Structure of lintnerized starch is related to X-ray diffraction pattern and susceptibility to acid and enzyme hydrolysis of starch granules

Acid-resistant residues (lintnerized starches, Ls) were prepared from starches showing A-, B- and C- X-ray diffraction patterns. Ls retained the same X-ray crystalline type as their native counterparts with an improvement in diffraction intensity. Fluorophore-assisted capillary electrophoresis (FACE) study indicated that structural characteristics of Ls were associated with X-ray diffraction patterns. Double helices originated from linear chains with an approximate average degree of polymerisation (DP) 14, 16, and 15 would span the entire length of crystalline lamellae of A-, B-, and C-type starches, respectively. The proportion of singly branched materials (SB) with DP 25 protected in Ls was higher for A-type Ls (10-17%) than for B-type Ls (4-6%) and C-type Ls (8%). The structures of SB were similar in which branched chain (DP 13-15) was longer than main chain (DP 10-12). The structural characteristics of Ls are discussed in relation to acid and enzymatic degradations of starch granules.     

Amylopectin molecular structure reflected in macromolecular organization of granular starch

For lintners with negligible amylose retrogradation, crystallinity related inversely to starch amylose content and, irrespective of starch source, incomplete removal of amorphous material was shown. The latter was more pronounced for B-type than for A-type starches. The two predominant lintner populations, with modal degrees of polymerization (DP) of 13-15 and 23-27, were best resolved for amylose-deficient and A-type starches. Results indicate a more specific hydrolysis of amorphous lamellae in such starches. Small-angle X-ray scattering showed a more intense 9-nm scattering peak for native amylose-deficient A-type starches than for their regular or B-type analogues. The experimental evidence indicates a lower contrasting density within the "crystalline" shells of the latter starches. A higher density in the amorphous lamellae, envisaged by the lamellar helical model, explains the relative acid resistance of linear amylopectin chains with DP > 20, observed in lintners of B-type starches. Because amylopectin chain length distributions were similar for regular and amylose-deficient starches of the same crystal type, we deduce that the more dense (and ordered) packing of double helices into lamellar structures in amylose-deficient starches is due to a different amylopectin branching pattern.     

Production of very‐high‐amylose cassava by post‐transcriptional silencing of branching enzyme genes

High amylose starch can be produced by plants deficient in the function of branching enzymes (BEs). Here we report the production of transgenic cassava (Manihot esculenta Crantz) with starches containing up to 50% amylose due to the constitutive expression of hair‐pin dsRNAs targeting the BE1 or BE2 genes. All BE1‐RNAi plant lines (BE1i) and BE2‐RNAi plant lines (BE2i) were grown up in the field, but with reduced total biomass production. Considerably high amylose content in the storage roots of BE2i plant lines was achieved. Storage starch granules of BE1i and BE2i plants had similar morphology as wild type (WT), however, the size of BE1i starch granules were bigger than that of WT. Comparisons of amylograms and thermograms of all three sources of storage starches revealed dramatic changes to the pasting properties and a higher melting temperature for BE2i starches. Glucan chain length distribution analysis showed a slight increase in chains of DP>36 in BE1i lines and a dramatic increase in glucan chains between DP 10‐20 and DP>40 in BE2i lines. Furthermore, BE2i starches displayed a B‐type X‐ray diffraction pattern instead of the A‐type pattern found in BE1i and WT starches. Therefore, cassava BE1 and BE2 function differently in storage root starch biosynthesis.     

Development of a low glycemic maize starch: preparation and characterization

A low glycemic index starch was developed by partial alpha-amylase treatment, and its fine structure responsible for slowly digestible and resistant properties was investigated. Different digestion rates were obtained for gelatinized, retrograded starch by varying the enzyme dosage and reaction time. Analysis by high performance size-exclusion chromatography (HPSEC) coupled with multiangle laser-light scattering indicated that the molecular weighs of amylopectin and amylose were reduced during the digestion, to less than 100 kDa. A debranched chain length study using high performance anion-exchange chromatography equipped with an amyloglucosidase reactor and a pulsed amperometric detector and HPSEC revealed that short chains of amylopectin and noncrystalline amylose were rapidly digested, while DPn 121 chains showed resistance, followed by DPn 46 chains. X-ray diffraction analysis revealed that the crystalline structure in the treated starches survived cooking. These starches not only have slowly digestible and resistant character, but also retain some branched structure for adequate functionality.     

A comparative study of edible canna (Canna edulis) starch from different cultivars. Part II. Molecular structure of amylose and amylopectin

Molecular structures of starches isolated from Japanese-green, Thai-green and Thai-purple cultivars of edible canna (Canna edulis Ker) were investigated. The absolute amylose content ranged from 19 to 25%. Degrees of polymerization (DPn) values of amylose determined by fluorescence-labeling method were 1590 for Thai-purple, 1620 for Japanese-green and 1650 for Thai-green cultivars. Mole% of branched fraction of amyloses from edible canna starches examined by a HPLC system after β-amylolysis of labeled amyloses was 13–16%. Branch chain-length distributions of amylopectin analyzed by HPSEC after debranching with isoamylase, followed by fluorescence-labeling of unit chain, showed bimodal distribution with the DPn range of 25–28. The amylopectin of edible canna starches contained high amounts of organic phosphorus (391–420 ppm). The distribution profile of phosphorylated chains, separated from non-phosphorylated chains by DEAE-Sephadex A-50 chromatography, indicated that the phosphate groups were located mostly in long B-chains of amylopectin molecules.     

Granule size affects the acetyl substitution on amylopectin populations in potato and sweet potato starches

Specific enzymatic degradation in combination with chromatographic and spectrometric techniques was used to understand acetyl group distribution over the amylopectin populations of differently sized granule fractions from potato and sweet potato starches. The hydrolysates obtained after -amylase, ß-amylase, pullulanase, and the combination of pullulanase, -amylase and amyloglucosidase treatment were investigated by high-performance size-exclusion chromatography (HPSEC), high-performance anion-exchange chromatography (HPAEC) and Maldi-Tof-MS (Matrix-Assisted Laser Desorption/Ionisation Time-Of-Flight Mass Spectrometry). The acetyl groups were found to be located near the branching point, in the external chain and in the internal chain regions. The acetyl group distributions were different for amylopectin from different granule size fractions. Higher DP (degree of polymerization) fragments were present in the digests of acetylated amylopectin populations of the small size granule starches. Our studies confirmed that acetyl groups were unevenly distributed over the amylopectin populations.     

Structure–function relationships in A and B granules from wheat starches of similar amylose content

Five wheat (Triticum aestivum L.) starches, from the varieties Sunco, Sunsoft, SM1118, and SM1028, with similar amylose content, and a waxy wheat were separated into large (A) and small (B) granules. The unfractionated starches, and isolated A and B granules, were characterized structurally and evaluated for their functional properties. The amylopectin chain length distribution revealed that A granules had a lower proportion of short chains with degree of polymerization (DP) 6-12 and a higher proportion of chains with DP 25-36 than B granules. X-ray diffraction (XRD) patterns showed predominantly A-type crystallinity for all of the starches. No differences in the crystallinity were found between unfractionated, A and B granules. Small-angle X-ray scattering (SAXS) patterns of the starches at 55% hydration showed that the lamellar repeat distance in A granules was larger than that of B granules for all the starches examined. However, the lamellar distances of both A and B granules from the waxy wheat were smaller than those of Sunco, Sunsoft, SM1118 and SM1028 starches. The swelling power of the B granules was greater than that of A granules from all five starches. The kinetics of digestion of A and B granules with α-amylase in vitro were complex, with B granules initially digested to a greater extent than A granules. After 4 h of incubation, A granules showed greater digestibility than B granules, except in the case of waxy starch where unfractionated and fractionated granules had similar in vitro digestibility. Correlations between structural and functional parameters were more significant for the isolated A and B granules than for the unfractionated starches. This study demonstrates that A and B granules differ in structure and functionality, and that some correlations between these properties could be masked in unfractionated starches with bimodal granule size distribution.     

Locations of hypochlorite oxidation in corn starches varying in amylose content

The general oxidation mechanism by hypochlorite on starch has been well studied, but the information on the distribution of the oxidation sites within starch granules is limited. This study investigated the locations where the oxidation occurred within corn starch granules varying in amylose content, including waxy corn starch (WC), common corn starch (CC), and 50% and 70% high-amylose corn starch (AMC). Oxidized corn starches were surface gelatinized by 13 M LiCl at room temperature to different extents (approximately 10%, 20%, 30%, and 40%). The surface-gelatinized remaining granules were separated and studied for structural characteristics including carboxyl content, amylose content, amylopectin chain-length distribution, thermal properties, and swelling properties. Oxidation occurred mostly at the amorphous lamellae. More carboxyl groups were found at the periphery than at the core of starch granules, which was more pronounced in oxidized 70% AMC. More amylose depolymerization from oxidation occurred at the periphery of CC. For WC and CC, amylopectin long chains (>DP 36) were more prone to depolymerization by oxidation. The gelatinization properties as measured by differential scanning calorimetry also supported the changes in amylopectin fine structure from oxidation. Oxidized starches swelled to a greater extent than their unmodified counterparts at all levels of surface removal. This study demonstrates that the locations of oxidation and physicochemical properties of oxidized starches are affected by the molecular arrangement within starch granules.     

Structural and thermodynamic properties of rice starches with different genetic background Part 2. Defectiveness of different supramolecular structures in starch granules

High-sensitivity differential scanning microcalorimetry (HSDSC), small-angle X-ray scattering (SAXS), light (LM) and scanning electronic (SEM) microscopy techniques were used to study the defectiveness of different supramolecular structures in starches extracted from 11 Thai cultivars of rice differing in level of amylose and amylopectin defects in starch crystalline lamellae. Despite differences in chain-length distribution of amylopectin macromolecules and amylose level in starches, the invariance in the sizes of crystalline lamellae, amylopectin clusters and granules was established. The combined analysis of DSC, SAXS, LM and SEM data for native starches, as well as the comparison of the thermodynamic data for native and annealed starches, allowed to determine the structure of defects and the localization of amylose chains in crystalline and amorphous lamellae, defectiveness of lamellae, clusters and granules. It was shown that amylose “tie chains”, amylose–lipid complexes located in crystalline lamellae, defective ends of double helical chains dangling from crystallites inside amorphous lamellae (“dangling” chains), as well as amylopectin chains with DP 6–12 and 25–36 could be considered as defects. Their accumulation can lead to a formation of remnant granules. The changes observed in the structure of amylopectin chains and amylose content in starches are reflected in the interconnected alterations of structural organization on the lamellar, cluster and granule levels.     

Changes in physicochemical properties of waxy corn starches at different stages of harvesting

Starches were isolated from immature waxy corn kernels harvested at 0, 2, 4 and 6 days after optimum stage (DAO) and from mature kernels at 16 DAO. The starch contents showed varied according to genotypes and harvesting stages. The accumulation of starches showed an increasing trend in relation to delayed harvesting time, from the optimum stage until the physiological maturity stage. Among all harvesting stages, medium granules had the highest contribution to the total starch volume (60.8–81.5%), followed by large (5.7–30.1%), and small granules (9.1–15.3%). Average chain length distribution of amylopectin ranged from DP 14.7 to 16.9 for KKU–UB, DP 16.9 to 17.4 for KKU–JD, and DP 5.7 to 30.1 for Violet white. The pasting behaviors of starches were greatly affected by harvesting times. The peak viscosity of starches increased with delayed harvesting until physiological maturity and then decreased until dried kernels at 35 days after pollination.     

Structural and physicochemical characterisation of rye starch

The gelatinisation, pasting and retrogradation properties of three rye starches isolated using a proteinase-based procedure were investigated and compared to those of wheat starch isolated in a comparable way. On an average, the rye starch granules were larger than those of wheat starch. The former had very comparable gelatinisation temperatures and enthalpies, but slightly lower gelatinisation temperatures than wheat starch. Under standardised conditions, they retrograded to a lesser extent than wheat starch. The lower gelatinisation temperatures and tendencies of the rye starches to retrograde originated probably from their higher levels of short amylopectin (AP) chains [degree of polymerisation (DP) 6–12] and their lower levels of longer chains (DP 13–24) than observed for wheat starch. The rapid visco analysis differences in peak and end viscosities between the rye starches as well as between rye and wheat starches were at least partly attributable to differences in the levels of AP short chains and in average amylose molecular weight. The AP average chain lengths and exterior chain lengths were slightly lower for rye starches, while the interior chain lengths were slightly higher than those for wheat starch.     

The effects of amylose content on the molecular size of amylose, and on the distribution of amylopectin chain length in maize starches

The amylose to amylopectin ratios in six maize starch samples of differing amylose contents were measured by enzymatic debranching, followed by high performance size exclusion chromatography (HPSEC). The molecular size of amyloses, estimated by -log K_wav, shows progressive decrease with the increase in amylose content in maize starches. The gel permeation chromatographs of the corresponding amylopectins, debranched with isoamylase, showed bimodal distributions containing long and short chains. The average chain length of amylopectin has a correlation with amylose content. The correlation coefficients between amylose content and average chain length, long chain length, weight ratio and the mole ratio of long and short chain length, were 0.97, 0.92, 0.96, 0.94 respectively. The maize starch with the highest amylose content has the lowest amylose molecular size and the longest chains, with a high ratio of long to short chains in its amylopectin fraction. Comparing the values of amylose content determined by HPSEC of starch or debranched starch with those of the iodinecomplex method, we conclude that long chains of amylopectin in high amylose starches contribute significantly to apparent amylose content.     

Starches from different botanical sources I: Contribution of amylopectin fine structure to thermal properties and enzyme digestibility

Fifteen starches from different botanical sources were selected to study the influence of structural features on thermal properties and enzyme digestibility. Morphological appearance, X-ray diffraction pattern, apparent amylose content, unit-chain length distribution of amylopectin, thermal properties and enzyme digestibility of starch varied with botanical source. It was demonstrated that the distribution of unit-chains of amylopectin significantly correlated with functional properties of the starches. Gelatinization temperature of native and retrograded starches decreased and increased with a relative abundance of unit-chains with an approximate degree of polymerization (DP) of 8–12 and 16–26, respectively (P<0.01). Similar unit-chain lengths also affected the enzyme digestibility of starch granules (P<0.01).     

Preparation of clear noodles with mixtures of tapioca and high-amylose starches

Ways to simulate the making of clear noodles from mung bran starch were investigated by studying the molecular structures of mung bean and tapioca starches. Scanning electron micrographs showed that tapioca starch granules were smaller than those of mung bean starch. X-ray diffraction patterns of mung bean and tapioca starch were A- and C_A-patterns, respectively. Iodine affinity studies indicated that mung bean starch contained 37% of apparent amylose and tapioca starch contained 24%. Gel permeation chromatograms showed that mung bean amylopectin had longer peak chain-length of long-branch chains (DP 40) than that of tapioca starch (DP 35) but shorter peak chain-length of short-branch chains (DP 16) than that of tapioca starch (DP 21). P-31 n.m.r. spectroscopy showed that both starches contained phosphate monoesters, but only mung bean starch contained phospholipids. Physical properties, including pasting viscosity, gel strength, and thermal properties (gelatinization), were determined. The results of the molecular structure study and physical properties were used to develop acceptable products using mixtures of cross-linked tapioca and high-amylose maize starches. Tapioca starch was cross-linked by sodium trimetaphosphate (STMP) with various reaction times, pH values, and temperatures. The correlation between those parameters and the pasting viscosity were studied using a visco/amylograph. Starches, cross-linked with 0.1% STMP, pH 11.0, 3.5 h reaction time at 25, 35, and 45°C (reaction temperature), were used for making noodles. High-amylose maize starch (70% amylose) was mixed at varying ratios (9, 13, 17, 28, 37, and 44%) with the cross-linked tapioca starches. Analysis of the noodles included: tensile strength, water absorption, and soluble loss. Noodle sensory properties were evaluated using trained panelists. Noodles made from a mixture of cross-linked tapioca starch and 17% of a high-amylose starch were comparable to the clear noodles made from mung bean starch.     

Structural basis for the slow digestion property of native cereal starches

Native cereal starches are ideal slowly digestible starches (SDS), and the structural basis for their slow digestion property was investigated. The shape, size, surface pores and channels, and degree of crystallinity of starch granules were not related to the proportion of SDS, while semicrystalline structure was critical to the slow digestion property as evidenced by loss of SDS after cooking. The high proportion of SDS in cereal starches, as compared to potato starch, was related to their A-type crystalline structure with a lower degree of perfection as indicated by a higher amount of shortest A chains with a degree of polymerization (DP) of 5-10. The A-type amorphous lamellae, an important component of crystalline regions of native cereal starches, also affect the amount of SDS as shown by a reduction of SDS in lintnerized maize starches. These observations demonstrate that the supramolecular A-type crystalline structure, including the distribution and perfection of crystalline regions (both crystalline and amorphous lamellae), determines the slow digestion property of native cereal starches.     

Chemical and physical properties of ginkgo (Ginkgo biloba) starch

Starch isolated from mature Ginkgo biloba seeds and commercial normal maize starches were subjected to α-amylolysis and acid hydrolysis. Ginkgo starch was more resistant to pancreatic α-amylase hydrolysis than the normal maize starch. The chain length distribution of debranched amylopectin of the starches was analyzed by using high performance anion-exchange chromatography equipped with an amyloglucosidase reactor and a pulsed amperometric detector. The chain length distribution of ginkgo amylopectin showed higher amounts of both short and long chains compared to maize starch. Naegeli dextrins of the starches prepared by extensive acid hydrolysis over 12 days demonstrated that ginkgo starch was more susceptible than normal maize to acid hydrolysis. Ginkgo dextrins also demonstrate a lower concentration of singly branched chains than maize dextrins, and unlike maize dextrin, debranched ginkgo shows no multiple branched chains. The ginkgo starch displayed a C-type X-ray diffraction pattern, compared to an A-type pattern for maize. Ginkgo starch and maize starch contained 24.0 and 17.6% absolute amylose contents, respectively.     

New starches: Physicochemical properties of sweetsop (Annona squamosa) and soursop (Anonna muricata) starches

Starch from the fruits of sweetsop (Anonna squamosa) and soursop (Anonna muricata) were isolated and purified and the fat, ash, phosphorus and protein contents measured. The amount of amylose present was determined spectrophotometrically and found to be very similar (19%) for both starches. Scanning electron microscopy showed very small indented and spherical granules from both with an average granule size of 4.84 μm and 4.72 μm, respectively. The physicochemical properties, namely the swelling power, solubility, pasting characteristics, paste clarity and freeze–thaw stability were studied to assess the functionality of the starch pastes as hydrocolloids. The sweetsop starch showed higher swelling power and solubility compared to soursop starch and had a lower gelatinization temperature indicating a weaker granular structure. Sweetsop starch exhibited a lower pasting temperature, higher viscosity peak, higher viscosity breakdown and lower setback, higher paste clarity and freeze–thaw stability compared to soursop starch. The low gelatinization temperature and high freeze thaw stability of sweetsop starch are comparable to that of waxy corn. The properties of sweetsop indicate that it has potential for application as a thickener in frozen foods.     

Structural characteristics and physicochemical properties of oxidized corn starches varying in amylose content

The effects of amylose content on the extent of oxidation and the distribution of carboxyl groups in hypochlorite-oxidized corn starches were investigated. Corn starches including waxy corn starch (WC), common corn starch (CC), and 50% and 70% high-amylose corn starches (AMC) were oxidized with NaOCl at three concentrations (0.8%, 2%, and 5%). Carboxyl and carbonyl content of oxidized starches increased with increasing NaOCl concentration. High-AMC (70%) had slightly higher carboxyl and carbonyl contents at 0.8% NaOCl, whereas WC had significantly higher carboxyl and carbonyl contents at 2% and 5% NaOCl levels. Carbohydrate profiles by high-performance size-exclusion chromatography indicate that amylose was more susceptible to depolymerization than amylopectin. Degradation of amylopectin long chains (DP >24) was more pronounced in WC and CC than in AMCs. The crystalline lamellae of WC started to degrade at 2% NaOCl, but those of the other corn starches remained intact even at 5% NaOCl level according to X-ray crystallinity. By using anion-exchange chromatography for separation and size-exclusion chromatography for characterization, carboxyl groups were found to be more concentrated on amylopectin than on amylose, particularly in AMCs. Oxidation decreased gelatinization temperature and enthalpy with WC showing the most decrease and 70% AMC showing the least. The gelatinization enthalpy of 50% AMC decreased significantly faster than those of CC and 70% AMC after 0.8% oxidation. Retrogradation of amylopectin slightly increased after oxidation with increasing oxidation level. The peak viscosities of oxidized WC and CC were higher than those of their native counterparts at 0.8% NaOCl, but this increase was not observed in AMCs. The setback viscosities of 2% NaOCl-oxidized 50% and 70% AMCs were much higher than those of the unmodified counterparts. The extent of oxidation and physicochemical properties of oxidized starches varied greatly with the amylase:amylopectin ratio of corn starches. Amylose was suggested to play an important role in controlling the oxidation efficiency.     

Origin of defects in assembled supramolecular structures of sweet potato starches with different amylopectin chain-length distribution

A combined approach of fluorophore-assisted capillary electrophoresis (FACEL), high-sensitivity differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS), small-angle X-ray scattering (SAXS), and light (LM) and scanning electron microscopy (SEM) was applied to study the effects of changes in amylopectin chain-length distribution on the assembly structures of sweet potato starches with similar amylose levels. It was shown that unlike ordinary sweet potato starch, starch extracted from Quick Sweet cultivar of sweet potato had anomalous high level of amylopectin chains with a degree of polymerization (DP) 6–12. Joint analysis of the obtained data revealed that amylopectin chains with DP 10–24 are, apparently, the dominant material for the formation of supramolecular structures in starch granules. In contrast, amylopectin chains with DP < 10 facilitated the formation of defects within crystalline lamellae. An increase in relative content of amylopectin chains with DP < 10 is accompanied by the correlated structural alterations manifested at all levels of starch granule organization (crystalline lamellae, amylopectin clusters, semi-crystalline growth rings, and granule morphology). Thus, the short amylopectin chains with DP < 10 were considered as an origin of the defectiveness in starch supramolecular structures.     

Physical, chemical and microscopic characterization of a new starch from chayote (Sechium edule) tuber and its comparison with potato and maize starches

In this work, the chayote tuber starch (CHS) was isolated and its chemical composition and its physical and microscopic characteristics were determined, and compared with potato (PS) and maize (MS) starches. The starch content in chayote tubers (728 g kg⁻¹ dry weight) was similar to potato tubers (700 g kg⁻¹ dry weight), with a high level of purity (>98%), while its phosphorous content was higher (0.15%) than PS (0.08%) and MS (0.01%). Starch granules were oval, irregular, truncated and rounded with sizes between 7 and 50 μm with smooth surfaces. CHS dispersions (1% and 4%, w/w) showed higher viscosity (75 and 1715 mPa s), than PS (350% and 50% lower) and MS (715% and 600% lower). The gelatinization temperature (65 to 74 °C) was similar in CHS and PS. The pasting properties (RVA) of the starches suggest that CHS showed better characteristics than the commercial potato and maize starches. Therefore, CHS could be used as a thickening agent and a substitute to PS in food dispersions where a high viscosity is needed.