Plant cell wall composition modulates the gut microbiota and metabolites in in-vitro fermentation.

This research investigated the effect of different types of plant cell wall fibres, including cereal (i.e., barley, sorghum, and rice), legume (i.e., pea, faba bean, and mung bean), and tuber (potato, sweet potato, and yam) cell wall fibres on in vitro faecal fermentation profiles and gut microbiota composition. The cell wall composition, specifically the content of lignin and pectin, was found to have a significant influence on the gut microbiota and fermentation outcomes. Compared with type I cell walls (legume and tuber) which have high pectin content, the type II cell walls (cereal) which are high in lignin but low in pectin had a lower fermentation rates and less short-chain fatty acid production. The redundancy analysis showed samples with similar fibre composition and fermentation profiles clustered together, and the principal coordinate analysis revealed separation among different types of cell walls and closer proximity among the same cell wall types. These findings emphasize the importance of cell wall composition in shaping the microbial community during fermentation and contribute to a better understanding of the relationship between plant cell walls and gut health. This research has practical implications for the development of functional foods and dietary interventions.

Salt adopted in soaking solution controls the yield and starch digestion kinetics of intact pulse cotyledon cells.

Intact cellular powders have gained attention as a functional ingredient due to their lower glycemic response and potential benefits in colon. The isolation of intact cells in the laboratory and pilot plant settings is mainly achieved through thermal treatment with or without the use of limited salts. However, the effects of salt type and concentration on cell porosity, and their impact on the enzymic hydrolysis of encapsulated macro-nutrients such as starch, have been overlooked. In this study, different salt-soaking solutions were used to isolate intact cotyledon cells from white kidney beans. The use of Na2CO3 and Na3PO4 soaking treatments, with high pH (11.5-12.7) and high amount of Na ion (0.1, 0.5 M), greatly improved the yield of cellular powder (49.6-55.5 %), due to the solubilization of pectin through ß-elimination and ion exchange. Intact cell walls serve as a physical barrier, significantly reducing the susceptibility of cell to amylolysis when compared to white kidney bean flour and starch counterparts. However, the solubilization of pectin may facilitate enzyme access into the cells by enlarging cell wall permeability. These findings provide new insights into the processing optimization to improve the yield and nutritional value of intact pulse cotyledon cells as a functional food ingredient.

Hard-to-cook phenomenon in common legumes: Chemistry, mechanisms and utilisation.

Future dietary protein demand will focus more on plant-based sources than animal-based products. In this scenario, legumes and pulses (lentils, beans, chickpeas, etc.) can play a crucial role as they are one of the richest sources of plant proteins with many health benefits. However, legume consumption is undermined due to the hard-to-cook (HTC) phenomenon, which refers to legumes that have high resistance to softening during cooking. This review provides mechanistic insight into the development of the HTC phenomenon in legumes with a special focus on common beans and their nutrition, health benefits, and hydration behaviour. Furthermore, detailed elucidation of HTC mechanisms, mainly pectin-cation-phytate hypothesis and compositional changes of macronutrients like starch, protein, lipids and micronutrients like minerals, phytochemicals and cell wall polysaccharides during HTC development are critically reviewed based on the current research findings. Finally, strategies to improve the hydration and cooking quality of beans are proposed, and a perspective is provided.

Starch retrogradation in potato cells: Structure and in vitro digestion paradigm.

The consumption of foods with intact cellular structure is recognized to lower the glycemic index. Studies have focused on the gelatinization and digestibility of starch in intact cells. However, the effects of intact cells on starch retrogradation and digestibility are still unclear. Mild acid and alkali soaking were used to isolate intact potato cells and hydrothermal treatment (73 °C for 30 min) followed by retrogradation (4 °C for 0-15 days). The retrogradation period of 0-15 days increased the crystallinity and enthalpy change, while decreased the digestion rate and extent in potato cell and starch ghost samples. Furthermore, the existence of cell wall (potato cell) lowered the starch digestion rate and extent than potato starch ghost. It was concluded that the ghost structure is the main factor that controls the starch retrogradation, whereas the intact cell wall structure provides the barrier for starch digestion and protects the ghost structure.

Wood hemicelluloses exert distinct biomechanical contributions to cellulose fibrillar networks.

Hemicelluloses, a family of heterogeneous polysaccharides with complex molecular structures, constitute a fundamental component of lignocellulosic biomass. However, the contribution of each hemicellulose type to the mechanical properties of secondary plant cell walls remains elusive. Here we homogeneously incorporate different combinations of extracted and purified hemicelluloses (xylans and glucomannans) from softwood and hardwood species into self-assembled networks during cellulose biosynthesis in a bacterial model, without altering the morphology and the crystallinity of the cellulose bundles. These composite hydrogels can be therefore envisioned as models of secondary plant cell walls prior to lignification. The incorporated hemicelluloses exhibit both a rigid phase having close interactions with cellulose, together with a flexible phase contributing to the multiscale architecture of the bacterial cellulose hydrogels. The wood hemicelluloses exhibit distinct biomechanical contributions, with glucomannans increasing the elastic modulus in compression, and xylans contributing to a dramatic increase of the elongation at break under tension. These diverging effects cannot be explained solely from the nature of their direct interactions with cellulose, but can be related to the distinct molecular structure of wood xylans and mannans, the multiphase architecture of the hydrogels and the aggregative effects amongst hemicellulose-coated fibrils. Our study contributes to understanding the specific roles of wood xylans and glucomannans in the biomechanical integrity of secondary cell walls in tension and compression and has significance for the development of lignocellulosic materials with controlled assembly and tailored mechanical properties.

Ring Shear Tester as an in-vitro testing tool to study oral processing of comminuted potato chips.

Oral processing of solid foods is an extremely dynamic and complicated activity that involves multiple processes in tandem such as comminution, mixing, dilution, hydration and enzymatic breakdown that gradually transform the food from a morsel or a bite to a bolus that is ready for swallowing. It is hypothesised that just after "first bite" and initial particle reduction and hydration of solid brittle foods, the response to deformation of food particles is analogous to studies on the flowability and cohesion of wetted powders, which are effectively characterised using a Ring Shear Tester (RST). We examine this hypothesis and determine whether the RST measures properties of solid snack foods (potato chips or crisps, PCs) that are relevant to their dynamic sensory response, which includes capturing the effect of hydration on comminuted PCs. The RST is found to differentiate PCs obtained from different manufacturing sources (e.g. baked versus fried), and its measurements of cohesion and friction can be considered in context of the structure and composition of the PCs as well as oral processing. Remarkably, RST measurements for this small set of PC samples correlate with several sensory attributes that arise during mastication, which includes Sharpness and Ease of Clearance. This study highlights the potential of the RST as a new tool for oral processing research.

Wall porosity in isolated cells from food plants: Implications for nutritional functionality.

Macronutrients in whole plant foods are enclosed inside cells. The metabolic response from these entrapped nutrients may depend on cell-wall porosity, by controlling the passage of digestive enzymes. As non-interacting size mimics of digestive enzymes, we investigated the diffusion of fluorescently-labelled probes across the walls of isolated plant cells from potato tuber, red kidney bean and banana. Diffusion properties of permeable probes, dextran (20-kDa and 70-kDa) and albumin, were quantified, using fluorescence recovery after photobleaching. The consistent reduction of diffusion rate in the presence of cell walls (around 40%) compared to free-diffusion rate was attributed to the limiting porosity of the wall matrix. A combination of the physical barrier effects demonstrated here and non-catalytic binding of enzymes to cell walls limits the hydrolysis of intracellular macronutrients. This and further understanding of the structural basis for the physical barrier properties would help to design foods from plant materials with enhanced nutrition.

Effects of palm oil on structural and in vitro digestion properties of cooked rice starches.

The glycemic potency of rice depends upon the rate and extent of starch hydrolysis by pancreatic amylase and intestinal alpha-glucosidases. However, complexation of starch molecules with lipids is known to reduce the enzymic hydrolysis. In this study, we elucidated the varietal effect of rice starches on the formation of amylose-lipid complex, after cooking with palm oil, a common cooking oil. The amount of complexed lipid followed the order of black (2.5%), brown (2.5%), white (1.5%) and waxy (0.5%) rice starches. After heating with palm oil, the relative crystallinity of all the rice starches were destroyed whilst a V-type peak at 20° 2θ was increased, indicating the formation of amylose-lipid complex. This is also suggested from the DSC data where the melting enthalpy increased significantly after cooking in palm oil for all rice samples. The formation of amylose-lipid complex reduced the in vitro starch digestibility, enhancing the resistance starch content whilst decreasing the rapid and slow digestion fractions of non-waxy varieties. The rate and extent of in vitro starch hydrolysis seems to be dependent on the presence or absence of amylose fraction. With the mechanistic details, the present study suggests the applicability of palm oil addition during the rice cooking to enhance its nutritional functionality.

Structural and physicochemical properties of granular starches after treatment with debranching enzyme.

The present study modified maize and potato granular starches by partial debranching treatment below the gelatinization temperature, and investigated their structural and physicochemical properties. Pullulanase was much effective (more than three times) on hydrolyzing potato starch compared to maize starch as measured from total carbohydrate values in the supernatant. The pullulanase hydrolysis decreased the amount of double helices as observed from DSC measurement. These effects were dependent upon the time of enzyme hydrolysis (24h>8h>1h) as well as type of starch (potato>maize). The pullulanase hydrolysis decreased the peak viscosity of the potato starch paste, whereas the effect was very less pronounced for maize starch. The current results showed that it is possible to achieve the starches with desired physicochemical properties by varying the starch type as well as modification process.

Rheological and microstructural properties of porcine gastric digesta and diets containing pectin or mango powder.

Hydrated polysaccharides and their assemblies are known to modulate gastric emptying rate due to their capacity to change the structural and rheological properties of gastric contents (digesta). In the present study, we investigated the rheological and microstructural properties of gastric digesta from pigs fed with diets incorporating mango powder or pectin, and compared results with those from hydrated diets of the same water content, in order to investigate the origins for rheological changes in the pig stomach. All of the hydrated diets and gastric digesta were particle-dominated suspensions, generally showing weak gel or more solid-like behavior with the storage modulus (G') always greater than loss modulus (G") under small deformation oscillatory measurements, and with small deformation viscosity greater than steady shear viscosity (i.e. non-Cox-Merz superposition). Although significant rheological differences were observed between the hydrated diets, rheological parameters for gastric digesta were similar for all diets, indicative of a rheological homeostasis in the pig stomach. Whilst the addition of gastric mucin (20mg/mL) to control and mango diets altered the rheology to match the gastric digesta rheology, the effect of mucin on the pectin-containing diet was negligible. The viscous effect of pectin also hindered the action of alpha amylase as observed from relatively less damaged starch granules in pectin digesta compared to mango and control digesta. Based on the experimental findings that the rheology of gastric digesta differs from hydrated diets of the same water content, the current study revealed composition-dependent complex behavior of gastric digesta in vivo, suggesting that the rheology of food products or ingredients may not necessarily reflect the rheological effect when ingested.

The interplay of α-amylase and amyloglucosidase activities on the digestion of starch in in vitro enzymic systems.

In vitro hydrolysis assays are a key tool in understanding differences in rate and extent of digestion of starchy foods. They offer a greater degree of simplicity and flexibility than dynamic in vitro models or in vivo experiments for quantifiable, mechanistic exploration of starch digestion. In the present work the influence of α-amylase and amyloglucosidase activities on the digestion of maize and potato starch granules was measured using both glucose and reducing sugar assays. Data were analysed through initial rates of digestion, and by 1st order kinetics, utilising logarithm of slope (LOS) plots. The rate and extent of starch digestion was dependent on the activities of both enzymes and the type of starch used. Potato required more enzyme than maize to achieve logarithmic reaction curves, and complete digestion. The results allow targeted design of starch digestion experiments through a thorough understanding of the contributions of α-amylase and amyloglucosidase to digestion rates.

Amylase binding to starch granules under hydrolysing and non-hydrolysing conditions.

Although considerable information is available about amylolysis rate, extent and pattern of granular starches, the underlying mechanisms of enzyme action and interactions are not fully understood, partly due to the lack of direct visualisation of enzyme binding and subsequent hydrolysis of starch granules. In the present study, α-amylase (AA) from porcine pancreas was labelled with either fluorescein isothiocyanate (FITC) or tetramethylrhodamine isothiocyanate (TRITC) fluorescent dye with maintenance of significant enzyme activity. The binding of FITC/TRITC-AA conjugate to the surface and interior of granules was studied under both non-hydrolysing (0 °C) and hydrolysing (37 °C) conditions with confocal microscopy. It was observed that enzyme binding to maize starch granules under both conditions was more homogenous compared with potato starch. Enzyme molecules appear to preferentially bind to the granules or part of granules that are more susceptible to enzymic degradation. The specificity is such that fresh enzyme added after a certain time of incubation binds at the same location as previously bound enzyme. By visualising the enzyme location during binding and hydrolysis, detailed information is provided regarding the heterogeneity of granular starch digestion.

Freeze-drying changes the structure and digestibility of B-polymorphic starches.

Starch granules both isolated from plants and used in foods or other products have typically been dried. Common food laboratory and industry practices include oven (heat), freeze, and ethanol (solvent-exchange) drying. Starch granules isolated from maize (A-type polymorph) and potato (B-type polymorph) were used to understand the effects of different dehydration methods on starch structure and in vitro digestion kinetics. Oven and ethanol drying do not significantly affect the digestion properties of starches compared with their counterparts that have never been dried. However, freeze-drying results in a significant increase in the digestion rate of potato starch but not maize starch. The structural and conformational changes of starch granules after drying were investigated at various length scales using scanning electron microscopy, confocal laser scanning microscopy, X-ray diffraction, FTIR spectroscopy, and NMR spectroscopy. Freeze-drying not only disrupts the surface morphology of potato starch granules (B-type polymorph), but also degrades both short- and long-range molecular order of the amylopectin, each of which can cause an increase in the digestion rate. In contrast to A-polymorphic starches, B-polymorphic starches are more disrupted by freeze-drying, with reductions of both short- and long-range molecular order. We propose that the low temperatures involved in freeze-drying compared with oven drying result in greater chain rigidity and lead to structural disorganization during water removal at both nanometer and micrometer length scales in B-type polymorphic starch granules, because of the different distribution of water within crystallites and the lack of pores and channels compared with A-type polymorphic starch granules.

Mechanism for starch granule ghost formation deduced from structural and enzyme digestion properties.

After heating in excess water under little or no shear, starch granules do not dissolve completely but persist as highly swollen fragile forms, commonly termed granule "ghosts". The macromolecular architecture of these ghosts has not been defined, despite their importance in determining characteristic properties of starches. In this study, amylase digestion of isolated granule ghosts from maize and potato starches is used as a probe to study the mechanism of ghost formation, through microstructural, mesoscopic, and molecular scale analyses of structure before and after digestion. Digestion profiles showed that neither integral nor surface proteins/lipids were crucial for control of either ghost digestion or integrity. On the basis of the molecular composition and conformation of enzyme-resistant fractions, it was concluded that the condensed polymeric surface structure of ghost particles is mainly composed of nonordered but entangled amylopectin (and some amylose) molecules, with limited reinforcement through partially ordered enzyme-resistant structures based on amylose (for maize starch; V-type order) or amylopectin (for potato starch; B-type order). The high level of branching and large molecular size of amylopectin is proposed to be the origin for the unusual stability of a solid structure based primarily on temporary entanglements.

Preparation and characterization of gelatinized granular starches from aqueous ethanol treatments.

In order to modify the properties of native starch granules, the formation of gelatinized granular forms (GGS) from normal, waxy, and high amylose maize, as well as potato and tapioca starches was investigated by treating granules with aqueous ethanol at varying starch:water:ethanol ratios and then heating in a rotary evaporator to remove ethanol. The modified starches were characterized using bright field, polarized and electron microscopy. Short/long range molecular order and enthalpic transitions on heating were also studied using infrared spectroscopy, X-ray diffractometry and differential scanning calorimetry respectively. A diffuse birefringence pattern without Maltese cross was observed for most GGS samples. Treatment with aqueous ethanol resulted in starch-specific changes in the surface of granules, most noticeably swelling and disintegration in waxy maize, surface wrinkling in normal maize and tapioca, swelling and opening-up in potato starches, and swelling and bursting in high amylose maize. The ratio of ethanol to water at which original granular order was disrupted also varied with starch type. GGS had less short range molecular order than native granules as inferred by comparing 1047/1022 wave number ratio from infrared spectroscopy. Similarly, A- and B-type diffraction reflections were either reduced or completely lost with evolution of V-type patterns in GGS.

Physicochemical and structural properties of maize and potato starches as a function of granule size.

Chemical composition, molecular structure and organization, and thermal and pasting properties of maize and potato starches fractionated on the basis of granule size were investigated to understand heterogeneity within granule populations. For both starches, lipid, protein, and mineral contents decreased and apparent amylose contents increased with granule size. Fully branched (whole) and debranched molecular size distributions in maize starch fractions were invariant with granule size. Higher amylose contents and amylopectin hydrodynamic sizes were found for larger potato starch granules, although debranched molecular size distributions did not vary. Larger granules had higher degrees of crystallinity and greater amounts of double and single helical structures. Systematic differences in pasting and thermal properties were observed with granule size. Results suggest that branch length distributions in both amylose and amylopectin fractions are under tighter biosynthetic control in potato starch than either molecular size or amylose/amylopectin ratio, whereas all three parameters are controlled during the biosynthesis of maize starch.