Submerged fermentation of lentil protein isolate and its impact on protein functionality, nutrition, and volatile profiles.

Fermentation of pulses as a clean processing technique has been reported to have a favorable impact on the functional and nutritional quality of the starting materials. Compared to commonly fermented pulses such as peas and chickpeas, limited information is available on the effect of fermentation on lentils, especially when using a high protein isolate (>80% protein) as compared to seeds or flours. Therefore, in the present work, lentil protein isolate was used as a feedstock for submerged fermentation with Aspergillus niger, Aspergillus oryzae, or Lactobacillus plantarum. After 48 h, the samples showed increased protein content with enhanced solubility and oil-holding capacity. Controlled fermentation, as opposed to spontaneous fermentation, maintained the high foaming capacity; however, all fermented samples had lower foam and emulsion stabilizing properties and reduced water-holding capacity compared to the control. The fermented proteins were also less digestible, possibly due to an increase in phenolics and saponins. New volatile compounds were identified in fermented samples that show promise for improved sensory attributes. Significant differences were observed in specific quality attributes depending on the microbial strain used. Further research is required to better understand the fermentative metabolism of microbial communities when provided high-protein lentil ingredients as growth substrates. PRACTICAL APPLICATION: Fermented lentil protein isolate has promising flavor profiles that may improve its sensory properties for food application.

Effect of roasting pulse seeds at different tempering moisture on the flour functional properties and nutritional quality.

Knowledge on the functional and nutritional properties of wet roasted pulses can increase the utilization of processed pulses as ingredients in food products. This study investigated the effects of tempering different pulse [chickpea (CP), green lentil (GL), navy bean (NB) and yellow pea (YP)] seeds to 20 or 30% moisture prior to roasting (160℃ for 30 min) on the functional properties and nutritional quality of their resulting flours. The surface charge of each pulse remained the same (p > 0.05) after wet roasting and there were no significant (p > 0.05) differences between the different raw pulse flours. The oil holding capacity (OHC) of GL (~2 g/g) was not improved by wet roasting (p > 0.05) whereas the other pulses generally had better OHC for one or both of the tempering moistures used prior to roasting. Foaming properties of all pulses decreased after heat treatment with the exception of both foaming capacity (107%) and stability (~71%) for GL tempered to 20% moisture prior to roasting (p > 0.05). Raw GL had inferior foaming properties compared to the other raw pulse flours (p < 0.001). Emulsion properties of the wet roasted pulses were similar to those of the control (raw flour) for each pulse. Solubility decreased with roasting regardless of the tempering moisture (p < 0.05) whereas in general the in vitro protein digestibility increased. Small improvements (2.4-6.9% increase) in the in vitro protein digestibility-corrected amino acid score were found for GL and NB tempered to 20% moisture before roasting and roasted YP at either moisture content (p < 0.05). Wet roasting increased (p < 0.05) the rapidly digestible starch content, more so with a tempering moisture of 30%. Overall the results from this study will allow for the utilization of wet roasted pulses as ingredients based on their functional properties and protein quality.

Effect of different levels of esterification and blockiness of pectin on the functional behaviour of pea protein isolate-pectin complexes.

This research examines changes to the functional (solubility, emulsifying and foaming) properties of pea protein isolate when complexed with commercial citrus pectin of different structural attributes. Specifically, a high methoxy (P90; degree of esterification: 90.0%; degree of blockiness: 64.5%; galacturonic acid content 11.4%) and low methoxy (P29; degree of esterification: 28.6%; degree of blockiness: 31.1%; galacturonic acid: 70%) pectin at their optimum mixing ratios with pea protein isolate (4:1 pea protein isolate to P90; 10:1 pea protein isolate to P29) were assessed at the pHs associated with critical structure forming events during the complexation process (soluble complexation (pHc), pH 6.7 and 6.1; insoluble complex formation (pHϕ1), pH 4.0 and 5.0; maximum complexation (pHopt), pH 3.5 and 3.8; dissolution of complexes, pH 2.4 and 2.1; for admixtures of pea protein isolate-P90 and pea protein isolate-P29, respectively). Pea protein isolate solubility was improved from 41 to 73% by the presence of P90 at pH 6.0 and was also moderately increased at pH 4.0 and pH 5.0 by P90 and P29, respectively. The emulsion stability of both pea protein isolate-pectin complexes was higher than the homogeneous pea protein isolate at all critical pHs except pHopt as well as pHc for pea protein isolate-P29 only. P90, with the higher level blockiness and esterification, displayed better foaming properties at the maximal complexation pH when complexed with pea protein isolate than pea protein isolate-P29 or pea protein isolate alone. However at pHϕ2, pea protein isolate-P29 admixtures produced foams with 100% stability, increasing pea protein isolate foam stability by 85%. The enhanced functionality of pea protein isolate-pectin complexes based on the type of pectin used at critical pHs indicates they may be useful biopolymer ingredients in plant protein applications.

Enzymatic cross-linking to improve the handling properties of dough prepared within a normal and reduced NaCl environment.

This research examines the use of three enzymes [glucose oxidase (GOX), hexose oxidase (HOX), and xylanase (XYL)] and their combinations [GOX-XYL and HOX-XYL] on the dough handling properties of CDC Plentiful and Stettler wheat cultivars prepared at reduced (1.0% wt. by flour) and normal (2.0% wt. by flour) NaCl levels. Properties investigated include dough rheology, stickiness, and ratio of resistance to extension and extensibility. The inclusion of XYL and its combinations with GOX and HOX increased the stickiness, yielded lower dough strength indicated by rheology, and reduced the ratio of resistance to extension and extensibility. The inclusion of oxidative enzymes yielded a stronger dough, where HOX addition to dough had the lowest stickiness values and highest |G*| values, whereas GOX addition led to the highest ratio of resistance to extension-extensibility. NaCl only had minor effects overall on dough strength and stickiness for the cultivars studied. Overall, superior dough handling properties were observed with oxidative enzyme addition (GOX and HOX) suggesting that the increased crosslinking that occurs could aid in improving low sodium bread dough properties.

The functional attributes of Peruvian (Kankolla and Blanca juli blend) and Northern quinoa (NQ94PT) flours and protein isolates, and their protein quality.

The overall goal of this research was to examine differences in the composition, functionality and protein quality between Peruvian (PQ) and Northern (NQ) quinoa flours, and their isolates prepared by alkaline extraction/isoelectric precipitation. In the case of the flours, PQ and NQ were comprised of 13.6% and 12.8% protein, respectively. Water hydration (mean value = 1.65 g/g) and oil holding capacities (mean value = 1.75 g/g) of both flours were similar, whereas solubility increased from pH 3 to 7 for both flours, but was higher for PQ. Flours were non-foaming at pH 3, but showed increased foam capacity as the pH increased from 5 to 7, but was higher for PQ. Similar foam stability was found for both flours. Emulsion stability (ES) was similar for both flours, and increased from pH 3 to pH 5/7. In the case of the isolates, water hydration capacity was greater for PQ (4.75 g/g) than NQ (2.85 g/g), whereas oil holding capacity was similar (mean value = 8.6 g/g). For both isolates, solubility was minimum at pH 5.0 and maximum at pH 3/7, with NQ being higher. Isolates showed 2-3 times the foam capacity as flours, the magnitude of which was cultivar and pH dependent. Foam stability was lower at pH 5 than at pH 3/7, whereas ES followed a similar pH effect. Tyrosine and phenylalanine were limiting in both flours, whereas threonine was limiting in both isolates. In vitro protein digestibility corrected amino acid scores for the flours was higher for PQ (56.8%) than NQ (47.7%); however, the reverse was observed for the isolates (NQ, 62.1%; PQ, 58.9%).

Effect of extrusion conditions on the physical properties of desi chickpea-barley extrudates and quality attributes of their resulting flours.

In this study, response surface methodology (RSM) was used to evaluate the effect of extrusion conditions on physical properties of chickpea:barley extrudates (60:40), and the resulting protein quality of their flours. Barrel temperature (150-170°C) and moisture content (16-20%) were chosen as independent variables to generate a central composite design. Hardness, expansion index, bulk density, and protein quality were analyzed as responses parameters. Expansion was found to be higher at lower temperatures and higher moisture for the 60:40 chickpea:barley blend; bulk density became reduced with increased moisture; and hardness was found to increase at higher temperatures and lower moistures. The protein quality of their resulting flours was found to be greater at moisture contents higher than 16%. The composition, protein quality, and functional attributes were also examined for raw and precooked flours of chickpea, barley, and their blend at the center point of the RSM design (18% moisture, 160°C). Extrusion also leads to improved water hydration capacities and reduced viscosities for precooked individual and blended flours relative to the raw. Moreover, extrusion also led to improved protein quality in the chickpea and chickpea-barley blend, but not the individual barley flour.

Protein quality and physicochemical properties of commercial cricket and mealworm powders.

The pressing need for protein supply growth gives rise to alternative protein sources, such as insect proteins. Commercial cricket and mealworm powders were examined for their protein quality, surface charge and functional attributes. Both insect powders had similar proximate compositions with protein and ash contents of ~ 66% db (dry weight basis) and 5% db, respectively, however cricket powder contained more lipid (16.1%, db) than mealworm powder (13.7%, db). Mealworm protein had an amino acid score of 0.71 and was first limiting in lysine, whereas cricket protein was first limiting in tryptophan with an amino acid score of 0.85. In vitro protein digestibility values of 75.7% and 76.2%, and in vitro protein digestibility corrected amino acid scores of 0.54 and 0.65, were obtained for mealworm and cricket powders, respectively. Zeta potential measurements gave isoelectric points near pH 3.9 for both insect powders. Mealworm and cricket powders had water hydration capacities of 1.62 g/g and 1.76 g/g, respectively, and oil holding capacities of 1.58 g/g and 1.42 g/g, respectively. Both insect proteins had low solubility (22-30%) at all pHs (3.0, 5.0, and 7.0) measured. Cricket powder had a foaming capacity of 82% and foam stability of 86%, whereas mealworm powder was non-foaming. Values for commercial pea and faba bean protein concentrates were reported for comparative purposes. The insect proteins had similar protein quality as the pulse proteins and had higher solubility at pH 5.0 but were much less soluble at pH 7.0.

Effect of glycerol on the physicochemical properties of films based on legume protein concentrates: A comparative study.

The overall goal of this research was to examine the mechanical, water vapor barrier properties and opacity of films prepared using legume protein concentrates (faba bean, pea, lupin, lentil, and soy) as a function of glycerol concentration (50, 75, or 100% [wt/wt]-relative to the protein weight). Overall, tensile strength (TS) decreased with increasing glycerol concentration, whereas tensile elongation (TE) and water vapor permeability (WVP) increased with increasing glycerol concentration. Film opacity was independent of glycerol concentration. The effect of protein-type varied considerably depending on the functional property of the film being measured; TS was greatest with faba bean and lowest with lupin, whereas TE was highest for pea, and lowest for soy. Lentil protein films had considerably higher WVP, at the 100% glycerol concentration, as compared to the other protein concentrates. Findings from this study indicate that legume protein concentrates are capable of forming biodegradable, edible films. Overall, pea protein concentrate films showed the most promise for application in terms of strength, elongation, and moisture barrier properties.

Effect of alkaline de-esterified pectin on the complex coacervation with pea protein isolate under different mixing conditions.

High methoxy citrus pectin (UM88) was saponified to produce modified pectin [M(72, 42, and 9)], with different levels of degree of esterification (DE), to investigate the complex coacervation of pea protein isolate (PPI) with pectin [UM88 and M(72, 42, and 9)]. Regardless of the DE value of pectin, the critical pH corresponding to when insoluble complexes form shifted to higher pH as the mixing ratio increased. The maximum amount of coacervates formed at a biopolymer-mixing ratio of 8:1, 8:1, 10:1 and 15:1 for PPI with UM88, M72, M42, and M9, respectively. Maximum interactions for the protein-pectin admixtures occurred between pH 3.70 and 3.85. PPI complexed with modified pectin displayed greater interactions under their optimal mixing conditions compared to the unmodified pectin. The de-esterification of pectin resulted in more rigid and stiffer pectin, which enhanced its interaction with PPI by shifting the critical parameters to a higher value.

Effect of Lactobacillus plantarum Fermentation on the Surface and Functional Properties of Pea Protein-Enriched Flour.

The effect of Lactobacillus plantarum fermentation on the functional and physicochemical properties of pea protein-enriched flour (PPF) was investigated. Over the course of the fermentation the extent of hydrolysis increased continuously until reaching a maximum degree of hydrolysis of 13.5% after 11 h. The resulting fermented flour was then adjusted to either pH=4 or 7 prior to measuring the surface and functional attributes as a function of fermentation time. At pH=4 surface charge, as measured by zeta potential, initially increased from +14 to +27 mV after 1 h of fermentation, and then decreased to +10 mV after 11 h; whereas at pH=7 the charge gradually increased from -37 to -27 mV over the entire fermentation time. Surface hydrophobicity significantly increased at pH=4 as a function of fermentation time, whereas at pH=7 fermentation induced only a slight decrease in PPF surface hydrophobicity. Foam capacity was highest at pH=4 using PPF fermented for 5 h whereas foam stability was low at both pH values for all samples. Emulsifying activity sharply decreased after 5 h of fermentation at pH=4. Emulsion stability improved at pH=7 after 5 h of fermentation as compared to the control. Oil-holding capacity improved from 1.8 g/g at time 0 to 3.5 g/g by the end of 11 h of fermentation, whereas water hydration capacity decreased after 5 h, then increased after 9 h of fermentation. These results indicate that the fermentation of PPF can modify its properties, which can lead towards its utilization as a functional food ingredient.

Effect of Fermentation on the Protein Digestibility and Levels of Non-Nutritive Compounds of Pea Protein Concentrate.

In order to determine the impact of fermentation on protein quality, pea protein concentrate (PPC) was fermented with Lactobacillus plantarum for 11 h and total phenol and tannin contents, protease inhibitor activity, amino acid composition and in vitro protein digestibility were analyzed. Phenol levels, expressed as catechin equivalents (CE), increased on dry mass basis from 2.5 at 0 h to 4.9 mg CE per 1 g of PPC at 11 h. Tannin content rose from 0.14 at 0 h to a maximum of 0.96 mg CE per 1 g of PPC after 5 h, and thereafter declined to 0.79 mg/g after 11 h. After 9 h of fermentation trypsin inhibitor activity decreased, however, at all other fermentation times similar levels to the PPC at time 0 h were produced. Chymotrypsin inhibitor activity decreased from 3.7 to 1.1 chymotrypsin inhibitory units (CIU) per mg following 11 h of fermentation. Protein digestibility reached a maximum (87.4%) after 5 h of fermentation, however, the sulfur amino acid score was reduced from 0.84 at 0 h to 0.66 at 11 h. This reduction in sulfur content altered the in vitro protein digestibility-corrected amino acid score from 67.0% at 0 h to 54.6% at 11 h. These data suggest that while fermentation is a viable method of reducing certain non-nutritive compounds in pea protein concentrate, selection of an alternative bacterium which metabolises sulfur amino acids to a lesser extent than L. plantarum should be considered.

Effect of the degree of esterification and blockiness on the complex coacervation of pea protein isolate and commercial pectic polysaccharides.

The complex coacervation of pea protein isolate (PPI) with commercial pectic polysaccharides [high methoxy citrus pectin (P90, 90 representing DE), apple pectin (P78) sugar beet pectin (P62), low methoxy citrus pectin (P29)] of different degrees of esterification (DE) [and galacturonic acid content (GalA)] and blockiness (DB), was investigated. The maximum amount of coacervates formed at a biopolymer weight mixing ratio of 4:1 for all PPI-pectin mixtures, with the exception of PPI-P29 where maximum coacervation occurred at the 10:1 mixing ratio. The pH at which maximum interactions occurred was pH 3.4-3.5 (PPI: P90/P78) and 3.7-3.8 (PPI: P62/P29). PPI complexed with pectins with high levels of DE (low levels of GalA) and DB displayed greater interactions at optimal mixing conditions compared to pectin having lower levels of esterification and blockiness. The addition of P78 to PPI greatly increased protein solubility at pH 4.5.

Effect of genotype on the physicochemical and functional attributes of faba bean (Vicia faba L.) protein isolates.

The functionality and surface characteristics of protein isolates prepared from seven genotypes of faba bean were investigated. All factors studied were independent of genotype. The average protein and isolate (~94% protein) yields were ~77% and ~25%, respectively, using an alkaline extraction process followed by isoelectric precipitation. The overall averages were: surface charge +22.1 mV, surface hydrophobicity 47.2 arbitrary units, and surface and interfacial tensions of 65.0 mN/m and 10.7 mN/m, respectively. The ratio of the major globulin fractions (legumin:vicilin) shifted from 3.8 for the flours to 4.5 in the isolates. Average values for foaming capacity and stability, emulsion capacity, creaming stability, oil holding capacity, emulsifying activity and stability indices, and solubility were 162.0%, 65.0%, 184.0 g/g, 94.0%, 5.7 g/g, 13.0 m2/g, 10.7 min, and 81.0%, respectively. The lack of significant varietal effects would be advantageous to food processors as the source of the feedstock would not affect ingredient functionality.

Nature of protein-protein interactions during the gelation of canola protein isolate networks.

The nature of interactions involved during the gelation of a canola protein isolate was investigated using rheology and fractal imaging at neutral pH as a function of protein concentration (5.0-9.0% w/w). The onset of denaturation and the denaturation temperature by differential scanning calorimetry for canola protein isolate (CPI; 98.2% protein) was 78.6°C and 87.1°C, respectively. Rheological testing determined the gelation temperature (Tgel) to be ~87-90°C for all concentrations. The log % strain at break increased from 1.70 to 1.80 as CPI concentration increased from 5.0 to 7.0% (w/w). Rheological testing of CPI in the presence of destabilizing agents, NaCl (0.1 and 0.5M), urea (0.1, 0.5, 1 and 5M) and 2-ß-mercaptoethanol (0.1 and 2%), was performed. Samples with NaCl and urea (0.1-1M) had similar temperature profiles and Tgel values to CPI alone whereas no gel was formed with the addition of 5M urea and 2-ß-mercaptoethanol reduced the strength of the gel network. Fractal dimension and lacunarity was analyzed using CLSM imaging. The fractal dimension value for all CPI concentrations was ~1.5. The lacunarity of the gel decreased from 0.62 to 0.41 as the concentration of CPI increased from 5 to 7% (w/w). Mechanistic understanding of CPI aggregation and network formation will enable the food industry to better tailor food structure when CPI is present as ingredient.