Solidification of concentrated pea protein-pectin mixtures as potential binder.

BACKGROUND: Binders in plant-based meat analogues allow different components, such as extrudate and fat particles, to stick together. Typically, binders then are solidified to transform the mass into a non-sticky, solid product. As an option for a clean-label binder possessing such properties, the solidification behavior of pea protein-pectin mixtures (250 g kg-1 , r = 2:1, pH 6) was investigated upon heating, and upon addition of calcium, transglutaminase, and laccase, or by combinations thereof. RESULTS: Mixtures of (homogenized) pea protein and apple pectin had higher elastic moduli and consistency coefficients and lower frequency dependencies upon calcium addition. This indicated that calcium physically cross-linked pectin chains that formed the continuous phase in the biopolymer matrix. The highest degree of solidification was obtained with a mixture of pea protein and sugar beet pectin upon addition of laccase that covalently cross-linked both biopolymers involved. All solidified mixtures lost their stickiness. A mixture of soluble pea protein and apple pectin solidified only slightly through calcium and transglutaminase, probably due to differences in the microstructural arrangement of the biopolymers. CONCLUSION: The chemical makeup of the biopolymers and their spatial distribution determines solidification behavior in concentrated biopolymer mixtures. In general, pea protein-pectin mixtures can solidify and therefore have the potential to act as binders in meat analogues. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Homogenization improves foaming properties of insoluble pea proteins.

Foams are essential in many food applications and require surface-active ingredients such as proteins for formation and stabilization. We investigated the influence of high-pressure homogenization on foaming properties of insoluble pea protein dispersions (5% w/w) at pH 3 and 5. Unhomogenized insoluble pea protein dispersions did not foam at either pH 3 or 5, as they consisted of large insoluble pea protein aggregates with limited surface activity. At pH 3, the homogenized pea protein dispersions generated foams due to higher protein solubility and surface activity through disruption of large protein aggregates into smaller particles. The foam stability decreased with increasing homogenization pressure and number of cycles due to a reduction in continuous phase viscosity. At pH 5, the insoluble pea proteins foamed when the homogenization resulted in formation of aggregates made of smaller protein entities, which was the case for homogenization ≥ 100 MPa and three cycles. In general, the foam capacity (amount of formed foam) was higher at pH 3 due to improved protein solubility and surface activity that facilitated incorporation of air, while the foam stability (resistance against foam collapse) was better at pH 5 because of the presence of larger protein aggregates that formed thicker and more viscous films around the air bubbles benefitting retention of gas bubbles. Overall, homogenization improved the foaming properties of insoluble pea proteins at pH 3 and 5. Practical Application Insoluble pea protein dispersions formed foams at pH 3 and 5 after homogenization highlighting the potential of this processing step for the food industry. The improvement in functionality of plant-derived ingredients helps to increase their use for consumer goods, thereby supporting the transition to more sustainable food system.

Comparison of Binding Properties of a Laccase-Treated Pea Protein-Sugar Beet Pectin Mixture with Methylcellulose in a Bacon-Type Meat Analogue.

A bacon-type meat analogue consists of different structural layers, such as textured protein and a fat mimetic. To obtain a coherent and appealing product, a suitable binder must glue those elements together. A mixture based on pea protein and sugar beet pectin (r = 2:1, 25% w/w solids, pH 6) with and without laccase addition and a methylcellulose hydrogel (6% w/w) serving as benchmark were applied as binder between textured protein and a fat mimetic. A tensile strength test, during which the layers were torn apart, was performed to measure the binding ability. The pea protein-sugar beet pectin mixture without laccase was viscoelastic and had medium and low binding strength at 25 °C (F ≤ 3.5 N) and 70 °C (F ≈ 1.0 N), respectively. The addition of laccase solidified the mixture and increased binding strength at 25 °C (F ≥ 4.0 N) and 70 °C (F ≈ 2.0 N), due to covalent bonds within the binder and between the binder and the textured protein or the fat mimetic layers. Generally, the binding strength was higher when two textured protein layers were glued together. The binding properties of methylcellulose hydrogel was low (F ≤ 2.0 N), except when two fat mimetic layers were bound due to hydrophobic interactions becoming dominant. The investigated mixed pectin-pea protein system is able serve as a clean-label binder in bacon-type meat analogues, and the application in other products seems promising.

Use of molecular interactions and mesoscopic scale transitions to modulate protein-polysaccharide structures.

Mixed protein-polysaccharide structures have found widespread applications in various fields, such as in foods, pharmaceuticals or personal care products. A better understanding and a more precise control over the molecular interactions between the two types of macromolecules leading to an engineering of nanoscale and colloidal building blocks have fueled the design of novel structures with improved functional properties. However, these building blocks often do not constitute the final matrix. Rather, further process operations are used to transform the initially formed structural entities into bulk matrices. Systematic knowledge on the relation between molecular structure design and subsequent mesoscopic scale transitions induced by processing is scarce. This article aims at establishing a connection between these two approaches. Therefore, it reviews not only studies on the underlying molecular interaction phenomena leading to either a segregative or associative phase behavior and nanoscale or colloidal structures, but also looks at the less systematically studied approach of using macroscopic processing operations such as shearing, heating, crosslinking, and concentrating/drying to transform the initially generated structures into bulk matrices. Thereby, a more comprehensive look is taken at the relationship between different influencing factors, namely solvent conditions (i.e. pH, ionic strength), biopolymer characteristics (i.e. type, charge density, mixing ratio, biopolymer concentration), and processing parameters (i.e. temperature, mechanical stresses, pressure) to generate bulk protein-polysaccharide matrices with different morphological features. The need for a combinatorial approach is then demonstrated by reviewing in detail current mixed protein-polysaccharide applications that increasingly make use of this. In the process, open scientific questions that will need to be addressed in the future are highlighted.

Primary resistance mechanism of the canine distemper virus fusion protein against a small-molecule membrane fusion inhibitor.

Morbilliviruses (e.g. measles virus [MeV] or canine distemper virus [CDV]) employ the attachment (H) and fusion (F) envelope glycoproteins for cell entry. H protein engagement to a cognate receptor eventually leads to F-triggering. Upon activation, F proteins transit from a prefusion to a postfusion conformation; a refolding process that is associated with membrane merging. Small-molecule morbilliviral fusion inhibitors such as the compound 3G (a chemical analog in the AS-48 class) were previously generated and mechanistic studies revealed a stabilizing effect on morbilliviral prefusion F trimers. Here, we aimed at designing 3G-resistant CDV F mutants by introducing single cysteine residues at hydrophobic core positions of the helical stalk region. Covalently-linked F dimers were generated, which highlighted substantial conformational flexibility within the stalk to achieve those irregular F conformations. Our findings demonstrate that "top-stalk" CDV F cysteine mutants (F-V571C and F-L575C) remained functional and gained resistance to 3G. Conversely, although not all "bottom-stalk" F cysteine variants preserved proper bioactivity, those that remained functional exhibited 3G-sensitivity. According to the recently determined prefusion MeV F trimer/AS-48 co-crystal structure, CDV residues F-V571 and F-L575 may directly interact with 3G. A combination of conformation-specific anti-F antibodies and low-resolution electron microscopy structural analyses confirmed that 3G lost its stabilizing effect on "top-stalk" F cysteine mutants thus suggesting a primary resistance mechanism. Overall, our data suggest that the fusion inhibitor 3G stabilizes prefusion CDV F trimers by docking at the top of the stalk domain.