Food Hydrocolloids Vol. 9 no. 4 pp.

317-332, 1995

Some physico-chemical aspects of protein processing in foods.

Multicomponent gels*

V.B. Tolstoguzov

NESTEC Ltd Research Centre, Vers-Chez-Les-Blanc, PO Box 44, CH-lOOO Lausanne 26, Switzerland

Abstract
The food industry produces an increasing variety of formulated foods. Processed food systems
usually contain proteins and polysaccharides performing both nutritional and structural functions.
Interactions between these macromolecular substances may result both in their thermodynamic
incompatibility and in complexing, thereby affecting the physico-chemical properties and structure
of many foods. Elucidation of these interactions is of particular importance for understanding
synergistic and antagonistic effects offood additives. This paper reviews the thermodynamic origin
of food hydrocolloid functionality.

Introduction
The main objective of this review is to discuss the
composition-structure-property relationships of food
hydrocolloid mixtures. The field is of special interest
because of both the increasing commercial importance of
hydrocolloid mixtures as functional food additives and their
constantly expanding range of applications.
Attention will be focused on three subjects:
(i) incompatibility between different hydrocolloid species
in aqueous media,
(ii) structure-property relationships of mixed gels, and
(iii) some features of hydrocolloid behaviour in multi-
component gels.
Technology for the production of a caviar analogue will be
given as an example of use of multicomponent gels.
The discussion will start with a brief survey of the most
important events in this field.
Thermodynamic incompatibility of food macromolecular
components
Incompatibility of biopolymers
The pioneers in the systematic investigation of phase
separation in biopolymer solution mixtures were:
(i) H.G.Bungenberg de Jong who studied mixed solutions
of gelatin and gum arabic (1-4) and
(ii) P. -0. Albertsson and H. Walter who investigated the
phase behaviour of aqueous mixed solutions of poly-
saccharides with synthetic polymers and proposed the
"Presented as part of the conference entitled 'Food Hydrocolloids, Ohio
94', Septemher 6-10, 1994.
first industrially important applications for two-phase
polymer aqueous systems in biotechnology (5,6).
Bungenberg de Jong was the first to show that on mixing
aqueous solutions of gelatin and gum arabic two different
types of phase behaviour can occur. He studied both and
gave them the names 'simple' and 'complex coacervations'.
Figure 1 shows some of his results. These phase diagrams
characterise phase equilibria under conditions of both
complex (Figure 1a) and simple coacervations (Figure
lb),
At pHs below gelatin's isoelectric point (IEP) and at a
low ionic strength (Figure la) the interaction of the
positively charged macro-ions of gelatin and the negatively
charged macro-ions of gum arabic can give rise to insoluble
electrostatic complexes. In this case both biopolymer
components are concentrated in the single complex co-
acervate phase.
Simple coacervation (Figure Ib) or thermodynamic
incompatibility takes place in solutions containing mixtures
of gelatin and gum arabic with a sufficiently high ionic
strength and at pH values above the IEP of gelatin. Under
these conditions biopolymer macro-ions have like net
charges. The mixed solution of biopolymers breaks down
into two liquid phases and each of the biopolymers
concentrates into one of the phases (1,4).
The phase diagram for a ternary system may be pre-
sented in the form of triangular co-ordinates. Figure lc
shows such a typical phase diagram for a ternary polymer-
I-polymer-2-solvent system, Each apex of the equilateral
triangle represents a pure component of the ternary system.
Each side of the triangle corresponds to a mixture of the

© Oxford University Press

318 V. B. Tolstoguzov
a b
water water
- 10
20
- 30
- 40
40 60 80
gum arabic
Figure 1 Phase diagrams for simple and complex coacervates for the gelatin-gum arabic-water system at the pH: (a) below and (b)
above the isoclectric point of gelatin [schematic representation, from H.G .Bungenberg de Jong (1952) In Kruyt,H.R . (cd .), Colloid
Science. Elsevier, New York, pp. 355-364) : (c) Generalities of triangular phase diagrams for a ternary biopolymer-I-biopolymer-2-
water system .
two components assigned to the two apexes. The three
sides refer to the three binary systems, namely: A-B , B-C
and C-A . The region of the triangle represents mixtures of
all three components. For instance, the mixture of three
components A, Band C corresponds to point D. In this
case the segments a, band c represent the proportion of
constituents in the mixture D. It should be noted that the
sum of the distances (a + b+c) to point D measured parallel
to each of the three sides is equal to the length of the side of
the equilateral triangle. Points along the line, e.g. a, which
is parallel to one of the three sides, A-C, represents
systems with a constant content of the component B. Any
point on a line going from an apex towards the opposite
side represents systems with a constant ratio of the two
other components (Figure la). Figure lc displays a polymer
pair (A and C) that is incompatible in the absence of a
solvent (B). This case seems to be representative of most
low moisture food systems (7-9).
The water content in the majority of food systems is
greater than the content of macromolecular compounds.
Therefore, rectangular co-ordinates are most often used to
describe the phase equilibrium in mixed biopolymer sol-
utions. This is similar to the system for reporting food
composition where only the quantities of the main con-
stituents are indicated, the rest is assumed to be water.
Investigation of protein-polysaccharide incompatibility,
complexing and functionality of complexes from the view
point of the food industry started about two decades ago
(10-14) .
Before this time , most studies on polymer incompatibility
were related to synthetic linear polymers. Both types of
phase separation in biopolymer solution mixtures had only
been studied for systems containing gelatin. However,
gelatin is not a typical protein. It has a higher excluded
volume than globular proteins. In aqueous solutions gelatin
behaves more like a polysaccharide or a synthetic polymer
than a protein (14).
Often, food systems contain native or denatured globular
c
Component B (e .g . water)
Component A 20 40 60 80 C Component
(e.g. Protein) (e.g. Polysaccharide)
proteins that have a compact molecular structure. Unlike
typical polymers, a native globular protein usually goes
over from a dilute to a semi-dilute solution within the
concentration region from 10 to 30% . Many of these
globular proteins are soluble in a limited range of pH values
only. When work started about 20 years ago it was not clear
whether mixed aqueous solutions of globular proteins and
mixed solutions of globular proteins and polysaccharides
could undergo phase separation . In other words the data on
thermodynamic incompatibility of gelatin with some poly-
saccharides were insufficient to gain insight into the phase
state of food systems and the behaviour of macromolecular
components in foods (10,12-14).
Systematic investigation on globulin-salt-water systems
began at Unilever Research about 25 years ago. The first,
original and very efficient application of these systems in
the food industry for producing protein fibres was devel-
oped by M.Tombs (15-18).
At the same time systematic investigations on complex-
ing and thermodynamic incompatibility in protein-poly-
saccharide-water systems containing the main classes of
food proteins and polysaccharides started at the laboratory
of novel food forms of the USSR Academy of Sciences (19-
24) . It was shown that incompatibility of macromolecular
components is typical for food systems and is of great
importance for structure formation and food properties
(10,12,13).
The first information about incompatibility in protein-l-
protein-2-water systems containing different proteins was
obtained during the last decade at the laboratory of novel
food forms of the USSR Academy of Sciences (25-28).
Two other events have been of special importance in this
field . The amylose-amylopectin-water system was the first
system of great significance for the food industry in which
the incompatibility of biopolymers was discovered and
studied. This research was performed at the Food Research
Institute in Norwich (29,30). The first large scale processing
of aqueous two-phase systems in the food industry was the
 Hydrocolloid interactions in mixed gels
production of low-fat spreads on the basis of a two-phase
gelatin-maltodextrin-water system. This process has been
recently developed by Unilever Research (31-35).
Now we will discuss some aspects of the phase behaviour
of mixed solutions of hydrocolloids which are of key
significance.
Phase diagrams
Normally, mixed solutions of chemically and/or structurally
different biopolymers are unstable. Figure 2 shows a typical
phase diagram for biopolymer-1-biopolymer-2-water
systems in rectangular co-ordinates. It exemplifies the
positions of binodal curve (EFGD), critical point (G),
phase separation threshold (F), tie-line (EAHD) and
rectilinear diameter (GH). At sufficiently high bulk con-
centrations of biopolymers, i.e. inside the binodal, a
mixture A, of homogeneous aqueous solutions Band C
converts into two new co-existing equilibrium phases (D
and E). These new phases may form water-in-water
emulsions. The tie-line connects binodal points (D and E)
representing the compositions of co-existing phases. Emul-
sion phases D and E can be separated by sedimentation and
analysed for protein and polysaccharide contents. The
results of the analysis of the phases are usually presented in
the form of a phase diagram as shown in Figure 2. This
diagram demonstrates that the initial ternary solution A
was obtained by mixing two binary solutions of composition
Band C in the volume ratio equal to the ratio of segments:
AB/AC. The unstable mixture A breaks down into two
B point, by the angle made by a tie-line with the concen-
,,
,,
,,
,,
,,
, Membraneless osmosis
\
,,
E
,, TIe-Une
\
,,
\ BINODAL
c of different concentrations: simple diffusion, osmosis and
Protein ,% wt.
Figure 2 Typical phase diagram for protein-polysaccharide-
water systems.
319
phases D and E. Binodal branches (GE and GD) represent
the compositions of these co-existing phases. The ratio (E/
D) of the co-existing phase volumes corresponds to the
ratio of segments AD/AE. When point A, representing the
system composition, is shifted along tie-line DE towards
either point D or point E, the volume ratio of co-existing
phase approaches either zero or a unit value at the binodal.
The dashed-line passing through the mid-tie-lines, e.g. at
point H, is called the rectilinear diameter. The rectilinear
diameter gives the composition of systems splitting into
phases of the equal volume. This line passes through the
critical point. The position of system critical point (G) can
be obtained as the point of intersection of the rectilinear
diameter with the binodal curve. The phase separation
threshold (F) is the minimal total concentration of biopoly-
mers (i.e. the sum of co-ordinates of point F) required for
phase separation. The point of contact between the binodal
curve and the straight line cutting segments of the same
length on both concentration axes gives the position of the
phase separation threshold.
There are significant differences in the compatibility
between various pairs of biopolymers. The lower the values
of both either critical point or the phase separation
threshold, the lower is the compatibility. The phase
separation threshold for biopolymer mixtures is in a
concentration region corresponding to the transition from a
dilute to a semi-dilute solution. This is the concentration
range where biopolymers start to overlap. The phase
separation threshold often decreases with increasing mol-
ecular weight of hydrocolloids (36-40).
An asymmetry of phase diagrams reflects the com-
petition between biopolymers for the solvent, water, and is
due to differences in hydrophilicity or in molecular weight.
The relative hydrophilicity of a biopolymer may be charac-
terised by the biopolymer concentration ratio at the critical
tration axis of one of the system components, or by the
length of the binodal curve segment (FG) between the
critical point and the phase separation threshold
(12,13,39,40).
Phase separation is accompanied by water transfer from an
initial aqueous solution to the solution of another biopoly-
mer. Water redistribution occurs until equilibrium is
established between the coexisting aqueous phases. This
phenomenon underlies the new process for concentrating
protein solutions that we call membraneless osmosis
(12,13,40,41). This process is of importance for our
considerations because it determines water partition
between food system phases. Figure 3 illustrates the three
types of processes for water transfer between two solutions
membraneless osmosis.
Simple diffusion takes place when a permeable mem-
brane separates two solutions of different concentrations.
Owing to thermal movement and membrane permeability
320 V.B. Tolstoguzov
Permeable Membrane
-)
Semipermeable Membrane
-)
Protein
and Polysaccharide
Solutions
Centrifugation
-)
Two-phase Tvvo Layers
System
Figure 3 Membraneless osmosis process.
for both the solvent and solutes a balancing of the solution
concentrations on both sides of the membrane occurs.
Conventional osmosis involves the passage of a solvent
through a semi-permeable membrane separating a solvent
and a solution, or two solutions of different concentrations.
There is the same tendency for concentrations of solutions
separated by a semi-permeable membrane to equalise.
However, a semi-permeable membrane is one through
which the solvent molecules can pass but macromolecules
cannot. The movement of the solvent, water, is unilateral in
this process.
Membraneless osmosis uses two immiscible aqueous
solutions of biopolymers. Since the biopolymers are not
miscible the interfacial surface separating them replaces the
semi-permeable membrane which is no longer necessary.
Thus, in membraneless osmosis we are dealing with the
diffusion of water through the interface between the two
immiscible solutions of hydrocolloids. We will now look at
gelation of hydrocolloid mixtures.
Mnlticomponent gels
Structure of multicomponent gels
Gels formed using more than one gelling agent can be
Simple
Diffusion
•
•
•
Osmosis
Membraneless
Osmosis
subdivided into three groups: complex, mixed and filled
gels (9-14,36-39,42-45). Figure 4 shows graphically the
compositions of mixed and filled gels.
Two characteristics of mixed solutions of biopolymers are
of great importance. These are the cosolubility limits (i.e.
the position of the binodal) and the critical concentrations
for gelation. Both quantities determine the boundary
conditions for the formation of mixed and filled gels.
A mixed gel has two or more relatively independent
three-dimensional networks. Mixed gels are formed when
the concentrations of biopolymers in a solution exceed the
critical concentration for gelation of these biopolymers.
When the bulk concentration (or cosolubility) of a biopoly-
mer in one of the phases is larger than that required for
gelation, the biopolymer forms a gel in that phase. In the
two-phase area, inside the binodal a gel filled with gel-like
or liquid dispersed particles is obtained. Filled gels may also
be formed at these concentrations from mixtures of a
gelling agent with a non-gel forming biopolymer.
As shown in Figure 4 (thick lines) the critical concen-
tration for gelation (or the gel-point) does not usually
exceed 0.1-0.3% for anionic polysaccharides, while for
proteins this value varies over a wider range, e.g. from 1%
for gelatin to 13% for the US broad bean globulin.
 Hydrocolloid interactions in mixed gels
Polysaccharide, (Ps) % wt.
1.5
~IPs------+----------+
1 15
Figure 4 State diagram for hydrocolloid mixtures. C' gPr and C' gps- critical concentration of gelation for protein and polysaccharide.
The phase separation occurs at slightly higher concen-
trations. The phase separation threshold is -2-4% for
mixtures of gelatin with linear anionic polysaccharides and
> 12% for mixtures of globular proteins. This means that
combinations of gelling agents can lead to the formation of
mixed gels on both sides of the binodal curve. A com-
position A in the two-phase region, will separate into the
phases B (rich in protein) and C (rich in polysaccharide).
These phases may form different microstructures: B dis-
persed in a continuous C-phase and C dispersed in a
continuous B-phase. Both phases of water-in-water emul-
sions can form mixed gels.
Causes of synergistic and antagonistic effects in biopolymer
mixtures
Most hydrocolloids are best used in mixtures. The syn-
ergistic and antagonistic effects resulting from blending
biopolymers are of great applied significance for the
improvement of many foods , and also for reducing their
cost-price.
The main cause of synergistic effects seems to be
thermodynamic incompatibility . Several types of synergy
can be assumed to occur. They concern hydrocolloid
mixtures with compositions both below and above the
binodal curve .
Excluded volume effects in single-phase mixed biopolymer
solutions
The first type of synergy takes place in a single-phase
region of the phase diagram. The reason is that macro-
molecules cannot occupy the same volume in the solution.
This means that each biopolymer can only use some part of
321
Protein, (Pr) % wt.
the volume of the mixed solution. In solutions incompatible
biopolymers mutually concentrate each other. Thus, each
incompatible biopolymer will behave as if it was more
concentrated (9 ,11,13,14,36-39). Since the shear modulus
of a gel is usually proportional to the square of its
concentration this means that small additions of a hydro-
colloid can increase the elastic modulus of a gel from a few
times to a several tens of times. Figure 5 shows that the
addition of a small amount of dextran to gelatin solutions
can increase several fold the elastic modulus of the gel.
Excluded volume effects also favour gelation of hydro-
colloids. For incompatible biopolymers in mixed solutions
the rate of gelation is higher and the critical concentration
for gelation is lower than for each of them individually
(Figure 5) . Excluded volume effects depend on the flexibil-
ity, shape and size of macromolecules as well as their bulk
concentration. It should also be stressed that because of
excluded volume effects , the range of compositions where
mixed gel formation occurs may be markedly larger than
that shown in Figure 4.
What is the cause of antagonistic effects of mixing
hydrocolloids at compositions below the binodal? There is
an apparent decrease in the concentration of gelling agent
due to the formation of electrostatic protein-anionic
polysaccharide or protein-protein complexes. Electrostatic
interbiopolymer complexes are noted for their compact
molecular structure and high critical concentration for
gelation. Presumably, an antagonistic effect results from
the formation of soluble and insoluble complexes that
cannot form an additional three-dimensional network
because of their low concentration.
322 V. B. Tolstogu zo v

Polysaccharide,%
Uquid Phase
Phase Diagram
4·
Gelatin, % wt
el-Iike Phase
4
Optical Rotation
10 20 30
Protein, %

Figure 6 Phase diagram for protein-polysaccharide-water

A system with gelation in the dispersed phase.

Gelation Rate
Days
other phase (Figures 2 and 6) (13,14,39,40,44,45) . This
produces two opposite effects.
1 2 3 4
The first is a strong (up to several fold) increase in the
Elastic modulus
concentration of one of the system phases. This increase
may be so substantial that the phase gels. Figure 6 shows
the case of casein gelation in the dispersed phase of the two-
20kD phase system obtained by adding gum arabic to skimmed
milk (41) . Tie-lines in the phase diagram converge into a
3OOkO
Dextran, "10 wt si.ngle point corresponding to the composition of a gel-like
d~spersed phase. Unlike the phase diagrams presented in
FIgures 2 and 4, where solvent equilibrium is reached in the
Figure 5 Effect of small additions of dextran on the rate of
coexisting phases , in this case osmosis is stopped by the
gelation and elastic properties of gelatin gel.
swollen gel applying a backpressure to the solution.
Concentration of the continuous phase of a water-in-
water emulsion can lead to synergistic effects . It should be
Interbiopolymer complexing and incompatibility are
noted , howe ver , that since food systems are not usual1y
mutually connected phenomena. They are two opposite
transparent the heterophase nature of aqueous food
consequences of non-specific interactions between bio-
systems is often not recognised and taken into account
poly~ers in aqueous media. Normal1y, conditions resulting
wh~n. their composition-property relationship is discussed .
In biopolymer immiscibility should inhibit the affinity
ThIS IS the case of the transition from homogeneous system
between dissimilar biopolymers. As a result of unfavour-
A to heterogeneous system AI with a continuous phase B 1
able interactions between segments of dissimilar biopoly-
produced by the addition of small amounts of gelling agent
mers each macromolecule shows a preference for a sur-
X. The phase diagram (Figure 7) shows that a significant
rounding of its own type . However under conditions of
(several-fold) increase in the concentration of gelling agents
compatibility dissimilar biopolymers may form soluble
within the continuous phase (B 1 compared A) of the system
complexes. The surface active complexes formed can
can cause a large increase in the elastic properties of the
conce~t~ate at the inte.rfaces at concentrations exceeding
gel. Because man y physico-chemical properties (e .g. rheo-
the critical concentration for gelation . Therefore these
logical) of disperse systems are determined by those of their
complexes affect the surface properties rather than the
continuous phase (7,8,39) and becau se the shear modulus
volume properties of gels. This phenomenon is illustrated
of a gel is proportional to the square of hydrocol1oid
in Figure 13 which shows the effect of smal1 additions of an
conc~ntration , an increase in the concentration of gel
anionic polysaccharide , pectin , on the surface properties of
continuous phase is of gre at importance for mechanical
gelatin gels.
proper~ies of gels. This is especially typical of heterophase
It should be emphasised that the importance of the
gels. WIth a continouus phase containing the 'strongest'
excluded volume effect leading to a synergistic gel is jam at
gelling agents , e .g. Y. According to Figure 7 hydrocolloid
compositions above the binodal where both phases are
Y with a lower critical concentration for gelation is a
saturated mixed solutions. However, this effect loses its
'stro nger' gelling agent than X. Differences in the mechan-
leading role when phase separation occurs .
ic~l pr?perties ~f g~l~ of different gelling agents depends
Concentration of the continuous phase of water-in-water pnmanly on their critical concentration for gelation since
emulsions as we ~lready mentioned, the shear modulus of ; gel i~
We mentioned above that phase separation is accompanied proportional to the square of hydrocolloid concentration .
by water redistribution between system phases. This can If water redistribution results in concentration of one
~ead to a large increase in the concentration of biopolymers phase the other phase will be correspondingly diluted. This
In one of the phases and a corresponding dilution in the results in a decrease in the modulus of elasticity of the gel.
 Hydrocolloid interactions in mixed gels 323

Hydrocolloid X, %
A

* Hydrocolloid Y,%
C9y

Elastic Modulus
Trends

Slngl.Phase 1
ex luded
ev ume .
Two-Phase
1: ~ontinuou~shase
Concentration
~ 7
Phase Inversion

: Liquid
: Filler

A H
Concentration of X, %

Figure 7 Possible mechanisms of synergistic effects. C*sx and C*gy-critical concentration of gelation for hydrocolloids X and Y.

This second and opposite effect of phase separation is due
to an increase in the dilution and the volume fraction of the
dispersed phase acting as a filler of the 'gel. The elasticity
modulus of gels filled with dispersed particles decreases as
the volume fraction of filler increases (Figure 5). Because
of incompatibility this is the case of low adhesion between
the filler and the gel matrix.
Figure 7 also shows possible mechanisms of synergistic
effects for two-phase systems. Minimal volume fraction of
the dispersed phase and a sufficiently large increase in the
concentration of the continuous phase seem to be import-
ant factors in synergistic effects. To successfully compete
for water from the continuous phase a second biopolymer
added to a stronger gelling agent should be of low
compatibility and should be relatively more- hydrophilic.
This means that the critical point co-ordinates must be as
low and as asymmetric as possible. Thus, for a synergistic
effect to occur when the phase separated system A I formed
by addition (A]-A) of a biopolymer X, gels the following
three favouring conditions must be united: first, the
continuous phase B] is strongly concentrated during phases
separation. Second, the continuous phase of this system
contains 'stronger' (C* g Y) gelling agent. Third, this
'stronger' gelling agent is also converted into the gel state as
the first. The last condition will be discussed later.
Phase inversion in water-in-water emulsions
Near the rectilinear diameter, where both phases of the
water-in-water emulsion have the same volume, inversion
of the phases can take place. Phase viscosities can strongly
affect the phase inversion. For instance, the addition of the
same biopolymer X may produce an 'antagonistic effect'
when we go over to the heterogeneous system A z with
continuous phase C z is formed. Continuous phase C z is
diluted and contains the 'weaker' gelling agent X or a non
gel-forming biopolymer. On the other hand, synergy may
be induced by an additive which is able to form insoluble
complexes with the biopolymer X thus returning system A z
to A, or even to A.
Other factors
Two other causes of synergy are the fractionation of
biopolymers and the partition of metal ions between
coexisting phases.
Phase separation in mixed biopolymer solutions is usually
324 V.B. Tolstoguzov

accompanied by the fractionation of biopolymers, e.g. Temperature, °c

polysaccharides. Polydispersity of polysaccharides in their
molecular weight, structure and chemical composition is of 70
great significance, due to effect of these characteristics on
cosolubility of hydrocolloids and their gelation. As an
50
example Figure 5 shows the effect of molecular weight on
the cosolubility of dextran with gelatin.
Many anionic polysaccharides are capable of forming gels 30
in the presence of certain metal ions; therefore the partition
of ions between the system phases is another important
-4-
factor influencing gel properties. Boundary conditions of Elasticity Modulus, E. 1 a Pa
gelation and gel properties are both dependent on the
nature and concentration of the anionic polysaccharide and
the ions used (11,46).
The binodalcurve usually lies closer to the concentration
axis of the lower molecular weight biopolymer. This means
that the higher molecular weight biopolymer expels the
lower molecular weight biopolymer from the solution. At
the phase separation threshold the system is therefore
enriched in the lower molecular weight component (20,40).
Generally, association between protein molecules in- 2
Gelatin
6

tensifies with concentration and leads to a decrease in

relative hydrophilicity of a given protein. This is the reason
for changes in the angle made by tie-lines with the
concentration axis of a protein when protein concentration
increases. This is a specific feature of phase diagrams for
many protein-polysaccharide-water and protein-protein-
water systems (37-40). Since these effects result in the
formation of ordered protein structures when protein-
protein interactions occur, the phase diagram asymmetry
can be higher for protein-l-protein-2-water systems than
for protein-polysaccharide-water systems.
Presumably, globular proteins with greatly differing
molecular weights can have higher cosolubility especially
proteins belonging to the same class within the Osborne
classification. For instance, the small size of molecules of
low molecular weight proteins, such as trypsin inhibitor,
seems to be necessary for an increase in their compatibility
with other proteins.
Gelatin-agarose mixed gels
Figure 8 shows experimental data (11,39,44,45) which
support suppositions made in the preceding sections. It
demonstrates the effect of changes in composition of a
gelatin-agarose mixed gel on its melting temperature and
elasticity modulus.
Addition of small amounts of agarose increase the
elasticity modulus of the gelatin gel due to the mutual
concentration effect of hydrocolloids (i.e. excluded volume
effects). At higher agarose concentrations the mixed
solution separates into two phases. Phase separation results
in particles of the incompatible polymer, agarose, dispersed
in the gelatin gel network which disrupts the three-
dimensional -gel network. However, this increase in the
concentration of agarose also increases the elasticity
modulus of the filled gelatin gel. Presumably, due to the
competitive effect of gelatin phase concentration (water
0.6 0.2
Agarose, % vvt
Figure 8 Composition dependence of melting point and the
modulus of elasticity of mixed gelatin-agarose gels.
redistribution) comes into play: the volume fraction of the
dispersed agarose phase and the concentration of the
continuous gelatin phase increase simultaneously. Up to a
certain degree super concentration of the gelatin network
can localise around the dispersed agarose particles and
contribute to the elasticity modulus of the gel and com-
pensate for the disruptive effect of the gel by the dispersed
particles. However, a further increase in volume fraction of
the dispersed phase (further addition of agarose) cannot be
compensated for, and gel elasticity decreases. To sum up,
volume fraction, size and size distribution of dispersed
phase particles as well as locally (around dispersed par-
ticles) increased concentration of the gelatin network,
contribute to the mechanical properties of gels. Com-
parison of the two curves in Figure 8 shows that the gel
melting point is strongly changed in the region of phase
inversion composition and that a maximum gel weakening
effect occurs at phase inversion, i.e. where the volume
fraction of the dispersed phase is maximal.
To summarise, the main factors contributing to the
formation of a 'synergistic gel' are the following. First, a
strong increase in the effective concentration of gelling
agents in single-phase mixed solutions. This is due to the
excluded volume effects of macromolecules of incompatible
 Hydrocolloid interactions in mixed gels
biopolymers. Second, a strong increase in the concentration
of gelling agents in the continuous phase resulting from
phase separation. This is due to the redistribution of water
and metal ions between the phases of water-in-water
emulsions. Third, the fractionation of polysaccharides and
the partition of metal ions between the phases. These are
the two principle factors which determine boundary con-
ditions in polysaccharide gelation and mechanical prop-
erties of gels. An antagonistic effect may also result from
phase inversion in a water-in-water emulsion , i.e. due to a
sudden strong change in the composition of the continuous
phase . The continuous (or matrix) phase of a filled gel is
primarily responsible for its mechanical properties.
One can see (Figure 8) that the gel melting temperature
changes from one virtually constant level to another. The
gel melting temperature depends mainly on the nature of
the system continuous phase, is weakly dependent on phase
composition and almost independent of phase volume
ratio. Within the intermediate range of gel compositions
the melting temperatures of gel vary proportionally with gel
composition . It is also characteristic for the range of
composition dependence of the elasticity modulus of
multicomponent gels corresponding to a wide minimum . In
both cases we are dealing with mechanical properties of gel
since gel melting temperature depends not only on the
thermodynamic parameter of breakdown of network cross-
links but also on temperature dependence of rheological
properties of a system under given conditions. Therefore, it
is also a characteristic feature of multicomponent gels that
both gel melting temperature and elasticity modulus are
linearly dependent on gel composition in a large inter-
mediate area. This large intermediate area has been
interpreted as corresponding to phase inversion (39,44,45).
It is noteworthy that the composition area corresponding
to phase inversion is quite extended. This relates to both
the range of gel melting temperature and the elasticity
modulus of multicomponent gels. Similar results were
obtained earlier when physico-chemical properties of
protein-polysaccharide two-phase systems, e.g. such as the
spinnability and mechanical properties of liquid and gel-like
protein-polysaccharide fibres as well as products of the
thermoplastic extrusion of protein-starch mixed melts were
studied (7-9,12,37-39). Within this large intermediate
range of gel compositions we are presumably dealing with
systems having two continuous phases which obey the
additivity low for rheological and other physico-chemical
properties. Such structural features, i.e. the presence of
two continuous phases within quite a large range of
compositions in protein-polysaccharide-water systems,
seem to be typical of water-in-water emulsions because of
low interfacial tension and high viscosity of coexisting
phases. This opens the way for controlling the structural
functions of hydrocolloids and functional properties of food
systems . For instance, it may be assumed that this feature
of water-in-water emulsions as well as the specific surface
properties of gelatin gels discussed below (Figure 13)
provide the basis for the formation of the continuous lipid
325
phase and the honeycomb-like structure of low fat spreads
(8,37,38).
The symmetry of the curve in Figure 8 reflects the fact
that we are dealing with relationships of a general nature.
The two incompatible gelling agents under investigation are
a protein and a polysaccharide, i.e. biopolymers of differ-
ent chemical nature. Nevertheless, the effect of their
addition to solutions of each other on the elastic modulus of
multicomponent gels is the same. An additional peak on
the right side of the curve (in Figure 8) corresponds to the
difference in the affinity for water between the
biopolymers.
Because biopolymer incompatiblity is of a general
nature, synergy in multicomponent gels is a widespread
phenomenon. This does not only relate to mixtures of
gelling agents, various combinations of gelling agents with
biopolymers which do not form gels also have synergistic
effects.
Now, we will dwell on the technology of a caviar
analogue in order to consider some other specific features
of multicomponent gels.
Granular caviar analogue
Basic ideas of the process
Caviar analogue is a granular food with taste and visual
appearance similar to the natural caviar of fish. Gelatin is
the main gelling agent, because it is easy to form granules of
gelatin gel and to fill them with nutrients. The gelatin
granules containing casein are covered by two membranes.
The first membrane is composed of gelatin and vegetable
tanning agents. This membrane is coloured black by
complexes of tanning agents with iron ions . Interbiopoly-
mer complexes are used for obtaining the second external
membrane and for stabilising the emulsion binding of the
granules. An oil-in-water emulsion is used to simplify the
addition and uniform distribution of all additives, to
prevent the product from drying and to increase the
adhesion between gel-like granules providing spreadability
of the product (47-51).
Production of a caviar analogue is given as an example of
processing of multicomponent gels. The following factors
determined the choice of technology for its production.
First, all three main types of gel, namely mixed , complex
and filled were used to develop this product. Second, both
the volume and the surface properties of gels were of
importance for producing the granules required . It is of
interest that the same raw materials (such as casein, pectin ,
vegetable oil and calcium salt) have been used for preparing
two, unlike ingredients of the product , namely gel-like
granules and an oil-in-water emulsion. The product , called
'protein granular caviar' , is the first protein food analogue
that has been commercially viable in the last 20 years . It is
an example of a functional food product developed for local
food preferences. This product has been available to
consumers since 1972.
The work was initiated by Professor A.N.Nesmejanov in
326 V. B. Tolstoguzov

1964. The technique for producing caviar analogue took
one year to develop. Then, about two more years were
needed for the development of pilot lines. The first
commercial plant started up in Moscow in 1972. A few
years later some other plants (e.g. in Tallinn, Estonia with
five lines and in Novokuznezk, Siberia with two lines)
began to manufacture the product. Each line produces
about 25 kg/h. The process and equipment for making
caviar has been patented (52-57). Large scale industrial use
of this technology during more than 20 years demonstrates
its high reliability.
There were several reasons for the development of this
product. First, the production of natural sturgeon caviar
had been limited and was decreasing continuously.
Amounts of the analogue comparable to that of natural
caviar could be produced quite rapidly to meet the demand.
Second, the aim was to extend the nutritional use of casein
as a high quality and low cost animal protein by processing
into an analogue of a highly attractive and popular food
product. And finally, since natural surgeon or salmon
caviar is esteemed by consumers to be a food product with a
very high reputation, it was assumed that production of a
sucessful analogue would facilitate the introduction of
further analogues.
Technology of the product
Figure 9 shows the outline of the product technology. First,
a mixed aqueous solution containing 12-20% casein and 6-
8% gelatin is prepared. This mixed solution is stable. It is
introduced dropwise at 50-60°C into vegetable oil cooled to

~'t Gelatin + 15% Casein

.:... ~., Liquid 45 - 70°C
O~I ranules
•

Tea Extract

First Membrane
Formation

First
Membrane
Coloring
~~

Pectin solution

Emulsion

Caviar Analogue

Figure 9 Pilot equipment for producing granular caviar analogue.

 Hydrocolloid interactions in mixed gels
2-1O°C. The denser aqueous drops sediment in the oil.
Surface tension results in a spherical drop shape. Liquid
drops gel upon cooling.
Gel-like granules are formed within the U-shaped
column. The two columns of liquids of different density,
namely vegetable oil and water, are balanced. Oil tempera-
ture and viscosity are controlled to ensure the necessary
cooling rate and gelation time of the granules. Oil tempera-
ture is maintained by cooling water in the jacket. Then
water is injected into the column to separate the gel-like
granules from the oil and to transport them into a collecting
vessel.
The granules are then treated with a cooled aqueous tea
extract. Under the action of tea tannins an elastic mem-
brane is formed on the surface of the gelatin granules.
Tanning increases the density of the surface layer of the
gelatin gel and decreases the diffusion rate of the tanning
agent into the bulk of the gel. Therefore the tanning agent
is located in the surface layer of the granules. Tea extracts
are prepared using by-products of tea processing, i.e. low
cost raw materials such as crude tea leaves, tea dust, etc.
Tea extracts are notable for their P-vitamin content and
bactericidal activity. Dark grey colouring of the membrane
is achieved by treatment of the granules with a dilute iron
salt (e.g. ferric chloride or lactate) solution. Interaction
between trivalent iron ions and tea phenols colours the
surface film. Granules are then successively treated with
aqueous solutions of sodium alginate or low-ester pectin
and with a solution of calcium acetate. As a result, a second
membrane of heat-resistant casein-Ca-alginate or -pectin-
ate gel is formed. Treatment of the granules in aqueous
media, including washing, tanning and colouring, is carried
out at I-10°C. Cod liver oil, herring juice and sodium
glutamate are used as flavouring ingredients. Granules
covered by two membranes are mixed with an emulsion
containing common salt, ascorbic and sorbic acids and
vegetable oil as well as dispersed fish products and cod liver
oil. We turn now to some physico-chemical aspects of the
technology.
Model systems to develop the process
Model systems studied for the development of the technol-
ogy included mixed solutions and mixed, complex and filled
gelatin gels containing other proteins and polysaccharides.
These liquid and gel-like systems were investigated under
conditions of complexing and incompatibility (10,11,14,19-
24,48-51).
Volume properties of gelatin gels
Gelation and washing of protein granules with water are of
prime significance for all other process stages, i.e. both
bulk and surface properties of freshly prepared gelatin gels
are of importance for the technology. Both these oper-
ations have to be done as completely and as rapidly as
possible. These conditions are necessary, first to form a
uniform membrane by tanning and colouring of the
327
granules and second to minimise swelling of the granules
during washing.
The formation of a casein-filled gelatin gel is complicated
by adsorption of the oil. This results in a reduced melting
point, increased swelling and decreased elasticity of the gel.
Fresh gelatin gel containing casein swells rapidly in water.
The presence of casein accelerates the gelation of gelatin
solutions. The rapid formation of a three-dimensional
network is obviously the same excluded volume effect as
that of dextran shown in Figure 5. According to the patents
(52,53) polysaccharides such as starch, dextrin, agar-agar,
or pectin may be added to the gelatin solution to increase
gelation rate and to improve mechanical properties of the
granules. However, the presence of casein and other
biopolymers intensifies the swelling of gelatin gels. For this
reason the use of water acidified to pH 4-6 has been
recommended for washing the granules. This partially
coagulates the casein and thereby decreases the swelling of
the granules.
The membrane that is formed by the treatment of
granules with vegetable tanning extracts, sharply reduces
the rate of swelling of the granules in water. This
membrane is unable, however, to preserve the spherical
shape of a granule after melting of the gelatin gel. Unlike
tea tannins, trivalent iron ions can diffuse into the volume
of the gelatin gel and cross-link protein macromolecules.
This increases the melting point of the gel. Iron ions also
contribute to cross-linking of the second gel-like
membrane.
Complex and mixed gels
Figures 10 and 11 show that mixtures of the same
biopolymers, namely of casein with pectin or alginate, can
display completely different phase behaviour depending on
the pH.
Biopolymers can form soluble and insoluble complexes
and complex gels. Figure 10 shows thermomechanical
properties of a casein-alginate complex gel containing
7.5% casein and 2.5% alginate at pH 4.5. This gives
information about the relative deformation of a gel sub-
jected to periodic loading carried out at constant selected
values of loading, loading time and heating or cooling rate.
The deformability of this complex gel is not significantly
changed with temperature. It does not melt.
To prepare the gel, sodium caseinate and sodium alginate
solutions were mixed at pH of ~7. Then the pH of the
mixed solution was adjusted to 5.5. Since casein is
precipitated at pHs <5.8, the stability of its mixed solutions
at pH 5.5 reflects the formation of soluble casein-anionic
polysaccharide complexes. This latter can be regarded as a
new biopolymer with a lower isoelectric point. At lower pH
values, e.g. 4.5, these soluble complexes form gels (Figure
10). It should be noted that neither casein nor sodium
alginate alone can gel under these conditions (14,42,43).
Since the formation of a complex gel results in a
completely new property of a mixed system it cannot be
treated as synergy. Strictly speaking, this term means an
328 V. B. Tolstoguzov

It) Anomalously high deformability of mixed gelatin-alginate

... ...
0 gels
I
Figure 12 shows the thermomechanical curves for several
til gels, namely gelatin gel, calcium alginate gel and a mixed
ll.
gel containing 10% gelatin and 0.5% calcium alginate.
S
c Curve 5 corresponds to re-heating of the last gel. To
.!!! prepare the gel a gelatin-alginate mixed solution was
Q.
cooled to room temperature. Then the gel was immersed in
E
0 ~ a solution of calcium acetate.
0
......
_-------------~.
25 45 65
Thermomechanical curves of mixed gels can be divided
into three segments. Below 30°C a mixed gel behaves like a
Tempera.ture. ~C gelatin gel. The deformability of gelatin gels increases
sharply at temperatures > 30°C. This reflects its melting
Figure 10 Thermomechanical curve for casein-sodium alginate temperature. Above 45°C the mixed gel behaves like a
complex gel. (pH 4.5; total concentration 10% wt). calcium alginate gel. Ca-alginate gel does not melt on
heating. Within an intermediate range of temperatures, i.e.
from 30 to 45°C, the gel is anomalously highly deformable.
Maximum gel compliance corresponds to the temperature
jot range from 34 to 38°C. The compliance maximum is
reversible and reproducible under cooling and re-heating of
'#- a gel. It depends on gel composition and on rates of heating
iN and cooling.
i The anomalously high deformability of gels can be
.~ T'" explained in terms of viscosity changes of the gel dispersion
Cl medium. These viscosity changes reflect the stepwise
1
<( nature of gelatin gel formation and melting processes. The
o 5 10 20
gelatin gel network is formed by aggregates of macro-
Casein %, wt. molecules. In an early stage of mixed gel heating the
network breaks down (at -30°C) into aggregates of gelatin
Figure 11 Phase diagram for the system sodium alginate-sodium macromolecules. Solutions of aggregates are notable for
caseinate-water at pH 7.2; 25°C (1) and pH 7.0; 20°C (2). their relatively low viscosity. Aggregates are melted by
heating 2::50°C. This causes a strong increase in viscosity,
increase (quantitative) of the effect of components working due to an increased concentration of solute particles and a
together where the combined action of the system com- change in their nature from rigid, large aggregates to very
ponents exceeds the sum of their individual actions. flexible, strongly interacting macromolecules of gelatin. In
Formally, the term 'synergy' seems to be more suitable for any thermomechanical study, non-equilibrium values of gel
describing composition-property relationships in mixed compliance are measured. These depend upon the dis-
and filled gels. persion medium viscosity. The decreased viscosity of the
Figure 11 shows phase diagrams for the same system. The dispersion medium of calcium alginate gel corresponds to
binodals (thick lines) correspond to pH 7.0 and 7.2. Tie-line an increase in its deformability. The increased viscosity
connects the casein-rich phase with another phase rich in results from the melting of macromolecular aggregates
alginate. leading to a decrease in gel compliance.
Phase separation occurs at pH values above the protein's On cooling the gelatin solution, the processes take place
isoelectric point and at an ionic strength >0.2. This can be in reverse order. Gelatin molecules are aggregated and
used for controlling volume properties of food gels. then their aggregates reform the gel network. Thus, the
Normally, complexing occurs at pH values below the network of the thermo-irreversible gel serves as a peculiar
protein isoelectric point. Addition of polyvalent cations viscometer of high sensitivity to a small deformation.
usually leads to widening of the range of pH values over
which complexes exist. Gelatin granules with iron ions Phase behaviour and cosolubility offood hydrocolloids in the
adsorbed in their surface layer were used as a protein gel state
macroreagent for complexing with alginate or pectinate. In The reproducibility of the compliance maximum under
other words anionic polysaccharides are precipitated onto cooling and re-heating of mixed gels shown in Figure 12
the granules and form a gel-like film of complex gel. Thus, means that neither gelation nor melting are accompanied
under sliglitly different conditions the same biopolymers by phase separation phenomena. Therefore, information
(Figures 10 and 11) can either form complexes or cannot be about compatibility of biopolymers in the gel state can be
mixed at all. Now we will consider some other important obtained.
features of multicomponent gels. It should be noted that gelation is always related to the
 Hydrocolloid interactions in mixed gels
-1 4
Compliance, Pa 10 ~_..,..,.
~=:::::::::502
-1 4
Compliance, Pa 10
25
-1 4
comPlianIL,_~_a_1_0
25
Figure /2 Thermomechanical curves for gels of I. 10% gelat in. 2, 0.5% Ca-alginate, 3,4. 10% gelatin + 0.5% alginate (heating and
cooling). 5. the same mixed gel. rehe ating. 6, 1% Ca- alginate + 5.8% ovalbumin , 7. 1% Ca-alginate + 10% potato starch .
aggregation of macromolecules and to a strong decrease in
the excluded volume effect of macromolecules. As a result
of gelatin gelation, its concentration in the gel dispersion
medium is reduced to a value below the critical concen-
tration for gelation. Therefore the dispersion medium of
gelatin gels may be a better solvent for polysaccharides and
casein than the initial gelatin solution . Also when food
substances are insoluble in a liquid mixed solution of gelling
agents they may still be dissolved after gelation. Pre sum-
ably, for this reason the formation of gelatin-alginate
mixed gels (Figure 12) is possible.
This also means that properties of a mixed gel depend
upon the sequence of gelation of individual gelling agents in
their mixtures . Favourable conditions for synergy include
that th e 'stronger' gelling component of a mixed solution
forms a gel network first.
Gel structure modification
It is necess ar y to stress that the mutual concentration of
biopolymers can give rise to several very different effects,
during the multistage gelation process. For instance, it can
be assumed that the excluded volume effect can lead to a
strong increase in the density and size of aggregates and a
corresponding decrease in the density of the three dimen-
sional network. This can result in an increase in both the
329
Temperature, °C
75
Temperature, °C
-=:::::==-
50 75
Temperature, °C
elastic modulus and the permeability of a multicomponent
gel. Presumably, this effect of protein incompatibility
contributes to increased casein losses during the washing of
the granules. The formation of multicomponent gels with a
more porous structure can also be favourable for enzymatic
attack (studied in vitro) throughout the whole volume of
the gel , not only at the surface of the gelatin granules.
Figure 12 gives some examples of the use of Ca-alginate
and Ca-pectinate gels as viscometers for studying the
behaviour of starches and globular proteins during gelation
and formation of mixed gels. These thermomechanical
curves have been employed for studying composition-
consistency relationships required for food formulation
(58) .
The anomalously high deformability of mixed gels at 34-
38°C is significant as this is the temperature of the mouth .
Anomalously high gel compliance at mouth temperature
ensures tender product consistency . Deformability of such
mixed gels is decreased both by heating and cooling the gel.
Thi s is important for product preservation (pasteurisation) .
The initial idea was to use these mixed gels for producing
granules of caviar analogue. However, this created some
problems related to the surface properties of the granules.
We will now concentrate on the surface properties of
gelatin gels containing other proteins and polysacharides.
330 V.B. Tolstoguzov
Wettability of gelatin gels
The main question facing us in the field of surface
properties was: how to clean the surface of the granules by
washing with water. Washing of proteingranules with water
is of prime significance for all other process stages. The aim
of washing is to remove the oil from the surface of the
granules. The efficiency of washing depends on the charac-
teristics of the gel surface. Figure 13 shows that gelatin gels
are not wetted with water. Gels are, however, perfectly
wetted with oils. They are more hydrophobic than any
other known material (59,60). Therefore, it is impossible to
separate oil from the surface of granules. The contact angle
of a water drop on the surface of a 10-20% gelatin gel is
-115-135°, depending on the gelatin preparation used.
Therefore, the problem was to make the surface of gelatin
gel hydrophilic.
It has been found that casein additives can make the
surface of gelatin gels hydrophilic. Figure 13 shows that
addition of several per cent (relative) of casein to a gelatin
solution results in a strong decrease, from 130 to 60-30°, of
the contact angle of water on the gel. Now, water can
rapidly expel the oil from the surface. This hydrophilising
effect of casein has been used for product formulation
development.
o incompatibility and complexing on surface properties of
o gels that, as we could see above, also affect volume
LO
,....
o
+ Pectin
C\J
,....
0.1 0.5 1.0%
Pectin, % wt.
3 5 10 15 17
Casein. % wt.
(Total Concentration of Proteins - 13%)
Figure 13 Wettability of gelatin gels. Effects of casein and low-
ester pectin.
Presumably, we are dealing with phase separation in the
mixed solution of gelatin and casein. Usually, either the less
hydrophilic protein or the disperse phase of a system
concentrates at the non-polar phase/water interface. Owing
to a relatively low hydrophilicity of casein compared to that
of gelatin phase separation results in the formation of a
two-phase gelatin-casein-water system with a relatively
small volume fraction of concentrated casein phase. This
latter expels the gelatin from the air/water interface and
makes the gel surface hydrophilic.
In the case of ovalbumin or bovine serum albumin the
contact angle of a water droplet on the surface of a gelatin
gel is only changed when the concentration of added
proteins exceeds 40% relative to gelatin. Unlike globular
proteins casein has an unordered molecular structure,
larger excluded volume and lower phase separation thres-
hold with polysaccharides. The difference in the effective
concentration of casein and globular proteins required
for making gel surfaces hydrophilic seems to be similar
to the difference in their phase separation threshold
with gelatin in their mixed solutions. Thus, incompatibility
of biopolymers determines both volume properties and
surface properties of many food systems (7-9,36-38).
Stable oil-in-water emulsions
The addition of 0.1-0.2% pectin can make the gelatin gel
hydrophobic once again (Figure 13). The restored high
hydrophobicity seems to be the result of the formation of
casein-Ca-anionic polysaccharide complexes which can
eject casein from the surface layer of a gelatin gel.
The data presented in Figure 13 can be regarded as
showing a synergistic effect of casein additives and an
antagonistic effect of pectin on gelatin gel surface prop-
erties. This reflects the opposite influence of biopolymer
properties of gels.
The hypothesis about the formation of protein-Ca-
polysaccharide complexes says that anionic polysaccharide
is concentrated at the non-polar phase/water interface. This
presumed effect has been supported by its successful use for
the development of a new way of stabilising emulsions. The
main principle of this method is illustrated in Figure 14.
In the first step, a calcium salt is dispersed in the oil. For
instance, an aqueous solution of calcium acetate, or dry
calcium gluconate. In the second step, the dispersion of
calcium salt-in-oil is introduced into a mixed water solution
containing 8-12% casein and 0.1-1.0% of low esterified
pectin or sodium alginate.
This method of emulsion stabilising has been used in
industry for producing the caviar analogue with desired
spreadability.
Concluding remarks
Emulsions stabilised by casein-Ca-pectinate complexes
were later used on a large industrial scale as meat
extenders.
 Hydrocolloid interactions in mixed gels
A alites et coacervates complex. 2. Coacervates auto-
of LIPID
......................
Dispersion Medium,
Mixed Solution, conta ining:
SODIUM CASEINATE
+ ALGINATE
Concentrated at the interface
AQUEOUS PHASE
Stabilising Layer
Figure /4 General scheme of emul sion stabilisation.
Similar problems of con sistency and flavour control in
the case of spherical protein granules used as fat replacers
has been recently solved in the same way by coverage of the
granules with thin lipid layers.
Later it was found that it is not necessary to use oils for
producing granules. Water-in-water emulsions are actually
spherical drops of a protein solution in a polysaccharide
solution and vice versa.
Granules can be formed and treated in aqueous media
using the phenomena of thermodynamic incompatibility
and membraneless osmosis . Because both of these
phenomena are of a general nature they presumably play an
important role not only in food processing but in food
digestion within the intestine (9). Phase separation and
control of substrate concentrations and substrate/enzyme
ratios could be responsible for interesting synergistic effects
in hydrolysis of food macromolecules.
The aim of this paper was to show, using some examples,
that phase behaviour of macromolecular components in
food s in both liquid and solid systems is of great importance
for controlling the stru ctur al functions of food hydro-
colloids.
Acknowledgements
The author is indebted to Mr Stephen G .Collyer and Mrs
Eliz abeth Prior for valuable discussions and editing of the
manuscript.
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Received on November 11. 1994, accepted on February 14,
1995