Altering structure and enzymatic resistance of high-amylose maize starch by irradiative depolymerization and annealing with palmitic acid as V-type inclusion compound.

This study explored a new physical modification approach to regulate enzymatic resistance of high-amylose starch for potentially better nutritional outcomes. High-amylose maize starch (HAMS) was subjected to chain depolymerization by electron beam irradiation (EBI), followed by inducing ordered structure through annealing in palmitic acid solution (APAS). APAS treatment significantly promotes the formation of ordered structure. Starch after the combinative modification showed up to 5.2 % increase in total crystallinity and up to 1.2 % increase in V-type fraction. The EBI-APAS modification led to increased gelatinization temperature (from 66.1 to 87.6 °C) and reduced final digested percentage under in vitro stimulated digestion conditions. The moderate extent of depolymerization resulted in higher enzymatic resistance, indicating that the extent of depolymerization is crucial in EBI-APAS modification. Pearson analysis showed a significant correlation between gelatinization onset temperature and digestion kinetic parameter (k1, rate constant of fast-phase digestion). Overall, the result suggests that ordered structures of degraded molecules induced by the combinative modification contribute to the enzymatic resistance of starch. This study sheds lights on future applications of EBI-APAS approach to regulate multi-scale structures and nutritional values of high-amylose starch.

Altered leaching composition of maize starch granules by irradiative depolymerization: The key role of degraded molecular structure.

The molecular composition of starch leachates from starch-based foods has been recently recognised as a crucial determinant of food properties. However, there is limited knowledge on the regulation of this composition through irradiative depolymerization of starch. This research investigates the leaching behaviour of maize starch depolymerized by electron beam irradiation, and the relationship between the composition of leached starch and structures of modified starch granules. The analysis using 1H NMR spectroscopy confirmed a decrease in the degree of branching (from 4.4 % to 2.8 %), while size-exclusion chromatography identified a newly-derived amylopectin fraction of a smaller hydrodynamic radius (approximately 60-80 nm). The structural properties of the starch granules were also analysed, revealing an increased BET-area of granules and reduced total crystallinity after depolymerization. In the leachates of swollen granules, the bimodal distribution of starch molecules evolves into unimodal with the increase of the irradiative dosage, while modified starch leached more starch molecules with Rh < 10 nm. The results of principal component analysis and Pearson correlation analysis indicate that the degree of branching of degraded starch molecules, as well as the newly-derived amylopectin fraction, significantly correlates (p < 0.01) with the molecular size of leached starch molecules (Rh < 10 nm). It is thus proposed that the cleavage of α-1,6 linkage may be a critical factor in controlling the leaching process of irradiated starch granules. This study highlights the potential of irradiative degradation to control the molecular composition and structure of starch leachates, thereby optimizing the properties of starch-based foods.

Enhancing enzymatic resistance of starch through strategic application of food physical processing technologies.

The demand for clean-label starch, perceived as environmentally friendly in terms of production and less hazardous to health, has driven the advancement of food physical processing technologies aimed at modifying starch. One of the key objectives of these modifications has been to reduce the glycaemic potency and increase resistant starch content of starch, as these properties have the potential to positively impact metabolic health. This review provides a comprehensive overview of recent updates in typical physical processing techniques, including annealing, heat-moisture, microwave and ultrasonication, and a brief discussion of several promising recent-developed methods. The focus is on evaluating the molecular, supramolecular and microstructural changes resulting from these modifications and identifying targeted structures that can foster enzyme-digestion resistance in native starch and its forms relevant to food applications. After a comprehensive search and assessment, the current physical modifications have not consistently improved starch enzymatic resistance. The opportunities for enhancing the effectiveness of modifications lie in (1) identifying modification conditions that avoid the intensive disruption of the granular and supramolecular structure of starch and (2) exploring novel strategies that incorporate multi-type modifications.

Physical modification of high amylose starch using electron beam irradiation and heat moisture treatment: The effect on multi-scale structure and in vitro digestibility.

This study explores a new strategy for manipulating the digestibility of high-amylose maize starch (HAMS) through combinative modifications, namely depolymerization via electron beam irradiation (EBI) followed by reorganizing glucan chains via heat moisture treatment (HMT). The results show that semi-crystalline structure, morphological features and thermal properties of HAMS remained similar. However, EBI increased branching degree of the starch at high irradiation dosage (20 kGy), resulting in more readily leached amylose during heating. HMT increased the relative crystallinity (3.9-5.4% increase) and V-type fraction (0.6-1.9% increase), without significant changes (p > 0.05) in gelatinization onset temperature, peak temperature and enthalpy. Under simulated gastrointestinal conditions, the combination of EBI and HMT either had no effect or negative effect on starch enzymatic resistance, depending on the irradiation dosage. These results suggest that the depolymerization by EBI predominantly affects the changes in enzyme resistance, rather than the growth and perfection of crystallites induced by HMT.

Amorphous packing of amylose and elongated branches linked to the enzymatic resistance of high-amylose wheat starch granules.

To elucidate starch structural features underlying resistant starch formation, wheat starch granules with three (A-, B- and C- type) crystalline polymorphisms and a range of amylose contents were digested in vitro. The changes in multi-level structure of digestion residues were compared. In the residues of A- and C-type starches, the molecular fine structure (distributions of chain length and whole molecular size), as analyzed by size exclusion chromatography (SEC), remained similar during digestion. In contrast, B-type high amylose wheat starch (HAWS) showed distinct changes in multi-level structures of digestion-resistant fractions: (1) the peak of longer amylopectin branches shifted to a lower degree of polymerization (40 DP); (2) production of α-limit dextrin (~2 nm hydrodynamic radius) in the residues; (3) a small increase of double helix content during digestion, in contrast to 6 % reduction for the A-type starch; (4) a decrease (6 °C lower) in the melting temperature of amylose-lipid complexes. The comparison suggests that elongated branches in B-type starch contribute to the formation of resistant fraction (including α-limit dextrin) against α-amylase. The amorphous packing of starch polymers with elongated branches together with the absence of surface pores and channels is proposed to be the basis for the enzymatic resistance of granular HAWS.

Modulating the glycemic response of starch-based foods using organic nanomaterials: strategies and opportunities.

Traditionally, diverse natural bioactive compounds (polyphenols, proteins, fatty acids, dietary fibers) are used as inhibitors of starch digestive enzymes for lowering glycemic index (GI) and preventing type 2 diabetes mellitus (T2DM). In recent years, organic nanomaterials (ONMs) have drawn a great attention because of their ability to overcome the stability and solubility issues of bioactive. This review aimed to elucidate the implications of ONMs in lowering GI and as encapsulating agents of enzymes inhibitors. The major ONMs are presented. The mechanisms underlying the inhibition of enzymes, the stability within the gastrointestinal tract (GIT) and safety of ONMs are also provided. As a result of encapsulation of bioactive in ONMs, a more pronounced inhibition of enzymes was observed compared to un-encapsulated bioactive. More importantly, the lower the size of ONMs, the higher their inhibitory effects due to facile binding with enzymes. Additionally, in vivo studies exhibited the potentiality of ONMs for protection and sustained release of insulin for GI management. Overall, regulating the GI using ONMs could be a safe, robust and viable alternative compared to synthetic drugs (acarbose and voglibose) and un-encapsulated bioactive. Future researches should prioritize ONMs in real food products and evaluate their safety on a case-by-case basis.

The relation between wheat starch properties and noodle springiness: From the view of microstructure quantitative analysis of gluten-based network.

Understanding the effect of starch properties on the gluten network during noodle-making will allow rational selection of ingredients. The microstructure and texture of dough sheets, dried noodles, and cooked noodles prepared by adding different wheat starches were compared. Increasing starch solubility and swelling power could reduce the compactness and homogeneity of the network in dough sheet. Lower swelling power and higher solubility of starch made the elastic modulus of dried noodles increase, leading to increasing branching rate along with decreasing average network width and length of cooked noodles. After cooking, noodles with increasing springiness showed higher branching rate of gluten network in dough sheet, greater elastic modulus of dried noodles, and lower average network length of cooked noodles. It was thus proposed that wheat starch with lower swelling power could enhance the springiness of noodles. The quantitative analysis of microstructure was an appropriate method to predict the noodle springiness.

Relation between adhesiveness and surface leachate rheological properties of cooked noodles: From the view of starch fine molecular structure.

Adhesiveness is one of the important sensory properties of noodles. A better understanding of the molecular basis of the sensory property would allow for a more reasonable selection of wheat ingredients. In this study, the adhesiveness of cooked noodles made from reconstituted wheat flour was evaluated using a texture profile analyzer. The rheological properties of dissolved solids from the noodle surface were measured. The composition and structure of leachate, as well as multi-structure of native starch were analyzed. The correlation analysis showed that adhesiveness positively correlates with the loss modulus parameters [log K' (consistency coefficient) and Ea (activation energy)]. Increased content of leached starch can ascend the rheological parameters (log K' and Ea). In addition, more short amylose chains with 100-500 degree of polymerization (DP) in leached starch also induced the increase of log K' (p < 0.01) and Ea (p < 0.01). The fraction of short amylose chains in leached starch was negatively correlated with the content of long amylopectin chains ranged from 37 to 100 DP in native starch (p < 0.05), as well as the median diameter of particles (p < 0.01). Multi-level structures of native starch granules largely determine the composition and molecular structure of noodle leachate, consequently contributing to surface rheological properties. It is thus proposed that wheat starch with more long amylopectin chains and large granules could be used to reduce noodle adhesiveness. Moreover, the rheological approach using noodle surface materials proposed here is useful to predict the noodle adhesiveness as an important sensory characteristic.

In vitro starch digestibility of buckwheat cultivars in comparison to wheat: The key role of starch molecular structure.

The objective of this study was to compare the in vitro digestibility of different buckwheat and wheat starch cultivars and establish the relationship between digestibility and structure of buckwheat starch. Structure of starches were analyzed with size exclusion chromatography and fluorophore-assisted capillary electrophoresis. Results showed that the amylose content of Tartary buckwheat starch (TBS) and common buckwheat starch (CBS) was 3-4% lower than that of wheat starch. However, no significant difference in the digestibility was found between them. The fast digestion rate coefficient of TBS was negatively correlated with the amount of long amylopectin chains (24 < DP ≤ 36), and the total digested starch percentage of CBS was negatively correlated with the amount of medium-long amylopectin chains (13 < DP ≤ 24). This suggests that the digestibility of fully gelatinized starch had no association with the botanical sources but may be more influenced by starch structure.

Improving the cold water swelling properties of oat starch by subcritical ethanol-water treatment.

The granular cold water swelling oat starch was prepared by subcritical ethanol-water, and the changes of properties and structure on oat starch were investigated. The oat starch was modified at the temperature of 95 °C and ethanol concentration of 48% and showed a higher cold water swelling ability of 22.58 g/g, whereas native oat starch was 6.73 g/g. Modified oat starch granule was kept intact, and it was swollen when dispersing in the water. The gelatinization enthalpy declined to 0 J/g. The surface of modified oat starch granules was honeycomb and porous observed by scanning electron microscope. The X-ray diffraction showed the A-type crystal decreased and the V-type crystal increased, and the result was quantitatively confirmed by solid-state 13C NMR spectroscopy. The ratio of 1047 cm-1/1022 cm-1 (determined by Fourier transform infrared spectroscopy) of modified oat starch was decreased. The molecular weight distribution of modified oat starch was slightly reduced, and the amylose content increased from 26.18% to 31.68%, and only a small amount of carbohydrates leached during the modification. Subcritical ethanol-water modification improved the cold water swelling ability of oat starch. The starch crystals changed from A-type to V-type provide a potential mechanism of subcritical ethanol-water modified oat starch.

pH-Responsive Smart Wettability Surface with Dual Bactericidal and Releasing Properties.

Biomaterial-associated infections caused by pathogenic bacteria have important implications on human health. This study presents the design and preparation of a smart surface with pH-responsive wettability. The smart surface exhibited synergistic antibacterial function, with high liquid repellency against bacterial adhesion and highly effective bactericidal activity. The wettability of the surface can switch reversibly between superhydrophobicity and hydrophobicity in response to pH; this controls bacterial adhesion and release. Besides, the deposited silver nanoparticles of the surface were also responsible for bacterial inhibition. Benefiting from the excellent liquid repellency, the surface could highly resist bacterial adhesion after immersing in a bacterial suspension for 10 s (85%) and 1 h (71%). Adhered bacteria can be easily eliminated using deposited silver nanoparticles during the subsequent treatment of alkaline bacterial suspension, and the ratio of deactivated bacteria was above 75%. After the pH returned to neutral, the deactivated bacteria can be easily released from the surface. This antibacterial surface showed an improved bacterial removal efficiency of about 99%. The results shed light on future antibacterial applications of the smart surface combining both bactericidal and adhesion-resistant functionalities.

Molecular-structure evolution during in vitro fermentation of granular high-amylose wheat starch is different to in vitro digestion.

This study investigates the evolution of the distributions of whole molecular size and of chain length of granular wheat starches (37 ~ 93% amylose content), subjected to in vitro fermentation with a porcine faecal inoculum or digestion with pancreatic enzymes. The results showed that the molecular structures of high-amylose starch (HAS) unfermented residues largely remained unchanged during the fermentation process, while wild-type starch (37% amylose content) showed a preferential degradation of the amylopectin fraction. In contrast, under simulated digestion conditions, the undigested residues of HAS showed structural changes, including a decrease in amylose content, a shift of amylose peak position towards lower degrees of polymerisation, and an enzyme-resistant fraction. These changes of starch structure are likely to be dependent on the different starch-degrading enzyme activities present in pancreatic vs. microbial systems. Molecular changes in response to fermentation metabolism revealed by size-exclusion chromatography can help understand the microbial utilization of resistant starch.

Protein-starch matrix plays a key role in enzymic digestion of high-amylose wheat noodle.

Wheat flour, consisting of a complex matrix of starch and protein, is used as a representative model of whole food here to investigate the binary interaction in relation to amylose level and hydrothermal treatment in noodles as a food exemplar. Noodle made of high-amylose wheat (HAW) flour showed an eight-fold higher resistant starch content, compared to the wild type. Protein removal under simulated intestinal digestion conditions resulted in higher starch digestion rate coefficients in raw and cooked flours. In cooked flours, the substrate becomes similarly accessible to digestive enzymes regardless of protein removal. The results indicate that the increased protein content in native HAW flour and thermal stability of starch in HAW noodles lead to higher food integrity and consequently enhance the resistance against α-amylase digestion. Overall, the study suggests that a diversity of starch-protein interactions in wheat-based food products underlies the nutritional value of natural whole foods.

Structural basis of wheat starch determines the adhesiveness of cooked noodles by affecting the fine structure of leached starch.

The relationship between the fine structure of original starch, leached starch during cooking, and the adhesiveness of noodles prepared by adding starches separated from different wheat cultivars was analyzed. The adhesiveness of noodles was primarily determined by the chain-length distributions of amylopectin rather than amylose. The adhesiveness of cooked noodles was positively correlated with the amount of short amylopectin chains with the degree of polymerization (DP) of 6-12, but negatively correlated with the amount of long chains with 25 < DP ≤ 36. The decrease of the proportion of short amylopectin chains and amylose chains and the increase of the amount of very long amylopectin chains with 37 < DP ≤ 100 in leachate led to decreased adhesiveness of cooked noodles. The reduction of the short-chain content in leached amylopectin caused by the increased proportion of long chains in original amylopectin is proposed to weaken the adhesiveness of cooked noodles.

Starch granular protein of high-amylose wheat gives innate resistance to amylolysis.

Granular protein is an important structural feature in determining starch digestibility. High-amylose wheat starch (HAWS) with >80% amylose content contains more granular protein than wild-type starch. As analyzed by mass spectrometry-based proteomics, granular-bound starch synthase (GBSS) is the major granular protein in isolated starch materials. GBSS content increases with amylose content (Spearman's correlation, p < 0.05), whereas the abundance relative to other proteins is similar among starches. Multiple amylase inhibitors were also identified. From Michaelis-Menten analysis, HAWS has a similar Km (Michaelis constant) as wild type, suggesting initial enzymatic binding is similar. After the pre-digestion of proteins, wild type had a greater change in starch digestibility than HAWS, probably due to the latter having 'thicker' granular-protein layers and higher enzymatic resistance of substrate per se. Overall, the study suggests that the greater granular protein content in HAWS is a factor that contributes to slower amylolysis compared to wild type.

A more general approach to fitting digestion kinetics of starch in food.

Two models for the digestion of starch in foods are considered: sequential, where two or more reactions follow one another, and parallel, where they occur simultaneously. Each reaction is assumed to be first-order, and to be characterized by a rate coefficient and a digestible fraction. For parallel kinetics, a new fitting method is developed; methods for sequential reactions are already available, based on the "logarithm of slopes" technique. Least-squares fitting methods for each are set out, which identify the uncertainties associated with each rate parameter, so that one can see if one has assumed too many or too few processes. The methods gave robust results with synthetic data, and were applied to real in vitro digestion data for wheat starches with up to 93% amylose. The combination of parallel and sequential models provides a general approach to investigating the kinetics of starch digestion in food.

Starch branching enzymes contributing to amylose and amylopectin fine structure in wheat.

Starch branching enzymes (SBEs) play a major role in determining starch molecular structure in cereal endosperm. This study investigates how SBEIIs contribute to the chain-length distributions (CLDs) of both amylopectin and amylose, obtained by enzymatic debranching of native starch. Wheat starches with low (37%) to high (93%) amylose content were obtained through altering SBEII in planta. Multiple components were detected in both amylose and amylopectin CLDs. Model fitting of these CLDs reveals a quantitative association between the enzyme activities in amylopectin and amylose. SBEIIa modifies shorter branches (degree of polymerization DP ≲ 12) in amylopectin and longer amylose chains with a CLD peak at ˜3000 DP. SBEIIb acts on longer branches (DP≲ 32) in amylopectin, while its effect on amylose fine structure is not significant. Using both the amylose and amylopectin models to analyze the CLD reveals connections between amylose and amylopectin in wheat starch biosynthesis.

Controlled gelatinization of potato parenchyma cells under excess water condition: structural and in vitro digestion properties of starch.

Modulation of the starch digestion rate and extent is a major target for increasing nutritional value of potato-based food. In order to understand the effect of controlled gelatinization on the structure and in vitro digestion properties of whole potato food, intact parenchyma cells were isolated as a model through soaking in mild acid and alkali solutions, and then hydrothermally treated at different incubation temperatures in excess water. The morphological and structural changes of entrapped starch granules as well as in vitro starch digestion properties were investigated. Three classes of starch digestion of potato cells are identified: (1) when the temperature of hydrothermal treatment was set at 55 °C or 60 °C, the efficient physical barrier of intact cell walls resulted in a limited starch digestion extent; (2) when the temperature is set at 65 °C, the cell wall structures apparently reduced the digestion rate but only slightly reduced the final extent compared to the starch counterpart, probably because starch swelling weakens the physical barrier; (3) at 70-95 °C, the damaged cell wall still reduced the digestion rate but did not reduce the digestion extent compared to the starch counterpart. The results suggest that controlling hydrothermal processing temperature is a viable approach to develop potato ingredients with enhanced nutritional functionality.

Autoclaved rice: The textural property and its relation to starch leaching and the molecular structure of leached starch.

Autoclave cooking is used to produce "convenience" rice. In this study, autoclaving effects on sensory properties are investigated, and mechanistic explanations in terms of the underlying molecular structure are explored by analyzing this structure by size-exclusion chromatography and fitting the results with models based on biosynthetic processes. Compared to steam cooking, autoclaving produces stickier texture, and slightly affects hardness. It is found that molecular sizes of leached starch of both autoclaved and steam cooked rice are similar, but significantly smaller than that of the parent grain starch; model fitting parameters of leached amylopectin and amylose structures between autoclaved rice and steam cooked rice display no large variations. The amount of leached amylopectin (an important texture-controlling parameter) of autoclaved rice is higher than that of steam cooked rice. Correlation analysis indicates that, compared to steam-cooked rice, the stickier texture of autoclaved rice is caused by more amylopectin leaching during autoclaving.

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.