Chain Length Distribution Research Articles

LIKE EARLY STARVATION 1 and EARLY STARVATION 1 promote and stabilize amylopectin phase transition in starch biosynthesis.

Starch, the most abundant carbohydrate reserve in plants, primarily consists of the branched glucan amylopectin, which forms semi-crystalline granules. Phase transition from a soluble to an insoluble form depends on amylopectin architecture, requiring a compatible distribution of glucan chain lengths and a branch-point distribution. Here, we show that two starch-bound proteins, LIKE EARLY STARVATION 1 (LESV) and EARLY STARVATION 1 (ESV1), which have unusual carbohydrate-binding surfaces, promote the phase transition of amylopectin-like glucans, both in a heterologous yeast system expressing the starch biosynthetic machinery and in Arabidopsis plants. We propose a model wherein LESV serves as a nucleating role, with its carbohydrate-binding surfaces helping align glucan double helices to promote their phase transition into semi-crystalline lamellae, which are then stabilized by ESV1. Because both proteins are widely conserved, we suggest that protein-facilitated glucan crystallization may be a general and previously unrecognized feature of starch biosynthesis.

What geometrically constrained models can tell us about real-world protein contact maps

The mechanisms by which a protein’s 3D structure can be determined based on its amino acid sequence have long been one of the key mysteries of biophysics. Often simplistic models, such as those derived from geometric constraints, capture bulk real-world 3D protein-protein properties well. One approach is using protein contact maps (PCMs) to better understand proteins’ properties. In this study, we explore the emergent behaviour of contact maps for different geometrically constrained models and compare them to real-world protein systems. Specifically, we derive an analytical approximation for the distribution of amino acid distances, denoted as P(s), using a mean-field approach based on a geometric constraint model. This approximation is then validated for amino acid distance distributions generated from a 2D and 3D version of the geometrically constrained random interaction model. For real protein data, we show how the analytical approximation can be used to fit amino acid distance distributions of protein chain lengths of L ≈ 100, L ≈ 200, and L ≈ 300 generated from two different methods of evaluating a PCM, a simple cutoff based method and a shadow map based method. We present evidence that geometric constraints are sufficient to model the amino acid distance distributions of protein chains in bulk and amino acid sequences only play a secondary role, regardless of the definition of the PCM.

Kinetic Monte Carlo Convergence Demands for Thermochemical Recycling Kinetics of Vinyl Polymers with Dominant Depropagation

As societal interest in recycling of plastics increases, modeling thermochemical recycling of vinyl polymers, e.g., via pyrolysis or reactive extrusion, becomes increasingly important. A key aspect remains the reliability of the simulation results with fewer evaluation studies regarding convergence as in the polymerization or polymer reaction engineering field. Using the coupled matrix-based Monte Carlo (CMMC) framework, tracking the unzipping of individual chains according to a general intrinsic reaction scheme consisting of fission, β-scission, and termination, it is however illustrated that similar convergence demands as in polymerization benchmark studies can be employed, i.e., threshold values for the average relative error predictions on conversion and chain length averages can be maintained. For this illustration, three theoretical feedstocks are considered as generated from CMMC polymer synthesis simulations, allowing to study the effect of the initial chain length range and the number of defects on the convergence demands. It is shown that feedstocks with a broader chain length distribution and a long tail require a larger Monte Carlo simulation volume, and that the head–head effects play a key role in the type of degradation mechanism and overall degradation rate. A minimal number of chains around 5 × 105 is needed to properly reflect the degradation kinetics. A certain degree of noise can be allowed at the higher carbon-based conversions due to the inevitable decrease in number of chains.

Characterization of starch extracted from seeds of Cycas revoluta.

Starch is major component in the big seeds of Cycas revoluta, however the characteristics of Cycas revoluta remain unknown. In this study, the physicochemical and structural properties of two starch samples extracted from Cycad revoluta seeds were systematically investigated, using various techniques. The amylose contents of the two samples were 34.3 % and 35.5%, respectively. The spherical-truncated shaped starch granules possessed A-type crystallinity, and had an average diameter less than 15 μm. Compared to most commonly consumed cereal and potato starch, Cycad revoluta starch showed distinctive characteristics. For physicochemical properties, in the process of gelatinization, the Cycad revoluta starch showed similar viscosity profile to starches of some potato varieties, but Cycad revoluta starch had higher gelatinization temperature. Upon cooling, Cycad revoluta starch formed harder gels than rice starch. For structure, the molecular weight (indexed by Mw, Mn and Rz values), branching degree and the branch chain length distribution were determined. The results suggested that Cycad revoluta starch were different in structure from the main-stream starches. Notable differences in some starch traits between the two samples were recorded, which could be attributed to environmental factors. In general, this study provides useful information on the utilization of Cycad revoluta starch in both food and non-food industries.

Exploring the roles of amylopectin in starch modification with Limosilactobacillus reuteri 121 4,6-α-glucanotransferase via developed methods

Limosilactobacillus reuteri 121 4,6-α-glucanotransferase (GtfBΔN) modifies starch by cleaving (α1 → 4) linkages and introducing non-branched (α1 → 6) linkages to produce functional starch derivatives. Research has mainly focused on GtfBΔN converting amylose (linear substrate), whereas the conversion of amylopectin (branched substrate) has not been studied in detail. In this study, we used GtfBΔN to understand amylopectin modification and performed a set of experiments to analyze this modification pattern. The donor substrates were segments from the non-reducing ends to the nearest branch point in amylopectin as shown from the results of the chain length distribution of GtfBΔN-modified starches. Decreased and increased contents of β-limit dextrin and reducing sugars, respectively, during the incubation of β-limit dextrin with GtfBΔN indicated that the segments from the reducing end to the nearest branch point in amylopectin act as donor substrates. Dextranase was involved in the hydrolysis of the GtfBΔN conversion products of three different substrates groups, maltohexaose (G6), amylopectin, and G6 plus amylopectin. No reducing sugars were detected, therefore, amylopectin was not used as an acceptor substrate, and no non-branched (α1 → 6) linkages were introduced into it. Thus, these methods provide a reasonable and effective approach to studying GtfB-like 4,6-α-glucanotransferase in analyzing the roles and contribution of branched substrates.

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.

Stochastic modelling of a three-dimensional glycogen granule synthesis and impact of the branching enzyme.

In humans, glycogen storage diseases result from metabolic inborn errors, and can lead to severe phenotypes and lethal conditions. Besides these rare diseases, glycogen is also associated to widely spread societal burdens such as diabetes. Glycogen is a branched glucose polymer synthesised and degraded by a complex set of enzymes. Over the past 50 years, the structure of glycogen has been intensively investigated. Yet, the interplay between the detailed three-dimensional glycogen structure and the related enzyme activity is only partially characterised and still to be fully understood. In this article, we develop a stochastic coarse-grained and spatially resolved model of branched polymer biosynthesis following a Gillespie algorithm. Our study largely focusses on the role of the branching enzyme, and first investigates the properties of the model with generic parameter values, before comparing it to in vivo experimental data in mice. It arises that the ratio of glycogen synthase over branching enzyme reaction rates drastically impacts the structure of the granule. We deeply investigate the mechanism of branching and parametrise it using distinct lengths. Not only do we consider various possible sets of values for these lengths, but also distinct rules to apply them. We show how combining various values for these lengths finely tunes glycogen macromolecular structure. Comparing the model with experimental data confirms that we can accurately reproduce glycogen chain length distributions in wild type mice. Additional granule properties obtained for this fit are also in good agreement with typically reported values in the experimental literature. Nonetheless, we find that the mechanism of branching must be more flexible than usually reported. Overall, our model provides a theoretical basis to quantify the effect that single enzymatic parameters, in particular of the branching enzyme, have on the chain length distribution. Our generic model and methods can be applied to any glycogen data set, and could in particular contribute to characterise the mechanisms responsible for glycogen storage disorders.

Effect of frozen storage on the quality and amylopectin structure of Chinese steamed bread made from different wheat cultivars

The physicochemical properties of wheat flour and the chain-length distributions of amylopectin from eleven cultivars were studied. Chinese steamed bread (CSB) was made by a one-step fermentation procedure using these flours to explore the effects of frozen storage on the qualities of CSB and starch molecular structures. The results indicated that the protein and wet gluten had significant effects on the fermentation rheological properties of wheat dough and CSB qualities. The amylose content and amylopectin structure had important influences on the pasting properties of wheat flour and the firming of CSB. Water loss and migration in CSB during freezing, accompanied by the starch retrogradation, depended on the amylose content and amylopectin structure. After 60-day of frozen storage at −18 °C, the amylopectin extracted from CSB showed decreased fractions of B1 and B2 chains while the long B3 chain and average chain length increased. Moreover, short amylopectin chains were negatively correlated with the hardness of CSB (P < 0.01). These results confirmed that the structure-property relationships of wheat flour have crucial influences on the qualities of CSB, which would benefit our selection of wheat materials for the frozen food industry and provide a reference for wheat breeding.

Physicochemical properties of starch in rice flour with different hardening rates of glutinous rice cake (mochi)

The hardening rate of glutinous rice cake (mochi) made by Kagura-mochi (Kag) was two times higher than that of Hakusanmochi (Hak). Protein in the waxy rice indicated a similar distribution and content; therefore, the textural properties of the rice product were dependent on the physicochemical properties of starch in the rice flour. Urea-induced gelatinization, differential scanning calorimetry, and rapid visco analysis revealed that starch in Kag was less easily gelatinized than that in Hak. The solubility and swelling power at 70 °C of starch in Kag rice flour were slightly lower than that in Hak. Relative crystallinities of starch in Kag and Hak rice flour were 55.1% and 49.3%, respectively. The chain length distribution of the short chain of amylopectin in Kag rice flour was lower than that in Hak. The low short-chain content in Kag may be related to the increase in the hardening rate of the mochi made by Kag.

Improved Route to Linear Triblock Copolymers by Coupling with Glycidyl Ether-Activated Poly(ethylene oxide) Chains.

Poly(ethylene oxide) block copolymers (PEOz BCP) have been demonstrated to exhibit remarkably high lithium ion (Li+) conductivity for Li+ batteries applications. For linear poly(isoprene)-b-poly(styrene)-b-poly(ethylene oxide) triblock copolymers (PIxPSyPEOz), a pronounced maximum ion conductivity was reported for short PEOz molecular weights around 2 kg mol-1. To later enable a systematic exploration of the influence of the PIx and PSy block lengths and related morphologies on the ion conductivity, a synthetic method is needed where the short PEOz block length can be kept constant, while the PIx and PSy block lengths could be systematically and independently varied. Here, we introduce a glycidyl ether route that allows covalent attachment of pre-synthesized glycidyl-end functionalized PEOz chains to terminate PIxPSy BCPs. The attachment proceeds to full conversion in a simplified and reproducible one-pot polymerization such that PIxPSyPEOz with narrow chain length distribution and a fixed PEOz block length of z = 1.9 kg mol-1 and a Đ = 1.03 are obtained. The successful quantitative end group modification of the PEOz block was verified by nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography (GPC) and differential scanning calorimetry (DSC). We demonstrate further that with a controlled casting process, ordered microphases with macroscopic long-range directional order can be fabricated, as demonstrated by small-angle X-ray scattering (SAXS), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). It has already been shown in a patent, published by us, that BCPs from the synthesis method presented here exhibit comparable or even higher ionic conductivities than those previously published. Therefore, this PEOz BCP system is ideally suitable to relate BCP morphology, order and orientation to macroscopic Li+ conductivity in Li+ batteries.

Image mapping of biological changes and structure-function relationship during rice grain development via Synchrotron FTIR spectroscopy

The spatial distribution of compositional elements in milky, dough, and mature rice, together with the structure-function relationship was studied using synchrotron-FTIR. The dough stage aleurone and endosperm had high lipid, protein (amide I and II), and hemicellulose. The amylopectin chain length distribution of starch exhibited small differences among developmental stages. Dough starch featured a greater portion of A and B1 chains, implying a more crystalline and high in resistant starch. The protein in milky-to-dough stage aleurone and endosperm had higher contents of α-helixes than β-sheets, resulting in higher protein digestibility and availability. There were high intensities of the glycosidic peaks of hemicellulose and cellulose, pectin, carbohydrates, and β 1–6 glucans in milky-to-dough stages. They corresponded to higher total, insoluble, and soluble dietary fiber contents in milky-to-dough stage than in mature stage. Young-stage rice was found to be more nutritious and could thus be a functional food source.

Improving the Resistance to Enzyme Digestion of Rice Debranched Starch via Narrowing Chain Length Distribution Combined with Oven Drying

AbstractIn this study, the effects of chain length distribution and drying methods on the structural properties and in vitro digestibility of resistant starch (RS) are investigated. Rice starch is debranched at 10% w/w solid concentrations, incubated at 60 °C, and further subjected to freeze drying and oven drying treatment to obtain debranched starch (DBS). The chain length distribution shows that DBS (the degree of polymerization [DP] = 19.52 ± 0.13) has a high proportion of B1 and B2 chains (DP13‐36) and a narrow chain length distribution. In addition, the results of scanning electron microscopy (SEM) show that the oven‐dried samples have denser structures and larger particle morphology compared to samples prepared by freeze‐drying. In FTIR, LCM‐Raman and X‐ray diffraction patterns, debranched starch supernatant fraction 1 (DBSS1) exhibit higher short‐range molecular order and crystallinity than other samples. Digestibility analysis shows that DBSS1 with narrow chain length distribution (DP13‐36) promotes the high content of RS formation and oven drying is more favorable for RS formation than freeze drying (RSDBSFS1 = 79.69 ± 5.09% and RSDBSOS1 = 84.17 ± 0.17%). This suggests that the proper narrow chain length distribution is conducive to the formation of ordered structure and an increase in resistance. Oven drying provides a larger starch particle size, which facilitates resistance to digestion.

Using starch structure to choose rices with an optimal combination of palatability and digestibility

Steamed white rices that are popular with consumers are usually quick to digest, which is however disadvantageous to metabolic health. However, varieties which are slow to digest are generally regarded as less palatable. Starch molecular structure is a major determinant of both properties. Starch structure includes distinct attributes, such as the chain-length distribution of debranched chains and the size distribution of (undebranched) whole molecules. While some structural attributes control both properties, it is found that there are some structural features which are significant determinants for one property and not the other. Here, a novel means of using this fact to overcome these competing demands of slow digestibility and acceptable palatability is explored. The chain-length distributions (CLDs) and in vitro digestibility of 98 varieties, and palatabilities of 9 market varieties, were measured using size-exclusion chromatography (SEC) for the CLDs, and human panelists for palatability. The CLDs were parameterized with biosynthesis-based models, one each for amylopectin and for amylose. Four structural features of the CLDs (βAp,3, hAm,2, hAm,1 and hAp,5), involving the amounts of short and long chains in various CLD regions, showed significant correlations, with correlation coefficients having opposite signs for both digestibility (0.597**, −0.451**,-0.350**, and −0.413, respectively) and palatability (0.769*, −0.396, −0.491 and −0.398, respectively), and which thus were the key factors in the conflicting demands of slower digestibility versus acceptable palatability. It was also found that different structural parameters were dominant for digestibility and palatability; for instance, some structural features of hAp,1 and βAp,5 were found to optimize palatability and digestibility independently (hAp,1 - digestibility, −0.547**; βAp,5 – palatability, 0.339). Hence optimizing these two separate sets of parameters provides a means of finding an overall set of structural parameters which can be used to give the best combination of lower digestibility and acceptable palatability. Because the genes controlling these CLDs have been established, this can help rice breeders to develop varieties which give the best compromise for simultaneously optimizing both properties.

The alterations in granule, shell, blocklets, and molecular structure of pea starch induced by ultrasound

Understanding the alterations to starch multi-scale structure induced by ultrasound treatment can help in determining the effective application of ultrasound in functional-starch preparation. This study aimed to comprehensively characterize and understand the morphological, shell, lamellae, and molecular structures of pea starch granules treated by ultrasound under different temperatures. Scanning electron microscopy and X-ray diffraction analyses showed that UT (ultrasound treatment) did not change C-type of crystalline, but caused a pitted surface and endowed a looser structure and higher enzyme susceptibility as the temperature increased above 35 °C for pea starch granules. Fourier transform infrared spectroscopy and small-angle X-ray scattering analyses revealed that UT reduced the short-range ordering and increased the thickness of semi-crystalline and amorphous lamellae by inducing starch chain depolymerization, which was manifested by molecule weight and chain length distribution analysis. The sample ultrasound-treated at 45 °C had the higher proportion of B2 chains compared with the other ultrasound-treated samples because the higher ultrasonic temperature altered the disruption sites of starch chains.

Interplay Between Doping, Morphology, and Lattice Thermal Conductivity in PEDOT:PSS

AbstractOrganic materials for thermoelectric (TE) applications have attracted a fair amount of attention in recent years due to remarkable advances achieved in terms of their figure of merit, ZT: a value of 0.42 has been reported by for poly(3,4‐ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) films treated with dimethyl sulfoxide, while 0.25 has been obtained for PEDOT:Tosylate. In this study various PEDOT morphologies are investigated, considering both neutral and doped (bipolaronic) samples, by means of classical molecular dynamics, by taking advantage of a recently developed all‐atom force field. In the case of bare PEDOT, it is found that changing the distribution of chain lengths affects the thermal conductivity of neutral and doped samples in a different way: longer chain lengths result in higher conductivities in the neutral scenario, whereas an intermediate chain length gives the highest value in the bipolaronic case. The role of PSS in the bipolaronic state, as compared to the simpler case where the counterion is Cl−, is discussed.

Performance of 3D-printed samples based on starch treated by radio frequency energy

In this study, we investigated changes in the chain length distribution, molecular weight distribution, amylose content, microstructures, thermal properties and rheological properties of potato starch following radio frequency (RF) electromagnetic fields exposure with varying moisture content (30, 40, 50, 60, 70%, w/w). We identified a RF-modified moisture content suitable for 3D printing. Our results showed that RF treatment roughened the starch surface and broke down the longer chains into shorter ones, which reduced the molecular weight, viscosity, and pasting temperature of the starch. The RF-treated starch gels showed better printing performance than untreated potato gels. In addition, changes in the physicochemical properties of the starch became more pronounced with increasing moisture content. Gels made from starch with 70% moisture content treated with RF energy provided the best morphological indicators, including clarity line uniformity and high retention rate. Overall, RF treatment of starch with varying moisture content is an effective method for preparing starch gels for 3D printing.

Effects of primary, secondary and tertiary structures on functional properties of thermoplastic starch biopolymer blend films

To better understand the correlation between structure and properties in thermoplastic starch biopolymer blend films, the effects of amylose content, chain length distribution of amylopectin and molecular orientation of thermoplastic sweet potato starch (TSPS) and thermoplastic pea starch (TPES) on microstructure and functional properties of thermoplastic starch biopolymer blend films were studied. After thermoplastic extrusion, the amylose contents of TSPS and TPES decreased by 16.10 % and 13.13 %, respectively. The proportion of the chains with the degree of polymerization between 9 and 24 of amylopectin in TSPS and TPES increased from 67.61 % to 69.50 %, and from 69.51 % to 71.06 %, respectively. As a result, the degree of crystallinity and molecular orientation of TSPS and TPES films increased as compared to sweet potato starch and pea starch films. The thermoplastic starch biopolymer blend films possessed a more homogeneous and compacter network. The tensile strength and water resistance of thermoplastic starch biopolymer blend films increased significantly, whereas thickness and elongation at break of thermoplastic starch biopolymer blend films decreased significantly.

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.

Structural changes of starch under different milling degrees affect the cooking and textural properties of rice

A long-term consumption of white rice increases the risk of T2D. Finding an appropriate milling degree (MD) of rice balancing nutrition and palatability benefits public health. This study investigated effects of MD-0 s, 5 s, 60 s on morphological, cooking and textural properties of rice. Texture profile analysis showed that milling decreased hardness, while increased adhesiveness of rice. SEM images showed that milling induced notches and structural damage, which facilitated gelatinization of rice determined by DSC. Leached starch was further analyzed by size exclusion chromatography and chain-length distribution. Pearson correlation analysis revealed that milling induced more leached shot-chain amylose and long-chain amylopectin, which decreased hardness and increased adhesiveness of cooked rice. Collectively, milling-induced changes of starch gelatinization and fine structure of leached starch were decisive factors of rice texture. Moderate processing improved the texture of brown rice and maintained nutrients. This would provide guidance for the health industry of whole grains.

Effect of Shearing and Heat Milling Treatment Temperature on the Crystallinity, Thermal Properties, and Molecular Structure of Rice Starch.

Rice flour is produced by various methods for use in the food industry, but little is known about how the structure of starch is affected during rice flour production. In this study, the crystallinity, thermal properties, and structure of starch in rice flour were investigated after treatment with a shearing and heat milling machine (SHMM) at different temperatures (10-150 °C). Both the crystallinity and gelatinization enthalpy of starch showed an inverse relationship with the treatment temperature; rice flour treated with the SHMM at higher temperatures showed lower crystallinity and gelatinization enthalpy than that treated at lower temperatures. Next, the structure of undegraded starch in the SHMM-treated rice flour was analyzed by gel permeation chromatography. A significant reduction in the molecular weight of amylopectin was observed at high treatment temperatures. Chain length distribution analysis showed that the proportion of long chains (degree of polymerization (DP) > 30) in rice flour decreased at temperatures ≥ 30 °C. By contrast, the molecular weight of amylose did not decrease. In summary, the SHMM treatment of rice flour at high temperatures resulted in starch gelatinization, and the amylopectin molecular weight decreased independently, due to the cleavage of amorphous regions connecting the amylopectin clusters.