Multi-scale structural insights on starch digestibility of instant rice.

Multi-scale structures were investigated to understand starch digestibility of instant rice. A wide range of maximum starch digested ratio, up to about 20%, was observed among instant rice prepared from different rice varieties. Instant rice with a smooth and densely packed cross-section showed slower starch digestibility than those with a porous and loosely packed structure. All samples displayed B + V type crystallinity, with V-type crystallinity negatively correlating with maximum starch digested percentage. After digestion, starch chain-length distributions were significantly altered: rapidly digested starch comprised long amylose and short amylopectin chains, while slowly digested starch comprised chains with a peak degree of polymerization (DP) around 130. These results indicate that instant rice with a compact microstructure, high V-type crystallinity, and DP 130 fractions during digestion can reduce starch digestibility. This study provides insights for food industry to develop instant rice products with slow starch digestibility, potentially improving human health.

Morphological, Molecular Structural and Physicochemical Characterization of Starch Granules Formed in Endosperm of Rice with Ectopic Overexpression of α-Amylase.

The objective of this study was to characterize the endosperm starch in rice that ectopically overexpressed the α-amylase. Transgenic rice plants, transformed with cauliflower mosaic virus 35S promoter driven AmyI-1 (35S::AmyI-1) and AmyII-4 (35S::AmyII-4), and 10 kDa prolamin promoter driven AmyI-1 (P10::AmyI-1), were cultivated under standard conditions (23 °C, 12 h in the dark/ 26 °C, 12 h in the light), and brown grains were subsequently harvested. Each grain displayed characteristic chalkiness, while electron microanalyzer (EPMA)-SEM images disclosed numerous small pits on the surface of the starch granules, attributable to α-amylase activity. Fluorescence labeling and capillary electrophoresis analysis of starch chain length distribution revealed no significant alterations in the starches of 35S::AmyI-1 and 35S::AmyII-4 transgenic rice compared to the wild-type. Conversely, the extremely short α-glucan chains (DP 2-8) exhibited a dramatic increase in the P10::AmyI-1 starch. Rapid visco-analyzer analysis also identified variations in the chain length distribution of P10::AmyI-1 starch, manifesting as changes in viscosity. Moreover, 1H-NMR analysis uncovered dynamic modifications in the molecular structure of starch in rice grain transformed with P10::AmyI-1, which was found to possess unprecedented structural characteristics.

Absolute Quantitative Lipidomics Reveals Different Granule-Associated Surface Lipid Roles in the Digestibility and Pasting of Waxy, Normal, and High-Amylose Rice Starches.

Granule-associated surface lipids (GASLs) and internal lipids showed different lipid-amylose relationships, contents, and distributions, suggesting their differing biological origins and functions, among waxy, normal, and high-amylose rice starch. The GASL content mainly depended on the pore size, while internal lipids regulated starch biosynthesis, as indicated by correlations of internal lipids with the chain length distribution of amylopectin and amylose content. Of the 1346 lipids detected, 628, 562, and 408 differentially expressed lipids were observed between normal-waxy, high-amylose-waxy, and normal-high-amylose starch, respectively. After the removal of GASLs, the higher lysophospholipid content induced greater decreases in the peak and breakdown viscosity and swelling power, while the highest digestibility increase was found with the highest triacylglycerol content. Thus, different GASL compositions led to different digestibility, swelling, and pasting outcomes. This study sheds new light on the mechanism of the role of GASLs in the structure and properties of starch, as well as in potential modifications and amyloplast membrane development.

Relationships among Structure, Physicochemical Properties and In Vitro Digestibility of Starches from Ethiopian Food Barley Varieties.

Studying diversity in local barley varieties can help advance novel uses for the grain. Therefore, starch was isolated from nine Ethiopian food barley varieties to determine starch structural, pasting, thermal, and digestibility characteristics, as well as their inter-relationships. The amylose content in the varieties significantly varied from 24.5 to 30.3%, with a coefficient of variation of 6.1%. The chain length distributions also varied significantly, and fa, fb1, fb2, and fb3 ranged from 26.3 to 29.0, 48.0 to 49.7, 15.0 to 15.9, and 7.5 to 9.5%, respectively. Significant variations were also exhibited in absorbance peak ratios, as well as thermal, pasting, and in vitro digestibility properties, with the latter two parameters showing the greatest diversity. Higher contents of amylose and long amylopectin fractions contributed to higher gelatinization temperatures and viscosities and lower digestibility. Structural characteristics showed strong relationships with viscosity, thermal, and in vitro digestibility properties. Cross 41/98 and Dimtu varieties are more suitable in functional food formulations and for bakery products. These results might inspire further studies to suggest target-based starch modifications and new product development.

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.

GBSS mutations in an SBE mutated background restore the potato starch granule morphology and produce ordered granules despite differences to native molecular structure.

Potato starch with mutations in starch branching enzyme genes (SBEI, SBEII) and granule-bound starch synthase gene (GBSS) was characterized for molecular and thermal properties. Mutations in GBSS were here stacked to a previously developed SBEI and SBEII mutation line. Additionally, mutations in the GBSS gene alone were induced in the wild-type variety for comparison. The parental line with mutations in the SBE genes showed a âˆ¼ 40 % increase in amylose content compared with the wild-type. Mutations in GBSS-SBEI-SBEII produced non-waxy, low-amylose lines compared with the wild-type. An exception was a line with one remaining GBSS wild-type allele, which displayed ∼80 % higher amylose content than wild-type. Stacked mutations in GBSS in the SBEI-SBEII parental line caused alterations in amylopectin chain length distribution and building block size categories of whole starch. Correlations between size categories of building blocks and unit chains of amylopectin were observed. Starch in GBSS-SBEI-SBEII mutational lines had elevated peak temperature of gelatinization, which was positively correlated with large building blocks.

Comparative analysis of granular starch hydrolysis and multi-structural changes by diverse α-amylases sources: Insights from waxy rice starch.

Three cultivars of waxy rice starch with different multi-scale structures were subjected to α-amylase hydrolysis to determine amylopectin fine structure, production of oligosaccharides, morphology, and crystallinity of the partially hydrolyzed starch granules. α-amylases hydrolyzed the amylopectin B2 chain during the initial stage of hydrolysis, suggesting that it is primarily located in the outer shell of the granules. For waxy rice starch with loose structure, α-amylases attacked the crystalline and amorphous regions simultaneously in the initial stage, while for starch granules with compact structure, the outer shell blocklet (crystalline structure) can be a hurdle for α-amylases to proceed to hydrolysis of the internal granule structure. The ability of α-amylases from porcine pancreatic α-amylases to attack the outer shell crystalline structure was lower than that of α-amylases from Bacillus amyloliquefaciens and Aspergillus oryzae. These results show that α-amylase source and rice cultivar combinations can be used to generate diverse structures in degraded waxy rice starch.

Prediction Models of Mechanical Properties of Jute/PLA Composite Based on X-ray Computed Tomography.

The tensile strength and modulus of elasticity of a jute/polylactic acid (PLA) composite were found to vary nonlinearly with the loading angle of the specimen through the tensile test. The variation in these properties was related to the fiber orientation distribution (FOD) and fiber length distribution (FLD). In order to study the effects of the FOD and FLD of short fibers on the mechanical properties and to better predict the mechanical properties of short-fiber composites, the true distribution of short fibers in the composite was accurately obtained using X-ray computed tomography (XCT), in which about 70% of the jute fibers were less than 300 µm in length and the fibers were mainly distributed along the direction of mold flow. The probability density functions of the FOD and FLD were obtained by further analyzing the XCT data. Strength and elastic modulus prediction models applicable to short-fiber-reinforced polymer (SFRP) composites were created by modifying the laminate theory and the rule of mixtures using the probability density functions of the FOD and FLD. The experimental measurements were in good agreement with the model predictions.

Variations in the effects of extrusion treatments and ferulic acid addition on starch digestibility with different botanical backgrounds.

In the current study, the effects of extrusion using a haake rheometer with a twin-roll mixer, with and without FA addition, on the structures and in vitro digestibility of starches from different sources were investigated. After extruding for 15 min at 90 °C with a moisture content of 40 %, no matter FA was added or not, lager Ap molecules were preferentially debranched, while Am with longer CL were depolymerized simultaneously, resulting to reduced averaged molecular size of Ap and shortened Am chains. Of all starches, regardless of their botanical backgrounds, although synergic effects were found between extrusion and FA addition on reducing their relative crystallinity and the ordered structures, distinctly different effects on the in vitro digestibility of these starches have also been observed especially regarding the digestion of starch branches with DP > 10 Particularly, the Am chains with DP 10-1000 was remaining undigested when FA was added. This study provides important information concerning how to adjust starch digestibility into a healthy range through altering the starch structures using extrusion technique with the addition of phytochemicals or not.

Starch molecular structure, physicochemical properties and in vitro digestibility of Ethiopian malt barley varieties.

Characterization of local varietal barley quality diversity can help boost further development of novel value-added utilization of the grain. Therefore, in this study starch was isolated from 11 Ethiopian malting barley varieties to determine starch structural, pasting, thermal and digestibility characteristics, and their inter-relationships. The varieties showed significant differences in all amylopectin chain length fractions, and the A, B1, B2 and B3 chains ranged from 25.4 to 30.1, 47.4-50.1, 14.3-16.0 and 7.8-9.0 %, respectively. The varieties also exhibited significant variation in amylose content, relative crystallinity, absorbance peak ratios, pasting and thermal properties. Moreover, on average about 83 % raw starch of the varieties was classified as slowly digestible and resistant, whereas after gelatinization this was reduced to 9 %. Molecular and crystalline structures were strongly related to pasting properties, thermal characteristics and in vitro digestibility of the starches. The study provides information on some starch quality characteristics and the inter-relationships among the parameters, and might inspire further studies to suggest possible target-based starch modifications, and future novel utilization of barley. More studies are required to investigate the association of starch quality parameters with malting quality attributes.

Importance of both leached and residual starch molecular structures in determining cooked rice texture at different rice-to-water cooking ratios.

Although texture of cooked rice has been investigated with regard to its relation to starch structure, it remains unclear how starch molecular structure and water content together affect its texture. Thus, the texture, and starch molecular structure of 10 rice varieties and their leachates during cooking were studied with a range of rice-to-water ratios. Although hardness of cooked rice decreased, no trend for the alteration of stickiness observed over the increase of water content. Generally, more amylopectin with DP 12-36 and amylose with DP 5000-20,000 in leachate, less starch molecules with DP > 36 in residual materials resulted in a higher stickiness of cooked rice, which is rationalized by their slower retrogradation tendency and higher amount of non-reducing ends available for binding to textural probe. Rice-to-water ratio was another crucial factor in determining relations between starch structures and cooked rice texture. For example, more leached amylopectin with DP 12-100 resulted in a higher stickiness of cooked rice, while this was not the case for rice cooked at rice-to-water ratios of 1: 1.2 and 1: 1.4. These results give new insights on the effects of starch molecular structure and water content on the cooked rice texture.

A novel method for obtaining high amylose starch fractions from debranched starch.

High amylose starch shows wide applications in food and non-food-based industries. Traditional complex-precipitation approach for the amylose fractionation required a large volume of organic reagents and was possibly risky for food safety. The object of this work was to establish a novel method to obtain starch fractions rich in amylose from debranch starch through repeated short-term retrogradation and centrifugation. Four starch fractions were obtained with the amylose content of 52.08% (C1), 62.28% (C2), 63.58% (C3), and 64.74% (C4). The thermograms of samples displayed that multiple endothermic peaks were detected in C1 and C2 and only one endothermic peak with melting temperature over 120 °C were observed in C3 and C4, indicating their differences in retrogradation behavior. The chain length distribution results of sample exhibited that C1 and C2 contained more short chains (DP ≤ 24), while C3 and C4 consisted of mainly long chains (DP ≥ 25). Accordingly, the differences in fine structures could provide more choices for these fractionated high amylose starch to utilize in practical applications.

Avocado seed starch: Effect of the variety on molecular, physicochemical, and digestibility characteristics.

Agro-industrial residues can increase environmental pollution owing to poor knowledge of the use of some components, such as dietary fiber, protein, starch, minerals, and bioactive compounds, which can be used in the food industry. This study compared the molecular, physicochemical, and digestibility characteristics of three avocado seed starches (Criolla, Fuerte, and Hass). Starch was extracted through successive washing and sedimentation. The morphology, size distribution, thermal properties, pasting properties, infrared spectra with Fourier transform, size distribution of amylopectin chains, and digestibility of the three avocado seed strains were analyzed. The starch grains were oval and spherical in shape. The average size of Criolla avocado starch (24.55 µm) was the largest, followed by Hass and Fuerte starches (21.37 µm). Higher gelatinization enthalpy (8.55 J/g), gelatinization temperature (75.28 °C), and pasting temperature (75.57 °C) were observed for Fuerte avocado starch, followed by Hass and Criolla starches. Hass avocado starch exhibited higher maximum (836.27 mPa.s), final (1407.37 mPa.s), setback (588.78 mPa.s), and breakdown (17.68 mPa.s) viscosities than Criolla and Fuerte avocado starches. In addition, the probed avocado starches exhibited high content of resistant starch (60.06-68.90%). Therefore, it was demonstrated that differences in the chemical composition and structure of avocado starch can affect the digestibility of native starch.

Critical examination of the characterization techniques, and the evidence, for the existence of extra-long amylopectin chains.

It has been suggested that amylopectin can contain small but significant amounts of extra-long chains (ELCs), which could affect functional properties, and also would have implications for the mechanism of starch biosynthesis. However, current evidence for the existence of ELCs is ambiguous. The amylose/amylopectin separation and the characterization techniques used for the investigation of ELCs are reviewed, problems in those techniques are examined, and studies of ELCs of amylopectin are discussed. A model for the biosynthesis of amylopectin chains in terms of conventional biosynthesis enzymes, which provides an excellent fit to a large amount of experimental data, is used to provide a rigorous definition of ELCs. In addition, current investigations of ELCs, involving separation, is hindered by the lack of a method to quantitatively separate all the amylopectin from starch without any traces of residual amylose (which would have long chains). Unambiguous evidence for the existence of ELCs can be obtained using two-dimensional (2D) characterization, these dimensions being the degree of polymerization of a chain and the size of the whole molecule. Available 2D data indicate that there are no ELCs present in currently detectable quantities in native rice starches. However, concluding this more rigorously requires improvements in the resolution of current 2D methods.

Physicochemical Properties and Tissue Structure of High Kernel Elongation Rice (Oryza sativa L.) Varieties as Affected by Heat Treatment.

Heat treatment could affect the structure and properties of rice varieties. The present study was conducted in order to determine the effects of heat treatment on the physicochemical properties and tissue structure of Mahsuri Mutan, Basmati 370 and MR219 rice varieties. The three rice varieties were subjected to heat treatment (ageing) at 90 °C, using an oven, for 3 h. After the heat treatment, the samples were cooled at room temperature (25 °C) for 1 h. Physicochemical properties, such as alkali digestion value, water uptake ratio, solids in cooking water, high kernel elongation ratio and amylose contents, were determined. The procedure used in determining both apparent and absolute amylose involved measuring the iodine affinity of defatted whole starch. Ahigh-performance anion-exchange chromatograph was used to analyse branch chain length distribution of amylopectin quantitatively. The starch structure of the rice samples was observed under a scanning electron microscope. Data collected on physicochemical traits, heat treatment and control (ageing and non-ageing) were subjected to an analysis of variance using SAS software version 9.4. In this study, Mahsuri Mutan and Basmati 370 showed superior high kernel elongation as compared to their respective rice progenies. This study also found that heat treatment directly affected the increasingly high kernel elongation for both populations. The phenotypic correlation co-efficient indicated that there was a high positive correlation between high kernel elongation and water uptake ratio, implying that selection for water uptake ratio would increase the high kernel elongation characteristic. The heat treatment showed significant difference in all the physicochemical traits of the varieties studied. Heat treatment also affected the very long branch chains of starch, such as amylose. Observation under an electron microscope showed that the samples subjected to heat treatment had more cracks on the tissue structure compared to normal rice samples. The hexagon structure in Mahsuri Mutan produced a greater elongation effect on its kernel. The findings from this study could be useful to breeders in the selection and development of a new high kernel elongation rice variety.

Effects of different altitudes on the structure and properties of potato starch.

The main element influencing the quality of potato starch is the environment. To investigate the effects of different altitude cultivation locations on the molecular structure and physicochemical properties of starch, two potato varieties, Jiusen No.1 B1 and Qingshu No.9 B2, were planted in three different altitude zones: A1 at low altitude (Chongzhou 450 m), A2 at middle altitude (Xichang 2800 m), and A3 at high altitude (Litang 3650 m). The results showed that the average volume, number, surface area diameter, average branched polymerization degree, crystallinity, and gelatinization temperature of two potato granules in high altitude areas were significantly lower than those in middle and low altitude areas were, and the gelatinization performance of potato starch was affected according to the correlation of starch structure characteristics. Potato starch with more short-branched chains and less long branched chains resulted in a lower gelatinization temperature in high altitude areas. The results showed that Jiusen No. 1 and Qingshu No. 9 were mainly affected by accumulated radiation and accumulated rainfall in Litang, a high altitude area, and by effective accumulated temperature in Xichang, a middle altitude area. This study quantified the influence of meteorological factors on the main starch quality of potato tubers. The results can be used as a theoretical basis for the scientific planting of high-quality potatoes.

Molecular structure of lotus seed amylopectins and their beta-limit dextrins.

Investigation on amylopectin molecular structure is gaining importance for understanding starch property. Lotus seeds are a novel starch source with high apparent amylose content. Current understanding on the molecular structure of amylopectin in lotus seed starch is scarce. This study compared the molecular structure of a range of lotus seed amylopectins with those of maize and potato amylopectins. Internal structures of these amylopectins were compared via investigating the chain length distribution of their ß-limit dextrins. The average lengths and molar compositions of unit chains in lotus seed amylopectins and their ß-limit dextrins fell generally between those of maize and potato. The average chain lengths of lotus seed, maize, and potato amylopectins were 19.95 (on average), 19.11, and 21.19 glucosyl residues, respectively. Lotus seed amylopectins had higher weight proportion of clustered unsubstituted chains (44.94 % on average) than those of potato (43.99 %) and maize amylopectins (42.95 %). Results of correlation analysis indicated that apparent amylose content of LS was related to structural characteristics of its amylopectin due to the presence of long external chains. The results of this study are of fundamental importance for the utilization of lotus seed starch as a novel starch source.

Molecular Weight Distribution of Whole Starch in Rice Endosperm by Gel-permeation Chromatography.

Starch is comprised of very large α-glucan molecules composed primarily of linear amylose and highly branched amylopectin. Most methods for analyses of starch structure use hydrolytic enzymes to cleave starch. When undegraded, whole starch structures can be analyzed by gel-permeation chromatography (GPC), but this typically yields a single peak each for amylopectin and amylose. The objective of this study was to stably separate amylopectins in whole starch based on their molecular weight using GPC, and to determine the structure of each peak. When alkali-gelatinized whole starch was applied to GPC columns (Toyopearl HW75S × 2, HW65S, and HW55S), it was separated into three peaks. Iodine staining and chain length distribution analyses of debranched samples showed that peaks were mainly composed of high-molecular weight (MW) amylopectin consisting of many clusters, low-MW amylopectin consisting of a small number of clusters, and amylose.

DeltaMSI: artificial intelligence-based modeling of microsatellite instability scoring on next-generation sequencing data.

BACKGROUND: DNA mismatch repair deficiency (dMMR) testing is crucial for detection of microsatellite unstable (MSI) tumors. MSI is detected by aberrant indel length distributions of microsatellite markers, either by visual inspection of PCR-fragment length profiles or by automated bioinformatic scoring on next-generation sequencing (NGS) data. The former is time-consuming and low-throughput while the latter typically relies on simplified binary scoring of a single parameter of the indel distribution. The purpose of this study was to use machine learning to process the full complexity of indel distributions and integrate it into a robust script for screening of dMMR on small gene panel-based NGS data of clinical tumor samples without paired normal tissue. METHODS: Scikit-learn was used to train 7 models on normalized read depth data of 36 microsatellite loci in a cohort of 133 MMR proficient (pMMR) and 46 dMMR tumor samples, taking loss of MLH1/MSH2/PMS2/MSH6 protein expression as reference method. After selection of the optimal model and microsatellite panel the two top-performing models per locus (logistic regression and support vector machine) were integrated into a novel script (DeltaMSI) for combined prediction of MSI status on 28 marker loci at sample level. Diagnostic performance of DeltaMSI was compared to that of mSINGS, a widely used script for MSI detection on unpaired tumor samples. The robustness of DeltaMSI was evaluated on 1072 unselected, consecutive solid tumor samples in a real-world setting sequenced using capture chemistry, and 116 solid tumor samples sequenced by amplicon chemistry. Likelihood ratios were used to select result intervals with clinical validity. RESULTS: DeltaMSI achieved higher robustness at equal diagnostic power (AUC = 0.950; 95% CI 0.910-0.975) as compared to mSINGS (AUC = 0.876; 95% CI 0.823-0.918). Its sensitivity of 90% at 100% specificity indicated its clinical potential for high-throughput MSI screening in all tumor types. Clinical Trial Number/IRB B1172020000040, Ethical Committee, AZ Delta General Hospital.

Variation of rice starch structure and physicochemical properties in response to high natural temperature during the reproductive stage.

Climate warming affects rice growth at different phenological stages, thereby increasing rice chalkiness and protein content and reducing eating and cooking quality (ECQ). The structural and physicochemical properties of rice starch played important roles in determining rice quality. However, differences in their response to high temperature during the reproductive stage have been rarely studied. In the present study, they were evaluated and compared between two contrasting natural temperature field conditions, namely, high seasonal temperature (HST) and low seasonal temperature (LST), during the reproductive stage of rice in 2017 and 2018. Compared with LST, HST significantly deteriorated rice quality, including increased grain chalkiness, setback, consistence, and pasting temperature and reduced taste values. HST considerably reduced the total starch and increased the protein content. Likewise, HST significantly reduced the short amylopectin chains [degree of polymerization (DP) <12] and increased the long amylopectin chains (DP > 12) and relative crystallinity. The starch structure, total starch content, and protein content explained 91.4%, 90.4%, and 89.2% of the total variations in pasting properties, taste value, and grain chalkiness degree, respectively. In conclusion, we suggested that rice quality variations were closely associated with the changes in chemical composition content (total starch and protein content) and starch structure in response to HST. These results indicated that we should improve the resistance of rice to high temperature during the reproductive stage to improve the fine structure of rice starch in further breeding and practice.