Mechanism of action of three different glycogen branching enzymes and their effect on bread quality.

Bread staling adversely affects the quality of bread, but starch modification by enzymes can counteract this phenomenon. Glycogen branching enzymes (GBEs) used in this study were isolated from Deinococcus geothermalis (DgGBE), Escherichia coli (EcGBE), and Vibrio vulnificus (VvGBE). These enzymes were characterized and applied for starch dough modification to determine their role in improving bread quality. First, the branching patterns, activity on amylose and amylopectin, and thermostability of the GBEs were determined and compared. EcGBE and DgGBE exhibited better thermostable characteristics than VvGBE, and all GBEs exhibited preferential catalysis of amylopectin over amylose but different degrees. VvGBE and DgGBE produced a large number of short branches. Three GBEs degraded the starch granules and generated soluble polysaccharides. Moreover, the maltose was increased in the starch slurry but most significantly in the DgGBE treatment. Degradation of the starch granules by GBEs enhanced the maltose generation of internal amylases. When used in the bread-making process, DgGBE and VvGBE increased the dough and bread volume by 9 % and 17 %, respectively. The crumb firmness and retrogradation of the bread were decreased and delayed significantly more in the DgGBE bread. Consequently, this study can contribute to understanding the detailed roles of GBEs in the baking process.

Resistant starch-enriched brown rice exhibits prebiotic properties and enhances gut health in obese mice.

Resistant starch serves as a prebiotic in the large intestine, aiding in the maintenance of a healthy intestinal environment and mitigating associated chronic illnesses. This study aimed to investigate the impact of resistant starch-enriched brown rice (RBR) on intestinal health and functionality. We assessed changes in resistant starch concentration, structural alterations, and branch chain length distribution throughout the digestion process using an in vitro model. The efficacy of RBR in the intestinal environment was evaluated through analyses of its prebiotic potential, effects on intestinal microbiota, and intestinal function-related proteins in obese animals fed a high-fat diet. RBR exhibited a higher yield of insoluble fraction in both the small and large intestines compared to white and brown rice. The total digestible starch content decreased, while the resistant starch content significantly increased during in vitro digestion. Furthermore, RBR notably enhanced the growth of four probiotic strains compared to white and brown rice, displaying higher proliferation activity than the positive control, FOS. Notably, consumption of RBR by high-fat diet-induced obese mice suppressed colon shortening, increased Bifidobacteria growth, and improved intestinal permeability. These findings underscore the potential prebiotic and gut health-promoting attributes of RBR, offering insights for the development of functional foods aimed at preventing gastrointestinal diseases.

Hydrothermal treatments of starch impact reaction patterns during subsequent chemical derivatization.

Differences in derivatization patterns (using a fluorescent reagent, fluorescein isothiocyanate) of wheat, pea, and potato starches between native granular (NAT) starches and their respective annealed (ANN) and heat-moisture treated (HMT) starches were investigated to reveal structural changes associated with starch hydrothermal treatments. Size-exclusion chromatography with fluorescence and refractive index detection assessed the reactivity of amylose (AM), intermediate chains (IM1 and IM2), and amylopectin branch chains (AP1, AP2, and AP3) within the different starches. Shifts in X-ray diffraction patterns of HMT starches and in the gelatinization properties of both ANN and HMT starches confirmed molecular rearrangement. The reaction homogeneity (wheat and pea) and the overall extent of reaction (pea and potato) increased for HMT starches compared to other starches. The lower reactivities of IM2 chains (HMT starch) and AP3 chains (ANN starch) relative to NAT starches, indicated their involvement in molecular rearrangements and improved double helical order. IM2 and AP branch chains in ANN pea starch also were less reacted than NAT starch chains, suggesting their co-crystallization. Molecular rearrangements in ANN and HMT starches led to altered swelling and pasting viscosities. Thus, changes in the relative crystallinity of individual starch branch chains induced by hydrothermal processing impact the final physical properties.

Structural characteristics of resistant starch-enriched rice during digestion and its effects on gut barrier function in high-fat induced obese mice.

Dietary supplementation with indigestible carbohydrates is known to improve the gut environment and prevent obesity and inflammatory diseases by modulating the gut microbiota. In previous work, we established a method for the production of resistant starch (RS)-enriched high-amylose rice (R-HAR) using citric acid. The present study aimed to evaluate changes in structural characteristics during digestion of R-HAR and its effects on the gut health. A three-step in vitro digestion and fermentation model was used, then, RS content, scanning electron microscopy, and branch chain length distribution were analyzed during in vitro digestion. During the digestion of R-HAR, the RS content increased, and the structure was predicted to have a greater impact on the gut microbiota and gut environment. To study the intestinal health effects of R-HAR, its anti-inflammatory and gut barrier integrity activities were assayed in HFD-induced mice. Intake of R-HAR suppressed colonic shortening and inflammatory responses induced by HFD. Furthermore, R-HAR exhibited gut barrier protective activity with an increase in tight junction protein levels. We determined that R-HAR may be a potentially beneficial intestinal environment improver, which may have various implications in the food industry as rice.

Impact of chemical modification by immersion with malic acid on the physicochemical properties and resistant starch formation in rice.

The effects of immersion time on the physicochemical properties and resistant starch (RS) formation of malic acid-treated rice were investigated. Malic acid treatment decreased the frequency of cracks within the rice kernel. The color (lightness) was significantly affected by the immersion time, reflecting the browning of rice. The degree of substitution gradually increased with the immersion time and reached a plateau after 12 h, and the intensity of the C=O bond peak detected in the Fourier-transform infrared spectroscopy showed a similar trend. However, the crystallinity of rice decreased as the immersion time increased, which was confirmed by the X-ray diffraction and thermal transition properties. A gradual increase in RS was observed as the immersion time and DS increased, ranging from 44.5% to 73.3%, reaching a maximum after 12 h of immersion. Therefore, 12 h was determined to be the optimal immersion time for maximizing RS content. This information about the structural characteristics and heat-stable properties of malic acid-treated rice in starch digestion can be used to develop a low-digestible food ingredient and lead to further application of the study. PRACTICAL APPLICATION: This study reported the preparation and physicochemical properties of malic acid-treated resistant starch with different immersion times. This information could contribute to the structural characterization of resistant starch and the development of low-calorie processed rice products.

Fabrication of citric acid-modified starch nanoparticles to improve their thermal stability and hydrophobicity.

Starch nanoparticles (SNPs) were reacted at 130 °C for 1.5 h in the presence of citric acid (30 %) to enhance their thermal stability and hydrophobicity. Citric acid content in SNP was controlled by washing with different concentrations of ethanol (95 %, 70 % and 60 %) for 2, 5 and 10 min and then subjected to heat treatment at 130 °C. After the modification, the peak at 1732 cm-1 representing ester bond was observed via FT-IR, and the intensity of the peak was decreased with a lower ethanol content in washing medium. For the 60 % ethanol condition, the granular structure was promptly fragmented into particles less than 50 nm in the aqueous solution. The modification enhanced the thermal stability and hydrophobicity of the SNPs. The modified SNPs was used as a nano-vehicle wall material for encapsulating beta-carotene as a model hydrophobic material. Approximately 80 % beta-carotene was encapsulated in the modified SNPs.

Modeling of in vitro digestion behavior of corn starches of different digestibility using modified log of slope (LOS) method.

This study aimed to further improve the previously described first-order equation representing in vitro digestion of starch by extensively explaining modified log of slope (LOS) plot method. Hydrolysis curves of various starches were analyzed using original and/or modified LOS plot methods. Some starches showed significant differences in the results from the two methods; specifically, the modified method better described the digestive behavior of starch with various digestion properties, supported by higher determination coefficient values and better estimation of the digestibility data over digestive phase. The digestion parameters obtained from the modified method provided multiple types of information, including amount and digestion rate of each starch fraction (rapidly digestible, slowly digestible, and resistant starch), supporting the concept of digestible fraction classification. Therefore, the modified LOS plot method described here can be applied as an effective tool for analyzing and describing the multi-scale in vitro digestion behavior of starch.

Impact of starch granule-associated channel protein on characteristic of and λ-carrageenan entrapment within wheat starch granules.

This study investigated the physicochemical characteristics of protease-treated wheat starch (PT-WST) to understand the role of starch granule-associated proteins (SGAPs) and the potential capability of PT-WST to provide a nutrient delivery system (NDS). Protease treatment was conducted at 4 °C and 37 °C (PT04 and PT37), respectively. A model delivery system was assessed with PT37 granules infiltrated by λ-carrageenan (λC) under variations of molecular size (λC hydrolysates produced from 0, 2.5, 100, and 500 mM HCl solution), agitation time, and temperature. Protein-specific (3-(4-carboxybenzyl)quioline-2-carboxaldehyde) or non-reactive (methanolic merbromin) fluorescent dye staining revealed that removal of SGAPs on surfaces and channels were more effective for PT37 than for PT04. Consistent amylose content, swelling, and gelatinization temperature before and after protease treatment suggested minimal impact on the starch structure. PT37 presented higher solubility and pasting viscosity than PT04. This resulted from excessive SGAP removal, which enhanced entrapment capacity. λC molecular size and agitation temperature showed a negative correlation with the content of λC entrapped within PT37, and this content depended on the interplay between the agitation time and λC molecular size. As λC molecular size decreased, the λC distribution became uniform throughout the granules, which confirmed the potential of PT-WST as a carrier for NDS.

Development of Freeze-Thaw Stable Starch through Enzymatic Modification.

The use of unmodified starch in frozen foods can cause extremely undesirable textural changes after the freeze-thaw process. In this study, using cyclodextrin glucanotransferase (CGTase) and branching enzymes, an amylopectin cluster with high freeze-thaw stability was produced, and was named CBAC. It was found to have a water solubility seven times higher, and a molecular weight 77 times lower, than corn starch. According to the results of a differential scanning calorimetry (DSC) analysis, dough containing 5% CBAC lost 19% less water than a control dough after three freeze-thaw cycles. During storage for 7 days at 4 °C, bread produced using CBAC-treated dough exhibited a 14% smaller retrogradation peak and 37% less hardness than a control dough, suggesting that CBAC could be a potential candidate for clean label starch, providing high-level food stability under repeated freeze-thaw conditions.

Effects of Citrus Peel Hydrolysates on Retrogradation of Wheat Starch.

Retrogradation is the principal cause for bread staling and, therefore, it has attracted a lot of interest from the food industry. In this study, the inhibitory effect of citrus peel hydrolysates (CPH) on retrogradation of wheat starch (WS) in the presence of sucrose was investigated. The pasting properties showed that further addition of CPH caused a lower setback value than the addition of sucrose alone. Hardness of the gel, retrograded at 4 °C for five days, showed a similar tendency, which was reduced more in CPH addition than WS itself or sucrose addition alone. The low retrogradation enthalpy of the CPH including starch gel also indicated the positive effect of CPH on retarding retrogradation. These results suggested that incorporation of CPH in starch-based foods would be effective for inhibiting retrogradation, preventing the deterioration of the quality of food products.

Impact of static and dynamic modes of semi-dry heat reaction on the characteristics of starch citrates.

The objective of this study was to demonstrate the validity of dynamic semi-dry heat reaction (SDHR) by investigating the effects of static and dynamic SDHR on the characteristics of starch citrates. The starch (normal and waxy corn)-citric acid (CA) mixture was heated in a convection oven (static mode) or a twin-screw extruder without a die (dynamic mode). The ester bonds of starch citrates were confirmed by FT-IR, and their molar degree of substitution did not differ between the reaction modes for the tested starch genotypes. Starch citrates by dynamic SDHR exhibited higher relative crystallinity, resistant starch content, solubility, swelling power, and gelatinization compared to those by static SDHR. Although static SDHR did not show a viscosity development during pasting, dynamic SDHR increased their pasting viscosities. Overall, these results suggest that dynamic SDHR could improve the defects (i.e., lack of solubility, swelling, and pasting attributes) of the traditional starch citrates.

Starch nanoparticles produced via acidic dry heat treatment as a stabilizer for a Pickering emulsion: Influence of the physical properties of particles.

To evaluate the effect of the starch particle size on the stability of a Pickering emulsion, starch particles with various mean diameters were prepared using dry heat treatment under mildly acidic conditions. As addition level of acid increased, mean diameter of starch particles decreased from 23,100 to 15.7 nm, and contact angle of water droplet on the starch film increased from12 to 42.7°. Confocal microscopy revealed that a cohesive layer of starch particles with diameter of 28.6 nm surrounded the oil droplets, which may stabilize the emulsion. For starch particles smaller than 33.5 nm, the emulsion stability index increased up to approximately 95 % when starch content was higher than 9.1 %. For 16.7 % starch content, starch particles (< 33.5 nm) could form a stable emulsion with oil content of up to 61 %. For the emulsion stabilized by starch with 28.6 nm, it was stable when subjected to heating and freeze-thawing treatment.

Impact of starch granule-associated surface and channel proteins on physicochemical properties of corn and rice starches.

The objective of this study was to investigate the impact of removing starch granule-associated proteins (SGAPs), especially starch granule-associated surface and channel proteins, on the overall characteristics of corn and rice starches. Protease treatment predominantly removed SGAPs on surfaces and in channels of the starches without significant damage, as evidenced by confocal laser scanning microscopy coupled with protein-specific and non-reactive fluorescent dye staining. Compared to untreated starches, protease-treated (PT) starches showed higher solubility and lower swelling power. However, there were no changes in their gelatinization and melting temperatures, despite their higher relative crystallinity. The stability of swollen starch granules during shearing was reduced following SGAP removal, reducing their peak, final and setback viscosities of the paste. Taken together these results indicate that SGAP removal predominantly affects the rheological properties of starch. Furthermore, the lower setback in PT-starch pastes suggests short-term retrogradation may be retarded by protease treatment of starch.

Characteristics of wheat starch-pectin hydrolysate complexes by dry heat treatment.

The objective of this study was to characterize dry heat-induced wheat starch-pectin hydrolysate (WST/PH) complexes to develop the retrogradation-retarded starch. Native (N-) and protease-treated (P-) WST were used as starch sources. Pectin hydrolysates were mixed independently with N-WST and P-WST to a mixing ratio of 49:1 (based on total solid contents), followed by drying below 10% moisture and dry heat treatment at 130 °C for 4 h. The molar degrees of substitution (MS) was higher for WST/PH complexes than its mixtures, and apparent amylose contents decreased with their MS. Relative to WST/PH mixtures, solubilities were higher for WST/PH complexes, while swelling powers didn't differ. WST/PH complexes showed the lower degree of retrogradation, setback viscosities, slowly gelling tendency, and syneresis. These phenomena were more pronounced in WST/PH mixtures and complexes prepared with P-WST. Overall results suggest that dry heat-induced WST/PH complexes could be a potential retrogradation-retarded starch to replace chemically-modified starches.

Role of maltogenic amylase and pullulanase in maltodextrin and glycogen metabolism of Bacillus subtilis 168.

The physiological functions of two amylolytic enzymes, a maltogenic amylase (MAase) encoded by yvdF and a debranching enzyme (pullulanase) encoded by amyX, in the carbohydrate metabolism of Bacillus subtilis 168 were investigated using yvdF, amyX, and yvdF amyX mutant strains. An immunolocalization study revealed that YvdF was distributed on both sides of the cytoplasmic membrane and in the periplasm during vegetative growth but in the cytoplasm of prespores. Small carbohydrates such as maltoheptaose and beta-cyclodextrin (beta-CD) taken up by wild-type B. subtilis cells via two distinct transporters, the Mdx and Cyc ABC transporters, respectively, were hydrolyzed immediately to form smaller or linear maltodextrins. On the other hand, the yvdF mutant exhibited limited degradation of the substrates, indicating that, in the wild type, maltodextrins and beta-CD were hydrolyzed by MAase while being taken up by the bacterium. With glycogen and branched beta-CDs as substrates, pullulanase showed high-level specificity for the hydrolysis of the outer side chains of glycogen with three to five glucosyl residues. To investigate the roles of MAase and pullulanase in glycogen utilization, the following glycogen-overproducing strains were constructed: a glg mutant with a wild-type background, yvdF glg and amyX glg mutants, and a glg mutant with a double mutant (DM) background. The amyX glg and glg DM strains accumulated significantly larger amounts of glycogen than the glg mutant, while the yvdF glg strain accumulated an intermediate amount. Glycogen samples from the amyX glg and glg DM strains exhibited average molecular masses two and three times larger, respectively, than that of glycogen from the glg mutant. The results suggested that glycogen breakdown may be a sequential process that involves pullulanase and MAase, whereby pullulanase hydrolyzes the alpha-1,6-glycosidic linkage at the branch point to release a linear maltooligosaccharide that is then hydrolyzed into maltose and maltotriose by MAase.

A study of the quality characteristics of frozen Korean rice cake by using different thawing methods.

The characteristics of frozen rice cakes after thawing them using different methods, such as standing at room temperature (NT), running water (RWT), pan-grill (PT), steam (ST), microwave (MWT), and superheated steam thawing (SHST), were compared. Frozen rice cakes treated by MWT or SHST showed the shortest thawing time of 3 min. The MWT treatment showed the largest thawing loss, while the ST treatment showed the highest moisture content. The ST, RWT, and MWT treatments showed the highest water activity values. The NT treatment exhibited the highest hardness values, whereas the ST treatment showed the lowest values, possibly due to the adverse effects of high temperature on them. Sensory evaluation showed differences in appearance, moistness, and tenderness according to the thawing method, but there was no significant difference in overall acceptability. This study suggests that the qualities of frozen rice cakes varied depending on the different thawing methods.

Reaction pattern differences impact physical properties of starches derivatized to the same extent in a model cross-linking system.

This study investigated the physical properties of maize (MS) and wheat (WS) starches derivatized with 5-(4,6-dichlorotriazinyl)aminofluorescein (model cross-linking system) to have the same overall fluorescence intensity on starch molecules, but reacted either more uniformly throughout granules (UD) or more at granule surfaces (SD). Both MS and WS derivatives had lower swelling powers (SP) at 90°C than their respective native starches. The UD derivatives had lower SP (90°C) and greater retrogradation enthalpies than did SD derivatives, consistent with their lower peak and higher setback pasting viscosities. Also, SD starches were less soluble and retained a greater degree of granular integrity than UD starches in time-lapse, hot-stage light microscopy studies (50-95°C), likely due to a greater concentration of cross-links at the granule surface. The results confirm that derivatization patterns impact the physical properties of modified starches. Thus, varying derivatization patterns can be a strategy to tailor modified starch properties.

Physicochemical interactions of cycloamylose with phenolic compounds.

The complex formation capability of cycloamylose (CA), having a degree of polymerization of 23-45, with phenolic compounds (PCs) was investigated using various physicochemical techniques. The fluorescence intensity of PCs increased and then reached a plateau at 10-20mM cyclodextrin, while it continued to increase at up to 60mM CA. Thermodynamic data of CA complexes with PCs revealed that the binding process was primarily enthalpy-driven and spontaneous. CA favored to form the most stable complex with chlorogenic acid (CHA) among all PCs. Chemical shift changes for the protons in interior and exterior of CA, as well as in PCs suggested a possible formation of both inclusion and extramolecular interactions between CA and PCs. The ROESY spectrum confirmed that the aromatic moieties of CHA were partially interacted with CA molecules through relatively weak binding. XRD, DSC, and SEM results also supported the complex formation by intermolecular interaction between CA and CHA.

Impact of reagent infiltration time on reaction patterns and pasting properties of modified maize and wheat starches.

The impact of granular and molecular reaction patterns on modified starch properties was investigated as a function of the length of time allowed for reagent to infiltrate starch granules. A fluorescent reagent [5-(4,6-dichlorotriazinyl)aminofluorescein] was dispersed in aqueous normal maize or wheat starch slurries (35%, w/v) for 0, 5, 10, 30, or 60min, after which reaction was initiated by increasing the pH to 11.5 and allowing reaction to proceed for 3h. With increasing lengths of infiltration, the reaction became increasingly homogeneous within the granule interior (matrix) and the AM:AP reactivity ratio increased (wheat starch), as assessed by confocal laser scanning microscopy (CLSM) and size-exclusion chromatography (refractive index and fluorescence detection), respectively. A longer reagent infiltration time also led to a more inhibited (i.e., cross-linked) pasting viscosity, suggesting that both granular and/or molecular reaction patterns were altered by varied reagent infiltration times to ultimately impact modified starch properties.

High-yield cycloamylose production from sweet potato starch using Pseudomonas isoamylase and Thermus aquaticus 4-α-glucanotransferase.

An optimal reaction condition for producing cycloamylose (CA) from sweet potato starch was investigated using a combination of isoamylase (from Pseudomonas sp.) and 4-α-glucanotransferase (from Thermus aquaticus, TAαGT). Starch was debranched by isoamylase for 8 h and subsequently reacted with TAαGT for 12 h. The yield and purity of CA products were determined using HPSEC and MALDI-TOFMS, respectively. Consequently, the maximum yield was 48.56%, exhibiting the highest CA production efficiency ever reported from starch. The CA products showed a wide range of the degree of polymerization (DP) with the minimum DP of 5. CA was also produced by simultaneous treatment of isoamylase and TAαGT. The yield was 3.31%, and the final products were contaminated by multiple branched and linear molecules. This result suggests that a former reaction condition (the sequential addition of isoamylase and TAαGT) is preferable for producing CA from sweet potato starch.