Structural properties of hydrolyzed high-amylose rice starch by α-amylase from Bacillus licheniformis

High-amylose cereal starch has a great benefit on human health through its resistant starch (RS) content. Enzyme hydrolysis of native starch is very helpful in understanding the structure of starch granules and utilizing them. In this paper, native starch granules were isolated from a transgenic rice line (TRS) enriched with amylose and RS and hydrolyzed by α-amylase. Structural properties of hydrolyzed TRS starches were studied by X-ray powder diffraction, Fourier transform infrared, and differential scanning calorimetry. The A-type polymorph of TRS C-type starch was hydrolyzed faster than the B-type polymorph, but the crystallinity did not significantly change during enzyme hydrolysis. The degree of order in the external region of starch granule increased with increasing enzyme hydrolysis time. The amylose content decreased at first and then went back up during enzyme hydrolysis. The hydrolyzed starches exhibited increased onset and peak gelatinization temperatures and decreased gelatinization enthalpy on hydrolysis. These results suggested that the B-type polymorph and high amylose that formed the double helices and amylose-lipid complex increased the resistance to BAA hydrolysis. Furthermore, the spectrum results of RS from TRS native starch digested by pancreatic α-amylase and amyloglucosidase also supported the above conclusion.     

Granule structure and distribution of allomorphs in C-type high-amylose rice starch granule modified by antisense RNA inhibition of starch branching enzyme

C-type starch, which is a combination of both A-type and B-type crystal starch, is usually found in legumes and rhizomes. We have developed a high-amylose transgenic line of rice (TRS) by antisense RNA inhibition of starch branching enzymes. The starch in the endosperm of this TRS was identified as typical C-type crystalline starch, but its fine granular structure and allomorph distribution remained unclear. In this study, we conducted morphological and spectroscopic studies on this TRS starch during acid hydrolysis to determine the distribution of A- and B-type allomorphs. The morphology of starch granules after various durations of acid hydrolysis was compared by optical microscopy, scanning electron microscopy, and transmission electron microscopy. The results showed that amorphous regions were located at the center part of TRS starch subgranules. During acid hydrolysis, starch was degraded from the interior of the subgranule to the outer surface, while the peripheral part of the subgranules and the surrounding band of the starch granule were highly resistant to acid hydrolysis. The spectroscopic changes detected by X-ray powder diffraction, 13C cross-polarization magic-angle spinning NMR, and attenuated total reflectance Fourier transform infrared showed that the A-type allomorph was hydrolyzed more rapidly than the B-type, and that the X-ray diffraction profile gradually changed from a native C-type to a CB-type with increasing hydrolysis time. Our results showed that, in TRS starch, the A-type allomorph was located around the amorphous region, and was surrounded by the B-type allomorph located in the peripheral region of the subgranules and the surrounding band of the starch granule. Thus, the positions of A- and B-type allomorphs in the TRS C-type starch granule differ markedly from those in C-type legume and rhizome starch.     

Effects of Milling and Cooking Conditions of Rice on In Vitro Starch Digestibility and Blood Glucose Response

The objective of this study was to investigate the effects of milling and cooking conditions of cooked rice prepared from cultivar Koshihikari on in vitro starch digestibility and in vivo glucose response in humans. In addition, compression and adhesiveness tests were conducted for texture analysis of the cooked rice. Brown rice (BR) and surface‐abraded BR (SABR, ≥99.5% of the original weight) were digested more slowly than white rice (91% of the original weight) when cooked rice grain was used for the in vitro test, but they were digested more rapidly in the initial stage of the reaction when cooked rice ground by a meat grinder was used. The increase in water added for cooking significantly increased the extent of starch digestion with BR and SABR. The changes in blood glucose levels after the ingestion of cooked rice were dependent on the sample type. The cooking conditions dramatically influenced the glucose response after the ingestion of BR. A significant correlation was found between blood glucose levels at 45 min and the extent of starch digestion with ground samples, whereas no relationship was found with cooked rice grain samples for in vitro digestibility.     

蒸煮工艺对米饭中淀粉消化性能的影响

为研究蒸煮工艺对米饭中淀粉消化特性的影响,为米饭的营养评价和新型电饭煲的开发提供试验数据,该文以3种大米为原料,采用不同加热工艺的机械煲、电脑煲、高压锅和微压力锅蒸煮米饭,用胰酶和糖化酶进行米饭的体外消化试验,研究酶解过程还原糖和葡萄糖含量,用指数模型模拟淀粉的酶解过程。结果表明:蒸煮工艺对米饭酶解后还原糖和葡萄糖的含量有较大影响。不同种类米饭的消化特性有差异,粳稻米饭较籼稻易于消化。采用具有较高的压力和较长焖饭时间的高压锅或微电脑煲制作的米饭的淀粉消化速度较快,酶解生成还原糖和葡萄糖最高,且消化中前期的快消化淀粉(RDS)和慢消化淀粉(SDS)含量较高。建立的基于葡萄糖含量的消化动力学指数方程具有较高的拟和精度。     

Effects of chemical modification on in vitro rate and extent of food starch digestion: an attempt to discover a slowly digested starch.

Differences in glycemic and insulinemic responses to dietary starch are directly related to the rate of starch digestion. Chemical modification of starch may allow for the production of a slowly digested starch that could be used for the treatment of certain medical modalities. An in vitro method was utilized to evaluate the effects of chemical modification on the rate and extent of raw and cooked starch digestion. The extent of starch digestion was significantly reduced by dextrinization, etherification, and oxidation. However, the rate of starch digestion was not significantly affected by chemical modification. For most modified starches, as the degree of modification increased, the extent of digestion decreased, suggesting an increase in the amount of resistant starch. The results of this study suggest that chemically modified starch has a metabolizable energy value of <16.7 kJ/g. Chemically modified starch ingredients may serve as a good source of resistant starch in human and animal diets.     

Plantain and banana starches: granule structural characteristics explain the differences in their starch degradation patterns

Different banana cultivars were used to investigate the influences of starch granule structure and hydrolases on degradation. The highest degrees of starch degradation were observed in dessert bananas during ripening. Scanning electron microscopy images revealed smooth granule surface in the green stage in all cultivars, except for Mysore. The small and round granules were preferentially degraded in all of the cultivars. Terra demonstrated a higher degree of crystallinity and a short amylopectin chain length distribution, resulting in high starch content in the ripe stage. Amylose content and the crystallinity index were more strongly correlated than the distribution of amylopectin branch chain lengths in banana starches. α- and β-amylase activities were found in both forms, soluble in the pulp and associated with the starch granule. Starch-phosphorylase was not found in Mysore. On the basis of the profile of α-amylase in vitro digestion and the structural characteristics, it could be concluded that the starch of plantains has an arrangement of granules more resistant to enzymes than the starch of dessert bananas.     

Resistance to α‐Amylase Digestion in Four Native High‐Amylose Maize Starches

Native and processed high‐amylose maize starch (HAMS) is an important source of resistant starch (RS). The objectives of this work were to use an in vitro procedure to estimate the RS content of native granules from a series of ae‐containing HAMS genotypes, and to examine the nature of the α‐amylase resistant starch (ARS). By the method of Englyst et al (1992), RS for ae V, ae VII, ae su2, and ae du were estimated to be 66.0, 69.5, 69.5, and 40.6%, respectively. By transmission electron microscopy, most of the residual granules from ae V, ae VII, and ae su2 showed little evidence of digestion. Partially digested granules had a radial digestion pattern in the interior and an enzyme‐resistant layer near the surface. Size and chain‐length profile of constituents of ARS were similar to those of the native HAMS (unlike type 3 RS), consistent with complete hydrolysis in susceptible granule regions. Between crossed polarizers, many iodine‐stained native and residual HAMS granules had blue centers and pink exteriors, which may be due to a difference in orientation of the amylose‐iodine complexes in the exterior. Four granule color types were observed for ae du, differing in enzyme resistance. The high‐enzyme resistance of native HAMS granules may result from altered granule organization, which appears to vary among and within granules from ae‐containing genotypes.     

Separation and Characterization of A- and B-Type Starch Granules in Wheat Endosperm

Mature wheat (Triticum aestivum L.) endosperm contains two types of starch granules: large A-type and small B-type. Two methods, microsieving or centrifugal sedimentation through aqueous solutions of sucrose, maltose, or Percoll were used to separate A- and B-type starch granules. Microsieving could not completely separate the two types of starch granules, while centrifuging through maltose and sucrose solutions gave a homogenous population for B-type starch granules only. Centrifuging through two Percoll solutions (70 and 100%, v/v) produced purified populations of both the A- and B-type starch granules. Analysis of starch granule size distribution in the purified A- and B-type granule populations and in the whole-starch granule population obtained directly from wheat endosperm confirmed that the purified A- and B-type starch granule populations represented their counterparts in mature wheat endosperm. Centrifugations through two Percoll solutions were used to purify A- and B-type starch granule populations from six wheat cultivars. The amylose concentrations and gelatinization properties of these populations were analyzed. All of the A-type starch granules contained higher amylose concentrations and had higher gelatinization enthalpies than did B-type starch granules. Although A- and B-type starch granules started to gelatinize at a similar temperature, B-type starch granules had higher gelatinization peak and completion temperatures than did A-type starch granules     

Fibrillar microfilaments associated with a high-amylose rice, goami 2, a mutant of ilpumbyeo, a high-quality japonica rice

The ultrastructure of cooked and malt-treated cooked rice of Ilpumbyeo (IP) and its mutant Goami 2 (G2), which have extreme contrasts in physicochemical properties, cooking quality, and ultrastructural characteristics in raw grains (1, 2), was compared. In cooked rice of IP, starch granules in endosperm cells were evenly coalesced, appearing as homogeneously smooth sheetlike matrix and/or globules, whereas those in G2 were a heterogeneously coarse matrix in which a novel structural feature, the microfilaments, was embedded. In malt-treated cooked rice of IP, most starch was hydrolyzed by the malt enzymes, appearing as empty vacuoles surrounded by the cell wall, whereas that in G2 was highly resistant to malt treatment, remaining as distinct structural features, the malt-resistant compound starch granules. The property of G2's compound starch granules, which are tolerant of mechanical and chemical treatments thereby retaining their structural integrity (2) and of cooking and malt treatment thereby retaining their physical hardness, appears to play a major role in determining the quality of cooked rice of G2.     

In Vitro Starch Digestibility and Expected Glycemic Index of Kodo Millet (Paspalum scrobiculatum) as Affected by Starch–Protein–Lipid Interactions

The effect of starch–protein–lipid interaction on the in vitro starch digestibility and expected glycemic index (eGI) of kodo millet flour (MF) was investigated. Debranned MF and the flour with lipid removed, protein removed, or both lipid and protein removed (MF‐L‐P) were subjected to digestion assays. The in vitro starch digestibility and eGI of the millet samples and millet starch were compared with rice or wheat flour. Rapidly digestible starch, slowly digestible starch, and resistant starch (RS) of the samples were also calculated. Protease treatment and defatting resulted in significant reduction (P < 0.05) in protein and lipid contents of samples. Significant increases in the in vitro starch digestibility and eGI of samples were observed after removal of protein, lipid, or both. The effect of lipid removal on in vitro starch digestibility of kodo millet was found to be more significant, compared with when proteins were removed. The eGI increased from 49.4 for cooked MF to 62.5 for MF‐L‐P. The eGI of cooked kodo millet starch was significantly lower than that of cooked rice flour. The RS (1.61%) of cooked rice was the least among the samples. The in vitro starch digestibility and eGI of rice were significantly higher than those of MF. Processes applied to kodo millet, such as decortication, that result in the removal of proteins, lipids, or both (especially lipids) would result in an increase in its in vitro starch digestibility and eGI. We therefore advocate for the development of acceptable products from whole millets to maintain its hypoglycemic property.     

Physiological and metabolic properties of a digestion-resistant maltodextrin, classified as type 3 retrograded resistant starch

There is a growing interest in highly fermentable dietary fibers having the potential to reduce risks of disease through the production of short-chain fatty acids (SCFA). Recently a digestion-resistant retrograded maltodextrin (RRM), classified as type 3 resistant starch was developed. Systematic work to determine its molecular and physiological properties was carried out to determine (1) the fraction resistant to digestion in vitro and in vivo, (2) its postconsumption effect on blood glucose in healthy volunteers, and (3) its in vitro fermentation pattern, at different ages, by use of pooled fresh human fecal inoculum. RESULTS: The digestion resistant fraction obtained in vivo from ileostomy patients (59.4%) is similar to that obtained by the AOAC method for measuring retrograded resistant starch (59.7%). The relative glycemic response after consumption of 50 g of RRM was 58.5% compared to glucose set as 100%. When exposed to colonic microbiota, in vitro obtained indigestible fractions behave similarly to those obtained in vivo in ileostomy patients. Fermentation of RRM and production of butyric acid is negligible during the first months of life but develops subsequently during weaning. In adults, RRM fermentation results in a high yield of SCFA, with butyrate representing 21-31 mol % of total SCFA. The high yield of SCFA during colonic fermentation, observed from weaning age on, as well as the potential to help reduce glycemic load may be of benefit to a number of health-related functions in the host. Further study on clear clinical end points is warranted.     

In Vitro Digestion of RS4‐Type Resistant Wheat and Potato Starches,and Fermentation of Indigestible Fractions

RS4‐type resistant wheat starch (RWS) and resistant potato starch (RPS) were subjected successively to in vitro digestion with pepsin and pancreatin‐bile, and the indigestible residues (82.1% db and 74.1% db, respectively) were recovered and subsequently fermented by in vitro techniques using fresh human fecal microbiota as inoculum. Scanning electron microscopy of the indigestible residues showed surface erosion on the residual granules. Total gas production during the in vitro fermentation increased almost linearly over time with the two resistant starches exhibiting similar gas production rates, as well as a similar rate of production of total short‐chain fatty acids (SCFA). The indigestible fractions from both starches produced acetate as the major SCFA and relatively higher levels of butyrate than propionate, but wheat starch tended to produce more butyrate over time than potato starch. Fractional molar ratios of acetate, propionate, and butyrate from the RWS and RPS were 0.586:0.186:0.228 and 0.577:0.200:0.223, respectively. The calculated caloric contributions of the RWS and RPS are ≈33% lower than for unmodified starch and are comparable to those reported in the literature for RS2 and RS3 high‐amylose maize starches.     

Starch properties of mutant rice high in resistant starch

As the staple food of over half the world's population, hot cooked rice high in resistant starch (RS) is of particular interest, which will have greater impact in the dietary prevention of diabetes and hyperlipidemia. A mutant rice high in RS in hot cooked rice, described as RS111, was comparatively studied with the wild type and common rice. Despite obviously low RS content in the raw milled rice, the RS content in the hot cooked rice of mutant RS111 was significantly higher than that of the wild type and common rice and, correspondingly, in vitro starch hydrolysis by porcine pancreatic alpha-amylase tends to be incomplete with low hydrolysis extent for the cooked mutant rice high in RS. Obvious differences in physicochemical properties, starch granule morphology, pasting properties, thermal properties, and X-ray diffraction pattern were observed among the mutant RS111, wild type, and common indica rice. The high-RS mutant was characterized by significantly higher apparent amylose content and crude lipid content, higher percentage of oval-shaped granules and bigger oval size, reduced paste viscosity, and low onset temperature, peak temperature, final temperature, enthalpy of gelatinization, and crystallinity.     

Effect of storage time on the retrogradation of banana starch extrudate

Starch was isolated from banana starch and the retrogradation phenomenon was studied using diverse techniques, including an enzymatic measurement. Wide-angle X-ray scattering (WAXS) showed that the sample stored for 7 h presented small peaks and when the storage time increased the peaks increased in intensity. The type of diffraction pattern found in banana extrudates is typical of the A-type crystal polymorph. The crystallinity index from the diffractograms, showed a plateau after approximately 20 h of storage. The short-range order measurement with Fourier transform infrared (FTIR) spectroscopy showed that banana starch retrogradation reached a maximum value at approximately 11 h of storage, a value that agrees with the results obtained with differential scanning calorimetry (DSC), because the maximum enthalpy value (approximately 5 J/g) was calculated in the stored sample for 8 h, without changes in the stored samples for more time. Retrograded resistant starch values did not change after 12 h of storage, obtaining the maximum starch retrogradation level. FTIR, DSC, and the enzymatic technique showed the changes at the molecular level in starch during storage; in the case of WAXS, they determine the long-range order that explains the differences found in the starch retrogradation pattern measurement in banana starch.     

Effects of resistant starch type III polymorphs on human colon microbiota and short chain fatty acids in human gut models

This study probed the possible effects of type III resistant starch (RS) crystalline polymorphism on RS fermentability by human gut microbiota and the short chain fatty acids production in vitro. Human fecal pH-controlled batch cultures showed RS induces an ecological shift in the colonic microbiota with polymorph B inducing Bifidobacterium spp. and polymorph A inducing Atopobium spp. Interestingly, polymorph B also induced higher butyrate production to levels of 0.79 mM. In addition, human gut simulation demonstrated that polymorph B promotes the growth of bifidobacteria in the proximal part of the colon and double their relative proportion in the microbiota in the distal colon. These findings suggest that RS polymorph B may promote large bowel health. While the findings are limited by study constraints, they do raise the possibility of using different thermal processing to delineate differences in the prebiotic capabilities of RS, especially its butryrogenicity in the human colon.     

Importance of Location of Digestion and Colonic Fermentation of Starch Related to Its Quality

For decades, quality of starch‐based foods has been associated with the in vivo measured glycemic index or the in vitro digestion rate‐based categories of rapidly digestible, slowly digestible, and resistant starch (RS). Glycemic index has been related to health‐based endpoints mostly through correlative or observational studies, with mechanisms proposed but not well established. Here, we bring forth the concept of locational delivery of glucose from dietary starches to the distal small intestine to elicit an ileal brake effect, as well as short‐chain fatty acid production from RS fermentation to cause a colonic brake. Both effects slow gastric emptying and, in turn, extend nutrient (i.e., energy) delivery to the body and may decrease appetite and promote weight management. Slowly digestible starches are currently a popular topic of research, although where they are digested and the released glucose is delivered in the small intestine is not known. A proposal is to further study and establish this mechanism of appetite and food intake regulation so that starch‐based ingredients and foods can be developed that promote both the ileal and colonic brake mechanisms.     

Starch characteristics and their influences on in vitro and pig prececal starch digestion

The main objective of this research was to study the characteristics of starch granules and their influences on in vitro and pig prececal starch digestion of corn, dehulled barley, wheat, and potato. Scanning electron microscopy was used to study the starch endosperm structure in the parent material as well as in vitro starch digestion. The results showed that corn starch granules were polyhedral, with a diameter ranging from 2 to 10 μm, whereas those of dehulled barley and wheat were spherical, with a diameter ranging from 5 to 20 μm. Potato had the largest starch granules among starch sources reported herein, with oval spheres of 10-50 μm in diameter. In vitro starch hydrolysis showed that starch granules of corn degraded faster than the starch of dehulled barley and wheat, with the potato starch being degraded the slowest. The in vivo digestibility trial using ileal-cannulated pigs confirmed the starch degradation of grains. The in vitro (x, %) and in vivo (y, %) digestibility were highly correlated [y = 6.5304x - 538.48 (R(2) = 0.9924)]. On the basis of the results, in vitro starch hydrolysis might be useful in predicting in vivo prececal starch digestibility. The digestion kinetic characteristics of different starch sources might be employed to evaluate the starch digestive rate at the pig ileum.     

α-Amylolysis of Large Barley Starch Granules

The α-amylolysis of large (volume average 16 μm) barley starch granules was studied by measuring the amount of carbohydrates solubilizing during hydrolysis, and the changes in morphology and molecular structure of the granule residues by scanning electron microscopy, particlesize analysis, size-exclusion chromatography, X-ray diffraction, and differential scanning calorimetry. X-ray diffraction showed that, in the earlier stages of α-amylolysis, both amorphous and crystalline parts of the granules were equally solubilized. More extensive hydrolysis caused a gradual decrease in A-type crystallinity and degradation of the granular structure. Scanning electron microscopy revealed that hydrolysis proceeded through pinholes, and pitted and partially hollow granule residues were formed. The lipid-complexed amylose was less susceptible to α-amylolysis than free amylose and amylopectin. Lipid-complexed amylose started leaching out of the granule residues only after half of the starch had solubilized due to the α-amylase treatment. Even though scanning electron microscopy indicated that there were intact granules left throughout the hydrolysis, the results obtained suggested that α-amylolysis of large barley starch granules proceeded rather evenly among the granules.     

Morphological Changes of Granules of Different Starches by Surface Gelatinization with Calcium Chloride

Native starch granules of 11 selected cultivars (potato, waxy potato, sweet potato, normal maize, high‐amylose maize, waxy maize, wheat, normal barley, high‐amylose barley, waxy barley, and rice) were treated with a calcium chloride solution (4M) for surface gelatinization. The surface‐gelatinized starch granules were investigated using light microscopy and scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). In general, those starches with larger granule sizes required longer treatment time to complete the gelatinization. The salt solution treatment of starch was monitored by light microscopy and stopped when the outer layer of the granule was gelatinized. The surface gelatinized starch granules were studied using scanning electron microscopy. On the basis of the gelatinization pattern from calcium chloride treatments, the starches could be divided into three groups: 1) starches with evenly gelatinized granule surface, such as normal potato, waxy potato, sweet potato, maize, and high‐amylose maize; 2) starches with salt gelatinization concentrated on specific sites of the granule (i.e., equatorial groove), such as wheat, barley, and high‐amylose barley; and 3) starches that, after surface gelatinization, can no longer be separated to individual granules for SEM studies, such as waxy barley, waxy maize, and normal rice. The morphology of the surface gelatinized starch resembled that of enzyme‐hydrolyzed starch granules.     

In vitro digestion characteristics of unprocessed and processed whole grains and their components

Chemical composition and in vitro digestion properties of select whole grains, before and after processing, and their components were measured. Substrates included barley, corn, oat, rice, and wheat. In addition to whole grain flours, processed substrates also were tested as were corn bran, oat bran, wheat bran, and wheat germ. Processing of most substrates resulted in higher dry matter and digestible starch and lower resistant starch concentrations. Dietary fiber fractions varied among substrates with processing. Digestion profiles for most substrates correlated well with their chemical composition. Corn bran and rice substrates were the least fermentable. Extrusion rendered barley, corn, and wheat more hydrolytically digestible and barley and oat more fermentatively digestible. Except for corn bran, all components had greater or equal fermentability compared with their native whole grains. Understanding digestion characteristics of whole grains and their components will allow for more accurate utilization of these ingredients in food systems.