Gum arabic glycoprotein contains glycomodules of both extensin and arabinogalactan-glycoproteins

Gum arabic glycoprotein (GAGP) is a large molecular weight, hydroxyproline-rich arabinogalactan-protein (AGP) component of gum arabic. GAGP has a simple, highly biased amino acid composition indicating a repetitive polypeptide backbone. Previous work (Qi, W., Fong, C., Lamport, D.T.A., 1991. Plant Physiology 96, 848), suggested small (approximately 11 residue) repetitive peptide motifs each with three Hyp-arabinoside attachment sites and a single Hyp-arabinogalactan polysaccharide attachment site. We tested that hypothesis by sequence analysis of the GAGP polypeptide after HF-deglycosylation. A family of closely related peptides confirmed the presence of a repetitive 19-residue consensus motif. However, the motif: Ser-Hyp-Hyp-Hyp-Thr-Leu-Ser-Hyp-Ser- Hyp-Thr-Hyp-Thr-Hyp-Hyp-Leu-Gly-Pro-His, was about twice the size anticipated. Thus, judging by Hyp-glycoside profiles of GAGP, the consensus motif contained six Hyp-arabinosides rather than three and two Hyp-polysaccharides rather than one. We inferred the glycosylation sites based on the Hyp contiguity hypothesis which predicts arabinosides on contiguous Hyp residues and arabinogalactan polysaccharides on clustered non-contiguous Hyp residues, i.e. the GAGP motif would consist of arabinosylated contiguous Hyp blocks flanking two central Hyp-polysaccharides. We predict this rigidifies the glycoprotein, enhances the overall symmetry of the glycopeptide motif, and may explain some of the remarkable properties of gum arabic.     

Gum arabic glycoprotein is a twisted hairy rope : a new model based on o-galactosylhydroxyproline as the polysaccharide attachment site

Separation of the wound exudate from Acacia senegal (L.) Willd., “gum arabic,” on a preparative Superose-6 column gave two major fractions: a high molecular weight gum arabic glycoprotein (GAGP) containing about 90% carbohydrate and a lower molecular weight heterogenous gum arabic polysaccharide fraction. Hydrogen fluoride-deglycosylation of GAGP gave a large (~400 residue) hydroxyproline-rich polypeptide backbone (dGAGP). Alkaline hydrolysis of GAGP showed that most of the carbohydrate was attached to the polypeptide backbone as small (~30 residue) hydroxyproline (Hyp)-polysaccharide substituents. After partial acid hydrolysis of the Hyp-polysaccharide fraction we identified O-galactosylhydroxyproline as the glycopeptide linkage, identical with that of hydroxyproline-rich arabinogalactan-proteins (AGPs). However, unlike the acidic alanine-rich AGPs, GAGP is basic and notably deficient in alanine. Thus, while the GAGP polypeptide backbone more closely resembles that of the Hyp-rich cell wall protein extensin, the GAGP polysaccharide sidechains resemble AGPs. Possibly all three proteins comprise a phylogenetically related extensin superfamily of extended rod-like macromolecules. The “wattle-blossom” model for AGP and gum arabic predicts a few large polysaccharide substituents along the polypeptide backbone of a spheroidal macromolecule. On the contrary, our data imply a rodlike molecule with numerous small polysaccharide substituents (attached to 24% of the Hyp residues), regularly arranged along a highly periodic polypeptide backbone based, hypothetically, on a 10 to 12 residue repetitive peptide motif. Thus, a simple statistical model of the gum arabic glycoprotein predicts a repeating polysaccharide-peptide subunit of about 7 kilodaltons. The small polysaccharide substituents will maximize intramolecular hydrogen bonding if aligned along the long axis of the molecule, forming in effect a twisted hairy rope. Electron micrographs of rotary shadowed GAGP molecules support that prediction and may also explain how such apparently large molecules can exit the cell by endwise reptation through the small pores of the primary cell wall.     

The lysine-rich arabinogalactan-protein subfamily in Arabidopsis: gene expression, glycoprotein purification and biochemical characterization

AtAGP17, AtAGP18 and AtAGP19 are homologous genes encoding three putative glycosylphosphatidylinositol (GPI)-anchored classical arabinogalactan-proteins (AGPs) in Arabidopsis. They are distinguished from other AGPs by a short, C-terminal lysine-rich region. Organ-specific expression of these genes was revealed by Northern blot analysis. AtAGP17 was strongly expressed in leaves and stems, and weakly expressed in flowers and roots; AtAGP18 was strongly expressed in flowers, and moderately expressed in roots, stems and young leaves; and AtAGP19 was strongly expressed in stems, moderately expressed in flowers and roots, and weakly expressed in young leaves. One of these genes, AtAGP17, was expressed and purified as a green fluorescent protein (GFP) fusion protein in transgenic tobacco cells using hydrophobic interaction chromatography, size exclusion chromatography and reverse phase high-performance liquid chromatography. The fusion (glyco)protein produced a characteristic AGP 'smear' with a molecular mass of 80-150 kDa when detected by Western blot analysis. Glycosyl composition and linkage analyses of purified GFP-AtAGP17 showed that carbohydrate accounted for approximately 86% of the molecule, with arabinose and galactose as major, and rhamnose and glucuronic acid as minor glycosyl residues and with 1,3,6-galactose, 1,4-glucuronic acid, 1,3-galactose and terminal arabinose as major linkages. GFP-AtAGP17 was also precipitated by beta-Yariv reagent, further confirming that AtAGP17 is a bona fide AGP. Confocal fluorescence microscopy of plasmolysed, transformed cells indicated that AtAGP17 is localized on the plasma membrane and in Hechtian strands. Hydroxyproline (Hyp) glycoside profiles of GFP-AtAGP17 in conjunction with the deduced protein sequence also served to corroborate the Hyp contiguity hypothesis, which predicts contiguous Hyp residues as attachment sites for arabinosides and clustered, non-contiguous Hyp residues as attachment sites for arabinogalactan polysaccharides.     

The amino acid composition of gum exudates from Prosopis species

The amino acid compositions of the proteinaceous components of the gum exudates from Prosopis alba, P. chilensis, P. glandulosa, P. laevigata, P. torreyana and P. velutina, and for a sample of commercial gum mesquite, are presented. In agreement with data published previously for the polysaccharide components of their gums, only minor differences in composition are shown by these species. The amino acid compositions are characterized by very high proportions of hydroxyproline and by high proportions of proline and serine; these three amino acids account for 62.5% of those present in the gum from Prosopis velutina. The amino acid compositions of these Prosopis gums are remarkably similar to that established recently for the gum from Acacia senegal (gum arabic).     

Structure of the exudate gum from Meryta sinclairii

A gum that exudes from the wounded trunk of the New Zealand native tree Meryta sinclairii has been isolated. The gum was completely precipitated by the β-glucosyl Yariv reagent and was thus determined to be an arabinogalactan-protein (AGP). It contained >95% w/w carbohydrate and only 2% w/w protein with a high level of hydroxyproline. SEC-MALLS showed that the gum had a weight-average molecular weight of 4.45×10⁶ Da compared with 6.02×10⁵ Da for gum arabic. Constituent sugar and linkage analyses were consistent with polymers comprised of a highly branched backbone of 1,3-linked galactopyranosyl (Galp) residues, with side-chains made up of arabinofuranose- (Araf) containing oligosaccharides, terminated variously by rhamnopyranosyl (Rhap), arabinopyranosyl (Arap), Galp and glucuronopyranosyl (GlcpA) residues. Analysis by one-dimensional and two-dimensional ¹H and ¹³C NMR experiments confirmed the linkage analyses. The structure of the gum is discussed in comparison with the structure of gum arabic and other AGPs.     

Preparation and characterization of polyacrylic acid/karaya gum and polyacrylic acid/tamarind seed gum adducts and utilization in textile printing

Polymerization of 20% neutralized acrylic acid (Na form), AA, in presence of Karaya gum, KG, or tamarind seed gum, TG, at AA/gum weight ratio of 1/1 and 2/1 results in PAA/KG1, PAA/KG2, PAA/TG1 and PAA/TG2 adducts, respectively (where the suffix 1 or 2 stands for AA/gum ratios of 1/1 or 2/1). Infra red spectra of adducts are examined. Aqueous pastes of adducts, native gums and GG are of non-Newtonian thixotropic flow within a shear rate range of 4–40 s⁻¹. Adduct pastes (7.5% w/v) are of higher apparent viscosities (η) than their native gums or GG, and pastes of TG adducts are of higher η than KG adducts. Except for PAA/TG2 adduct, the power law does not correlate well to the other pastes. Preliminary trials showed that adducts are excellent thickeners for reactive and acid printing on wool, silk and nylon 6. Prints by adducts are of higher color strength than those by native gums or GG. GG paste was completely destroyed after storing for 7 days, whereas η of pastes of adducts and native gums were noticeably decreased upon storing.     

Affinity chromatography of two sets of isomeric antibodies having specificity for different oligosaccharide units of gum arabic

Two sets of antibodies directed against different carbohydrate units of gum arabic were isolated from the sera of rabbits immunized intramuscularly with gum arabic and Freund's complete adjuvant. The isolation was effected by affinity chromatography on two columns attached in series and containing an absorbent of AH-Sepharose 4B with ligands of partially hydrolyzed gun arabic in the first column and an adsorbent of AH-Sepharose 4B with ligands of native gum arabic in the second column. The two populations of anti-gum arabic antibodies were obtained and have been designated as Set 1 and Set 2 on the basis of their mobilities on agar diffusion. The antibodies of Set 1 consisted of 4 isomeric antibodies and those of Set 2 consisted of 11 isomeric antibodies. Native gum arabic samples were oxidized with periodate or reduced with sodium borohydride and carbodiimide under standard conditions and the modified samples were totally inactive in the precipitin test. On the basis of methylation data and immunological results it was concluded that terminal disaccharide moieties of the gum having the structure beta-D-glucosyluronic acid-(1----6)-D-galactose and alpha-L-arabinofuranosyl-(1----4)-D-glucuronic acid were the immunodeterminant groups for Set 1 and Set 2 antibodies, respectively.     

Composition and structure of whey protein/gum arabic coacervates

Complex coacervation in whey protein/gum arabic (WP/GA) mixtures was studied as a function of three main key parameters: pH, initial protein to polysaccharide mixing ratio (Pr:Ps)(ini), and ionic strength. Previous studies had already revealed under which conditions a coacervate phase was obtained. This study is aimed at understanding how these parameters influence the phase separation kinetics, the coacervate composition, and the internal coacervate structure. At a defined (Pr:Ps)(ini), an optimum pH of complex coacervation was found (pH(opt)), at which the strength of electrostatic interaction was maximum. For (Pr:Ps)(ini) = 2:1, the phase separation occurred the fastest and the final coacervate volume was the largest at pH(opt) = 4.0. The composition of the coacervate phase was determined after 48 h of phase separation and revealed that, at pH(opt), the coacervate phase was the most concentrated. Varying the (Pr:Ps)(ini) shifted the pH(opt) to higher values when (Pr:Ps)(ini) was increased and to lower values when (Pr:Ps)(ini) was decreased. This phenomenon was due to the level of charge compensation of the WP/GA complexes. Finally, the structure of the coacervate phase was studied with small-angle X-ray scattering (SAXS). SAXS data confirmed that at pH(opt) the coacervate phase was dense and structured. Model calculations revealed that the structure factor of WP induced a peak at Q = 0.7 nm(-1), illustrating that the coacervate phase was more structured, inducing the stronger correlation length of WP molecules. When the pH was changed to more acidic values, the correlation peak faded away, due to a more open structure of the coacervate. A shoulder in the scattering pattern of the coacervates was visible at small Q. This peak was attributed to the presence of residual charges on the GA. The peak intensity was reduced when the strength of interaction was increased, highlighting a greater charge compensation of the polyelectrolyte. Finally, increasing the ionic strength led to a less concentrated, a more heterogeneous, and a less structured coacervate phase, induced by the screening of the electrostatic interactions.     

Solution properties of conventional gum arabic and a matured gum arabic (Acacia (sen) SUPER GUM)

Dilute solution properties of two specially matured gum arabic samples (EM1 and EM2) were compared to the conventional gum (EM0) using static light scattering. The apparent molar mass (M(w,app) and radius of gyration (R(g,app)) for the three samples showed unusual concentration dependence. These data were satisfactorily interpreted by a simple association model that takes into account the repulsive interaction among clusters, which allowed us to obtain the true molar mass (Mw(0)) and radius of gyration (Rg(0)). A common power law relation was observed between Mw(0) and Rg(0) , giving a somewhat higher exponent than expected for linear and branched polymers in a good solvent. Mw(0) and Rg(0) obtained for the three gums do not differ significantly from each other. However, the data showed clearly a constant increase of the association from EM0 to EM2 with increasing concentration. This is in accordance with the previously observed improved functional properties for the matured products.     

Differences among captive callitrichids in the digestive responses to dietary gum

Although many members of the Callitrichidae, a monophyletic family of small, New World monkeys, have been observed to feed on plant exudates, available field data support the generalization that pygmy and common marmosets (Cebuella pygmaea and Callithrix jacchus) feed on gums to a greater extent than most other callitrichids. Because microbial fermentation is required for vertebrates to digest gums, gum-feeding primates may react differently to dietary gum from their relatives that eat little gum. To test this hypothesis, digestion trials were conducted on animals from the two marmoset species, two tamarin species (Saguinus fuscicollis and S. oedipus), and a species of lion tamarin (Leontopithecus rosalia). These species span the range of body sizes within the Callitrichidae. All animals were fed two variations of a homogeneous diet, which differed only in that gum arabic was added to one. Transit time of digesta (TFA) and digestive efficiency (as measured by the coefficients of apparent digestibility of dry matter and energy [ADDM and ADE, respectively]) were compared between diets for each individual. As predicted, the digestive responses of marmosets differed from the responses of the other study species. In marmosets, TFA tended to be longer when gum was added to the diet, while TFA did not change in the other three species. Digestive efficiency decreased in tamarins and lion tamarins with the addition of gum to the diet; marmoset digestive efficiency was unaffected by diet. The results of this research are consistent with the hypothesis that marmosets have digestive adaptations that aid in the digestion of gum that other callitrichids lack. © 1996 Wiley-Liss, Inc.     

Oxidation of gum arabic by soluble colloidal MnO2

In the present work, the oxidative degradation of gum arabic by colloidal manganese dioxide (MnO2) was carried out. Monitoring the disappearance of the MnO2 spectrophotometrically at 375 nm was used to follow the kinetics. The oxidation obeyed fractional-order kinetics with respect to the [gum arabic]. Effect of various experimental parameters such as the initial colloidal [MnO2], [HClO4], temperature, and complexing agents (P2O7(4-), F-, and Mn2+) for the oxidation of gum arabic was studied. The reaction was acid catalyzed. Addition of P2O(7)4-, F-, and Mn2+ ions enhances the rate of oxidation significantly. Gum arabic adsorbs onto the surface of the colloidal MnO2 through the equatorial -OH groups of the rhamnose moiety, and the complex breaks down into products. The Arrhenius equation was valid for the oxidation kinetics between 40 and 60 degrees C. To explain the observed kinetic results, a suitable mechanism and rate law for the reaction taking place at the surface of the colloidal particle has been proposed. The reducing nature of gum arabic is found be due to the presence of -OH group in the skeleton.     

Exudate gums: occurrence,production, and applications

This paper presents a review of the industrially most relevant exudate gums: gum arabic, gum karya, and gum tragacanth. Exudate gums are obtained as the natural exudates of different tree species and exhibit unique properties in a wide variety of applications. This review covers the chemical structure, occurrence and production of the different gums. It also deals with the size and relative importance of the various players on the world market. Furthermore, it gives an overview of the main application fields of the different gums, both food and non-food.     

Phase separation induced molecular fractionation of gum arabic—Sugar beet pectin systems

This paper investigates the phase separation and phase separation-induced fractionation of gum arabic (GA)/sugar beet pectin (SBP) mixed solutions. A phase diagram, including cloud and binodal curves, was established by visual observation and phase composition analysis. The deviation of the binodal curve from the cloud curve was a result of phase separation-induced fractionation of polydisperse GA and SBP molecules. Fractionation of GA increased the content of arabinogalactan-protein complex (AGP) from ca. 13% to 27%. The fractionated GA (FGA) showed improved emulsifying functionality, whereas the fractionated SBP (FSBP) had a reduced emulsifying functionality. The changes in emulsifying efficiency can be explained by interfacial adsorption behaviors at the oil–water interface as indicated by interfacial tension measurements.     

Synergistic interaction between xanthan and guar gum

The non-Newtonian behavior and dynamic viscoelasticity of a series of aqueous mixtures of xanthan and guar gum were measured with a rheogoniometer. At a concentration of 0.2% of total gums, gelation did not occur at room temperature but occurred at a low temperature (0°). A much stronger interaction was observed with a mixture of deacetylated xanthan than that with native xanthan. The maximum dynamic modulus was obtained when the ratio of xanthan to guar gum was 2:1. The transition temperatures of dynamic viscoelasticity for mixtures with native and deacetylated xanthan were observed at 25 and 30°, respectively. It was concluded that the side chains of the guar gum molecular prevent an intermolecular interaction with the side chains of the xanthan molecule. An intermolecular interaction between xanthan and guar gum at low temperature might be promoted between the periphery of the side chains of the xanthan molecule and the backbone of the guar gum molecule and dissociation takes place at the transition temperature.     

Arabinogalactan proteins improve plant regeneration in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) anther culture

Androgenesis-based methods of doubled haploid (DH) production show considerable variation in efficiency in different barley genotypes. Arabinogalactan proteins (AGPs) have been shown to play a key role in several developmental processes, including embryogenesis, in different plant species. In this study we investigated the effect of exogenous AGPs from gum arabic on androgenesis and the regeneration efficiency in barley anther culture. Supplementation of the induction medium with 10 mg l^?1 gum arabic increased the total plant regeneration rate up to 2.8 times; when exposure to GA was extended to also include the pretreatment step, the regeneration rate was up to 6.6-times higher than in control. The effect of gum arabic was reversed by the Yariv reagent, an AGPs antagonist. This suggests a direct involvement of AGPs in androgenic development from barely microspores. Addition of gum arabic reduced cell mortality, increased the frequency of mitotic divisions of microspores and the number of multicellular structures (MCSs) when compared to control. The positive effect of gum arabic also included reduction in time required for the androgenic induction and substantially improved the quality of formed embryos. Observations made in this study imply a complex role of AGPs during androgenic development and confirmed the usefulness of gum arabic in production of barley androgenic plants.     

Engineering ‘designer’ glycomodules for boosting recombinant protein secretion in tobacco hairy root culture and studying hydroxyproline‐O‐glycosylation process in plants

The key technical bottleneck for exploiting plant hairy root cultures as a robust bioproduction platform for therapeutic proteins has been low protein productivity, particularly low secreted protein yields. To address this, we engineered novel hydroxyproline (Hyp)‐O‐glycosylated peptides (HypGPs) into tobacco hairy roots to boost the extracellular secretion of fused proteins and to elucidate Hyp‐O‐glycosylation process of plant cell wall Hyp‐rich glycoproteins. HypGPs representing two major types of cell wall glycoproteins were examined: an extensin module consisting of 18 tandem repeats of ‘Ser‐Hyp‐Hyp‐Hyp‐Hyp’ motif or (SP4)₁₈ and an arabinogalactan protein module consisting of 32 tandem repeats of ‘Ser‐Hyp’ motif or (SP)₃₂. Each module was expressed in tobacco hairy roots as a fusion to the enhanced green fluorescence protein (EGFP). Hairy root cultures engineered with a HypGP module secreted up to 56‐fold greater levels of EGFP, compared with an EGFP control lacking any HypGP module, supporting the function of HypGP modules as a molecular carrier in promoting efficient transport of fused proteins into the culture media. The engineered (SP4)₁₈ and (SP)₃₂ modules underwent Hyp‐O‐glycosylation with arabino‐oligosaccharides and arabinogalactan polysaccharides, respectively, which were essential in facilitating secretion of the fused EGFP protein. Distinct non‐Hyp‐O‐glycosylated (SP4)₁₈‐EGFP and (SP)₃₂‐EGFP intermediates were consistently accumulated within the root tissues, indicating a rate‐limiting trafficking and/or glycosylation of the engineered HypGP modules. An updated model depicting the intracellular trafficking, Hyp‐O‐glycosylation and extracellular secretion of extensin‐styled (SP4)₁₈ module and AGP‐styled (SP)₃₂ module is proposed.     

Development of an optimal formulation for oxidative stability of walnut-beverage emulsions based on gum arabic and xanthan gum using response surface methodology

The susceptibility of lipids to oxidation is one of the most fundamental problems in oil-in-water emulsions. A response surface methodology 5-level-3-factor central-composite rotatable design was applied to study the effects of key formula ingredients including walnut oil (WO, 3-6%, w/w), gum arabic (GA, 5-10%, w/w) and xanthan gum (XG, 0.05-0.15%, w/w) on lipid oxidation in walnut-beverage emulsions. During 30 days’ storage, the oxidation process was monitored by evaluating the peroxide value, anisidine value and total oxidation (Totox) value in different emulsion formulations. Use of XG as a stabilizer at high concentrations considerably inhibited the oxidation of WO in the prepared emulsions. The experimental data were satisfactorily fitted to quadratic models using multiple regression analysis. The optimum conditions to obtain the minimum peroxide (0.923 mequiv. O₂/kg oil), anisidine (0.500) and Totox (2.347) values are met when a walnut-beverage emulsion is formulated with 3% WO, 10% GA and 0.12% XG.     

Effect of Different Gums on Features of 3D Printed Object Based on Vitamin-D Enriched Orange Concentrate

This study examined the effects of different gums viz. gum arabic (GA), guar gum (GG), k-carrageenan gum (KG), and xanthan gum (XG) on rheological and 3D printing characteristics of vitamin D (Vit D) enriched orange concentrate (OC) wheat starch (WS) blends. The textural and microstructural properties of printed objects from above mixture were evaluated and compared. The addition of gums induced an increase in apparent viscosity, storage modulus (G′), and loss modulus (G″) of the OC-WS mixtures, while GA decreased the apparent viscosity and G′. Nuclear magnetic resonance (NMR) analysis of 3D printed samples revealed that the movement of transverse time (T₂) toward closer to 0 ms indicated an increase in immobilized and bound water populations suggesting the gel formation. The slight shift toward shorter wavelength in FT-IR results for the broadband centered around 3400 cm^?1 after addition of gums possibly caused an increase of G′ and load bearing capacity of the blends. 3D printing characteristics revealed that the objects printed using KG containing blend possessed maximum fidelity to the target geometry and good loading bearing capacity, preventing collapsing over time due to the proper G′ value. At tanδ of 0.238, OC-WS-KG mixture achieved the best printing condition. Higher tanδ of GA (0.038) containing samples led to an unwanted collapse of the printed constructs. The objects printed using KG also exhibited the smoothest visible surface as well as microstructure and best mastication properties. Considering the studied features, vitamin D enriched OC with WS-KG was found to be the best match for orange fruit concentrate-based 3D food printing. This work demonstrates the novel ways to develop fortified 3D printed foods.     

Interaction of gum arabic, maltodextrin and pullulan with lipids in emulsions

The interaction of gum arabic, maltodextrin and pullulan with lipids in emulsion systems was investigated. Interfacial tension and interfacial viscosity measurements revealed that only gum arabic could adsorb and form a viscoelastic film at the oil-water interface. Good emulsifying activity was demonstrated for gum arabic, whereas fine emulsions could not be produced from the other polysaccharide solutions and oil. Frequency-dependent increases in the storage and loss moduli were observed for all the polysaccharide solutions. Such rheological behavior did not substantially change when maltodextrin and pullulan were mixed with oil to form emulsions. However, the frequency-dependence of the dynamic moduli disappeared in the gum arabic-stabilized emulsion, suggesting the formation of a network structure in which oil droplets could form junctions with gum arabic chains. The results on the inhibition of lipid oxidation by polysaccharides suggest that gum arabic protected lipids from the attack of lipoxygenase and free radicals by adsorbing at the oil droplet surface.     

Complex coacervation of whey proteins and gum arabic

Mixtures of gum arabic and whey protein (whey protein isolate, WP) form an electrostatic complex in a specific pH range. Three phase boundaries (pH(c), pHphi(1), pHphi(2)) have been determined using an original titration method, newly applied to complex coacervation. It consists of monitoring the turbidity and light scattering intensity under slow acidification in situ with glucono-delta-lactone. Furthermore, the particle size could also be measured in parallel by dynamic light scattering. When the pH is lowered, whey proteins and gum arabic first form soluble complexes. This boundary is designated as pH(c). When the interaction is stronger (at lower pH), phase separation takes place (at pHphi(1)). Finally, at pHphi(2) complexation was suppressed by the charge reduction of the gum arabic. The major constituent of the whey protein preparation used was beta-lactoglobulin (beta-lg), and it was shown that beta-lg was indeed the main complex-forming protein. Moreover, an increase of the ionic strength shifted the pH boundaries to lower pH values, which was summarized in a state diagram. The experimental pH(c) values were compared to a newly developed theory for polyelectrolyte adsorption on heterogeneous surfaces. Finally, the influence of the total biopolymer concentration (0-20% w/w) was represented in a phase diagram. For concentrations below 12%, the results are consistent with the theory on complex coacervation developed by Overbeek and Voorn. However, for concentrations above 12%, phase diagrams surprisingly revealed a "metastable" region delimited by a percolation line. Overall, a strong similarity is seen between the behavior of this system and a colloidal gas-liquid phase separation.