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Characterization and modulation of technofunctional properties of pea proteins
(2023) Moll, Pascal Bernd; Weiss, Jochen
Plant-derived ingredients for food formulation have gained increasing interest in recent years as animal products pose a higher burden on the environment. Among plant proteins, those from pea (Pisum sativum L.) are of particular interest because of their low allergenicity, low cost, high availability, and good reputation among consumers. However, the technofunctionality of pea proteins is often inferior to animal-derived proteins limiting a more widespread use in food products. These technofunctional properties include - among others - foaming, gelling, and binding of other ingredients and it depends on the food product, which functionality food scientists must utilize and optimize. Cost effective approaches to improve the technofunctionality of pea proteins are therefore desirable and would allow the industry to further implement the use of sustainable ingredients in foods. In line with these overall goals, the aim of the first section of this thesis was to characterize a commercial pea protein isolate and to modulate the physicochemical and technofunctional properties through homogenization for foaming application. The main goal of the second section was to mix pea proteins with pectin to obtain a suitable binder with desired properties for the application in meat alternatives. The mixing approach was based on previous research data that had shown that interacting protein-polysaccharide systems display a synergistic behaviour in terms of their functional properties. First section: Foams are two phase systems consisting of gas bubbles that are stabilized by surface-active ingredients such as proteins in the discontinuous, aqueous phase. The physico-chemical properties of proteins such as their solubility determines foaming performance. In Chapter I, a commercial pea protein isolate was fractionated into a water-soluble and a water-insoluble fraction for characterization. Although the two fractions were similar in protein composition, they showed distinct differences in physicochemical properties. For instance, the particle size of soluble pea proteins was around 40-50 µm at acidic pH (3-5), while no measurable particles were detected at neutral The insoluble pea proteins were large at pH 3 and 7 (> 80 µm) and ca. 40-50 µm close to their isoelectric point at pH 5. The results suggest that commercial pea protein isolates consisted of several fractions with differences in their physico-chemical properties. The yield of the water-insoluble fraction was higher and therefore used in Chapter II, where experimental results illustrated that dispersions of insoluble pea protein aggregates (5% w/w, pH 7) could be disrupted from 180 ± 40 µm (control) to 0.2 ± 0.0 µmm upon homogenization at pressures ≥ 125 MPa. This was attributed to a cleavage of intermolecular interactions such as disulphide bonds, hydrogen bonds, and hydrophobic interactions. The decrease in insoluble pea protein aggregate size was accompanied by an increase in solubility from 23 ± 1% to ≥ 80% that may be beneficial for its technofunctionality. Consequently, homogenization was applied to the same material at pH 3 and 5 with the aim of investigating its foaming performance in Chapter III. In general, unhomogenized dispersions of pea protein aggregates (5% w/w, pH 3 or 5) did not foam at both tested pH values due to large pea protein aggregates with low solubility and surface activity. At pH 3, the dissociation of pea protein aggregates into smaller, more soluble, and more surface-active proteins was responsible for a high foam capacity (FC = 360-520%) with medium foam stability as measured by drainage (FS = 19-30 min). Only a limited particle size reduction upon homogenization was observed at pH 5, which was close to the isoelectric point of the pea proteins. Nevertheless, the still large aggregates consisted of re-aggregated smaller protein particles that were able to form a smaller amount of rather stable foams with thick interfacial films (FC = 213-246%, FS = 32-42 min). Overall, homogenization of insoluble pea protein aggregates was shown to change its physicochemical properties thereby benefitting technofunctional properties such as foaming. Second section: Another technofunctionality of interest is binding of different structural elements in e.g., meat alternatives. For this, the binder must be i.) sticky to glue heterogeneous components together and ii.) able to readily solidify upon further processing thereby ensuring a coherent bulk matrix. In Chapter IV, the influence of pH (3.50, 4.75, 6.00) and biopolymer concentration (17.5-50.0% w/w) on the stickiness of a pea protein isolate – apple pectin mixture (mixing ratio r = 6:1) was investigated. It was found that biopolymer concentrations of 17.5-20.0% w/w led to low stickiness due to a lack of cohesive forces (WoA = 0.29-0.51 mJ). At high biopolymer concentrations of 40-50% w/w, the biopolymer mixtures were also not sticky because of adhesion being limited (WoA = 0.02-0.05 mJ). There was a good balance of adhesion and cohesion that facilitated a high stickiness (WoA = 0.48-0.65 mJ) at intermediate concentrations of 25-30% w/w, which was also indicated by a viscoelastic behavior (G’ ≈ G’’). At those concentrations, the mixtures at pH 6 were stickier due to increased swelling of the pea proteins. The importance of viscoelasticity for stickiness of biopolymer mixtures was confirmed in Chapter V, where pea protein isolate and apple pectin (25% w/w, pH 6) were mixed in different ratios r. Mixtures of pea protein and apple pectin and particularly the sample with r = 2:1 possessed high stickiness due to the development of a multiphase morphology that allowed for a good balance of adhesion and cohesion with distinct frequency dependency. Pea protein alone (r = 1:0, c = 25% w/w) had an elastic but soft texture with low stickiness due to limited viscous properties, whereas a sample solely consisting of apple pectin (r = 0:1, c = 25% w/w) was also not sticky because of its high cohesion and stiffness. The results of Chapter VI revealed that pea protein homogenization prior to mixing with apple pectin led to smaller protein particles in the blend that contributed to a higher cohesive strength. Interestingly, vacuum-dried pea proteins resulted in a higher network strength as this drying method prevented reaggregation of small protein particles to a higher extent as compared to freeze-drying. Overall, the mixture with homogenized and vacuum-dried pea proteins was nearly twice as sticky as the mixture with untreated pea proteins. In Chapter VII, sticky mixtures of different pea protein preparations (soluble, homogenized and unhomogenized pea proteins) and pectin (25% w/w, pH 6, r = 2:1) were tested for their ability to solidify upon different treatments, namely heating as well as the addition of transglutaminase, laccase, calcium, and combinations thereof. Calcium was found to facilitate crosslinking of pectin chains and thus induced solidification of the mixtures. For instance, the consistency coefficient K’ increased from 2800 ± 1000 Pasn for pea protein isolate – apple pectin mixtures to around 19000 Pasn when calcium was added. Heat treatment and transglutaminase did not lead to solidification indicating that pectin made up the continuous phase. Furthermore, laccase led to the highest degree of solidification when sugar beet pectin was used (K’ > 30000 Pasn) due to ferulic acid and pea protein tyrosine crosslinking. Consequently, the sticky mixture of pea protein and sugar beet pectin (25% w/w, pH 6, r = 2:1) with the addition of laccase for solidification was identified as the most suitable binder for a bacon type meat analogue, which was the object of the study carried out in Chapter VIII. This binder had the highest binding strength (W = 2.0-4.3 mJ) between textured protein, fat mimic, and both layers at 25 °C due to the introduction of covalent bonds by laccase within the binder and between the binder and the adherends. A control sample without laccase addition had lower binding properties (W = 0.7-1.0 mJ) and the binding strength of a methylcellulose hydrogel (6% w/w) serving as benchmark was only higher between two fat mimics at 70 °C (W = 1.8 ± 1.1 mJ) due to increased hydrophobic forces. Finally, the pea protein – sugar beet pectin binder (22.5% w/w, pH 6, r = 2:1) was tested in burger patty type meat analogues to glue textured vegetable protein and fat particles together (Chapter IX). The binder system did not influence the hardness of the burger patties suggesting that this property was governed by the structural elements and not the binder. However, the cohesiveness as determined by sensory analysis was found to be superior when the pea protein – sugar beet pectin binder was used (-0.7 ± 0.2) as compared to the methylcellulose benchmark (-2.9 ± 0.3). This was attributed to the sticky character of the biopolymer mixture that enabled improved binding of the different structural elements. Overall, this novel binder based on plant-derived ingredients was shown to be applicable in different meat alternatives. Last, Chapter X reviewed the functionality and binding mechanism of currently used binders in foods and showed that stickiness, hardening/solidification, and water holding capacity are of great importance. In many food products, the binder transitions from a sticky food glue to a solid matrix triggered by different process operations that depend on the characteristics of the applied binder. From the presented results, it can be concluded that pea proteins are useful functional ingredients in various application scenarios. The desired technofunctionality can be improved through different process operations such as fractionation, homogenization, or mixing with other plant-derived ingredients. For this, knowledge regarding structure-function relationship and other influential factors is needed. In some cases – such as in binders – process operations must be well orchestrated to induce structural transitions and therefore changes in functionality at the desired time during manufacturing. Overall, the results of this thesis contributed to a better understanding for a more widespread use of pea proteins to promote a more sustainable food system. The appended graphical abstract summarizes the key steps undertaken in this thesis to come to this conclusion.
Pflanzenbauliche Untersuchungen zum ökologischen Anbau von Körnerleguminosen an sommertrockenen Standorten Südwestdeutschlands
(2007) Poetsch, Jens; Claupein, Wilhelm
Grain legumes, as nitrogen fixing crop, protein rich animal feed and marketable product are of great importance for organic agriculture. Due to staged abolition of the possibility to add non-organic products in organic animal feeding, the EU?s demand for organically produced protein feed is further increasing. Field bean (Vicia faba) and field pea (Pisum sativum) are large-scale crops but feature a limited feeding value. Lupin species (Lupinus spp.) excel by protein contents of up to 40% in the seed and higher protein value. At warmth favoured locations in southwestern Germany the valuable soybean (Glycine max) can be grown successfully and obtain above-average proceeds in natural food industry. Constraints of yield stability of grain legumes result amongst other things from frequently high weed infestation in organic cropping systems and suboptimal water supply at summer-dry locations. For lupins, moreover, particular soil requirements and the seed-borne fungal disease anthracnosis are problematic. Nitrogen residues after harvest are relevant for subsequent crop as well as groundwater protection. The presented work aimed at defining preconditions and developing cropping strategies to optimise yield stability and level of organically grown grain legumes with a main focus on summer-dry locations, to increase diversity of cultivatable crops and provide information on disposition of nitrogen residues. For this purpose from 2003 to 2005 trials at several locations as well as in greenhouse and laboratory were accomplished. Field trials on organic weed control in soybean as well as white and narrow-leafed lupin (Lupinus albus und L. angustifolius) were conducted at organically managed commercial sites in the upper rhine valley. At the same time agronomic measures for optimisation of competitiveness and machinery implementation were varied. Early high soil coverage and crop height contributed considerably to grain legumes? competitiveness. Delayed sowing at elevated temperatures supported rapid juvenile development and allowed for pre-sowing weed control. At optimum sowing date these effects may be used without yield depression or maturity problems. Reduced row distance was beneficial for optimum space utilisation and early crop closure, but effectiveness of mechanical means was highest at high row distance and large areal proportion for interrow cultivation. As an optimum compromise for grain legumes row distances of 30 - 35 cm are recommended. Optimum impact of mechanical means against weeds was achieved by combining interrow cultivation with harrow or fingerweeder. Forgoing interrow cultivation may be considered in strongly competitive crops like field bean. Lupin species appeared rather poor in competitiveness compared to other crops. Field trials on effects of cultivar and cropping strategy on overwintering and yield performance of autumn-sown field bean, field pea and white lupin were conducted at three locations. Summer drought caused substantial yield advantages of autumn-sown compared to spring-sown cultivars due to superior water supply at earlier flowering. With sufficient water supply a head start was not yield effective. Differing coincidence with pests and diseases could account for advantages (head start on aphid infestation) or disadvantages (fungal infections during winter period) of autumn-sown cultivars. Overwintering was excellent for winter field bean and good for winter field pea. For winter white lupin further trials are required. Temperatures down to -12°C were well endured by all of the three crops. The most important cropping parameter was the sowing date. Winter field bean permitted a relatively wide sowing window. Winter white lupin required strong development before winter and preferably early sowing. Sowing date of winter field pea presented an optimisation problem, because sowing too early leads to overdevelopment and reduced cold-tolerance, while sowing too late may reduce yield potential. Optimum sowing dates for southwestern Germany according to experimental results are in the range of early September (winter white lupin), mid-October (winter field bean) and late October (winter field pea). Water use efficiency may gain significantly in importance in the future. A two-year trial on cultivation prospects and yield performance of the notably drought tolerant chickpea (Cicer arietinum) in the upper rhine valley resulted in successful crop development, but problems with empty pods and inadequate grain quality. Further trials are considered promising. A field trial with white and narrow-leafed lupin confirmed that anthracnosis of lupin spreads less rapidly and yield effectively at summer-dry locations, and narrow-leafed lupin frequently stays unaffected. Laboratory studies for optimising detection methodology of the causative organism Colletotrichum lupini showed advantages of using sectioned petri dishes (quad plates), which confined propagation of disturbing organisms. A trial on seed storage under different temperatures, seed moisture contents and CO2-atmosphere produced no distinct treatment effect, but could confirm the general decrease of seed infection by storage. According to literature hot air (approx. 4 days at 65°C) also reduces seed infection effectively. Thus, storage or hot air treatment of basic seed and propagation at summer-dry locations appear as a viable over-all strategy. Difficult soil requirements of white and narrow-leafed lupin were studied by a pot trial as well as a comprehensive literature analysis. It is concluded that the so-called lime chlorosis is caused by HCO3--induced inactivation of physiologically relevant Fe(II) in the plant. Accumulation of HCO3- is basically caused by insufficient soil aeration and promoted by the presence of lime in the clay fraction. Furthermore, especially in narrow-leafed lupin, disturbances of root development are caused by high Ca-content or high and at the same time strongly buffered pH of soil solution. These conditions are often but not necessarily caused by lime. Analyses of harvest residues and soil were consulted for estimation of nitrogen dynamics. Immobilisation due to degradation of residues with high C:N ratio as well as uptake by catch crops contributed substantially to nitrogen conservation. Risk of leaching is predominantly site dependent. The over-all nitrogen balance of grain legumes when exporting the seed may be low or even negative. In conclusion, results of the presented work indicate that site adapted cropping systems with agronomic measures in the areas of crop rotation, choice of cultivar, sowing date or space allocation can still contribute considerably to yield stability in organic cultivation of grain legumes.