Barley Starch Granules Research Articles

Physicochemical properties of A- and B-type granules isolated from waxy and normal hull-less barley starch

Large A-type and small B-type starch granules separated from waxy and normal hull-less barley starches were investigated for their physicochemical properties. Hull-less barley starch granules were covered by a membrane composed mainly of phospholipids. Channels of waxy A- and B-type granules were rich in proteins and phospholipids. Compared with A-type starch, B-type starch exhibited higher specific surface area, volume and average diameter of mesopores. Waxy A-type granules exhibited the higher peak, breakdown, final and setback viscosity than did B-type granules, while normal A-type granules showed the lower peak, breakdown, final viscosity and the higher setback viscosity than did B-type granules. B-type starch gels with lower storage modulus exhibited a less elastic gel network structure and retrograded more slowly. Moreover, in vitro hydrolysis of starch showed that the B-type granules exhibited a higher hydrolysis extent and rate than the A-type granules in the first stage, which was consistent with higher initial α-amylase binding ability of B-type granules. The study showed that the A-type and B-type starch separated from waxy and normal hull-less barley exhibited very different physicochemical properties.

Morphological, functional and thermal characteristics of hydroxypropylated-crosslinked barley starches

The present study investigated effects of three different levels of crosslinking using a mixture of sodium trimetaphosphate (STMP) and sodium tripolyphosphate (STPP) on hydroxypropylated barley starches. Hydroxypropylated barley starches crosslinked with 1%, 1.5% and 2% mixture of STMP and STPP were coded as HPCL(1), HPCL(1.5) and HPCL(2.0), respectively. However, the level of hydroxypropylation employed was same for all the modifications i.e. 6%. The results showed that increase in level of crosslinking increased swelling power, solubility and water holding capacity of starches. HPCL(2.0) starch demonstrated noticeably lower percent transmittance, higher percent retrogradation and elevated peak viscosity. Harder gels were produced by dual modified barley starches. A decline was observed in thermal transition temperatures after dual modification. FTIR was unable to detect much difference among different samples. However, peak at 1414 cm−1 associated with hydroxypropylation was detected. Hydroxypropylation followed by crosslinking led to roughness and grooves formation on the surface of barley starch granules.

Different gelatinization characteristics of small and large barley starch granules impact their enzymatic hydrolysis and sugar production during mashing

This study investigates the impact of different gelatinization characteristics of small and large barley starch granules on their enzymatic hydrolysis and sugar production during mashing. Therefore, a barley malt suspension was consecutively incubated at 45, 62, 72 and 78 °C to monitor starch behavior and enzymatic starch hydrolysis and sugar production. The combination of microscopic and rapid visco analyses showed that small starch granules persisted longer in the system and were present non-gelatinized at temperatures higher than 62 °C. HPAEC-PAD analysis showed that 8% of the total amount of starch, predominantly small granules, gelatinized at temperatures between 62 °C and 78 °C. Due to their delayed gelatinization in this system, their enzymatic hydrolysis yielded relatively more dextrins compared to what was observed for large granules. It was concluded that small granules should be taken into account when optimizing enzymatic hydrolysis of barley starch, like in brewing, distilling or bio-ethanol production.

In vitro digestibility, pasting, and structural properties of starches from different cereals

ABSTRACTStarches isolated from seven different cereals were evaluated for their composition, physicochemical, in vitro digestibility, structural, morphological, and pasting properties. The in vitro starch digestion rate and estimated glycemic index (GI) of cereal starches were evaluated along with the impact of cooking on starch digestion. The cooking of starch slurries increased the rapidly digestible starch content from a range of 34.7–54.4% to a range of 60.5–78.5%. On the basis of hydrolysis index, the GI ranged from 83.6 to 91.8 and after cooking it increased from 95.1 to 98.6 for different cereal starches. Both the swelling power and solubility showed an increasing trend with rising temperature. Paste clarity of starches negatively correlated with fat content. The amylose content of various starches ranged from 17.7 to 24.7% and was negatively correlated to crystalline index (r = -0.975, p ≤ 0.05). XRD pattern revealed A-type pattern of crystalline starch, where crystallinity index ranging between 28.2 to 44.9%. FTIR revealed slight differences among chemical bonding of starches from different cereals. From scanning electron micrograph observations, wheat and barley starch granules proved smoother as compared to other cereal granules. Barley contained the highest (27.5 µm) and rice had the lowest (10.2 µm) size starch granules. The pasting properties were significantly (p ≤ 0.05) different for different cereal starches. Peak, breakdown, and final viscosities were the highest for maize starch (1725, 384, and 2112 mPa.s, respectively), whereas rice and oats exhibited the highest trough and setback viscosities (1420 and 954 mPa.s, respectively).

Influence of diurnal photosynthetic activity on the morphology, structure, and thermal properties of normal and waxy barley starch

This study investigated the influence of diurnal photosynthetic activity on the morphology, molecular composition, crystallinity, and gelatinization properties of normal barley starch (NBS) and waxy barley starch (WBS) granules from plants cultivated in a greenhouse under normal diurnal (16h light) or constant light photosynthetic conditions. Growth rings were observed in all starch samples regardless of lighting conditions. The size distribution of whole and debranched WBS analyzed by gel-permeation chromatography did not appear to be influenced by the different lighting regimes, however, a greater relative crystallinity measured by wide-angle X-ray scattering and greater crystalline quality as judged by differential scanning calorimetry was observed under the diurnal lighting regime. NBS cultivated under the diurnal photosynthetic lighting regime displayed lower amylose content (18.7%), and shorter amylose chains than its counterpart grown under constant light. Although the relative crystallinity of NBS was not influenced by lighting conditions, lower onset, peak, and completion gelatinization temperatures were observed in diurnally grown NBS compared to constant light conditions. It is concluded that normal barley starch is less influenced by the diurnal photosynthetic lighting regime than amylose-free barley starch suggesting a role of amylose to prevent structural disorder and increase starch granule robustness against environmental cues.

Impact of full range of amylose contents on the architecture of starch granules*

The effects of amylose deposition on crystalline regions of barley starch granules were studied in granules containing zero to 99.1% amylose using “waxy” (WBS, 0% amylose), normal (NBS, 18% amylose) and amylose-only barley lines (AOS, 99.1% amylose). The effects were probed after hydrolysis of amorphous regions of starch granules in dilute HCl generating lintners, which typically represent the crystalline lamella of starch granules. Compared to NBS and WBS, AOS granules exhibited an irregular, multilobular morphology with a rough surface texture. AOS displayed lower rates of acid hydrolysis than WBS, and AOS reached a plateau at ∼45wt% acid hydrolysis. High-performance anion-exchange chromatography of lintners at equivalent levels of hydrolysis (45wt%) revealed the average degree of polymerization (DP) of AOS lintners was 21, substantially smaller than that of NBS and WBS (DP 42). AOS lintners contained the lowest number of chains (NC) per molecule (1.1) compared to NBS (2.8) and WBS (3.3) and the average chain length of AOS, NBS and WBS lintners was 19, 15 and 13, respectively. Hence, both NC and the average chain length correlated with amylose content. The size distribution profile of AOS lintners revealed a repeat motif in the molecules corresponding to 5–6 glucose residues.

Second harmonic generation microscopy investigation of the crystalline ultrastructure of three barley starch lines affected by hydration.

Second harmonic generation (SHG) microscopy is employed to study changes in crystalline organization due to altered gene expression and hydration in barley starch granules. SHG intensity and susceptibility ratio values (R'SHG ) are obtained using reduced Stokes-Mueller polarimetric microscopy. The maximum R'SHG values occur at moderate moisture indicating the narrowest orientation distribution of nonlinear dipoles from the cylindrical axis of glucan helices. The maximum SHG intensity occurs at the highest moisture and amylopectin content. These results support the hypothesis that SHG is caused by ordered hydrogen and hydroxyl bond networks which increase with hydration of starch granules.

Starch Microstructure and Starch Hydrolysis in Barley and Oat Tempe During In Vitro Digestion

Various botanical and structural characteristics of starchy foods are considered to modify the rate of starch digestion and the glycaemic responses in humans. The main objective of the study was to examine the impact of fermented barley and oat microstructure on the rate of in vitro starch hydrolysis. A dynamic gastrointestinal model was used to estimate the degree of starch hydrolysis during in vitro digestion of fermented whole grain cereal meals. Light microscopy and confocal laser scanning microscopy were used to study the microstructural changes. In parallel to the in vitro studies, the impact of fermented barley and oats on postprandial plasma glucose responses was evaluated in a human study. Micrographs were taken during in vitro digestion experiments with fermented whole grains and compared with micrographs of boiled barley (undigested). Images showed that most of the oat starch granules were degraded after 120 min of digestion, whereas barley starch granules were less degraded, even after 180 min of digestion. The findings were confirmed by faster starch hydrolysis from the fermented oat meal, measured as maltose generated during in vitro digestion. The area under the curve (AUC) was calculated from the plotted maltose curves of the meals. AUC for barley tempe (266 ± 33) was 40 % of the AUC for oat tempe (663 ± 8) and significantly different (p < 0.007) from AUC oat tempe. The in vitro data closely resembled the AUCs for plasma glucose from the parallel human study. In terms of glucose response, the mean AUC for barley tempe was 46 % of the AUC for oat tempe in the human study. The agreement between the in vitro and in vivo data indicates the potential of the in vitro method as a tool to predict the rate of starch degradation of cereal products.

Morphological, Thermal, Pasting, and Rheological Properties of Barley Starch and Their Blends

Native barley starch, as well as its blends with corn, wheat, and rice starch at different ratios of 75:25, 50:50, 25:75 were examined in terms of morphology, thermal, pasting, rheological, and retrogradation properties. Amylose content varied between 10.9–41.4% in rice, corn, wheat, and barley while it ranged from 18.02–38.40% in blends of barley starch with rice, corn, and wheat. A rapid visco analyzer showed that barley starch and its blends having low amylose content exhibited higher peak viscosity, breakdown, and setback than the high-amylose-containing starches and their blends. Amylose content was found to be negatively correlated with swelling power while it exhibited nonlinear relationship with solubility index. The transmittance of starch suspension stored at 4°C decreased during storage up to 6 days. Barley starch granules were largest (<110 μm) in size followed by wheat (<30 μm), corn (<25μm) and rice (<20μm) starches. Gelatinization temperatures (To, Tp, Tc) and enthalpies of gelatinization (ΔHgel) of starches from different sources also differed significantly. Corn and rice starches showed higher transition temperatures in general than those from wheat and barley; however, they showed higher ΔHgel values. Barley starch showed a higher tendency towards retrogradation than the cereal starches. Barley starch showed highest peak G′, G″ and lower tan ð than corn, rice and wheat starches during the heating cycle. This study showed that the magnitude of changes in their properties during blending depends on the amylase content and morphological characteristics.

Relationship Between the Ratio of Large and Small Starch Granules Determined by Gravitational Field‐Flow Fractionation and Malting Quality of Barley Varieties

The ratio of large and small barley starch granules was determined by gravitational field‐flow fractionation (GFFF) under optimized conditions. The optimization was aimed to reach the elution of both large and small starch granules in the focusing elution mode and long‐term reproducibility. Kernels of twelve spring and winter barley varieties grown in three testing stations during three years were used for isolation and size determination of starch granules, i.e., monitoring was conducted with 108 samples. The two‐row winter varieties had the highest ratios of large and small starch granules A/B. On the other hand, the two‐row varieties of spring malting barley had the lowest ratios A/B. The ratios of large and small starch granules A/B of six‐row winter varieties were between the previous groups. The obtained relationship between the ratio of large and small starch granules and malting properties for this set of barley varieties is opposite to previous expectations, assuming that spring malting barley varieties have higher ratios of large and small starch granules than winter ones. GFFF was shown as a suitable technique for the determination of the ratio A/B starch granules. From the point of view of the results obtained in this study, it is evident that not only the ratio A/B of large and small starch granules is important for malting properties, but other factors are also important, e.g., activity of hydrolytic enzymes.

High‐Speed Separation and Size Characterization of Wheat and Barley Starch Granules by Lift‐Hyperlayer Asymmetrical Flow Field‐Flow Fractionation in Synergy with SPLITT Fractionation

AbstractThe complementary employment of two separation techniques, one analytical‐scale, the other preparative‐scale, i.e. the lift‐hyperlayer asymmetrical flow field‐flow fractionation (AsyFlFFF) and the split flow‐thin cell (SPLITT) is presented for the first time as a methodology to size‐separate and rapidly characterize starch granules of different botanical origins. Very rapid separation and size characterization is achieved by AsyFlFFF and the techniques are mutually used to characterize and confirm data produced by the other. Light microscopy (LM) was used to test the quality of the performed SPLITT separations and consequently to evaluate the AsyFlFFF results. The procedure was applied to wheat and barley starch.

The activity of barley α-amylase on starch granules is enhanced by fusion of a starch binding domain from Aspergillus niger glucoamylase

High affinity for starch granules of certain amylolytic enzymes is mediated by a separate starch binding domain (SBD). In Aspergillus niger glucoamylase (GA-I), a 70 amino acid O-glycosylated peptide linker connects SBD with the catalytic domain. A gene was constructed to encode barley α-amylase 1 (AMY1) fused C-terminally to this SBD via a 37 residue GA-I linker segment. AMY1-SBD was expressed in A. niger, secreted using the AMY1 signal sequence at 25 mg × L −1 and purified in 50% yield. AMY1-SBD contained 23% carbohydrate and consisted of correctly N-terminally processed multiple forms of isoelectric points in the range 4.1–5.2. Activity and apparent affinity of AMY1-SBD (50 nM) for barley starch granules of 0.034 U × nmol −1 and K d = 0.13 mg × mL −1, respectively, were both improved with respect to the values 0.015 U × nmol −1 and 0.67 mg × mL −1 for rAMY1 (recombinant AMY1 produced in A. niger). AMY1-SBD showed a 2-fold increased activity for soluble starch at low (0.5%) but not at high (1%) concentration. AMY1-SBD hydrolysed amylose DP440 with an increased degree of multiple attack of 3 compared to 1.9 for rAMY1. Remarkably, at low concentration (2 nM), AMY1-SBD hydrolysed barley starch granules 15-fold faster than rAMY1, while higher amounts of AMY-SBD caused molecular overcrowding of the starch granule surface.

Starch from hull-less barley: IV. Morphological and structural changes in waxy, normal and high-amylose starch granules during heating

Morphological and ultrastructural changes of waxy, normal and high-amylose barley starch granules heated in water to various temperatures were examined by scanning electron microscopy and transmission electron microscopy. Swelling and gelatinization/solubilization patterns of waxy hull-less barley (HB) starch were different from those of non-waxy HB starches. Waxy HB starch granules were split into two halves with a large cavity in the central region of the granule and started melting at the equatorial grooves during heating in water to 55 °C, whilst only a small amount of molecules leached out of granules. Heating resulted in rapid fragmentation of granules at temperatures exceeding 55 °C. However, with increasing heating temperature, the extent of release of the exudate became more pronounced while swollen starch granules remained intact in non-waxy HB starches. Knobs/protrusions (200–400 nm diameter) were present on the granule surfaces of non-waxy HB starches during gelatinization, but not present on the granule surface of gelatinised waxy HB starch. Ghost structure was formed and embedded in exudate/soluble matrix with a web network within ghosts at 90–100 °C. Bundles of brush-like strands in the exudate of high-amylose starch were formed after heating. Fine particles (∼10 nm diameter) arranged into chains were present on the granule remnants of all three types of barley starches. MALDI-MS analysis showed that distribution of branched chains in leached (at 65 °C) amylopectin was similar in both waxy and non-waxy HB starches. An increase in heating temperature from 65 to 100 °C increased the molecular size and branch chain length of the leached amylopectin fraction from high-amylose HB starch. The chain length distribution of amylopectin was identical in both the soluble and insoluble fractions obtained by heating high-amylose HB starch in water to 100 °C.

Barley Starch Granules Subject to SPLITT Cell Fractionation and Sd/StFFF Size Characterization

The coupling of two separation techniques is proposed as a methodology able to fractionate, starch granules of different botanical origins, both on preparative and analytical scale. The split flow thin cell (SPLITT cell) was optimized to produce a binary separation of the starch granules into two different size classes with a 5 μm cutoff point. The size separation was always better than 80%. The Sedimentation Steric Field Flow Fractionation (Sd/StFFF) provides sample particle size distribution (PSD) and is used here to control the SPLITT fractions. The proposed techniques, especially the former, are gentle and easy to use. Both the SPLITT and Sedimentation Fractionation results were checked by scanning electron microscopy (SEM). The procedure was originally applied to two samples from barley. The bimodal diameters were computed.

Adhesion of bifidobacteria to granular starch and its implications in probiotic technologies.

Adhesion of 19 Bifidobacterium strains to native maize, potato, oat, and barley starch granules was examined to investigate links between adhesion and substrate utilization and to determine if adhesion to starch could be exploited in probiotic food technologies. Starch adhesion was not characteristic of all the bifidobacteria tested. Adherent bacteria bound similarly to the different types of starch, and the binding capacity of the starch (number of bacteria per gram) correlated to the surface area of the granules. Highly adherent strains were able to hydrolyze the granular starches, but not all amylolytic strains were adherent, indicating that starch adhesion is not a prerequisite for efficient substrate utilization for all bifidobacteria. Adhesion was mediated by a cell surface protein(s). For the model organisms tested (Bifidobacterium adolescentis VTT E-001561 and Bifidobacterium pseudolongum ATCC 25526), adhesion appeared to be specific for alpha-1,4-linked glucose sugars, since adhesion was inhibited by maltose, maltodextrin, amylose, and soluble starch but not by trehalose, cellobiose, or lactose. In an in vitro gastric model, adhesion was inhibited both by the action of protease and at pH values of < or =3. Adhesion was not affected by bile, but the binding capacity of the starch was reduced by exposure to pancreatin. It may be possible to exploit adhesion of probiotic bifidobacteria to starch granules in microencapsulation technology and for synbiotic food applications.

Stability and function of interdomain linker variants of glucoamylase 1 from Aspergillus niger.

Several variants of glucoamylase 1 (GA1) from Aspergillus niger were created in which the highly O-glycosylated peptide (aa 468--508) connecting the (alpha/alpha)(6)-barrel catalytic domain and the starch binding domain was substituted at the gene level by equivalent segments of glucoamylases from Hormoconis resinae, Humicola grisea, and Rhizopus oryzae encoding 5, 19, and 36 amino acid residues. Variants were constructed in which the H. resinae linker was elongated by proline-rich sequences as this linker itself apparently was too short to allow formation of the corresponding protein variant. Size and isoelectric point of GA1 variants reflected differences in linker length, posttranslational modification, and net charge. While calculated polypeptide chain molecular masses for wild-type GA1, a nonnatural proline-rich linker variant, H. grisea, and R. oryzae linker variants were 65,784, 63,777, 63,912, and 65,614 Da, respectively, MALDI-TOF-MS gave values of 82,042, 73,800, 73,413, and 90,793 Da, respectively, where the latter value could partly be explained by an N-glycosylation site introduced near the linker C-terminus. The k(cat) and K(m) for hydrolysis of maltooligodextrins and soluble starch, and the rate of hydrolysis of barley starch granules were essentially the same for the variants as for wild-type GA1. beta-Cyclodextrin, acarbose, and two heterobidentate inhibitors were found by isothermal titration calorimetry to bind to the catalytic and starch binding domains of the linker variants, indicating that the function of the active site and the starch binding site was maintained. The stability of GA1 linker variants toward GdnHCl and heat, however, was reduced compared to wild-type.

Optimization of experimental conditions for the separation of small and large starch granules by gravitational field-flow fractionation

Separation of small and large barley starch granules by gravitational field-flow fractionation was investigated from the point of view of sample pre-treatment, amount of injected sample, and elution conditions. The sample pre-treatment study resulted in the conclusion that it is reasonable to soak the starch granules for at least 24 h prior to separation. The experiments with different amounts of injected sample show that it is possible to increase as well as decrease twofold the sample amount usually used without any change in retention ratios. The implementation of flow-rate gradients for elution of the starch granules reduced total separation time. However, the applied flow-rate gradients did not improve the resolution of peaks A and B compared with the generally used constant flow-rate. Thus, for barley starch granules, the constant flow-rates within the range from 0.8 to 1.0 ml/min seem to provide the best compromise of total separation time, peak resolution and instrumental expense.

Fine Structures of Amylose and Amylopectin from Large, Medium, and Small Waxy Barley Starch Granules

ABSTRACTAmylose and amylopectin were prepared from large, medium, and small granule starches of classified waxy barley flour, and their fine structures were investigated. The amylose content had a wide distribution range (≈1.4–9.4%). Number‐average degrees of polymerization (DPn) of the amyloses were similar among the samples (≈1,200–1,300). But number of chains per molecule (NC) decreased from the surface to the center (≈6–10 chains). DPn of the amylopectins varied from 4,657 to 14,604; decreased in the order of large, medium, and small granules in same fractions of the grain; and increased from the surface layer to the center. Longest chains (LC) were not found in any of the amylopectin molecules. The large amylopectin molecule had more long chains and fewer A chains than the small molecule. The amylose content had definite effects on the transition temperature range and crystal formation of the starch granules. There were positive correlations between DPn of the amylopectin and relative crystallinity (γ = +0.69) and enthalpy value (γ = +0.80), respectively. These findings may help to elucidate biosynthesis mechanism of starch.

Comparison of Size Characterization of Barley Starch Granules Determined by Electron and Optical Microscopy, Low Angle Laser Light Scattering and Gravitational Field-Flow Fractionation

Several methods were used for the characterization of starch granules isolated from barley kernels. A procedure based on a combination of alkaline digestion, toluene treatment and filtration over sieves with pore diameters of 70 and 40 μm was used for isolation and purification of starch granules from kernels. The released starch granules were characterized by various methods: scanning electron microscopy (SEM), image analysis of optical microscopy data (IAOM), low angle laser light scattering (LALLS), and gravitational field-flow fractionation (GFFF). All methods showed the bimodal size distribution of the isolated starch granules, however, they differed in the ratio of large and small starch granules. LALLS and GFFF were also used for determination of the ratio of large and small starch granules (ratio A/B) isolated from two malting barley cultivars Kompakt and Akcent. Both techniques determined the higher ratio A/B for the cultivar Akcent. SEM was also used to examine the extent of digestion. The micrographs indicate that a significant proportion mainly of small granules are still embedded into residues of endosperm and a more extensive digestion must be performed to release all starch granules from barley kernels.

Effect of Partial Gelatinization and Lipid Addition on α‐Amylolysis of Barley Starch Granules

ABSTRACTThe effect of partial gelatinization with and without lipid addition on the granular structure and on α‐amylolysis of large barley starch granules was studied. The extent of hydrolysis was monitored by measuring the amount of soluble carbohydrates and the amount of total and free amylose and lipids in the insoluble residue. Similarly to the α‐amylolysis of native large barley starch granules, lipid‐complexed amylose (LAM) appeared to be more resistant than free amylose and amylopectin. Partial gelatinization changed the hydrolysis pattern of large barley starch granules; the pinholes typical of α‐amylase‐treated large barley starch granules could not be seen. Lipid addition during partial gelatinization decreased the formation of soluble carbohydrates during α‐amylolysis. Also free amylose remained in the granule residues and mostly amylopectin hydrolyzed into soluble carbohydrates. These findings indicate that lysophospholipid (LPL) complexation with amylose occurred either during pretreatment or after hydrolysis, and free amylose was now part of otherwise complexed molecules instead of being separate molecules. Partial gelatinization caused the granules to swell somewhat less during heating 2% starch‐water suspensions up to 90°C, and lipid addition prevented the swelling completely. α‐Amylolysis changed the microstructure of heated suspensions. No typical twisting of the granules was seen, although the extent of swelling appeared to be similar to the reference starch. The granules with added LPL were partly fragmented after hydrolysis.