Starch Research Articles

Effect of the Presence of Physiological Disorder Recognized as “yellow berry” on the Quality of Starch in Bread Wheat (Triticum aestivum) and Durum Wheat (Triticum durum)

AbstractGenetic and environmental factors influence wheat grain quality, including vitreousness, protein, and starch content. Yellow berry (YB), a physiological condition, adversely affects wheat quality, particularly in irrigated fields with limited nitrogen application, leading to starch‐rich, low‐protein grains. This study examines common wheat, used in baking industries, and durum wheat, primarily employs for pasta production. The focus is on investigating how the physicochemical properties of these wheat types change with the presence of YB, that impacts protein content negatively, increasing total starch content. Fourier‐transform infrared spectroscopy reveals structural changes in YB‐affected grains, while X‐ray diffraction indicates varying crystallinity. In determination of amylose, an increase of 6% in bread wheat with YB and 3% in durum wheat with YB is observed. In the case of RVA (Rapid Visco Analyzer) analysis, a notable increase in viscosity is evident in the treatments that included the yellow berry. This finding suggests that YB is associated with substantial modifications in starch properties, such as amylose‐to‐amylopectin ratio, chain length, and degree of branching, among several other components, which may have important implications for the texture and quality of food products. This study calls for further research to mitigate YB's impact and enhances the quality of wheat‐derived foods.

Mass Transfer Resistance and Reaction Rate Kinetics for Carbohydrate Digestion with Cell Wall Degradation by Cellulase.

This paper introduces an enzymatic approach to estimate internal mass-transfer resistances during food digestion studies. Cellulase has been used to degrade starch cell walls (where cellulose is a significant component) and reduce the internal mass-transfer resistance, so that the starch granules are released and hydrolysed by amylase, increasing the starch hydrolysis rates, as a technique for measuring the internal mass-transfer resistance of cell walls. The estimated internal mass-transfer resistances for granular starch hydrolysis in a beaker and stirrer system for simulating the food digestion range from 2.2 × 107 m-1 s at a stirrer speed of 100 rpm to 6.6 × 107 m-1 s at 200 rpm. The reaction rate constants for cellulase-treated starch are about three to eight times as great as those for starch powder. The beaker and stirrer system provides an in vitro model to quantitatively understand external mass-transfer resistance and compare mass-transfer and reaction rate kinetics in starch hydrolysis during food digestion. Particle size analysis indicates that starch cell wall degradation reduces starch granule adhesion (compared with soaked starch samples), though the primary particle sizes are similar, and increases the interfacial surface area, reducing internal mass-transfer resistance and overall mass-transfer resistance. Dimensional analysis (such as the Damköhler numbers, Da, 0.3-0.5) from this in vitro system shows that mass-transfer rates are greater than reaction rates. At the same time, SEM (scanning electron microscopy) images of starch particles indicate significant morphology changes due to the cell wall degradation.

Effect of copper and temperature on the photosynthetic physiological characteristics of Ulva linza under elevated CO2 concentrations

Copper (Cu) is vital for macroalgae's functions, but high concentrations can be toxic. Rising CO2 levels affect algal growth and Cu bioavailability. In this study, the results reveal that at 5 °C, low Cu increased Ulva linza growth, while high Cu and elevated CO2 decreased growth. At 10 °C, low Cu and elevated CO2 enhanced growth, but high Cu did not have a significant impact. At 15 °C, high Cu reduced growth, but elevated CO2 offset this effect. Furthermore, under elevated CO2 conditions, the chloroplast structure of the algae appeared to be denser, accompanied by a large amount of starch granules, compared to low CO2 conditions. These results emphasize that lower temperatures, in conjunction with elevated CO2 concentration, could intensify the toxic effects of high Cu concentrations on thalli. However, at higher temperatures, elevated CO2 concentration appeared to be capable of mitigating the detrimental effects of heavy metals on algae.

The maltose-related starch degradation pathway promotes the formation of large and spherical transitory starch granules.

Previously, in Arabidopsis thaliana, we found atypical spherical starch granules in dpe2ss4 and dpe2phs1ss4. However, the mechanism of such abnormal morphogenesis is still obscure. By tracking starch granule length and thickness with leaf ageing, we reported that the starch granules in dpe2phs1ss4 gradually change to a spherical shape over time. In comparison, Col-0 and the parental line ss4 did not exhibit macroscopic morphological alteration. In this study, firstly, we specify that the additional lack of DPE2 resulted in the gradual alteration of starch granule morphology over time. Similar gradual morphological alterations were also found in dpe2, mex1, and sex4 but not in the other starch degradation-related mutants, such as sex1-8, pwd, and bam3. The gradual alteration of starch morphology can be eliminated by omitting the dark phase, suggesting that the particular impaired starch degradation in dpe2- and mex1-related mutants influences starch morphology. Secondly, we observed that spherical starch morphology generation was accompanied by prominent elevated short glucan chains of amylopectin and an increased amylose proportion. Thirdly, the interplay between soluble starch synthase 2 and branching enzymes was affected and resulted in the formation of spherical starch granules. The resulting spherical starch granules allow for elevated starch synthesis efficiency. Fourthly, the starch phosphate content at the granule surface correlated with the morphology alteration of the starch granules. Herewith, we propose a model that spherical starch granules, accumulated in mutants with a misbalance of the starch degradation pathway, are result of elevated starch synthesis to cope with overloaded carbohydrates.

Relationship between internal structure and in vitro digestibility of A- and B-type wheat starch revealed by chemical surface gelatinization

In this study, the properties of remaining starch granules obtained with different degrees of exfoliation were explored by removing the outer layers of A- and B-type wheat starch (AWS and BWS) granules with chemical surface gelatinization. SEM images revealed significant morphological variations with increasing exfoliation. CLSM and amylose content analysis indicated a predominance of lipid complexes in the outer granule layers, particularly in BWS. The structural characteristics of AWS and BWS were analyzed using PLM, XRD, FT-IR and DSC, verifying the conclusion of the alternation of starch crystalline and amorphous zone. And the amorphous regions are proportionally higher in the inner starch layer. Moreover, raw AWS and BWS granules were more easily digested from the outside in, with the RS content decreasing from 80.65 % to 66.92 % and 49.06 % to 45.01 %, respectively. The RS content of cooked WS were affected by the internally structures, particularly lipid content (11.46 % – 19.09 %) in BWS outer layers and amylose content (13.59 % – 19.43 %) in the inner layers. These results revealed the internal radial structural differences and digestibility patterns of AWS and BWS granules.

Exploring oligosaccharide effects on wheat starch gelatinization: Mechanisms and oligosaccharide structural perspectives

Controlling the gelatinization of starch is vital for achieving desired food design, and sugars and oligosaccharides (OSs) have a significant impact on the gelatinization temperature (Tgel) of starch. This study delved into the effects of sucrose, maltotetraitol, and ten glucose-based OSs with different glycosidic linkage patterns, degrees of polymerization (DP), and linear or cyclic structures on the Tgel of wheat starch. The objective of this study was to uncover the underlying mechanisms by which OSs alter Tgel to support the design and use of OSs for sugar reduction in baked goods. Findings underscore the notable impact of both OS concentration and type on Tgel. Increases in OS concentration resulted in Tgel elevation, closely linked to changes in water activity and effective water volume fraction (φw,eff). Among the same DP OSs, a higher effective number of hydroxyl groups available for hydrogen bonding per unit volume of the solute (NOH,S/Vs) corresponded to increased Tgel, indicating higher hydroxyl density in OSs leads to an increased number of hydrogen bond formation between OSs and starch that stabilizes the structure. Of particular note is that isomaltose (α-1,6 linkage) significantly elevated the Tgel more than the other DP2 OSs, trehalose (α-1,1 linkage), kojibiose (α-1,2 linkage), and maltose (α-1,4 linkage), even with low NOH,S/Vs. This underscores the importance of OS structural flexibility in antiplasticizing starch. The study enriches our understanding of how OSs influence starch Tgel relative to the effects of sucrose, emphasizing the roles of intermolecular hydrogen bonding and OS structural flexibility in elevating starch Tgel in a manner which is beneficial for maintaining the texture and structural properties of reduced-sugar baked goods.

The evolution of granular surface structure and functional properties in rice starch during grain filling

The developmental changes in the granular surface structure and functional properties of starch during the entire grain filling period of rice (around 40 days) were investigated. The specific surface area of rice starch significantly decreased firstly then stabilized during growth due to increasing granular size. The pore volume decreased from 5.40 cm3/g at 6th day after anthesis (DAA-6) to 3.02 cm3/g (DAA-46). More starch granule-associated proteins (SGAPs) accumulated on the surface and in channels. Swelling power decreased by 46 %, whereas the flow behavior index (n) decreased by 32 % in upward curve during starch development from DAA-6 to DAA-30. Tan δ first dropped then remained steady at DAA 22–34 and lightly rebounded at the final stage, indicating that starch in the middle stage tended to have greater viscoelastic gel behavior at all sweeps. Mature starch showed lower in vitro hydrolysis rate and exhibited stronger enzymatic resistance. The results showed that granular surface features of rice starch may be an essential factor in determining rheological behavior and resistance to hydrolysis.

Performance and reaction mechanism of montmorillonite/α-Fe2O3/starch bio-nanocomposite as high-efficiency photocatalytic degradation of acetaminophen: Characterization, feasibility, and pathway

The worldwide challenge of eliminating pharmaceutical contaminants requires immediate attention. Developing bio-based catalysts that are eco-friendly, reusable, and high-performance, employing starch (ST) and montmorillonite (MMT) as support, holds tremendous promise as a novel biocatalyst for pharmaceutical waste removal. In this study, a montmorillonite/α-Fe2O3/starch (MMT/α-Fe2O3/ST) bio-nanocomposite photocatalyst was successfully synthesized and used for acetaminophen (ACT) degradation under UVA-LED irradiation. The influence of operational factors, such as catalyst, ACT concentrations, and solution pH, on photocatalytic activity was examined in detail; catalyst: 0.75 g/L, pH: 7.1, leading to total ACT (10 mg/L) removal in ∼80 min. MMT/α-Fe2O3/ST showed excellent durability due to negligible Fe leaching. After four successive degradation cycles, ACT and TOC elimination efficiencies remained over 91 and 42.7 %. Compared to other anions studied, carbonate ions suppressed the most ACT degradation. Based on the radical scavenger experiments, hydroxyl and superoxide radicals and holes were involved in the MMT/α-Fe2O3/ST system. LC-MS results were used to propose ACT degradation pathways. This work illuminated the significance of biocatalysts in removing emerging pollutants from wastewater.

How does high hydrostatic pressure treatment improve the esterification of quinoa (Chenopodium quinoa Willd.) starch?

High hydrostatic pressure (HHP) treatment was combined with octenyl succinic anhydride (OSA) modification of quinoa starch (QS) to improve esterification efficiency. The modified QS was used as a stabilizer to prepare a Pickering emulsion. The results showed that the HHP treatment disrupted the morphology and crystalline structure of QS, exposed numerous hydrophilic hydroxyl groups, and added esterification reaction sites. The degree of substitution (DS) and esterification efficiency (RE) of OSA-HHP-QS were significantly (p < 0.05) increased compared with OSA-QS. In addition, the short-range ordering, relative crystallinity, and thermal stability of OSA-HHP-QS decreased with increasing treatment pressure. Except for OSA-HHP200-QS, the starch granules treated at 200 MPa were annealed, resulting in molecular recrystallization. The Contact angle, emulsion stability index (ESI), and emulsion activity index (EAI) indicated that the emulsions stabilized with OSA-HHP-QS were highly stable. Therefore, HHP can be used as a novel technology to assist OSA modified starches in stabilizing Pickering emulsions.

Unraveling cereal physical barriers composed of cell walls and protein matrix: Insights from structural changes and starch digestion

Physical barriers composed of cell walls and protein matrix in cereals, as well as their cooking changes, play important roles in starch digestion. In this study, the physical barriers of native and cooked highland barley (HB), brown rice (BR), and oats (OA) kernels and their contribution to starch digestion were investigated. The resistant starch content was similar in cereal flours, but varied among cooked kernels (HB > BR > OA: 45.05 %, 10.30 %, and 24.71 %). The water adsorption, gelatinization enthalpy, and decrease in hardness of HB kernels were lower than those of OA and BR kernels. Microstructural observations of native kernels showed that HB had the thickest cell walls. After cooking, the lowest cell wall deformation and a dense continuous network developed from the protein matrix were observed in HB kernels. During digestion, undigested starch granules encapsulated by the stable cell walls and strong protein network were observed in HB kernels, but not in BR or OA kernels. Furthermore, the heavily milled HB kernels still had more resistant starch than the intact OA and BR kernels. Therefore, the physical barriers of HB kernels exhibited stronger inhibition of starch gelatinization and digestion. Differences in cereal physical barriers led to various inhibitory effects.

Effect of the Ratio of Soluble and Insoluble Dietary Fiber from Rice Bran with Extrusion Cooking Treatment on Pasting, Thermal, and Structural Properties of Rice Starch Gel

AbstractThis study aims to investigate the effect of different ratios of soluble and insoluble dietary fiber (0/100, 25/75, 40/60, 55/45, 70/30, 85/15, 100/0) in rice bran treated with or without extrusion cooking on the physicochemical and structural properties of rice starch. As the ratio of soluble dietary fiber (SDF) increases, the PV (peak viscosity), FV (final viscosity), SB (setback value), and BD (breakdown value) of starch are significantly decreased, indicating that a higher SDF ratio can delay the short‐term retrogradation of starch. Compared with the unextruded rice bran SDF/IDF starch gel, the content of weak bond water and free water in the extruded rice bran SDF/IDF starch gel is higher while the content of strong bond water in the starch gel is lower at the same SDF/IDF ratio. Meanwhile, for the results X‐ray diffraction (XRD) and fourier transform infrared spectrum (FTIR), the rice starch with the extruded rice bran SDF/IDF ratio of 25:75 shows the lowest relative crystallinity of 3.48% and the lowest peak to height ratio of FTIR (1047 cm−1/1022 cm−1) of 1.0746, suggesting that the inhibition effect on rice starch retrogradation is better. This may be beneficial for the utilization of different ratios of SDF/IDF from extruded rice bran and physicochemical properties improvement of rice starch gel.

The Influence of Different Polyphenols on the Digestibility of Various Kinds of Starch and the Value of the Estimated Glycemic Index

Considering the prevalence of diet-related diseases, new ways of preventing them are being sought. One of them is the addition of polyphenols to high-starch products to inhibit their digestibility and reduce their glycemic index. Therefore, this study aimed to investigate the differences between polyphenols popular in food ((+)catechin, epigallocatechin gallate, quercetin, kaempferol, naringenin, hesperidin, trans-ferulic acid, and p-coumaric acid), in terms of their impact on wheat, rice, potato, and maize starch digestibility. Polyphenols were added to starch separately, before and after its pasting, in one of the following doses: 5, 10, and 20 mg. Starch was digested in the presence of single polyphenols to measure RDS (rapidly digestible starch), SDS (slowly digestible starch), RS (resistant starch), and TS (total starch) content. On that basis, the SDI (starch digestion index) was calculated, and the GI (glycemic index) was estimated. The results show that polyphenols inhibit starch digestion at different levels depending on the type of tested starch and the time of polyphenol addition. However, in terms of RDS, TS, and eGI (estimated glycemic index), the greatest impact was observed for epigallocatechin gallate in a dose of 20 mg most frequently, independently of the kind of tested starch and the time of polyphenol addition.

Avocado oil used to modify rice starch and prepare gluten‐free cakes: physical–chemical, microbiological, sensory, and nutritional evaluations

SummaryThis study aimed to use modified rice starch by heat‐moisture treatment with avocado oil (AO) in gluten‐free cakes. Four cake formulations were prepared with native or modified rice starch with AO or soybean oil (control). The cake's sensory, microbiological, technological, nutritional, and bioactive compound characteristics were analysed. All formulations were sensorily acceptable (acceptance index: 72.44% to 79.44%) and microbiologically safe. The cake formulated with modified starch and AO exhibited the best textural properties, softness, and colour, and demonstrated the lowest in vitro digestibility (81.52 ± 2.75%) and glycaemic index (88.04 ± 3.83). AO resulted in higher levels of phenolic compounds in the cake (112.91 ± 1.8 to 237.38 ± 4.1 mg EAG.100 g−1). The use of modified rice starch and AO showed greater potential in the application of gluten‐free cake than those with native starch, resulting in improved product characteristics and good technological and sensory properties. It has also become a promising alternative in gluten‐free food products.

Synthesis and characterization of etherified cationic starch flocculant derived from Manihot esculenta peel with varying degrees of substitution

Cationic Manihot esculenta (ME) peel starch was synthesized through etherification method using 3-chloro-2-hydroxypropyl trimethylammonium chloride (CHPTAC) as cationizing monomer. The optimization of the main factors influencing the degree of substitution (DS) was conducted using central composite design (CCD) and response surface methodology (RSM). The factors assessed include CHPTAC concentration, catalyst sodium hydroxide (NaOH) concentration, and reaction time. The DS values of the cationic starches were obtained between 0.39 and 0.99. The maximum DS value was up to 0.99 at 0.615 mol/L of CHPTAC, 30 % (w/v) NaOH, and a reaction time of 5 h. The finding based on the optimization using RSM reflected that CHPTAC and NaOH concentrations are the key variables determining the DS value, while reaction time has a negligible impact on the etherification process. Furthermore, the chemical composition, morphology, and structure of the cationic ME peel starch were characterized by scanning electron microscopy with energy dispersive X-ray spectrometry (SEM-EDX), Fourier transform infrared spectroscopy (FTIR), X-ray diffractometry (XRD) and nuclear magnetic resonance spectroscopy (1H NMR). It was confirmed that the modifying monomers penetrated the surface layer of the starch granules and attached to the starch backbone.

Effect of dual modification on structural, functional and dielectric properties of cassava starch

The combined effect of hydrocolloid addition and microwave heating on the structural, physical and functional properties of cassava starch remains underexplored, despite individual studies on these modification methods. The dielectric properties, important for the microwave treatment of starch, have received little attention in recent studies. Cassava starch with and without addition of hydrocolloids (guar and xanthan gums) was modified by microwave heating for two different exposure times, and its structural, functional, and dielectric properties were studied. The dual modification enhanced the swelling power and oil holding capacity of the starch, reduced its gelatinization temperature, and did not alter its crystallinity. Notably, xanthan-modified starch microwaved for 6 min exhibited higher color variation (ΔE*) and increased granules agglomeration, while the microwave treatment alone roughened the surface of the starch granules. The mean diameters D[4,3] of starch increased with xanthan gum addition, microwave treatment (6 min), and their combination. The relative permittivity (ε’), loss factor (ε”) and penetration depth (Dp) were lower for samples modified by the combined method. Considering the type of hydrocolloid and the microwave exposure time, the combined modification significantly influenced the properties of starch, offering a promising approach to producing starch with enhanced functional properties.

Structure and digestibility changes of Indica and japonica waxy rice starch during in vitro pre-digestion

The digestion of starch have been of great interest, yet little is known about the structure changes and structure-digestibility relationships of waxy rice starch during digestion. In this study, waxy rice starch from Indica and Japonica cultivars were in vitro pre-digested for different times, and the changes in their structure and properties were investigated, including granule morphology, chain length distribution, short-range ordered structure, crystallinity, thermal properties, and digestibility. Pre-digested Indica and Japonica waxy rice starch had the characteristics of porous starch, showing similar surface erosion and pores. With the prolongation of pre-digestion time, the amylose content decreased by 0.74 %–2.69 %, the proportion of amylopectin short A chain (DP6–12) and B1 chain (DP13–24) decreased, and the proportion of long B2 (DP25–36) and B3 chain (DP ≥ 37) increased, especially in pre-digested Indica waxy rice starch. The short- and long-range ordered structure of pre-digested starch increased, manifested by an increase in the absorbance ratio at 1047/1022 cm−1, a decrease at 1022/995 cm−1, and an increase in relative crystallinity, leading to higher gelatinization temperature and enthalpy. Pre-digested waxy rice starch had a reduced rapidly digestible starch of 18.27 %–33.93 % and an increased resistant starch of 29.51 %–41.32 %, which will be applied in functional starch and healthy starchy foods.

Multi-omics analyses reveal mechanism for high resistant starch formation in an indica rice SSIIIa mutant

Soluble starch synthase IIIa (SSIIIa) is a key enzyme involved in amylopectin biosynthesis in rice, and deficiency of SSIIIa results in high content of resistant starch, which is benefit to human health. However, little is known about metabolic differences and carbon re-allocation in the seeds of the indica rice ss3a mutant. We found that SSIIIa deficiency impaired the storage of starch, but increased the soluble sugars, free amino acids and lipids. By multi-omic analyses, we found inactivation of SSIIIa triggered carbon repartitioning by downregulating sucrose synthase, grain incomplete filling 1, fructokinase and hexokinase (HK), and promoted the accumulation of soluble sugars. Meanwhile, the downregulation of HK and upregulation of plastidic phosphoglucomutase reduced the carbon flow through glycolysis and promoted glycogenesis. The downregulation of OsbZIP58 and the deleterious effect on ribosome formation might result in the reduction of storage protein synthesis and increased free amino acids content in ss3a. The higher levels of amylose and lipids could form more amylose-lipid complexes (starch phospholipids), resulting in a higher resistant starch content. Taken together, our study unraveled a functional cross talk between starch, protein and lipids in rice endosperm during seed development of ss3a, providing new insights for formation of high resistant starch in rice.

Regulating heat-induced fibrous whey protein-wheat starch composite emulsion gels as dysphagia food by preheating temperature: Insights from protein-starch interactions

Heat-induced processes significantly affect the gel behavior of protein and starch, but the mechanism by which preheating temperature affects the quality characteristics of the gel is unclear. Herein, to explore the feasibility of developing texture-modified foods based on proteins, carbohydrates and lipids, the effects of preheating temperatures (40–80 °C) on the texture and starch digestibility of fibrous whey protein (FWP)-wheat starch composite emulsion gels (CEGs) under two-step heating condition were investigated. The rheological and texture properties, protein-starch interactions and microstructure of CEGs were characterized. The results showed that preheating temperature synchronously regulated the texture and starch digestibility of heat-induced CEGs. When preheating temperature is below 60 °C, two-step heated CEGs showed bicontinuous networks with harder texture, similar as one-step heated gels. While temperature exceeds 60 °C, preheating altered FWP conformation and promoted protein-starch interactions, forming protein-starch complexes or protein aggregates. This changed protein distribution and disrupted bicontinuous network, forming more hydrophobic interaction, yielding a soft-textured CEGs. In this case, SDS content increased and RS content decreased. Notably, two-step heating process with high preheating temperature (>60 °C) is universal for softening fibrous whey protein-starch-based composite gels. This study enhances the understanding of traditional heat-induced protein-starch gelation processes and provides a simple and feasible method for developing texture-modified foods.

Effects of soy protein isolate interaction with brown rice starch on the multiscale structure of brown rice bread.

Gluten-free bread (GFB) has technical bottlenecks such as hard texture, rough taste and low nutrition in practical production. In order to solve these problems, this study used germinated brown rice starch as the main raw material, and investigated the effects of soybean isolate protein (SPI) on the multiscale structure of germinated brown rice starch and bread quality. A gluten-free rice bread process simulation system was established, and the interaction between SPI and starch in the simulation system was characterized. The result shows that the interaction forces between SPI and germinated brown rice starch were mainly represented by hydrogen bonds, and with the addition of SPI, the crystallinity of starch showed a downward trend. At the same time, when the amount of SPI was 3%, the appearance quality was the best and the specific volume of bread was 1.08 mL g-1. When the amount of SPI was 6%, the texture quality was the best. Compared with the bread without SPI, the hardness of the bread with 6% SPI was reduced by 0.13 times, the springiness was increased by 0.03 times, the color was the most vibrant, the L* value being 1.02 times the original, and the baking loss was reduced to 0.98 times the original. The interaction force between SPI and germinated brown rice starch and its effect on bread quality were clarified, and these results inform choices about providing a theoretical basis for the subsequent development of higher-quality GFB. © 2024 Society of Chemical Industry.

Impact of waxy protein deletions on the crystalline structure and physicochemical properties of wheat V-type resistant starch (RS5)

This study investigated the effects of waxy (Wx) protein on wheat V-type resistant starch (RS5) formation, molecular structure, and physicochemical properties. We discovered that waxy protein deletions led to a rise in B- and C-type starch granules, while reducing A-type starch granules, amylose, and slowly digestible starch contents. Further, dodecyl gallate (DG) addition significantly increased RS5 content, and molecular dynamics simulations indicated that amylose and DG can form stable complexes. Molecular docking indicated that DG could potentially aid in protecting wheat starch from digestion by human α-glucosidase. RS5 content was significantly reduced by waxy protein deletions. X-ray powder diffraction, Fourier-transform infrared spectroscopy, and laser confocal microscopy-Raman analyses revealed that waxy protein deletions decreased long-range crystalline structures and relative crystallinity and increased short-range crystalline structures，and full width at half maximum at 480 cm−1 of RS5. Pearson correlation analysis showed that RS5 content was highly correlated with its crystal structure, functional characteristics, and digestive characteristics. Principal component analysis revealed that five parameters (amylopectin, long-range crystalline structures, amylose, relative crystallinity, and RS5 content) had significant effect on the crystalline structure and functionality of RS5.