolloids 19 (2005) 549–556

groups interrupted by insertion of rhamnose residues 2. Materials and methods

carrying neutral sugars as side chains (Lau, Mc Neil,
Darvill, & Albertsheim, 1985). LM pectins with a degree of 2.1. Materials
methoxylation below 50% show a high affinity for calcium
ions leading to the formation of a gel on cooling (Voragen, The LM pectin sample, kindly given by Degussa Food
Thibault, Pilnik, Axelos, & Renard, 1995). Several studies Ingredients (Baupte, France), had a degree of esterification
have been performed on globular protein/LM pectin of 28.1%. Pectin powder was purified by washing with acidic
systems and showed that mixtures formed at 20 8C without ethanol in order to eliminate ions in excess and to obtain the
heating stay homogeneous and translucent even after polysaccharide under an acidic form. LM pectin solutions
centrifugation. These biopolymers are therefore considered were prepared by dissolving LM pectin powder in deionized
as compatible when globular protein is in the native state water. BSA (98–99 wt% high purity), was purchased from
(Dumay, Laligant, Zasypkin, & Cheftel, 1999; Semenova ICN Biomedicals (Aurora, Ohio). The protein powder was
et al., 1991; Takada & Nelson, 1983). Influence of heat- defatted with n-pentane. b-Lg (97.5 wt% purity) used in this
induced globular protein gelation on the properties of study was a gift from Lactalis (Rennes, France). Solutions of
anionic polysaccharide/globular protein mixtures has been globular protein were prepared by adding protein powder to
the subject of many studies (Cai & Arntfield, 1997; Dumay deionized water under gentle magnetic stirring at 4 8C
et al., 1999; Neiser, Draget, & Smidsrød, 1998; Syrbe, overnight. Traces of insoluble matter were eliminated by
Bauer, & Klostermeyer, 1998; Tolstoguzov, 2003; Turgeon centrifugation (16,000!g, 20 min). Calcium content of both
& Beaulieu, 2001; Wang & Qvist, 2000). Most of the time, biopolymer powders was determined by atomic absorption
the polysaccharide was non-gelling or employed in non- spectroscopy. Results were 2.3 mmol gK1 of defatted BSA
gelling conditions. Fewer studies are found on mixed powder, 15 mmol gK1 of b-Lg powder and 34.1 mmol gK1 of
systems on heat-induced gelation of a globular protein in the purified LM pectin powder. pH was adjusted at 6.8 for each
presence of a gelling polysaccharide (Beaulieu, Turgeon, & solution with NaOH. Sodium azide (0.02 wt%) was added to
Doublier, 2001; Bernal, Smajda, Smith, & Stanley, 1987; biopolymer solutions to prevent from bacteria
Ndi, Swason, Smith, & Stanley, 1996; Neiser, Draget, & contamination.
Smidsrød, 1999). A variety of structures and of rheological
properties have been reported depending on the nature of 2.2. Preparation of biopolymer mixtures
biopolymers and solvent conditions.
The present study is part of investigations we have Protein and LM pectin solutions were mixed at room
undertaken on BSA/LM pectin mixtures. In a first study temperature. It was verified that the pH after mixing was
(Donato, Garnier, Novales, Durand, & Doublier, 2004a), we still 6.8. Blends were then stirred at 50 8C before addition of
investigated rheological and structural properties of the a NaCl and/or CaCl2 warm solution. The final concen-
mixtures in comparison with the properties of individual trations in the mixture were 8 wt% of globular protein and
biopolymers in 0.1 M NaCl at pH 6.8. The phase separation 0.85 wt% of LM pectin, in water or in 0.1 M NaCl, and in
process was clearly evidenced by confocal laser scanning the presence or not of 3 mM CaCl2.
microscopy observations. Image analysis allowed us to
discriminate unambiguously the structures. We found 2.3. Dynamic oscillatory measurements
various structures and rheological properties depending on
biopolymer concentrations and on the presence or not of Time sweep oscillatory measurements were performed at
CaCl2 (3 mM) in the medium. On the same basis (Donato, a frequency of 1 rad sK1, for a strain amplitude of 1% using
Garnier, Novales, & Doublier, 2004b), we then explored the a controlled-strain rheometer (AR2000, TA Instruments)
influence of different concentrations of NaCl in presence or equipped with a Peltier temperature controller and with a
not of calcium ions. The presence of NaC in the medium cone-plane device (40 mm diameter, 48 angle). Warm
and the NaC/Ca2C ratio was shown to be determining protein/LM pectin mixtures were poured at 50 8C on the
parameters of the mechanical and structural properties of rheometer. The temperature was increased from 50 to 80 8C
the mixtures. The present study was aimed at better at 6 8C/min, then was kept at 80 8C for 30 min and decreased
understanding the role of cations (NaC and/or Ca 2C) in from 80 to 20 8C at 10 8C/min. Temperature was then
the properties of globular protein/LM pectin mixtures maintained for 1 h at 20 8C. A frequency sweep test was
by comparing their effect with two globular proteins then performed. A strain sweep test was realised to check
(b-lactoglobulin or bovine serum albumin), known to that measurements had been done within the linearity limits
display different interactions with these cations. Conditions of the viscoelastic behaviour.
of this study were close to those of the previous
investigations. The rheological and structural properties of 2.4. CLSM observations
mixtures were studied in water and in 0.1 M NaCl, in
presence or not of 3 mM CaCl2 at pH 6.8 and compared to CLSM was used in the fluorescence mode. Observa-
the properties of protein only. tions were made with a Carl Zeiss LSM 410 Axiovert
 L. Donato et al. / Food Hydrocolloids 19 (2005) 549–556 551

(Le Pecq, France) with a laser at a wavelength of 543 nm. close to this optimum. It has been suggested that specific
As proteins do not exhibit intrinsic fluorescence at this ionic interactions can occur between calcium and native or
wavelength, proteins were stained by adding rhodamine B pre-heated b-Lg by the formation of intramolecular ion
isothyocyanate (RITC) to the protein solution under bridges between charged groups of b-Lg and Ca2C
magnetic stirring during 1 h. Mixtures of RITC–protein/LM (Matsudomi et al., 1991). Jeyarajah and Allen (1994)
pectin were prepared as described above and poured showed that this interaction induced small structural
between a concave slide and a coverslip, then hermetically changes in the protein that led to increased hydrophobicity.
sealed. The same heat treatment as for rheological The present results, with a dramatic increase in gel strength,
measurements was applied using a thermostated stage are consistent with these descriptions.
(Linkam PE 60). It was verified that labelling did not Addition of 0.85 wt% LM pectin to b-Lg in absence of
change the rheological behaviour of the systems (results not calcium did not change the shape of the G 0 traces. However,
shown). a weaker gel (G 0 Z310 Pa) than for protein only was
yielded. For the mixture with calcium, G 0 was lower during
heat treatment and a larger increase occurred during the
3. Results and discussion cooling step resulting in a stronger gel (G 0 Z510 Pa).
Contribution of LM pectin gel to final gel strength could
3.1. Gelation of globular protein/LM pectin systems explain this result. However, the mechanical spectra (not
in water shown) obtained for the mixtures in presence or not of
calcium ions displayed a pattern similar to that of proteins
3.1.1. b-Lg based systems alone with a slight G 0 (u) and G 00 (u) dependence suggesting
Variations of the storage modulus (G 0 ) as a function of the predominant role of proteins in the viscoelastic behaviour
time for 8 wt% b-Lg and for mixtures in water, in presence (tan dw0.09 for uZ1$ rad. sK1 for mixtures in presence of
or not of calcium ions are shown in Fig. 1. For protein only calcium and w0.11 for protein only in both cases).
in water, G 0 increased strongly during the plateau at 80 8C The corresponding microstructures observed by CLSM
and a smoother increase occurred during the cooling step. are shown in Fig. 2. For protein only in presence or not of
After 1 h at 20 8C, G 0 was nearly constant (G 0 Z2900 Pa) calcium ions, the fluorescence was regularly distributed in
and the system was stable, a strong gel being formed. Same the medium. As gelation has been shown by rheology, it
evolution of G 0 was obtained in presence of calcium ions but could be concluded that the protein network cannot be seen
G 0 was four times higher (G 0 Z11750 Pa) resulting in a at this scale of observation. For the mixtures, inhomogene-
stronger gel. From the literature, it has been shown that ities in the distribution of fluorescence were evidenced by
there is an optimum Ca2C content until the gel is
strengthened. Matsudomi et al. (1991) showed that for 5%
b-Lg gel at pH 8, this optimum in CaCl2 concentration is
between 2 and 5 mM whereas Mulvihill and Kinsella (1988)
found this optimum at 10 mM CaCl2 for 10% b-Lg gel at the
same pH. Therefore, we can suppose that our conditions are

Fig. 1. Variations of G 0 during the thermal process (dashed line: Fig. 2. CLSM micrographs of RITC–b-Lg (A) and RITC–b-Lg/LM pectin
temperature profile) in water for b-Lg (,) and b-Lg/LM pectin mixture mixtures (B) after the thermal treatment, without (first raw) and with
(B) without (empty symbols) and with (filled symbols) 3 mM CaCl2 in (second raw) 3 mM CaCl2 in water at pH 6.8. Proteins appear in bright.
water at pH 6.8. Scale bar is 25 mm.
552 L. Donato et al. / Food Hydrocolloids 19 (2005) 549–556

the presence of fluorescent beads, more or less connected,

dispersed in a continuous dark phase. For systems without
calcium, the size of the beads was approximatively 5 mm.
Addition of calcium resulted in a less homogeneous
distribution of the beads, which appeared smaller
(w3 mm) and less connected. From double localization
experiments using LM pectin labelled with fluorescein
amine and protein labelled with RITC, we checked that the
protein was located in the beads while the polysaccharide
was in the continuous phase (results not shown). Of course,
these observations provide evidence that phase separation
has occurred in the mixed system. Therefore, this system
can be described as spherical particles of b-Lg embedded in
the pectin phase. Phase separation was so pronounced that
protein did not form any true network and it can be supposed
that phase separation occurred prior to protein gelation. This
type of structure was also observed for whey protein/HM
Fig. 3. Variations of G 0 during the thermal process (dashed line:
pectin mixed systems by Syrbe et al. (1998). Since the temperature profile) in water for BSA (,) and BSA/LM pectin mixture
protein beads can be regarded as colloidal particles, a (B) without (empty symbols) and with (filled symbols) 3 mM CaCl2 in
further depletion–flocculation mechanism of the b-Lg water at pH 6.8.
containing microspheres caused by the presence of the
LM pectin chains in the medium may have occurred, as it proposed to explain the influence of calcium ions on protein
has been described by Tuinier, Dhont, and de Kruif (2000) gelation. However, the BSA gel strength was lower than
in the case of aggregated whey protein colloidal particles b-Lg gel (G 0 w104 Pa).
mixed with an exocellular polysaccharide from a lactic acid Mastudomi et al. (1991) measured the optimum concen-
bacterium. tration at pH 8 for 10% BSA between 5 and 10 mM CaCl2.
From rheological properties and microscopic obser- In our conditions, we determined this optimum concen-
vations, we can suppose that in absence of calcium, tration at 4 mM CaCl2 (unpublished results). A specific
considering that the protein is the only gelling component, ionic affinity between BSA and calcium could take place, as
phase separation results in a decreased connectivity of b-Lg was reported by Powell-Baker and Saroff (1965). However,
network and a weaker gel than protein only is obtained. In it has been shown that calcium binding to BSA is not so
presence of calcium, both biopolymers can form a gel. From strong as to b-Lg, which displays a high affinity for calcium
rheological results, it is showed that a stronger gel is ions (Dufour, 2004). Since the reinforcement of protein
obtained for the mixture with calcium. This suggests that gel by addition of calcium is more noticeable for BSA than
LM pectins gel in the continuous phase, and thus contributes b-Lg gel and b-Lg gels were stronger than BSA gel, we can
to the final strength of the mixture. LM pectin gelation is suppose that interactions between proteins and calcium do
probably due to a concentrating phenomena of the not involve the same structural modification.
polysaccharide phase induced by phase separation process. Addition of 0.85 wt% LM pectin to BSA in absence and
Indeed, LM pectin only formed very weak gel in water in presence of calcium resulted in quite similar gelation
(result not shown) in presence of calcium. This is probably profiles that were comparable to that of BSA in presence of
due to a screening effect of calcium ions to prevent calcium. The only difference was in the G 0 values, the
electrostatic repulsions between pectin chains that occurred mixture without calcium being less rigid while in the
in absence of other salts in the medium prior to the presence of calcium the gel strength of the mixture was
formation of specific linkage between pectin chains slightly higher than for protein only. As shown for b-Lg
(Garnier, Axelos, & Thibault, 1993). based systems, same types of mechanical spectra were
measured for mixtures in presence or not of calcium ions
3.1.2. BSA-based systems than with protein only.
Variations of the storage modulus (G 0 ) as a function of The corresponding microstructures of BSA and BSA/LM
time for 8 wt% BSA and for the mixture are shown in Fig. 3. pectin systems are showed in Fig. 4. As for b-Lg gel, the
For BSA only, G 0 increased slightly during the thermal network was not visible at this scale of observations. For
process and an extremely weak gel was formed after 1 h at BSA/LM pectin mixtures in absence of calcium, inhomo-
20 8C (final G 0 !1 Pa). Addition of calcium to BSA resulted geneities in the distribution of fluorescence were observed.
in a rapid rise of G 0 during the plateau at 80 8C and a As for b-Lg mixtures, bright zones containing the protein
smoother increase during the cooling step. After 1 h at appeared with a spherical shape and a lower size (!1 mm)
20 8C, G 0 did not increase significantly and stayed at that was hardly visible because observations were probably
w3000 Pa. Same conclusion as for b-Lg gel can be made at the limit of scale observation. The size distribution
 L. Donato et al. / Food Hydrocolloids 19 (2005) 549–556 553

Fig. 5. Variations of G 0 during the thermal process (dashed line:

temperature profile) in 0.1 M NaCl at pH 6.8 for b-Lg (,) and b-Lg/LM
pectin mixture (B) without (empty symbols) and with (filled symbols)
3 mM CaCl2.

This reinforcement of the gel can be related to the decrease

Fig. 4. CLSM micrographs of RITC–BSA (A) and RITC–BSA/LM pectin of electrostatic repulsions between proteins by the screening
mixtures (B) at 20 8C after the thermal treatment, without (first raw) and
with (second raw) 3 mM CaCl2 in water at pH 6.8. Proteins appear in bright.
charges brought by NaCl at the surface of the protein
Scale bar is 25 mm. (Relkin, 1996). Addition of calcium in 0.1 M NaCl had an
opposite effect to that experienced in water and resulted in a
of the protein microspheres appeared more homogeneous weakening effect on b-Lg gel. This result is consistent with
compared to b-Lg/LM pectin. the findings of Foegeding, Kuhn, and Hardin (1992) where
The phase separation may result in an increase in protein same behaviour was observed in presence of both salts but
local concentration that can explain the strong reinforce- in different conditions (0.1 M NaCl, 20 mM CaCl2 at pH 7
ment of BSA gel induced by the presence of LM pectin. with 7% b-Lg).
Similar results have been shown by Beaulieu et al. (2001) on Addition of LM pectin in 0.1 M NaCl did not change the
8 wt% whey protein/1 wt% LM pectin gel at pH 6. overall G 0 profile but the gels were much weaker than for
When calcium was present in the mixture, the fluor- protein only. The presence of calcium in the mixed systems
escence was more regularly distributed in the medium and resulted in a similar gelation pattern. However, the G 0
the microstructure was almost similar to that of the BSA gel. increase during the step at 80 8C was slower although the
This feature is to be linked to the fact that the gelation final gel strength was of the same order of magnitude
profiles of the protein and of the mixture with calcium ions (102 Pa). By comparison with Fig. 1, addition of NaCl to the
are almost superimposed. These overall results suggest that mixture decreased gel strength in the presence or not of
calcium ions play the same role in both systems by inducing calcium ions.
BSA gelation and have more affinity for BSA than for LM The corresponding microscopic observations are pre-
pectin. Therefore, pectin gelation does not seem to influence sented in Fig. 6. For protein only in absence of calcium, the
the properties of the mixture whose gelation is mostly same structure as in water was observed, the gel network
governed by the protein, which was not the case with b-Lg being not visible in the picture. Addition of calcium in the
systems. system resulted in inhomogeneities in the fluorescence
distribution with bright small beads, which appeared
3.2. Gelation of globular protein/LM pectin systems in interconnected and dark zones devoid of proteins that
0.1 M NaCl could be assimilated as pores containing the solvent. A
decrease in connectivity of protein aggregates due to the
3.2.1. b-Lg based systems presence of calcium ions may explain the decrease of the gel
The kinetics of gel formation of b-Lg/LM pectin strength observed by rheology. It may be suggested that
mixtures in 0.1 M NaCl in the presence or not of CaCl2 addition of sodium to the system decreases the optimum salt
are illustrated in Fig. 5 and compared to that of b-Lg only. concentration needed to strengthen the protein gel and in
By comparison with Fig. 1, the G 0 profile for protein only this case addition of both salts to protein enhances
in 0.1 M NaCl was similar to that in water, but at the coagulation rather than gelation (Mulvihill & Kinsella,
end a stronger gels was obtained (G 0 Z3.5!104 Pa). 1988; Matsudomi et al., 1991).
554 L. Donato et al. / Food Hydrocolloids 19 (2005) 549–556

Fig. 7. Variations of G 0 during the thermal process (dashed line:

temperature profile) for BSA (,) and BSA/LM pectin mixture (B) in
0.1 M NaCl at pH 6.8 without (empty symbols) and with (filled symbols)
3 mM CaCl2 in 0.1 M NaCl at pH 6.8.

the G 0 profile was the same as for b-Lg gel but a weaker gel
Fig. 6. CLSM micrographs of RITC–b-Lg (A) and RITC–b-Lg/LM pectin
mixtures (B) at 20 8C after the thermal treatment, without (first raw) and was obtained (G 0 w5!103 Pa instead of w3.5!104 Pa for
with (second raw) 3 mM CaCl2 in 0.1 M NaCl at pH 6.8. Proteins appear in b-Lg). By comparison with Fig. 3, the presence of NaCl
bright. Scale bar is 25 mm. reinforced the BSA gel as it was observed for the b-Lg gel.
Addition of calcium had a slighter strengthening effect than
For the mixed systems in 0.1 M NaCl, fluorescent in water, in opposition with b-Lg. This may suggest that the
particles were dispersed in a continuous matrix (dark total salt concentration in BSA systems is still below the
zones). Again phase separation took place with protein
optimum salt concentration after which the gel is weakened.
microspheres (in bright) dispersed in a LM pectin medium
The G 0 profile for the mixture was similar to the one in
(dark zones). By comparison with Fig. 2B, the presence of
water (Fig. 3) but a weaker gel was obtained in presence of
NaCl in the system seems to decrease the size of protein
NaCl. In presence of calcium, the kinetics was different:
beads (w!2 mm). Addition of calcium to the mixed systems
during the step at 80 8C, G 0 increased slowly but steadily
had the same effect on the structural properties as in water.
and a sharp G 0 increase occurred only during the cooling
The beads tended to aggregate yielding larger dark zones.
step below 60 8C, this being followed by a levelling at
These observations can be related to the rheological
20 8C. Final gel strength (G 0 w400 Pa) was close to that of
properties of the systems particularly from the mechanical
spectra obtained with the final gel. In absence of calcium, the systems without calcium (G 0 w300 Pa) but as already
the viscoelastic behaviour (mechanical spectrum) of the noticed for b-Lg based systems the mechanical spectra were
mixture was similar to that of the protein gel only whereas in different. In these conditions, the gels obtained for the
presence of calcium, the final structure seemed to result mixtures were weaker than for protein gel only but these
from the superposition of LM pectin and protein gels. BSA-based systems were stronger than corresponding b-Lg
Therefore, even if final gel strengths of the mixtures with based systems.
and without calcium are similar, gelation of the system As illustrated in Fig. 8A, similarly to b-Lg gel, the BSA
involved different mechanisms. Without calcium ions network was not visible in 0.1 M NaCl and contrary to b-Lg
added, a protein network is formed with a low connectivity gel, addition of calcium did not clearly change the structure
of protein aggregates likely resulting from a depletion– of the network. A coarsening occurred, which is hardly
flocculation mechanism due to LM pectin chains. When visible here but was identified by image analysis (Donato
calcium is present in the mixture, a mixed gel is formed with et al., 2004a). This may explain the slight increase in G 0
a LM pectin continuous network (evidenced from rheology) showed in Fig. 7.
reinforced by the presence of protein aggregates (visible by The microscopic observations of BSA/LM pectin gels
CLSM observations). (Fig. 8B) were quite similar to the previous one. Without
calcium, the protein beads seemed larger (w!2 mm)
3.2.2. BSA-based systems than in water (w!1 mm). Addition of calcium resulted
The variations of G 0 for BSA and BSA/LM pectin gels in in a decrease in the apparent size of the protein beads
presence or not of CaCl2 are shown in Fig. 7. For BSA only, and the system became slightly more homogeneous.
 L. Donato et al. / Food Hydrocolloids 19 (2005) 549–556 555

different in water, depending on the nature of the protein,

but similar behaviours were obtained in 0.1 M NaCl for b-
Lg and BSA mixed gels. Addition of calcium to the
mixtures induced pectin gelation, which was favoured in
the presence of sodium, the pectin network contributing to
the formation of a mixed gel. A balance between pectin and
protein gelation appears to govern the gelation of the system
depending on solvent conditions and biopolymers/ions
interactions.

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