The effects of chain-length distributions on starch-related properties in waxy rices.

Normal rice starch consists of amylopectin and amylose, whose relative amounts and chain-length distributions (CLDs) are major determinants of the digestibility and rheology of cooked rice, and are related to metabolic health and consumer preference. Here, the mechanism of how molecular structural features of pure amylopectin (waxy) starches affect starch properties was explored. Following debranching, chain-length distributions of seven waxy varieties were measured using size-exclusion chromatography, and parameterized using biosynthesis-based models, which involve breaking up the chain-length distribution into contributions from five enzyme sets covering overlapping ranges of chain length; structure-property correlations involving the fifth set were found to be statistically significant. Digestibility was measured in vitro, and parameters for the slower and longer digestion phase quantified using non-linear least-squares fitting. The coefficient for the significant correlation involving amylopectin fine structure for the fifth set was -0.903, while the amounts of amylopectin short and long chains were found to dominate breakdown viscosity (correlation coefficients 0.801 and - 0.911, respectively). This provides a methodology for finding or developing healthier starch in terms of lower digestion rate, while also having acceptable palatability. As rice breeders can to some extent control CLDs, this can help the development of waxy rices with improved properties.

Molecular insights on the origin and development of waxy genotypes in major crop plants.

Starch is a significant ingredient of the seed endosperm with commercial importance in food and industry. Crop varieties with glutinous (waxy) grain characteristics, i.e. starch with high amylopectin and low amylose, hold longstanding cultural importance in some world regions and unique properties for industrial manufacture. The waxy character in many crop species is regulated by a single gene known as GBSSI (or waxy), which encodes the enzyme Granule Bound Starch Synthase1 with null or reduced activity. Several allelic variants of the waxy gene that contribute to varying levels of amylose content have been reported in different crop plants. Phylogenetic analysis of protein sequences and the genomic DNA encoding GBSSI of major cereals and recently sequenced millets and pseudo-cereals have shown that GBSSI orthologs form distinct clusters, each representing a separate crop lineage. With the rapidly increasing demand for waxy starch in food and non-food applications, conventional crop breeding techniques and modern crop improvement technologies such as gene silencing and genome editing have been deployed to develop new waxy crop cultivars. The advances in research on waxy alleles across different crops have unveiled new possibilities for modifying the synthesis of amylose and amylopectin starch, leading to the potential creation of customized crops in the future. This article presents molecular lines of evidence on the emergence of waxy genes in various crops, including their genesis and evolution, molecular structure, comparative analysis and breeding innovations.

Examining starch metabolism in lotus roots (Nelumbo nucifera Gaertn.) during post-harvest storage at different temperatures.

This study explores the impact of postharvest storage temperatures (4 °C and 25 °C) on starch metabolism and textural attributes of glutinous lotus root. While starch metabolism is a well-known factor influencing texture, changes in powdery and sticky qualities have remained unexplored. Our research reveals that storing lotus roots at 4 °C delays water dissipation, amylopectin reduction, and the decline in textural elements such as hardness, adhesiveness, springiness, gumminess, and resilience. Lower temperatures postpone amylopectin reduction and sugar interconversion, thereby preserving the sticky texture. Additionally, they suppress starch formation, delay starch metabolism, and elevate the expression of genes involved in starch metabolism. The correlation between gene expression and root texture indicates the critical role of gene regulation in enzyme activity during storage. Overall, low-temperature storage extends lotus root preservation by regulating metabolite content, enzyme activities, and the corresponding genes involved in starch metabolism, preserving both intrinsic and external root quality.

Insights into multiscale structure and digestive characteristic of starch from two cultivars of chestnut during kernel development.

Multiscale structure and digestive characteristic of starch during kernel development of Castanea henryi ('Jinzhui' (YS) and 'Baiyan No.1' (WS)) were investigated in this study. Structural analysis revealed that the surface of starch granules became smooth, the amylopectin content decreased (from 71.32 % to 70.47 %, from 71.44 % to 68.37 %, respectively), the chain length distribution of amylopectin reduced (the proportion of B1 chain decreased from 52.35 % to 50.60 %, from 52.22 % to 50.59 %, respectively) while the amorphous and semi-crystalline lamellae of starch increased during development, which was consistent with the decreasing relative crystallinity (from 28.79 % to 24.11 %, from 29.57 % to 23.66 %, respectively) and short-range ordering degree. The degradation of ordered structure further resulted in the increase of digestibility, especially in the late developmental stage, supported by a significant decrease of resistant starch content (from 70.21 % to 61.70 % and from 73.58 % to 58.86 %, respectively). Transcriptome analysis and RT-qPCR were performed to explore the possible molecular mechanisms affecting starch structure. The high expression of several key genes including AGPase, GBSS, SBE, SSS, ISA and PUL in late development stage might be the reason of structural changes during development. The results provided valuable information for starch accumulation during kernel development of Castanea henryi.

Persimmon leaf polyphenols as potential ingredients for modulating starch digestibility: Effect of starch-polyphenol interaction.

The interaction mode between persimmon leaf polyphenols (PLP) and corn starch with different amylose content and its effect on starch digestibility was studied. Results of iodine binding test, TGA, and DSC revealed that PLP interacted with starch and reduced the iodine binding capacity and thermal stability of starch. High amylopectin corn starch (HAPS) interacted with PLP mainly via hydrogen bonds, since the FT-IR of HAPS-PLP complex showed higher intensity at 3400 cm-1 and an obvious shift of 21 cm-1 to shorter wavelength, and the chemical shifts of protons in 1H NMR and the shift of C-6 peak in 13C NMR of HAPS moved to low field with the addition of PLP. Results of 1H NMR also showed the preferential formation of hydrogen bonds between PLP and OH-3 of HAPS. Different from HAPS, PLP formed V-type inclusion complex with high amylose corn starch (HAS) because XRD of HAS-PLP complex showed characteristic feature peaks of V-type inclusion complex and C-1 signal in 13C NMR of PLP-complexed HAS shifted to low field. Interaction with PLP reduced starch digestibility and HAS-PLP complex resulted in more resistant starch production than HAPS-PLP complex. To complex PLP with starch might be a potential way to prepare functional starch with slower digestion.

Molecular Basis of the Granular Characteristics of Small-Granule Starch: A Comparative Study.

Small-granule starches (SGSs) have technological advantages over starches of conventional sizes for many applications. The study compared the granular characteristics of three SGSs (from amaranth, quinoa, and taro) with those of maize and potato starches and revealed their molecular basis. The results indicated that the supramolecular architecture of starch granules was not necessarily correlated with granule size. Acid hydrolysis of amaranth and quinoa starches was fast due to not only their small granule sizes but also the defects in the supramolecular structure, to which short external and internal chain lengths of amaranth and quinoa amylopectins contributed. By comparison, the granular architecture of taro starch granules was more stable partly due to the longer external chain length of taro amylopectin. Comparison of the molecular composition of branched subunits (released by using α-amylase of Bacillus amyloliquefaciens) in amylopectins and that in lintnerized starches suggested a significant heterogeneous degradation of amaranth and quinoa starches at supramolecular levels.

Effects of different extrusion temperatures on the physicochemical properties, edible quality and digestive attributes of multigrain reconstituted rice.

Multigrain reconstituted rice, as a nutritious and convenient staple, holds considerable promise for the food industry. Furthermore, highland barley, corn, and other coarse cereals are distinguished by their low glycemic index (GI), rendering them effective in mitigating postprandial blood glucose levels, thereby underscoring their beneficial physiological impact. This study investigated the impact of extrusion temperature on the physicochemical properties, edible quality, and digestibility of multigrain reconstituted rice. The morphology revealed that starch particles that are not fully gelatinized in multigrain reconstituted rice are observed at an extrusion temperature range of 60 °C-90 °C. As the extrusion temperature increased, the degree of gelatinization (DG) increased, while the contents of water, protein, total starch, and amylopectin decreased substantially. Concurrently, the relative crystallinity, orderliness of starch, and heat absorption enthalpy (ΔH) decreased significantly, and water absorption (WAI) and water solubility (WSI) increased markedly. Regarding edible quality, sensory evaluation displayed an initial increase followed by a decrease. In terms of digestibility, the estimated glycemic index (eGI) increased from 61.10 to 70.81, and the GI increased from 60.41 to 75.33. In addition, the DG was significantly correlated with both eGI (r = 0.886**) and GI (r = 0.947**). The results indicated that the ideal extrusion temperature for multigrain reconstituted rice was 90 °C. The findings underscored the pivotal role of optimal extrusion temperatures in the production of multigrain reconstituted rice, which features low GI and high nutritional quality.

Enhancing starch functionality through synergistic modification via sequential treatments with cold plasma and electron beam irradiation.

The impact of dual sequential modifications using radio-frequency (RF) plasma and electron beam irradiation (EBI) on starch properties was investigated and compared with single treatments within an irradiation dose range of 5-20 kGy. Regardless of sequence, dual treatments synergistically affected starch properties, increasing acidity, solubility, and paste clarity, while decreasing rheological features with increasing irradiation dose. The molecular weight distribution was also synergistically influenced. Amylopectin distribution broadened particularly below 10 kGy. Amylose narrowed its distribution across all irradiation doses. This was due to dominating EBI-induced degradation and molecular rearrangements from RF plasma. With the highest average radiation-chemical yield (G) and degradation rate constant (k) of (2.12 ± 0.14) × 10-6 mol·J-1 and (3.43 ± 0.23) × 10-4 kGy-1, respectively, upon RF plasma pre-treatment, amylose underwent random chain scission. In comparison to single treatments, dual modification caused minor alterations in spectral characteristics and crystal short-range order structure, along with increased granule aggregation and surface irregularities. The synergistic effect was dose-dependent, significant up to 10 kGy, irrespective of treatment sequence. The highest synergistic ratio was observed when RF plasma preceded irradiation, demonstrating the superior efficiency of plasma pre-treatment in combination with EBI. This synergy has the potential to lower costs and extend starch's technological uses by enhancing radiation sensitivity and reducing the irradiation dose.

Insights into wheat-starch biosynthesis from two-dimensional macromolecular structure.

Starch structure is often characterized by the chain-length distribution (CLD) of the linear molecules formed by breaking each branch-point. More information can be obtained by expanding into a second dimension: in the present case, the total undebranched-molecule size. This enables answers to questions unobtainable by considering only one variable. The questions considered here are: (i) are the events independent which control total size and CLD, and (ii) do ultra-long amylopectin (AP) chains exist (these chains cannot be distinguished from amylose chains using simple size separation). This was applied here to characterize the structures of one normal (RS01) wheat and two high-amylose (AM) mutant wheats (an SBEIIa knockout and an SBEIIa and SBEIIb knockout). Absolute ethanol was used to precipitate collected fractions, then size-exclusion chromatography for total molecular size and for the size of branches. The SBEIIa and SBEIIb mutations significantly increased AM and IC contents and chain length. The 2D plots indicated the presence of small but significant amounts of long-chain amylopectin, and the asymmetry of these plots shows that the corresponding mechanisms share some causal effects. These results could be used to develop plants producing improved starches, because different ranges of the chain-length distribution contribute independently to functional properties.

Creating a zero amylose barley with high soluble sugar content by genome editing.

Amylose biosynthesis is strictly associated with granule-bound starch synthase I (GBSSI) encoded by the Waxy gene. Mutagenesis of single bases in the Waxy gene, which induced by CRISPR/Cas9 genome editing, caused absence of intact GBSSI protein in grain of the edited line. The amylose and amylopectin contents of waxy mutants were zero and 31.73%, while those in the wild type were 33.50% and 39.00%, respectively. The absence of GBSSI protein led to increase in soluble sugar content to 37.30% compared with only 10.0% in the wild type. Sucrose and ß-glucan, were 39.16% and 35.40% higher in waxy mutants than in the wild type, respectively. Transcriptome analysis identified differences between the wild type and waxy mutants that could partly explain the reduction in amylose and amylopectin contents and the increase in soluble sugar, sucrose and ß-glucan contents. This waxy flour, which showed lower final viscosity and setback, and higher breakdown, could provide more option for food processing.

Amylopectin chain length distributions and amylose content are determinants of viscoelasticity and digestibility differences in mung bean starch and proso millet starch.

This study aimed to reveal the underlying mechanisms of the differences in viscoelasticity and digestibility between mung bean starch (MBS) and proso millet starch (PMS) from the viewpoint of starch fine molecular structure. The contents of amylopectin B2 chains (14.94-15.09 %), amylopectin B3 chains (14.48-15.07 %) and amylose long chains (183.55-198.84) in MBS were significantly higher than PMS (10.45-10.76 %, 12.48-14.07 % and 70.59-88.03, respectively). MBS with higher amylose content (AC, 28.45-31.80 %) not only exhibited a lower weight-average molar mass (91,750.65-128,120.44 kDa) and R1047/1022 (1.1520-1.1904), but also was significantly lower than PMS in relative crystallinity (15.22-23.18 %, p < 0.05). MBS displayed a higher storage modulus (G') and loss modulus (G'') than PMS. Although only MBS-1 showed two distinct and discontinuous phases, MBS exhibited a higher resistant starch (RS) content than PMS (31.63-39.23 %), with MBS-3 having the highest RS content (56.15 %). Correlation analysis suggested that the amylopectin chain length distributions and AC played an important role in affecting the crystal structure, viscoelastic properties and in vitro starch digestibility of MBS and PMS. These results will provide a theoretical and scientific basis for the development of starch science and industrial production of low glycemic index starchy food.

Effects of cooking methods on the physical properties and in vitro digestibility of starch isolated from Chinese yam.

The objective of this study was to compare the structural and functional attributes of Chinese yam starches obtained via different domestic cooking methods. Cooking changed the crystalline type from the C type to the CB type, and disrupted the short- and long-range molecular order of Chinese yam starch. The average chain length of amylopectin in BOS (boiling starch) was the smallest at 22.78, while RWS had the longest average chain length, reaching 24.24. These alterations in molecular structure resulted in variations in functional properties such as solubility, swelling power (SP), pasting characteristics, and rheological properties. Among these alterations, boiling was the most effective method for increasing the water-binding capacity and SP of starch. Specifically, its water holding capacity was 2.12 times that of RWS. In vitro digestion experiments indicated that BOS has a higher digestion rate (k = 0.0272 min-1) and lower RDS (rapidly digestible starch), which may be related to its amylopectin chain length distribution. This study can guide us to utilize yam starch through suitable cooking methods, which is relevant for the processing and application of Chinese yam starch.

Rheological properties of indica rice determined by starch structure related enzymatic activities during after-ripening.

The processing quality of indica rice must undergo ripening after harvest to achieve stability and improvement. However, the mechanism underlying this process remains incompletely elucidated. Starch, the predominant component in indica rice, plays a crucial role in determining its properties. This study focused on analyzing the rheological properties and starch fine structure, as well as the related biosynthetic enzymes of indica rice during the after-ripening process. The results showed that after-ripened rice exhibited increased elastic modulus (G') and viscous modulus (G″), accompanied by a decrease in the loss tangent (Tan δ), indicating an enhancement in viscoelasticity and the gel network structure. Moreover, the proportions of amylopectin super long chains (DP 37-60) decreased, while those of medium chains (DP 13-24 and DP 25-36) or short chains (DP 6-12) of amylopectin increased. Additionally, the activities of starch branching enzyme (SBE) and starch debranching enzyme (DBE) declined over the after-ripening period. Pearson correlation analysis revealed that the rheological properties of after-ripened rice were correlated with the chain length distribution (CLD) of starch, which, in turn, was associated with its related endogenous enzymes. These findings provied new insights into understanding the quality changes of after-ripened indica rice.

Impact of soluble soybean polysaccharide on the gelatinization and retrogradation of corn starches with different amylose content.

Polysaccharides have a significant impact on the physicochemical properties of starch, and the objective of this study was to examine the effect of incorporating soluble soybean polysaccharide (SSPS) on the gelatinization and retrogradation of corn starches (CS) with varying amylose content. In contrast to high-amylose corn starch (HACS), the degree of gelatinization of waxy corn starch (WCS) and normal corn starch (NCS) decreased with the addition of SSPS. The inclusion of SSPS resulted in reduced swelling power in all CS, and led to a decrease in gel hardness of the starches. The intermolecular forces between SSPS and CS were primarily hydrogen bonding, and a gel network structure was formed, thereby retarding the short-term and long-term retrogradation of CS. Scanning electron microscopy results revealed that the addition of SSPS in starches led to a loose network structure with larger poles and a reduced ordered structure after retrogradation, as observed from the cross-section of formed gels. These findings suggested that SSPS has great potential for applications in starchy foods, as it can effectively retard both gelatinization and retrogradation of starches.

A preparation of debranched waxy maize starch derivatives: Effect of drying temperatures on crystallization and digestibility.

The impact of recrystallization conditions and drying temperatures on the crystallization and digestibility of native waxy maize (Zea mays L.) starch (NWMS) was explored. This study involved subjecting NWMS to concurrent debranching and crystallization at 50 °C for up to 7 days. Samples were collected by oven-drying at 40, 60, and 80 °C for 24 h. This simultaneous debranching and crystallization process increased the resistant starch (RS) content by approximately 48 % compared to the native starch. The drying temperatures significantly influenced the RS content, with samples dried at 60 °C exhibiting the lowest digestibility. X-ray diffraction (XRD) analysis revealed that most crystals demonstrated a characteristic A-type arrangement. Debranching and crystallization processes enhanced the crystallinity of the samples. The specific crystal arrangement (A- or B-type) depended on the crystallization conditions. A 15 min heating of NWMS in a boiling water bath increased the digestible fraction to over 90 %, while the samples subjected to debranching and crystallization showed an increase to only about 45 %. A linear correlation between starch fractions and enthalpy was also observed.

V-type granular starches prepared by maize starches with different amylose contents: An investigation in structure, physicochemical properties and digestibility.

V-type granular starches (VGSs) were prepared via an ethanol-alkaline (EA) method using maize starch with different amylose contents, specifically, high amylose (HAM), normal maize starch (MS), and waxy maize starch (WS). The X-ray diffraction pattern of the native starch was completely transformed into a V-type pattern after the EA treatment, indicating a structural change in the starch granules. The VGSs prepared by HAM had highest relative crystallinity (31.8°), while the VGSs prepared by WS showed amorphous diffraction pattern. Excessive NaOH, however, would disrupt the formation of V-type structures and cause granular shape rupture. The quantity of double-helical structures, particularly those formed by amylopectin at the starch granules' periphery, significantly decreased. Conversely, single-helical structures formed by amylose increased. A notable rise in the relative crystallinity of V crystals. Four VGS samples, characterized by granular integrity, were chosen for the next investigation of physicochemical and digestive properties. VGS prepared from HAM exhibited higher granular integrity, lower cold-water swelling extent (59.0 and 161.0 cP), improved thermal stability (the value of breakdown as lower as 57.67 and 186.67 cP), and higher resistance to digestion (RS content was up to 10.38 % and 9.00 % higher than 5.86 % and 5.66 % of VGS prepared from WS and MS). The results confirmed that amylose content has a substantial impact on the microstructural and physicochemical properties of VGSs.

Enhancing the anti-thixotropic properties of waxy maize starch modified by different α-amylases and its underlying molecular mechanism.

The large thixotropy of the starch-thickened foods is often unfavorable in many applications. This study examined the contribution of the proportion of amylopectin chain length to time-dependence of starch gels. The α-amylase (AM) from Bacillus stearothermophilus and maltogenic α-amylase (MA) from Bacillus subtilis were used to trim amylopectin in different reaction patterns. HPLC, HPAEC and IBC data suggested AM attacked B-chains (DP 12-36), causing an increment in number of the chains with DP 6-12, whereas MA primarily trimmed the short B-chains (DP 12-18) and partial A-chains (DP 9-12) to generate short chains with DP 6-9. Interestingly, the recovery of AM-gels was faster than MA-gels at the same degree of hydrolysis when subjected to shear according to the linear correlation analysis. When releasing the same mass of sugar, shortening of the long internal chains played an important role in reducing time dependence of starch gel rather than the external side chains. Possible models were proposed to illustrate the differences in the mechanism of rapid-recovery caused by different side-chain distributions. The outcome provided a new perspective to regulate the thixotropy behavior of starch through enzyme strategies in the granular state.

Physicochemical properties and in vitro digestibility of ginseng starches under citric acid-autoclaving treatment.

In this study, the effects of citric acid-autoclaving (CA-A) treatment on physicochemical and digestive properties of the native ginseng starches were investigated. The results showed that ginseng starch exhibited a B-type crystal structure with a low onset pasting temperature of 44.23 ± 0.80 °C, but high peak viscosity and setback viscosity of 5897.34 ± 53.72 cP and 692.00 ± 32.36 cP, respectively. The granular morphology, crystal and short-range ordered structure of ginseng starches were destroyed after CA-A treatment. The more short-chain starches were produced, resulting in the ginseng starches solubility increased. In addition, autoclaving, citric acid (CA) and CA-A treatment promoted polymerization and recrystallization of starch molecules, increased the proportion of amylopectin B1, and B3 chains, and improved molecular weight and resistant starch (RS) content of ginseng starches. The most significant multi-scale structural change was induced by CA-A treatment, which reduced the relative crystallinity of ginseng starch from 28.26 ± 0.24 % to 2.75 ± 0.08 %, and increased the content of RS to 54.30 ± 0.14 %. These findings provided a better understanding of the structure and properties of Chinese ginseng starches and offered new ideas for the deep processing of ginseng foods.

Effects of purple potato anthocyanins on the in vitro digestive properties of starches of different crystalline types.

This study explored the potential of purple potato anthocyanins (PPAs) in regulating the digestive properties of starches of various crystalline types. In vitro digestion experiments indicated that PPAs inhibit the hydrolysis of rice starch (A-type) better than that of garden pea starch (C-type) and potato starch (B-type). Further structural assessment of different PPA-starch systems showed that PPAs and starch likely interact through non-covalent bonds, resulting in structural changes. Microstructural changes observed in the starches were consistent with the in vitro digestion results, and the chain length and proportions of short/long chains in amylopectin molecules affected the binding strengths and interaction modes between PPAs and starch. Hence, the three starches differed in their PPA loading efficiency and digestibility. These discoveries contribute to a deeper understanding of the mechanisms underlying the inhibition of starch digestibility by PPAs. They can aid the formulation of value-added products and low-glycemic-index foods.

Dynamics of starch formation and gene expression during grain filling and its possible influence on grain quality.

In rice, grain filling is a crucial stage where asynchronous filling of the pollinated spikelet's of the panicle occurs. It can influence both grain quality and yield. In rice grain, starch is the dominant component and contains amylose and amylopectin. Amylose content is the chief cooking quality parameter, however, rice varieties having similar amylose content varied in other parameters. Hence, in this study, a set of varieties varying in yield (04) and another set (12) of varieties that are similar in amylose content with variation in gel consistency and alkali spreading value were used. Panicles were collected at various intervals and analysed for individual grain weight and quantities of amylose and amylopectin. Gas exchange parameters were measured in varieties varying in yield. Upper branches of the panicles were collected from rice varieties having similar amylose content and were subjected to gene expression analysis with fourteen gene specific primers of starch synthesis. Results indicate that grain filling was initiated simultaneously in multiple branches. Amylose and amylopectin quantities increased with the increase in individual grain weight. However, the pattern of regression lines of amylose and amylopectin percentages with increase in individual grain weight varied among the varieties. Gas exchange parameters like photosynthetic rate, stomatal conductance, intercellular CO2 and transpiration rate decreased with the increase in grain filling period in both good and poor yielding varieties. However, they decreased more in poor yielders. Expression of fourteen genes varied among the varieties and absence of SBE2b can be responsible for medium or soft gel consistency.