Pulse seeds as promising and sustainable source of ingredients with naturally bioencapsulated nutrients: Literature review and outlook.

Pulse seeds are nutritious and sustainable matrices with a high level of intrinsic microstructural complexity. They contain high-quality plant-based protein and substantial amounts of slowly digestible starch and dietary fiber. Starch and protein in pulses are located inside cotyledon cells that survive cooking and subsequent mechanical disintegration, hence preserving natural nutrient bioencapsulation. In this context, several authors have explored a number of techniques to isolate individual cotyledon cells from these seeds, aiming to unveil their digestive and physicochemical properties. In recent years, isolated pulse cotyledon cells are also being highlighted as promising novel ingredients that could improve the nutritional properties of traditionally consumed food products. Even more, they could enable to implement a strategy for increasing pulse intake in populations where these seeds have not been traditionally consumed. This review mainly focuses on the reported digestive, physicochemical, and technofunctional properties of pulse cotyledon cells isolated through different techniques, preceded by a descriptive summary of the nutritional properties, structural organization, and traditional process chain of pulse seeds. It also offers an outlook of research directions to take, based on the identified research gaps. All in all, it is clear that isolation of pulse cotyledon cells using diverse techniques constitutes a promising strategy for the development of pulse-based ingredients where natural bioencapsulation of macronutrients is preserved. However, much more research is needed at the level of ingredient characterization to better understand the effect of starting pulse seed material, isolation technique, and isolation conditions on the nutritional and functional properties of the finished product(s) where the isolated cells are (to be) used.

How postharvest variables in the pulse value chain affect nutrient digestibility and bioaccessibility.

Pulses are increasingly being put forward as part of healthy diets because they are rich in protein, (slowly digestible) starch, dietary fiber, minerals, and vitamins. In pulses, nutrients are bioencapsulated by a cell wall, which mostly survives cooking followed by mechanical disintegration (e.g., mastication). In this review, we describe how different steps in the postharvest pulse value chain affect starch and protein digestion and the mineral bioaccessibility of pulses by influencing both their nutritional composition and structural integrity. Processing conditions that influence structural characteristics, and thus potentially the starch and protein digestive properties of (fresh and hard-to-cook [HTC]) pulses, have been reported in literature and are summarized in this review. The effect of thermal treatment on the pulse microstructure seems highly dependent on pulse type-specific cell wall properties and postharvest storage, which requires further investigation. In contrast to starch and protein digestion, the bioaccessibility of minerals is not dependent on the integrity of the pulse (cellular) tissue, but is affected by the presence of mineral antinutrients (chelators). Although pulses have a high overall mineral content, the presence of mineral antinutrients makes them rather poorly accessible for absorption. The negative effect of HTC on mineral bioaccessibility cannot be counteracted by thermal processing. This review also summarizes lessons learned on the use of pulses for the preparation of foods, from the traditional use of raw-milled pulse flours, to purified pulse ingredients (e.g., protein), to more innovative pulse ingredients in which cellular arrangement and bioencapsulation of macronutrients are (partially) preserved.

How Cooking Time Affects In Vitro Starch and Protein Digestibility of Whole Cooked Lentil Seeds versus Isolated Cotyledon Cells.

Lentils are sustainable sources of bioencapsulated macronutrients, meaning physical barriers hinder the permeation of digestive enzymes into cotyledon cells, slowing down macronutrient digestion. While lentils are typically consumed as cooked seeds, insights into the effect of cooking time on microstructural and related digestive properties are lacking. Therefore, the effect of cooking time (15, 30, or 60 min) on in vitro amylolysis and proteolysis kinetics of lentil seeds (CL) and an important microstructural fraction, i.e., cotyledon cells isolated thereof (ICC), were studied. For ICC, cooking time had no significant effect on amylolysis kinetics, while small but significant differences in proteolysis were observed (p < 0.05). In contrast, cooking time importantly affected the microstructure obtained upon the mechanical disintegration of whole lentils, resulting in significantly different digestion kinetics. Upon long cooking times (60 min), digestion kinetics approached those of ICC since mechanical disintegration yielded a high fraction of individual cotyledon cells (67 g/100 g dry matter). However, cooked lentils with a short cooking time (15 min) showed significantly slower amylolysis with a lower final extent (~30%), due to the presence of more cell clusters upon disintegration. In conclusion, cooking time can be used to obtain distinct microstructures and digestive functionalities with perspectives for household and industrial preparation.

Size exclusion chromatography to evaluate in vitro proteolysis: A case study on the impact of microstructure in pulse powders.

Cellular pulse ingredients are increasingly being studied but little knowledge on their proteolysis patterns upon digestion is available. This study investigated a size exclusion chromatography (SEC) approach to study in vitro protein digestion in chickpea and lentil powders, providing novel insights into proteolysis kinetics and the evolution of molecular weight distributions in the (solubilized) supernatant and (non-solubilized) pellet fractions. For the quantification of proteolysis, SEC-based analysis was compared to the commonly used OPA (o-phthaldialdehyde) approach and nitrogen solubilized upon digestion, leading to highly correlated proteolysis kinetics. Generally, all approaches confirmed that microstructure dictated proteolysis kinetics. However, SEC analysis delivered an additional level of molecular insight. For the first time, SEC revealed that while bioaccessible fractions reached a plateau in the small intestinal phase (around 45-60 min), proteolysis continued in the pellet, forming smaller but mostly insoluble peptides. SEC elutograms showed pulse-specific proteolysis patterns, unidentified using other current state-of-the-art methods.

In vitro digestion of protein and starch in sponge cakes formulated with pea (Pisum sativum L.) ingredients.

This study investigated the in vitro digestion of purified pea fractions (protein isolate and starch) in sponge cakes when compared to unrefined pea flour and to the whole wheat flour and purified maize starch commonly used in the food industry. Proteins in the wheat cake were hydrolysed more rapidly than those in cakes made with either pea flour or a combination of pea proteins and purified starch. In absolute terms, however, more readily bioaccessible protein was released from these pea cakes (by around 40%). By contrast, cakes containing wheat flour or maize starch were more susceptible to amylolysis compared to those based on pea starch in the form of the purified ingredient or whole flour. This could be attributed to a higher proportion of amylose and resistant starch in the pea cakes as well as structural characteristics that might have decelerated enzyme-substrate interactions. Interestingly, similar digestion patterns were observed regarding the purified pea ingredients and unrefined whole pea flour. It was therefore concluded that pea ingredients, and particularly the less purified and thus more sustainable whole pea flour, are promising plant-based alternatives for use in gluten-free baked products.

Utilizing Hydrothermal Processing to Align Structure and In Vitro Digestion Kinetics between Three Different Pulse Types.

Processing results in the transformation of pulses' structural architecture. Consequently, digestion is anticipated to emerge from the combined effect of intrinsic (matrix-dependent) and extrinsic (processed-induced) factors. In this work, we aimed to investigate the interrelated effect of intrinsic and extrinsic factors on pulses' structural architecture and resulting digestive consequences. Three commercially relevant pulses (chickpea, pea, black bean) were selected based on reported differences in macronutrient and cell wall composition. Starch and protein digestion kinetics of hydrothermally processed whole pulses were assessed along with microstructural and physicochemical characteristics and compared to the digestion behavior of individual cotyledon cells isolated thereof. Despite different rates of hardness decay upon hydrothermal processing, the pulses reached similar residual hardness values (40 N). Aligning the pulses at the level of this macrostructural property translated into similar microstructural characteristics after mechanical disintegration (isolated cotyledon cells) with comparable yields of cotyledon cells for all pulses (41-62%). We observed that processing to equivalent microstructural properties resulted in similar starch and protein digestion kinetics, regardless of the pulse type and (prolonged) processing times. This demonstrated the capacity of (residual) hardness as a food structuring parameter in pulses. Furthermore, we illustrated that the digestive behavior of isolated cotyledon cells was representative of the digestion behavior of corresponding whole pulses, opening up perspectives for the incorporation of complete hydrothermally processed pulses as food ingredients.

Effect of processing and microstructural properties of chickpea-flours on in vitro digestion and appetite sensations.

Nowadays, pulse flours are ingredients that are more and more used as substitutes in traditional staples (i.e., pasta, bread). In this study, cellular chickpea-flour was used as an ingredient to replace conventional raw-milled chickpea-flour in suspensions and semi-solid purees. The contribution of cellular integrity on in vitro macronutrient digestion and the subsequent effect on in vivo appetite sensations were investigated. Alternating the flour preparation sequence by interchanging hydrothermal treatment and mechanical disintegration (thermo-mechanical treatment) resulted in three chickpea-flours with distinct levels of cellular integrity, and thus nutrient accessibility. The study showed that cellular integrity in chickpea-flours was preserved upon secondary hydrothermal treatment and led to significant attenuation of in vitro macronutrient digestion as compared to conventional chickpea-flour. In a randomized crossover design, significant increase of mean in vivo subjective appetite sensations satiety and fullness along with decreases in hunger, desire to eat, and prospective food consumption were achieved when cellular integrity was kept without an effect on palatability and appearance of the purees (n = 22). In vitro digestion along with microstructural assessment confirmed the importance of cellular integrity for attenuating macronutrient digestion and thereby contributing to enhanced subjective satiety and fullness in pulses. Overall, this study highlights the promising potential of altarenating the flour preparation sequence resulting in macronutrient and energy-matched flours with different nutrient encapsulation which lead to different in vitro digestion kinetics and in vivo appetite sensations.

In vitro protein and starch digestion kinetics of individual chickpea cells: from static to more complex in vitro digestion approaches.

Attention has been given to more (semi-)dynamic in vitro digestion approaches ascertaining the consequences of dynamic in vivo aspects on in vitro digestion kinetics. As these often come with time and economical constraints, evaluating the consequence of stepwise increasing the complexity of static in vitro approaches using easy-to-handle digestion set-ups has been the center of our interest. Starting from the INFOGEST static in vitro protocol, we studied the influence of static gastric pH versus gradual gastric pH change (pH 6.3 to pH 2.5 in 2 h) on macronutrient digestion in individual cotyledon cells derived from chickpeas. Little effect on small intestinal proteolysis was observed comparing the applied digestion conditions. Contrary, the implementation of a gradual gastric pH change, with and without the addition of salivary α-amylase, altered starch digestion kinetics rates, and extents by 25%. The evaluation of starch and protein digestion, being co-embedded in cotyledon cells, did not only confirm but account for the interdependent digestion behavior. The insights generated in this study demonstrate the possibility of using a hypothesis-based approach to introduce dynamic factors to in vitro models while sticking to simple and cost-efficient set-ups.

In vitro starch and protein digestion kinetics of cooked Bambara groundnuts depend on processing intensity and hardness sorting.

When pulse seeds from a single batch are cooked, considerable variability of hardness values in the population is usually observed. Sorting the seeds into hardness categories could reduce the observed diversity and increase uniformity. Therefore, we investigated the effect of processing intensity whether or not combined with sorting into hardness categories on the in vitro starch and protein digestion kinetics of cooked Bambara groundnuts (cooking times 40 min and 120 min). The average hardness values were 89 ± 32 N and 42 ± 20 N for 40 min and 120 min cooking time, respectively. The high standard deviation of hardness for each cooking time revealed a high level of diversity amongst the seeds. Individual cells were isolated from (non-)sorted seeds before simulating digestion. The estimated lag phase describing the initial phase of starch digestion was not significantly different despite the processing intensity or the hardness category, implying that cell wall barrier properties for these samples were not majorly different. However, the rate constants and the extents of starch digestion of samples cooked for 40 min were significantly higher for the low hardness (50-65 N) compared to the high hardness (80-95 N) category (0.71 vs 1.02 starch%/min and 63 vs 77%, respectively). Kinetic evaluation of digested soluble protein (after acid hydrolysis of the digestive supernatant) showed that low hardness samples were digested faster than high hardness samples (0.037 vs 0.050 min-1). The faster protein hydrolysis in the low hardness samples was accompanied by faster starch digestion, indicating the possible role of the protein matrix barrier. Individual cells of comparable hardness obtained from the two different processing times had similar starch and protein digestion kinetics. Our work demonstrated that, beyond cooking time, hardness is a suitable food design attribute that can be used to modulate starch and protein digestion kinetics of pulse cotyledon cells.