Amylolysis of wheat starches. II. Degradation patterns of native starch granules with varying functional properties

Scanning electron microscopy (SEM) and X-ray diffraction (XRD) were employed to investigate degradation patterns of native starch granules from wheat (Triticum aestivum L.) by different starch-degrading enzymes. The starches examined were from a waxy wheat and four varieties with slightly elevated amylose content, but with different functional properties. Differences in the digestion patterns after partial α-amylolysis of starch granules were noted between the starches. The waxy starch seemed to be degraded by endocorrosion, whereas the amylose-rich starches followed a slower mode of hydrolysis starting from the granular surface. X-ray diffractograms of the amylose-rich starches were not significantly altered by 2 h of α-amylolysis, whereas partial hydrolysis of the waxy starch decreased scattering intensity at higher 2θ angles, consistent with a different mode of attack by α-amylase in the initial digestion stages of granules of waxy and amylose-rich starches. We propose these differences are due to the combined effects of the change in packing density and partial preference for hydrolysis of amorphous material. The native starch granules were also attacked by beta-amylase, isoamylase and amyloglucosidase, which indicates that α-amylase is not the only starch-degrading enzyme that is able to initiate starch hydrolysis of native granules.     

Physicochemical properties and amylopectin fine structures of A- and B-type granules of waxy and normal soft wheat starch

This work fractionated waxy and normal wheat starches into highly purified A- and B-type granule fractions, which were representative of native granule populations within parent native wheat starches, to accurately assess starch characteristics and properties of the two granule types. Wheat starch A- and B-type granules possessed different morphologies, granule specific surface area measurements, compositions, relative crystallinities, amylopectin branch chain distributions, and physical properties (swelling, gelatinization, and pasting behaviors). Within a genotype, total and apparent amylose contents were greater for A-type granules, while lipid-complexed amylose and phospholipid contents were greater for B-type granules. B-type (relative to A-type) granules within a given genotype possessed a greater abundance of short amylopectin branch chains (DP_n < 13) and a lesser proportion of intermediate (DP_n 13–33) and long (DP_n > 33) branch chains, contributing to their lower relative crystallinities. Variation in amylose and phospholipid characteristics appeared to account for observed differences in swelling, gelatinization, and pasting properties between waxy and normal wheat starch fractions of a common granule type. However, starch granule swelling and gelatinization property differences between A- and B-type granules within a given genotype were most consistently explained by their differential amylopectin chain-length distributions.     

Performance of Granular Starch with Controlled Pore Size during Hydrolysis with Digestive Enzymes

Studies on porous starch have been directed to explore different industrial applications as bio-adsorbents of a variety of compounds. However, the analysis of starch digestibility is essential for food application. The objective of this study was to determine the impact of porous structure on in vitro starch digestibility. Porous starches were obtained using a range of concentrations of amyloglucosidase (AMG), α-amylase (AM), cyclodextrin-glycosyltransferase (CGTase) or branching enzyme (BE). Porous starches exhibited major content of digestible starch (DS) that increased with the intensity of the enzymatic treatment, and very low amount of resistant starch (RS). Porous starches behaved differently during in vitro hydrolysis depending on their enzymatic treatment. AMG was the unique treatment that increased the digestive amylolysis and estimated glycemic index, whereas AM, CGTase and BE reduced them. A significant relationship was found between the pore size and the severity of the amylolysis, suggesting that a specific pore size is required for the accessibility of the digestive amylase. Therefore, pore size in the starch surface was a limiting factor for digestion of starch granules.     

Effects of endosperm texture and cooking conditions on the in vitro starch digestibility of sorghum and maize flours

The effects of endosperm vitreousness, cooking time and temperature on sorghum and maize starch digestion in vitro were studied using floury and vitreous endosperm flours. Starch digestion was significantly higher in floury sorghum endosperm than vitreous endosperm, but similar floury and vitreous endosperm of maize. Cooking with 2-mercaptoethanol increased starch digestion in both sorghum and maize, but more with sorghum, and more with vitreous endosperm flours. Increasing cooking time progressively reduced starch digestion in vitreous sorghum endosperm but improved digestibility in the other flours. Pressure-cooking increased starch digestion in all flours, but markedly more in vitreous sorghum flour; probably through physical disruption of the protein matrix enveloping the starch. Irrespective of vitreousness or cooking condition, the alpha-amylase kinetic constant (k) for both sorghum and maize flours remained similar, indicating that differences in their starch digestion were due to factors extrinsic to the starches. SDS-PAGE indicated that the higher proportion of disulphide bond-cross-linked prolamin proteins and more extensive polymerisation of the prolamins on cooking, resulting in polymers of M_r>100k, were responsible for the lower starch digestibility of the vitreous sorghum endosperm flour.     

The composition of floury and vitreous endosperm affects starch digestibility kinetics of the whole maize kernel

Maize grain starch is the major energy source in animal nutrition, and its high digestion and utilization largely depend on endosperm traits and the structure of the starch-lipoprotein matrix. The aim of this work was to determine floury and vitreous endosperm traits and its relation to starch digestibility rate. In total, kernels of 30 hybrids were manually dissected, and amylose, total zein and starch and non-starch lipids were determined in both vitreous and floury endosperm. Starch digestibility of the whole kernel was determined based on glucose released during a two-step in vitro pig model of enzymatic digestion, and starch digestibility rate was calculated according to the first-order kinetics. The vitreous endosperm of tested hybrids had higher contents of amylose (204.6 vs 190.4 g/kg), zein (63.2 vs 40.4 k/kg) and starch lipids (5.6 vs 4.9 g/kg), and lower content of non-starch lipids (7.3 vs 9.6 g/kg) than floury endosperm. Digestibility coefficients varied among hybrids, and starch digestibility rate varied from 0.73 to 1.63 1/h. Lipids in both vitreous and floury endosperm negatively correlated with the most of digestion coefficients, whereas zein correlated in vitreous and amylose in the floury endosperm (P < 0.05). Starch digestibility rate negatively correlated with all traits, except amylose content in vitreous endosperm. As a result, a linear regression model with four variables including contents of zein and starch lipids in vitreous and zein and amylose in floury endosperm can predict more than 65% variability of starch digestibility rate of tested hybrids.     

Partial-degradation and heat-moisture dual modification on the enzymatic resistance and boiling-stable resistant starch content of corn starches

Normal corn, Hylon V and Hylon VII starches were partially degraded by acid-ethanol treatment and applied to heat-moisture treatment (HMT) for improving the enzymatic resistance of starch. The weight-average degree of polymerization (DP_w) of acid-ethanol-treated (AET) corn starches ranged from 6.75 × 10⁵ to 181, 4.48 × 10⁵ to 121, and 1.94 × 10⁵ to 111 anhydrous glucose units for normal corn, Hylon V and Hylon VII starches, respectively. Starch retained its granular structure after AET and HMT, recovery of starch granules after modifications were higher than 92%. Resistant starch (RS) content and boiling-stable RS content of corn starch increased after dual modification, and the increment increased with increasing duration of AET. The boiling-stable RS content of dual-modified starch increased from 1.5 to 9.2, 12.2 to 24.1, and 18.0 to 36.2% for normal corn, Hylon V and Hylon VII starches, respectively. Increments of RS content and boiling-stable RS content of dual-modified starches were significantly correlated (r² > 0.700) with DP_w of starch, revealing that the enzymatic resistance of dual-modified corn starch granules increased with decreasing molecular size of starch. Result also suggested that starch granules partially degraded with AET could improve the molecular mobility and ordering during the consequent HMT.     

Study on melting and crystallization of short-linear chains from debranched waxy starches by in situ synchrotron wide-angle X-ray diffraction

In situ melting and crystallization of short-linear α-1,4-glucan (short-chain amylose, or SCA) from debranched waxy starches were investigated by synchrotron wide-angle X-ray diffraction. Amorphous SCA was prepared by dissolving completely debranched waxy starches in alkaline solution and neutralized by hydrochloric acid. When hydrated with 50% water at 25 °C, all amorphous SCA crystallized immediately and gave a B-type structure. The SCA from debranched waxy potato starch had a longer average chain length and a higher melting temperature but relatively lower crystallinity upon hydration; it was not completely melted at 100 °C and retained its original B-type structure during rapid cooling. In contrast, the SCA from debranched waxy wheat and waxy maize starches had a large portion of low molecular weight fractions, a higher crystallinity upon hydration, and a lower melting temperature. These differences suggest that amylopectin short chains crystallized more readily but their crystals were weaker than those of long chains. After the B-type crystals of hydrated SCA from waxy wheat and waxy maize starches melted, they reformed into the A-type polymorph upon rapid cooling. The thermal properties showed that the A-type polymorph of debranched waxy wheat and waxy maize starches had a higher melting temperature than their B-type structure.     

Starch Digestibility and Functional Properties of Rice Starch Subjected to Gamma Radiation

This study investigated the effect of gamma radiation on the digestibility and functional properties of rice starch. Rice cultivars IRGA417 and IAC202 were used for isolation of starch by the alkaline method. Starch samples were irradiated with 1, 2 and 5 kGy doses of ⁶⁰Co at a rate of 0.4 kGy/h. A control sample, which was not irradiated, was used for comparison. Irradiated and control starches were characterized by in vitro starch digestibility, total dietary fiber, color, water absorption index, water solubility index, syneresis, swelling factor, amylose leaching, pasting properties and gel firmness. Irradiations changed starch digestibility differently in either cultivar. Increasing radiation doses promoted increase in the color parameter b* (yellow), elevation in the capacity to absorb water, and solubility in water as well as the amylose leached from granules for both cultivars. Pasting properties showed a decrease that was proportional to the dose applied, caused by the depolymerization of starch molecules. Gel firmness of the starch from IAC202 was inversely proportional to the radiation dose applied, whereas for IRGA417, there was a reduction at 5 kGy dose. Rice starches can be modified by irradiation to exhibit different functional characteristics and they can be used by the food industries in products such as soups, desserts, flans, puddings and others.     

Molecular Structure of a Low-Amylopectin Starch and Other High-Amylose Maize Starches

The molecular structure of two commercially available high-amylose maize starches, HYLON® V starch and HYLON® VII starch, and of a newly developed low-amylopectin starch (LAPS) was examined. These high-amylose starches give three apparent fractions as determined by gel-permeation chromatography: a high-molecular weight (mol.wt) amylopectin fraction, a low-mol.wt amylose fraction, and an intermediate-mol.wt fraction which contains both linear and branched components. The low-mol.wt amylose fraction increases from 9·4% in HYLON V starch to 17·7% in HYLON VII starch and 26·5% in LAPS, whereas the high-mol.wt amylopectin fraction decreases from 31·1% in HYLON V starch to 13·8% in HYLON VII starch and 2·5% in LAPS. The percentage of linear components in HYLON V starch, HYLON VII starch, and LAPS are 42, 54, and 80%, respectively. High-amylose starches have a large proportion of long chains in their branched fractions compared to waxy-maize and normal-maize starch. Both HYLON VII starch and the LAPS give B plus V-type X-ray diffraction patterns, but the LAPS has even a higher gelatinization temperature, lower swelling power in hot water, and is more resistant to acid digestion. With the lack of amylopectin, amylose accounts for at least part of the double helical structure in the LAPS granules.     

Functionality of chemically modified wild-type,partial waxy and waxy starches from tetraploid wheats

Partial waxy (reduced-amylose) and fully waxy (amylose-free) tetraploid durum wheats (Triticum turgidum L. var durum) were developed by introgression of null alleles at the Wx-A1 and Wx-B1 loci from common hexaploid wheat (Triticum aestivum L.). Purified starches were obtained from each genotype, and chemically modified by: 1) cross-linking with phosphorus oxychloride, 2) substitution with propylene oxide, and 3) sequential cross-linking with phosphorus oxychloride followed by substitution with propylene oxide. Functional properties were compared to blends of waxy and wild-type durum starches of known amylose contents. Significant differences in functionality were observed amongst the genotypes and blends after each modification. Waxy (0% amylose) and wild-type (30% amylose) typically were at the extremes of the observed ranges of functional properties. In general, the functional properties of the chemically modified starches were dependent upon amylose content. Starches from Wx-B1 null lines (24% amylose), were an exception. After substitution, such starches had the significantly highest value for RVA final viscosity, and generally performed in a manner similar to starch blends of 12–18% amylose.     

Thermal dissolution of maize starches in aqueous medium

Starches are insoluble in neutral water at room temperature. However, if they are heated beyond the boiling point of water in a closed container, they eventually dissolve. The dissolution profile depends on the type of starch. The dissolution processes of maize starches were monitored in real time by measuring the transmittance of starch dispersion/solution while it was heated. It was found that the dissolution of starches in water results from the degradation of the two components of starches, amylopectin and amylose. Starches from both normal maize and waxy maize were fully dissolved in water when their aqueous solution was heated to approximately 174 °C. Upon cooling to room temperature, amylopectin remained in a solubilized state while amylose molecules were retrograded. For starch solutions that have been heated to higher than 191 °C, the color turned brown and the retrogradation of amylose was not observed.     

Glass transition temperature of starches with different amylose/amylopectin ratios

The glass transition temperatures (T_g) of starch with different amylose/amylopectin ratios were systematically studied by a high-speed DSC. The cornstarches with different amylose contents (waxy 0; maize 23, G50 50 and G80 80) were used as model materials. The high heating speed (up to 300 °C/min) allows the weak T_g of starch to be visible and the true T_g was calculated by applying linear regression to the results from different heating rates. It is confirmed for the first time, that the higher the amylose content is, the higher the T_g is for the same kind of starch. The sequence of true T_g of cornstarch is G80 > G50 > maize > waxy when samples contain the same moisture content, which corresponds to their amylose/amylopectin ratio. It was found that T_g was increased from about 52 to 60 °C with increasing amylose content from 0 to 80 for the samples containing about 13% moisture. The microstructure and phase transition were used to explain this phenomenon, in particular the multiphase transitions that occur in high-amylose starches at higher temperatures, and the gel-ball structure of gelatinized amylopectin.     

Physicochemical characterization and in-vitro digestibility of extruded rice noodles with different amylose contents based on rheological approaches

The physicochemical properties and in-vitro digestibility of extruded rice noodles with different amylose contents were characterized from a rheological point of view. Thermo-mechanical measurements showed that the rice flour with higher amylose contents exhibited greater stability to dual-mixing and higher degrees of starch gelatinization and retrogradation. In addition, greater elastic properties were clearly observed in the high amylose rice samples. The use of high amylose rice flour produced noodles with a harder texture, consequently contributing to reduced cooking loss. Furthermore, the rheological changes of extruded rice noodles were monitored in real time during the in-vitro starch digestion. The rice noodle digesta with higher amylose contents exhibited greater viscosities throughout the simulated oral-gastric-intestinal digestion steps. The flow behaviors of the rice noodle digesta consisted of the Power-law region and infinite shear plateau that were satisfactorily characterized by the Sisko model (R² > 0.99).     

Hydrothermal modification of wheat starch. Part 2. Thermal characteristics of pasting and rheological properties of pastes

Water suspensions of starch (with the concentration of 8 g/100 g) were prepared in a measuring vessel of a Brabender viscograph and heated to temperatures of 74, 76.5, 79, 81.5, 84, 86.5, 89, 91.5 or 94 °C under continuous stirring. The resultant solution was cooled and frozen, and then defrosted. Thermal characteristics of re-pasting, rheological properties of produced pastes, starch solubility in water and swelling power were determined.The heating and freezing of the wheat starch suspension induced changes in its properties, with tendency and extent of these changes depending on temperature of pre-heating. The thermal characteristics of the analyzed starches revealed three peaks that corresponded to transitions proceeding during solubilization of retrograded amylopectin and retrograded amylose and solubilization of amylose–lipid complexes. Retrogradation of amylose induced by starch pre-heating followed by its freezing affected also the consistency coefficient and yield stress of the pastes formed by the analyzed starches. Values of these rheological parameters were higher at higher temperatures of pre-heating, compared to the pastes prepared from native starch, and were changing accordingly to the determined second order polynomial function. Amylose retrogradation occurring during the production of starch preparations diminished their solubility in water and increased their swelling power compared to native starch.     

Functionality of native tetraploid wheat starches: Effects of waxy loci alleles and amylose concentration in blends

Partial waxy (reduced amylose) and fully waxy (amylose-free) tetraploid durum wheats (Triticum turgidum L. var. durum) were used to investigate the relationships between both intra- and inter-granular variation in amylose concentration and starch functionality. Starches isolated from each genotype (intra-granular amylose variants) were compared to those of commercially available hexaploid wild-type and waxy starches, and functionalities compared to blends (inter-granular variants) of durum waxy and wild-type starches of 0, 6, 12, 18, 24, & 30% amylose content. Starch particle size distributions were not significantly different amongst partial waxy and wild-type genotypes; waxy samples had significantly smaller mean starch granule size. Few significant differences for crystallite melting and related intrinsic heat as determined by differential scanning calorimetry (DSC) were observed. The detected differences in starch gel color or gel strength generally were observed for the waxy samples relative to wild-type. Numerous significant differences were observed via Rapid-Visco Analysis (RVA). Pasting peak viscosity and breakdown were inversely proportional to % amylose. Wx-B1 null final viscosity differed significantly from that of all other blends and genotypes, demonstrating that genotypic differences exist amongst the partial waxy types, independent of amylose concentration per se.     

Effect of instantaneous controlled pressure drop (DIC) on physicochemical properties of wheat, waxy and standard maize starches

Standard maize starch (SMS), waxy maize starch (WMS) and wheat starch (WTS) were hydrothermally treated by the Instantaneous Controlled Pressure Drop (DIC) process. This process consists in a short pressurisation obtained by the injection of saturated steam at fixed pressure during a predetermined time followed by a sudden pressure drop towards vacuum. The effects of DIC conditions on thermal characteristics, enzyme susceptibility, pasting (Brabender) and rheological properties of treated starches were investigated. For treated starches, an increase of transition temperatures (T_o and T_p), a narrowing of the width of gelatinization endotherms and a decrease of the gelatinization enthalpies (ΔH) were observed as the severity of processing conditions increased. WMS, SMS and WTS showed a significant increase in enzymatic hydrolysis after treatment. The saccharification yield showed an increase from 19% (native) to 44%, 21% (native) to 59% and 55% (native) to 79% for SMS, WMS and WTS, respectively. The study suggests that the structural modifications due to the previous DIC treatment influence the in-vitro hydrolysis and the access to the ultrastructure of starch granules; the susceptibility to hydrolysis increases from SMS to WMS and WTS. For all treated starches, the decrease in peak viscosity and in apparent viscosity was related to the processing conditions.     

A Multi-stages Biosynthetic Pathway in Starch Granules Revealed by the Ultrastructure of Maize Mutant Starches

Our previous work indicated that starches containing B-type crystallites show low susceptibility to amylolysis and suggested that B-type crystallites have an effect on starch granule organisation. To elucidate granular ultrastructure, double wxae and aedu maize mutant starches containing A- (30 and 50% respectively) and B-type (70 and 50% respectively) crystallites were treated with porcine pancreatic alpha -amylase. The surface structure of the native and degraded starches was studied by scanning electron microscopy, and the internal ultrastructure by transmission electron microscopy after staining with PATAg reagents. The results confirm the influence of B-type crystallites on granule organisation and indicate that starches containing B-type crystallites show an amylolysis attack pattern with minor exocorrosion and major endocorrosion. The granule organisation of A- and B-type starches proposed is not consistent with an onion ring model¹and may account for the different behaviour of these starches to amylolysis. Transmission electron microscopy showed that most native wxae and aedu starch granules are composed of a core with a classical alternating structure and a peripheral ring. The peripheral ring in wxae starch was ordered and resistant to amylolysis, whereas that of aedu was disordered and degradable. It is proposed that these two specific forms of granule organisation are attributable to variations in the enzymatic activities of specific starch synthesising enzymes during the course of starch biosynthesis.     

An innovative approach for significantly increasing enzyme resistant starch type 3 content in high amylose starches by using extrusion cooking

In this study effects of extrusion cooking on enzyme resistant starch (RS) formation in high amylose corn starches (Hylon V and VII) and the functional properties of RS preparations were investigated. Native starches were extruded at 50, 60, 70% feed moisture contents, at constant screw speed (100 rpm) and barrel temperature (140 °C). Among these samples, the highest RS contents were observed at 60% feed moisture. Therefore, feed moisture in the second and third extrusion cycles was set at 60%. There were significant increases in RS contents of both Hylon V and Hylon VII after the second extrusion cycle (p < 0.05). After the third extrusion, the RS levels reached to 40.0 and 45.1% for Hylon V and Hylon VII, respectively. Substantial loss of birefringence in these samples indicated that the increases in RS were mainly due to RS3 formation. The RS samples produced by extrusion did not have high emulsion capacity, but the ones produced from Hylon VII had high emulsion stability. Although, decreases in L* and increases in b* values of extruded samples were significant as compared to respective native starches, the changes were not substantial. Therefore, their incorporation is not expected to cause major changes in the colour of end-products.     

不同直链淀粉含量小麦淀粉理化性质与消化特性研究

为深入理解小麦直链淀粉含量与淀粉理化性质及消化特性之间的关系,以4份直链淀粉含量差异较大的小麦品种为材料,分析了其淀粉理化性质以及消化特性方面的差异。结果表明,随着直链淀粉含量的增加,回生值呈增加趋势;在体外消化前期,4个小麦品种的淀粉消化速率差异显著,西农836最慢,陕糯1号最快。这种差异与直链淀粉含量及回生值密切相关,说明直链淀粉比支链淀粉更易于回生,糊化后的冷却过程中更容易发生分子间部分重排和再结晶,重新组装成有序的聚集体,导致供试材料之间快速消化淀粉含量及总体消化程度不同。直链淀粉含量高的品种具有更多的B型小淀粉粒,但是大量B型小淀粉粒的存在并没有导致淀粉消化速率以及淀粉总体消化程度的提高,说明淀粉充分糊化后,粒度的差异并不改变淀粉消化特性。     

Texture determinants in cooked, parboiled rice. I: Rice starch amylose and the fine stucture of amylopectin

The constituent starches from eleven cultivars of non-waxy rice with different degrees of cooking hardness were subjected to detailed analysis. No significant differences were observed in the granule morphology, crystallinity and size distribution of the starch granules that could be correlated with the textures of the cooked, parboiled rices. Differences in the contents of amylose and the fine structures of amylopectin were detected, however, which did correlate with texture. It appeared that the texture of the rices was critically controlled by the proportion of the longest (DP 92–98) and shortest (DP&lE25) amylopectin chains but not the intermediate (DP 43–68) chains. Hard cooking rice tended to have a higher amylose content (or amylose:amylopectin ratio) and more longer chain amylopectin than soft cooking rice, which feature is thought to encourage more extensive intra and/or inter molecular interactions with other components in rice grain, such as protein, lipid and non-starch polysaccharides and results in a firmer texture. The different amylopectin structures may explain why rices that possess similar amylose contents can have different textural properties. Such a correlation suggests that the structure of the starches determined from the SE–HPLC profile of the debranched amylopectin could be useful as a sensitive screening method in the classification of cooked rice texture.