Arno G.B. Wouters

Relevance of the Functional Properties of Enzymatic Plant Protein Hydrolysates in Food Systems

Proteins play a crucial role in determining texture and structure of many food products. Although some animal proteins (such as egg white) have excellent functional and organoleptic properties, unfortunately, they entail a higher production cost and environmental impact than plant proteins. It is rather unfortunate that plant protein functionality is often insufficient because of low solubility in aqueous media. Enzymatic hydrolysis strongly increases solubility of proteins and alters their functional properties. The latter is attributed to 3 major structural changes: a decrease in average molecular mass, a higher availability of hydrophobic regions, and the liberation of ionizable groups. We here review current knowledge on solubility, water- and fat-holding capacity, gelation, foaming, and emulsifying properties of plant protein hydrolysates and discuss how these properties are affected by controlled enzymatic hydrolysis. In many cases, research in this field has been limited to fairly simple set-ups where functionality has been assessed in model systems. To evolve toward a more widely applied industrial use of plant protein hydrolysates, a more thorough understanding of functional properties is required. The structure-function relationship of protein hydrolysates needs to be studied in depth. Finally, test model systems closer to real food processing conditions, and thus to real foods, would be helpful to evaluate whether plant protein hydrolysates could be a viable alternative for other functional protein sources.

Foam fractionation as a tool to study the air-water interface structure-function relationship of wheat gluten hydrolysates

Enzymatic hydrolysis of wheat gluten protein improves its solubility and produces hydrolysates with foaming properties which may find applications in food products. First, we here investigated whether foam-liquid fractionation can concentrate wheat gluten peptides with foaming properties. Foam and liquid fractions had high and very low foam stability (FS), respectively. In addition, foam fractions were able to decrease surface tension more pronouncedly than un-fractionated samples and liquid fractions, suggesting they are able to arrange themselves more efficiently at an interface. As a second objective, foam fractionation served as a tool to study the structural properties of the peptides, causing these differences in air-water interfacial behavior. Zeta potential and surface hydrophobicity measurements did not fully explain these differences but suggested that hydrophobic interactions at the air-water interface are more important than electrostatic interactions. RP-HPLC showed a large overlap between foam and liquid fractions. However, a small fraction of very hydrophobic peptides with relatively high average molecular mass was clearly enriched in the foam fraction. These peptides were also more concentrated in un-fractionated DH 2 hydrolysates, which had high FS, than in DH 6 hydrolysates, which had low FS. These peptides most likely play a key role in stabilizing the air-water interface.

Exploring the Relationship between Structural and Air–Water Interfacial Properties of Wheat (Triticum aestivum L.) Gluten Hydrolysates in a Food System Relevant pH Range

The relationship between structural and foaming properties of two tryptic and two peptic wheat gluten hydrolysates was studied at different pH conditions. The impact of pH on foam stability (FS) of the samples heavily depended on the peptidase used and the degree of hydrolysis reached. Surface dilatational moduli were in most, but not all, instances related to FS, implying that, although the formation of a viscoelastic protein hydrolysate film is certainly important, this is not the only phenomenon that determines FS. In contrast to what might be expected, surface charge was not a major factor contributing to FS, except when close to the point-of-zero-charge. Surface hydrophobicity and intrinsic fluorescence measurements suggested that changes in protein conformation take place when the pH is varied, which can in turn influence foaming. Finally, hydrolyzed gluten proteins formed relatively large particles, suggesting that protein hydrolysate aggregation probably influences its foaming properties.

Impact of casein and egg white proteins on the structure of wheat gluten‐based protein‐rich food

BACKGROUND There is a growing interest in texturally and nutritionally satisfying vegetable alternatives to meat. Wheat gluten proteins have unique functional properties but a poor nutritional value in comparison to animal proteins. This study investigated the potential of egg white and bovine milk casein with well-balanced amino acid composition to increase the quality of wheat gluten-based protein-rich foods. RESULTS Heating a wheat gluten (51.4 g)-water (100.0 mL) blend for 120 min at 100 °C increased its firmness less than heating a wheat gluten (33.0 g)-freeze-dried egg white (16.8 g)-water (100.0 mL) blend. In contrast, the addition of casein to the gluten-water blend negatively impacted firmness after heating. Firmness was correlated with loss of protein extractability in sodium dodecyl sulfate containing medium during heating, which was higher with egg white than with casein. Even more, heat-induced polymerization of the gluten-water blend with egg white but not with casein was greater than expected from the losses in extractability of gluten and egg white on their own. CONCLUSION Structure formation was favored by mixing gluten with egg white but not with casein. These observations were linked to the intrinsic polymerization behavior of egg white and casein, but also to their interaction with gluten. Thus not all nutritionally suitable proteins can be used for enrichment of gluten-based protein-rich foods.

Wheat (Triticum aestivum L.) lipid species distribution in the different stages of straight dough bread making

Although wheat endogenous lipids strongly impact bread quality, knowledge on their detailed distribution throughout the different stages of straight dough bread making is lacking. We here compared the lipid populations in hexane [containing free lipids (FLs)] and water-saturated butanol extracts [containing bound lipids (BLs)] of wheat flour, freshly mixed and fermented doughs, and bread crumb using high-performance liquid-chromatography [for nonpolar lipids, i.e. mainly free fatty acids (FFA) and triacylglycerols] and electrospray ionization tandem mass spectrometry (for polar lipids). Freshly mixed doughs had lower FL and higher BL levels than flour, a phenomenon referred to as lipid-binding. Furthermore, probably due to the disintegration of flour particles, the overall extractability of nonpolar lipids was higher in freshly mixed dough than in flour. Dough fermentation decreased the extractability of glycolipids, but increased that of nonpolar lipids and phospholipids. We hypothesize that these phenomena result from stretching of the gluten network due to gas cell expansion, which leads to the replacement of some lipids associated with gluten proteins by others. Baking increased the extractability of bound lysophospatidylcholine (LPC) levels, but decreased that of free FFA. This is probably due to in situ dissociation of amylose-LPC inclusion complexes and formation of amylose-FFA inclusion complexes during bread baking and cooling, respectively. The approach and ESI-MS/MS methodology we developed provided valuable insights regarding the distribution of lipids at the different stages of bread making. Hence, it opens perspectives for future efforts to relate differences in lipid composition between wheat cultivars to their bread making quality.

Enzymatically Hydrolyzed Wheat Gluten as a Foaming Agent in Food: Incorporation in a Meringue Recipe as a Proof-of-Concept: Gluten hydrolysates in a meringue recipe…

There is a growing interest in substituting animal proteins with plant protein sources in food systems. A notable example is the replacement of hen egg white (EW) protein, which is used in a wide range of food products because of its excellent foaming characteristics. Here, enzymatically hydrolyzed wheat gluten, which has greater solubility and better foaming properties than wheat gluten itself, was prepared and incorporated in a classical meringue recipe to investigate its potential as a foaming agent. Meringues based on gluten hydrolysates (GHs) had batters with lower density and greater apparent viscosity than those based solely on EW protein. Furthermore, after baking, these GH containing meringues had greater specific volume than those based on EW protein alone and no notable differences in color or texture between the different samples were noted. These outcomes were related to basic insights in the air-water interfacial behavior of GHs obtained in earlier studies. More specifically, the greater foaming capacity of GH than of EW protein solutions was related to their superior meringue batter (density and apparent viscosity) and product (specific volume) properties. While EW protein solutions had better foam stability than GH solutions (in the absence of sugar), this was apparently less relevant for meringue properties, probably due to the very high viscosity of the sugar rich batter, which could obscure differences in the intrinsic foam stabilizing ability of the samples. PRACTICAL APPLICATION Replacing animal proteins with plant protein sources in the food industry is desirable from an economic and environmental perspective. Enzymatic hydrolysis serves as a tool to improve the foaming properties of water-insoluble wheat gluten proteins. We conclude that wheat gluten hydrolysates can be a valid functional alternative for egg white proteins in meringues, and possibly other food systems.

Electrical resistance oven baking as a tool to study crumb structure formation in gluten-free bread

Gradientless baking by means of ohmic heating was used for the first time in gluten-free (GF) bread making. Combination thereof with in-line measurements of batter height, viscosity and carbon dioxide (CO2) release proved to be powerful for studying structure formation in GF breads. GF breads studied here were based on (i) a mixture of potato and cassava starches and egg white powder (C/P-S+EW), (ii) rice flour (RF) or (iii) a mixture of RF and egg white powder (RF+EW). The work revealed that bread volume and crumb structure rely heavily on the balance between the moment of CO2 release from batter during baking and that of crumb setting. At the moment of CO2 release, C/P-S+EW bread crumb had already (partly) set, while this was not the case for RF bread crumb, resulting in a collapse and thus low volume of the latter. When a part of RF was replaced by egg white powder, the moment of CO2 release was postponed and the batter collapse was less pronounced, leading to a higher volume and a finer crumb. The presence of egg white proteins in C/P-S+EW or RF+EW batters improved gas cell stabilization. Thus, increasing batter stability or altering the moment of crumb setting results in GF breads with higher volume and a finer crumb structure.

Relating the composition and air/water interfacial properties of wheat, rye, barley, and oat dough liquor

Gas cell stabilization in dough by its aqueous phase constituents is arguably more important in non-wheat than in wheat dough due to weaker protein networks in the former. Dough liquor (DL), a model for the dough aqueous phase, was isolated from fermented wheat, rye, barley, and oat doughs by ultracentrifugation. DL composition (protein, lipid, arabinoxylan, β-glucan) and air/water interfacial functionality [foaming, viscosity, surface tension, surface dilatational modulus (E)] were related to bread quality. Poor foaming and low E of wheat DL were ascribed to lipids and proteins co-occurring at the interface. Nonetheless, the presence of a gluten network resulted in high-quality wheaten breads. Homogeneous and heterogeneous crumb structures of rye and barley breads, respectively, were attributed to high and low E values of their respective DLs. High lipid content and low surface tension of oat DL indicated a lipid-dominated interface, which may explain the heterogeneous crumb structure of oat breads.