Normal and abnormal glycogen structure - A review.

Glycogen, a complex branched glucose polymer, is found in animals and bacteria, where it serves as an energy storage molecule. It has linear (1 â†’ 4)-α glycosidic bonds between anhydroglucose monomer units, with branch points connected by (1 â†’ 6)-α bonds. Individual glycogen molecules are referred to as ß particles. In organs like the liver and heart, these ß particles can bind into larger aggregate α particles, which exhibit a rosette-like morphology. The mechanisms and bonding underlying the aggregation process are not fully understood. For example, mammalian liver glycogen has been observed to be molecularly fragile under certain conditions, such as glycogen from diabetic livers fragmenting when exposed to dimethyl sulfoxide (DMSO), while glycogen from healthy livers is much less fragile; this indicates some difference, as yet unknown, in the bonding between ß particles in healthy and diabetic glycogen. This fragility may have implications for blood sugar regulation, especially in pathological conditions such as diabetes.

Using molecular fine structure to identify optimal methods of extracting fungal glycogen.

Glycogen is a highly branched glucose polymer that is an energy storage material in fungi and animals. Extraction of glycogen from its source in a way that minimizes its molecular degradation is essential to investigate its native structure. In this study, the following extraction methods were compared: sucrose gradient density ultracentrifugation, thermal alkali, hot alcohol and hot water extractions. Molecular-size and chain-length distributions of glycogen were measured by size-exclusion chromatography and fluorophore-assisted carbohydrate electrophoresis, respectively. These two fine-structure features are the most likely structural characteristics to be degraded during extraction. The results show that the thermal alkali, hot alcohol and hot water extractions degrade glycogen molecular size and/or chain-length distributions, and that sucrose gradient density ultracentrifugation with neither high temperature nor alkaline treatment is the most suitable method for fungal glycogen extraction.

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.

Liver glycogen fragility in the presence of hydrogen-bond breakers.

Glycogen, a complex branched glucose polymer, is responsible for sugar storage in blood glucose homeostasis. It comprises small ß particles bound together into composite α particles. In diabetic livers, α particles are fragile, breaking apart into smaller particles in dimethyl sulfoxide, DMSO; they are however stable in glycogen from healthy animals. We postulate that the bond between ß particles in α particles involves hydrogen bonding. Liver-glycogen fragility in normal and db/db mice (an animal model for diabetes) is compared using various hydrogen-bond breakers (DMSO, guanidine and urea) at different temperatures. The results showed different degrees of α-particle disruption. Disrupted glycogen showed changes in the mid-infra-red spectrum that are related to hydrogen bonds. While glycogen α-particles are only fragile under harsh, non-physiological conditions, these results nevertheless imply that the bonding between ß particles in α particles is different in diabetic livers compared to healthy, and is probably associated with hydrogen bonding.

Formation mechanism of α particles in glycogen: Testing the budding hypothesis by Monte-Carlo simulation.

Glycogen, a complex branched glucose polymer and a blood-sugar reservoir in animals, comprises small ß particles joined together into composite α particles. In diabetic animals, α particles fragment more easily than those in healthy animals. Finding evidence for or against postulated mechanisms for α-particle formation is thus important for diabetes research. Insight into this is obtained here using Monte-Carlo simulations, including addition and loss of glucose monomer, branching and debranching, based on earlier simulations which were in acceptable agreement with experiment [Zhang et al., Int J Biol Macromolecules 2018, 116, 264]. One postulated mechanism for α-particle formation is "budding": occasionally a glucan chain temporarily protrudes from the particle, and if its growing end is sufficiently far from its parent particle, it propagates to a new linked particle. We tested this by simulations in which an "artificial" bud (a chain extending well outside the average particle radius) is added to a glycogen molecule in a dynamic steady state, and the system allowed to evolve. In some simulations, the particle reached a new steady state having an irregular dumbbell shape: a rudimentary α particle. Thus 'budding' is a possible mechanism for α particles to form. If no simulations had shown this behaviour, it would have refuted the postulate.

Influence of Storage Temperature on Starch Retrogradation and Digestion of Chinese Steamed Bread.

Chinese steamed bread (CSB), which is widely consumed in East Asia, usually undergoes storage before consumption, but it is unclear how different storage temperatures affect CSB starch retrogradation and digestion properties, which are important for consumers. CSB was stored for 2 days at 25 °C, 4 °C, -18 °C, 4 °C/25 °C temperature cycling (i.e., 24 h at 4 °C, followed by 24 h at 25 °C) and -18 °C/ 25 °C temperature cycling. The results revealed for the first time that more orderly starch double helices are formed when CSB was stored at 4 °C or 4 °C/25 °C. Storage under -18 °C produced lower amounts of, but more heterogenous, starch double helices, with fewer B-type, but more V-type, crystallites. Compared to other storage temperatures, more long-range intermolecular interactions formed between the starch and protein at 4 °C or 4 °C/25 °C. CSB samples showed the slowest starch digestibility when stored at 4 °C. The impact of storage temperature on the starch retrogradation properties and digestibility of CSB also depended on the wheat variety, attributed to differences in the starch molecular structure. These results have significance and practical applications to help the CSB food industry to control starch retrogradation and digestibility. For example, CSB could be stored at 4 °C for 2 days in order to reduce its starch digestibility.

Subtle structural variations of resistant starch from whole cooked rice significantly impact metabolic outputs of gut microbiota.

While cooked rice is widely consumed as a whole food, the specific characteristics and impact of its resistant starch (RS) on gut microbiota are largely unexplored. In this study, three rice varieties with distinct starch molecular structures were used to prepare RS from cooked rice. All three types of RS had a crystalline structure characterized as B + V type, with the V type being the predominant crystalline polymorph. Distinct differences in chain-length distributions were observed among different RSs, with rapidly fermentable starch fractions comprising short amylopectin and long amylose chains, while the degrees of polymerization (DPs) âˆ¼ 10, 37, 65, and 105 fractions comprised the slowly fermentable starch. Jasmine rice RS showed the highest proportion of this slowly fermentable starch fraction, which appeared to be specifically utilized by Megasphaera_elsdenii_DSM_20460 OTU198. The fermentation of Jasmine RS resulted in the highest production of butyrate after 24 h, which was positively correlated with the relative abundance of Megasphaera_elsdenii_DSM_20460 OTU198. These findings collectively indicate that RS in cooked rice with a higher V type crystallinity and DPs âˆ¼ 10, 37, 65, and 105 fractions promote butyrate production and stimulate the growth of butyrate-producing bacteria in the human gut, thereby conferring beneficial effects on gut health.

Development of a model for granule-bound starch synthase I activity using free-energy calculations.

Starch is a branched polymer of glucose with two components, both of which have (1 â†’ 4)-α linear links and (1 â†’ 6)-α branch points: amylopectin, of high molecular weight with many short branches, and amylose, of lower molecular weight and only a few long-chain branches. Granule-bound starch synthase I (GBSSI) is one of the main enzymes controlling amylose synthesis and chain-length distribution. As production of different GBSSI mutants is time-consuming and laborious, molecular dynamics (MD) simulations are used here to predict the binding of different GBSSI mutants to a representative amylose fragment. The simulations were atomistic, with explicit solvent and docking, a method successfully used to understand the binding of wild-type GBSSI to amylose fragments. The binding of GBSSI to G5 (a pentasaccharide amylose fragment) is combined with free-energy calculations employing a thermodynamic integration method to predict the effects of mutations on enzyme activity. Ten GBSSI mutants with different enzyme activities were analyzed to find the structural and energy changes among different single amino-acid mutants and their possible relationship to starch characteristics. Comparing the structural changes and the relative binding free energy of G5 to the wild type GBSSI and GBSSI mutants, it was found that mutants with negative binding energy (lower than -2.0 kcal/mol) are more likely to have higher enzyme activity and amylose content compared to the wild type. This theoretical paper used simulations and robust free energy calculations to interpret in planta data with potential predictions as to what mutants might be generated to give desired properties. This study can be used to help develop grains with improved functional properties.

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.

Effects of the Molecular Structure of Starch in Foods on Human Health.

Starch provides approximately half of humans' food energy, and its structural features influence human health. The most important structural feature is the chain length distribution (CLD), which affects properties such as the digestibility of starch-containing foods. The rate of digestion of such foods has a strong correlation with the prevalence and treatment of diseases such as diabetes, cardiovascular disease and obesity. Starch CLDs can be divided into multiple regions of degrees of polymerization, wherein the CLD in a given region is predominantly, but not exclusively, formed by a particular set of starch biosynthesis enzymes: starch synthases, starch branching enzymes and debranching enzymes. Biosynthesis-based models have been developed relating the ratios of the various enzyme activities in each set to the CLD component produced by that set. Fitting the observed CLDs to these models yields a small number of biosynthesis-related parameters, which, taken together, describe the entire CLD. This review highlights how CLDs can be measured and how the model-based parameters obtained from fitting these distributions are related to the properties of starch-based foods significant for health, and it considers how this knowledge could be used to develop plant varieties to provide foods with improved properties.

Effects of storage temperatures on the starch digestibility of whole rice with distinct starch fine molecular structure.

Although the retrogradation of rice starch has been extensively investigated, there remain questions as to how storage temperature affects starch inter- and intramolecular interactions in cooked white rice, and the relationship of these interactions with the digestion rate. To this end, micromorphology, crystallinity polymorphisms, molecular interaction patterns and in vitro starch digestibility of 3 rice varieties kept under 5 different storage temperature programs (room temperature (RT), 4 °C, -18 °C, 4 °C/RT (4 °C for 24 h and then RT for 24 h), -18 °C/RT (-18 °C for 24 h and then RT for 24 h)) were investigated. As expected, a significant variance in starch digestibility was observed for samples after storage at different temperatures. Overall, storage at 4 °C could most effectively decrease the starch digestibility of retrograded rice. The digestion rate constant was for the first time found to be determined by short-range amylopectin intermolecular interactions rather than long-range starch molecular interactions, for all different storage conditions. Furthermore, the digestion extent was determined by both inter- and intramolecular interactions among starch molecules as well as by the long-range order of the retrograded double helices. These results could prove useful to devise storage regimes which use retrogradation to produce cooked rice with lower glycemic index.

Formation, Structural Characterization, and Functional Properties of Corn Starch/Zeaxanthin Composites.

Zeaxanthin is a natural xanthophyll carotenoid and the main macular pigment that protects the macula from light-initiated oxidative damage, but it has poor stability and low bioavailability. Absorption of this active ingredient into starch granules as a carrier can be used to improve both zeaxanthin stability and controlled release. Optimization using three variables judged important for optimizing the system (reaction temperature of 65 °C, starch concentration of 6%, and reaction time of 2 h) was conducted for incorporation of zeaxanthin into corn starch granules, aiming for high zeaxanthin content (2.47 mg/g) and high encapsulation efficiency (74%). Polarized-light microscopy, X-ray diffraction, differential scanning calorimetry, and Fourier transform infrared spectroscopy showed that the process partially gelatinized corn starch; additionally, it showed the presence of corn starch/zeaxanthin composites, with the zeaxanthin successfully trapped in corn starch granules. The half-life time of zeaxanthin in corn starch/zeaxanthin composites increased to 43 days as compared with that of zeaxanthin alone (13 days). The composites show a rapid increase in zeaxanthin release with in vitro intestinal digestion, which is favorable for possible use in living systems. These findings could have application in designing effective starch-based carriers of this bioactive ingredient with enhanced storage stability and improved intestines-targeted controlled-release delivery.

The Effects of Starch Molecular Fine Structure on Thermal and Digestion Properties of Rice Starch.

Whole white rice is a major staple food for human consumption, with its starch digestion rate and location in the gastrointestinal tract having a critical role for human health. Starch has a multi-scale structure, which undergoes order-disorder transitions during rice cooking, and this structure is a major determinant of its digestibility. The length distributions of amylose and amylopectin chains are important determinants of rice starch gelatinization properties. Starch chain-length and molecular-size distributions are important determinants of nucleation and crystal growth rates, as well as of intra- and intermolecular interactions during retrogradation. A number of first-order kinetics models have been developed to fit starch digestograms, producing new information on the structural basis for starch digestive characteristics of cooked whole rice. Different starch digestible fractions with distinct digestion patterns have been found for the digestion of rice starch in fully gelatinized and retrograded states, the digestion kinetics of which are largely determined by starch fine molecular structures. Current insights and future directions to better understand digestibility of starch in whole cooked rice are summarized, pointing to ways of developing whole rice into a healthier food by way of having slower starch digestibility.

Mechanistic Understanding of the Effects of Pectin on In Vivo Starch Digestion: A Review.

Obesity and type II diabetes are closely related to the rapid digestion of starch. Starch is the major food-energy source for most humans, and thus knowledge about the regulation of starch digestion can contribute to prevention and improved treatment of carbohydrate metabolic disorders such as diabetes. Pectins are plant polysaccharides with complex molecular structures and ubiquitous presence in food, and have diverse effects on starch digestion. Pectins can favorably regulate in vivo starch digestion and blood glucose level responses, and these effects are attributed to several reasons: increasing the viscosity of digesta, inhibiting amylase activity, and regulating some in vivo physiological responses. Pectins can influence starch digestion via multiple mechanisms simultaneously, in ways that are highly structure-dependent. Utilizing the multi-functionalities of pectin could provide more ways to design low glycemic-response food and while avoiding the unpalatable high viscosity in food by which is commonly caused by many other dietary fibers.

Effects of amylose and amylopectin molecular structures on starch electrospinning.

The role of amylose content in electrospinning starch nanofibres is well understood, but that is not the case for the roles of the molecular structures of amylose and amylopectin. Here, correlations between starch molecular-structure parameters and electrospinnability evaluation indices (average droplet number, average bead number, and average fibre diameter) and dope properties (shear viscosity, conductivity, and surface tension) were examined. Starches with lower amounts of short amylopectin chains, higher amounts of either/or long amylopectin chains and/or lower degree of branching showed decreased viscosity of the electrospinning dopes, and resulted in a reduced average droplet number of electrospun fibre mats. The molecular sizes of amylose and whole starch, and the average degree of polymerization for amylose chains, all correlated with the shear viscosity and surface tension of dopes, and thus influenced the average fibre diameter. This expands the current understanding between amylopectin molecular structure and starch electrospinning, thereby assisting a better choice of starches for desired electrospinnability properties.

The fragility of liver glycogen from humans with type 2 diabetes: A pilot study.

Liver glycogen is a highly branched glucose polymer found as ß particles (~20 nm in diameter), which can bind together into larger composite α particles. Hepatic α particles have been shown to be structurally fragile (breaking up into smaller particles in certain solvents) in mouse models of diabetes; if occurring in vivo, the resulting small glycogen particles could exacerbate the poor blood-sugar homeostasis characteristic of the disease. Here we tested if this α-particle fragility also occurred in liver glycogen obtained from humans with diabetes. It was found that liver glycogen from diabetic humans was indeed more fragile than from non-diabetic humans, which was also seen in the mouse experiments we ran in parallel. Proteomic analysis revealed three candidate proteins from differentially expressed glycogen proteins (Diabetes/ Non-diabetes) in both human and mouse groups. Identifying these proteins may give clues to the binding mechanism that holds together α particles together, which, being different in diabetic glycogen, is relevant to diabetes prevention and management.

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

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

Testing the Linearity Assumption for Starch Structure-Property Relationships in Rices.

Many properties of starch-containing foods are significantly statistically correlated with various structural parameters. The significance of a correlation is judged by the p-value, and this evaluation is based on the assumption of linear relationships between structural parameters and properties. We here examined the linearity assumption to see if it can be used to predict properties at conditions that are not close to those under which they were measured. For this we used both common domesticated rices (DRs) and Australian wild rices (AWRs), the latter having significantly different structural parameters and properties compared to DRs. The results showed that (1) the properties were controlled by more than just the amylopectin or amylose chain-length distributions or amylose content, other structural features also being important, (2) the linear model can predict the enthalpy ΔHg of both AWRs and DRs from the structural parameters to some extent but is often not accurate; it can predict the ΔHg of indica rices with acceptable accuracy from the chain length distribution and the amount of longer amylose chains (degree of polymerization > 500), and (3) the linear model can predict the stickiness of both AWRs and DRs to acceptable accuracy in terms of the amount of longer amylose chains. Thus, the commonly used linearity assumption for structure-property correlations needs to be regarded circumspectly if also used for quantitative prediction.

Starch molecular structural differences between chalky and translucent parts of chalky rice grains.

Chalky rice has an undesirable appearance and reduced commercial value. To understand the relationship between starch structural characteristics and chalkiness, a comprehensive investigation was conducted of molecular structural differences between starch in chalky and translucent parts of the same chalky grains (three Japonica and two Indica rices), this strategy being such as to minimize genetic and environmental effects. Compared to translucent parts, chalky parts had a larger ratio of large to small branched molecules and more short amylopectin chains (degree of polymerization < 35), but fewer longer chains, which affect higher-level starch structures, such as crystallinity. No significant differences in amylose structure were observed. White-belly and white-core chalky grains showed distinguishable starch characteristics, suggesting studying different chalkiness types separately. These findings extend understanding of chalkiness from the perspective of starch structure, and control of this structure can in the future help breeders to develop strategies against the formation of chalkiness.

Combined effects of starch fine molecular structures and water content on starch digestibility of cooked white rice.

Although the starch digestibility of cooked white rice has been investigated with regard to its relation to starch structure, it is not yet clear how starch molecular structure and water content affect its digestion rate. To investigate this, the in vitro starch digestibility and molecular structure of 10 rice varieties with a range of rice-to-water cooking ratios were investigated. As expected, starch digestibility varied with different conditions. Typically, a higher amylose content resulted in a lower maximum digestion extent for a given water content. Having relatively more and longer amylopectin intermediate chains caused a slower starch digestion rate, but only with rice-to-water ratios between 1:1 and 1:1.2. These results could prove useful to find combinations of starch fine molecular structures and water contents to produce cooked rice with low glycemic index.