Enzymatic Hydrolysis of Barley Starch Lipids

The efficiency of phospholipase and lipase preparations in the hydrolysis of lysophospholipids of native and gelatinized barley starch was examined. The degree of hydrolysis was analyzed by determination of the amount of released fatty acids by an enzymatic method. Thermal and structural properties of the enzyme-treated starch were studied by differential scanning calorimetry and light microscopy. Lysophospholipids of the gelatinized barley starch were easily hydrolyzed, in contrast to the lipids of the granular starch. The maximum degree of hydrolysis achieved for the gelatinized starch was 80% and for the native starch ≈20%. Gelatinization enthalpies and micrographs indicated that even though the amount of the released fatty acids from the native starch was small, formation of free fatty acids inhibited swelling and gelatinization of starch granules.     

Effects of β-amylolysis on the resistant starch formation of debranched corn starches

Retrograded amylose is resistant to digestion by amylolytic enzymes, which is known as resistant starch type III (RS3). In this study we investigated the effect of β-amylase hydrolysis on the formation and physicochemical properties of RS3 from debranched corn starches. Three types of corn starch (Hylon VII, Hylon V, and common corn) were first gelatinized and then hydrolyzed using β-amylase to varying degrees. The resultant hydrolyzed starch was debranched with isoamylase and then exposed to temperature cycling to promote RS formation. A broad endotherm from approximately 45 to 120 °C and a small endotherm above 150 °C were noted for all retrograded starches. All three corn starches had increased RS contents after moderate β-amylolysis, with Hylon V having the highest RS content at 70.7% after 4 h of β-amylolysis. The results suggest that RS3 formation is affected by the starch composition as well as the starch structure and can be increased by moderate β-amylolysis.     

Influence of Botanical Source and Processing on Formation of Resistant Starch Type III

Influence of botanical source and gelatinization procedure (autoclaving or boiling) on resistant starch (RS) formation was investigated in starches from wheat, corn, rice, and potato. RS yields did not vary within the same sample but differed among samples with different starch botanical sources. Differences also existed in RS contents in native and retrograded starches. Slight or minor variations in RS values were found after both gelatinization procedures, although no clear pattern was found in the behavior of samples based on gelatinization procedure. The degree of polymerization (DP) of retrograded samples was assigned using high-performance anion exchange chromatography with pulsed amperometric detector (average DP 50–60), with no differences between autoclaved and boiled samples.     

Physicochemical Properties of Starch in Extruded Rice Flours

The effects of extruding temperatures and subsequent drying conditions on X‐ray diffraction patterns (XRD) and differential scanning calorimetry (DSC) of long grain (LG) and short grain (SG) rice flours were investigated. The rice flours were extruded in a twin‐screw extruder at 70–120°C and 22% moisture, and either dried at room temperature, transferred to 4°C for 60 hr, or frozen and then dried at room temperature until the moisture was 10–11%. The dried materials were milled without the temperature increasing above 32°C. XRD studies were conducted on pellets made from extruded and milled flours with particle sizes of 149–248 μm; DSC studies were conducted from the same material. DSC studies showed that frozen materials retrograded more than the flours dried at room temperature. The LG and SG samples had two distinct XRD patterns. The LG gradually lost its A pattern at >100°C, while acquiring V patterns at higher temperatures. SG gradually lost its A pattern at 100°C but stayed amorphous at the higher extruding temperatures. DSC analysis showed that retrograded flours did not produce any new XRD 2θ peaks, although a difference in 2θ peak intensities between the LG and SG rice flours was observed. DSC analysis may be very sensitive in detecting changes due to drying conditions, but XRD data showed gradual changes due to processing conditions. The gradual changes in XRD pattern and DSC data suggest that physicochemical properties of the extruded rice flours can be related to functional properties.     

Resistant Starch Escaped from Ethanol Production: Evidence from Confocal Laser Scanning Microscopy of Distiller's Dried Grains with Solubles (DDGS)

The origin of resistant starch (RS) in distiller's dried grains with solubles (DDGS) of triticale, wheat, barley, and corn from dry‐grind ethanol production was studied. A considerable portion of starch (up to 18% in DDGS) escaped from either granular starch hydrolysis or conventional jet‐cooking and fermentation processes. Confocal laser scanning microscopy revealed that some starch granules were still encapsulated in cells of grain kernel or embedded in protein matrix after milling and were thus physically inaccessible to amylases (type RS1). The crystalline structures of native starch granules were not completely degraded by amylases, retaining the skeletal structures in residual starch during granular starch hydrolysis or leaving residue granules and fragments with layered structures after jet‐cooking followed by the liquefaction and saccharification process, indicating the presence of RS2. Moreover, retrograded starch molecules (mainly amylose) as RS3, complexes of starch with other nonfermentable components as RS4, and starch–lipid complexes as RS5 were also present in DDGS. In general, the RS that escaped from the granular starch hydrolysis process was mainly RS1 and RS2, whereas that from the jet‐cooking process contained all types of RS (RS1 to RS5). Thus, the starch conversion efficiency and ethanol yield could be potentially affected by the presence of various RS in DDGS.     

Phase transitions in the starch-water system studied by adiabatic scanning calorimetry

We report high-precision measurements of phase transitions in the starch-water system by using for the first time adiabatic scanning calorimetry (ASC). Potato starch and nixtamalized corn flours were studied by this technique as a function of the moisture content. We calculated the percentage of gelatinized starch granules, as a function of the temperature, for both flours. For bi-phasic behavior in potato starch, at intermediate water contents, we propose an explanation based on previous hypotheses and on the experimental behavior as deduced from ASC measurements. ASC results suggest, in the case of nixtamalized corn flour, that uniform moisture is present in corn starch granules for all the considered moistures. A shift to higher temperatures of the peak temperature is observed when comparing the c(p)(T) curves with the classical DSC endotherms as measured for this flour. For both flours we estimated from the ASC results the melting temperatures of the starch granules, at zero moisture content, by using Flory's equation.     

Functionality of Rice and Sorghum Flours in Baked Tortilla and Corn Chips

The effects of raw and gelatinized sorghum and rice flours on the structure and texture of baked corn and tortilla chips were evaluated. Dry masa flour was hydrated into masa, sheeted, and cut. Corn chips were baked in an air-impingement oven, and tortilla chips were baked first in a three-tier oven and then in an air-impingement oven. Baked tortilla chips required significantly greater force to break and were less susceptible to breakage during handling than baked corn chips. Raw and gelatinized, normal and waxy rice and sorghum flours significantly changed the texture and structure of baked chips. Waxy rice and sorghum flours reduced peak force and work, increased chip thickness, and improved overall acceptability (as assessed by a taste panel), but waxy rice and sorghum chips were more fragile and had a greater number of large central air cells. Waxy rice was more beneficial than waxy sorghum flour. Gelatinization of waxy flours increased thickness of baked chips, whereas gelatinization of nonwaxy flours had no improvement over waxy flours alone. Gelatinization of sorghum flour significantly decreased the peak force and work values for baked tortilla chips when compared with the control chips. Gelatinized rice flour tortilla chips were not significantly different than the control chips but were significantly harder than the other baked tortilla chips. The complex interactions that occur in baked corn and baked tortilla chips suggest that each ingredient acts differently in the two products. Thus, each ingredient must be evaluated for specific products and processes.     

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.     

Effects of Cultivar and Processing Condition on Physicochemical Properties and Starch Fractions in Parboiled Rice

Starch can be classified into rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) according to its resistance to amylolytic enzymes. This study investigated the effects of cultivar and feedstock under varying parboiling conditions on the physicochemical properties and starch fractions of parboiled rice. Rice (rough or brown) was soaked, steamed under pressure, dried immediately or stored at room temperature for 24 hr prior to drying, and then treated with or without a repeated steam cycle prior to milling. The storage treatment significantly increased the retrograded amylopectin enthalpy and amylose‐lipid complex melting temperature of parboiled rice. Parboiled rice samples prepared from brown rice feedstock had higher peak melting temperatures but lower enthalpy values of retrograded amylopectin than samples prepared from rough rice after the storage treatment. The pasting viscosity of parboiled rice was most affected by the repeated autoclaving treatment and cultivar. Starch fractions in parboiled rice were significantly affected by cultivar and storage and by the interactions of cultivar and parboiling conditions. The storage treatment significantly increased SDS and generally decreased RDS in parboiled rice. Parboiled rice with different SDS and RS contents can be produced by varying rice cultivar and parboiling conditions.     

Effect of gelatinization and hydrolysis conditions on the selectivity of starch hydrolysis with alpha-amylase from Bacillus licheniformis

Enzymatic hydrolysis of starch can be used to obtain various valuable hydrolyzates with different compositions. The effects of starch pretreatment, enzyme addition point, and hydrolysis conditions on the hydrolyzate composition and reaction rate during wheat starch hydrolysis with alpha-amylase from Bacillus licheniformis were compared. Suspensions of native starch or starch gelatinized at different conditions either with or without enzyme were hydrolyzed. During hydrolysis, the oligosaccharide concentration, the dextrose equivalent, and the enzyme activity were determined. We found that the hydrolyzate composition was affected by the type of starch pretreatment and the enzyme addition point but that it was just minimally affected by the pressure applied during hydrolysis, as long as gelatinization was complete. The differences between hydrolysis of thermally gelatinized, high-pressure gelatinized, and native starch were explained by considering the granule structure and the specific surface area of the granules. These results show that the hydrolyzate composition can be influenced by choosing different process sequences and conditions.     

Pulsed Nuclear Magnetic Resonance (PNMR) Study of Rice Starch Retrogradation

Although pulsed NMR (PNMR) has been used for qualitative study of starch retrogradation in selected systems, validation is necessary for its application to new systems. PNMR was used to analyze the retrogradation of rice starches in purified form, in rice flour, and in cooked rice grains. The standard curves between the relative solid content (S′, %) by PNMR and the percentage of gelatinized starch (GS, %) were determined for common rice flour, common rice starch, and waxy rice starch at different moisture contents. The coefficients of linear regression for these curves (R²) were all >0.997. Starches with different amylose contents were tested for S′ values at the stages of freshly gelatinized, retrograded (4°C, 18 days), and reheated (90°C, 20 min). The S′ of reheated starch (S′_reheat) was similar to the S′ of freshly gelatinized starch (S′₀), so we concluded that the increase in S′ during storage corresponded to amylopectin retrogradation. The effect of moisture content on retrogradation of rice starch, rice flour, and cooked rice grains was studied by PNMR, and the data were interpreted using the Avami equation. Decreasing the moisture content increased the rate of retrogradation and led to a higher parameter k and a lower parameter n. For moisture content in the range studied, PNMR can be used to follow amylopectin retrogradation of different rice starch systems.     

Comparison of MALDI-TOF mass spectrometric to enzyme colorimetric quantification of glucose from enzyme-hydrolyzed starch

Successful quantification of the glucose produced by enzyme hydrolysis of starch was achieved by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protocol, using sorbitol as an internal standard. The starch contents measured by MALDI-TOF MS of corn starch, fiber-enriched oat flour derivatives, oat and barley flours, and barley flour/corn starch composites were evaluated in comparison to a widely accepted and validated method of starch determination, which relies on enzyme colorimetry (EC). The average starch content measured in a series of corn starch samples of different masses was 93 and 101% for EC and MALDI-TOF MS, respectively, values that represent the estimated purity of the sample. There was an agreement of 99% between the starch contents determined by the two analytical methods for complex flour-derived samples. Starch values estimated by MALDI-TOF MS consistently showed a greater degree of variability than those determined by EC, but this limitation was readily compensated by rapid acquisition of multiple mass spectra. This study is the first to report the quantification of glucose by MALDI-TOF MS, and it offers new perspectives into the potential utility of MALDI-TOF MS as a definitive tool for monosaccharide analysis and rapid starch determination in complex samples.     

Effects of Different Mill Types on Ethanol Production Using Uncooked Dry‐Grind Fermentation and Characteristics of Residual Starch in Distiller's Dried Grains (DDG)

This study aimed to investigate impacts of milling methods on ethanol production using an uncooked dry‐grind (cold fermentation) process and characterize residual starch in the distiller's dried grains (DDG) coproduct. Four corn lines with different chemical compositions were ground with cyclone, ultra‐centrifugal, or hammer mills equipped with a screen of 0.5 mm opening and used for the cold fermentation process. Greater starch hydrolysis and ethanol yield were obtained from cyclone‐milled corn, resulting from larger damaged starch contents and smaller particle sizes of the ground corn. Corn grains and ground corn after five‐month storage showed less starch hydrolysis than the freshly ground counterpart. Residual starch (2.8–8.0%) with large proportions of intact amylopectin contents (up to 42.5%) was found in the DDG from all types of milling. The results suggested that the entrapment of starch granules in ground corn and a low activity of amylolytic enzymes at a high ethanol concentration were accountable for the remaining of starch in the DDG.     

Effect of Extrusion Cooking on the Primary Structure and Water Solubility of β‐Glucans from Regular and Waxy Barley

Water‐soluble β‐glucan from native and extrusion‐cooked barley flours of two barley cultivars, Candle (a waxy starch barley) and Phoenix (a regular starch barley), was isolated and purified. The purity of β‐glucan samples was 85–93% (w/w, dry weight basis) for Candle and 77–86% (w/w, dry weight basis) for Phoenix. The water solubility of β‐glucan (at room temperature, 25°C) in the native and extruded flours (primary solubility) was different from that of the purified β‐glucan samples (secondary solubility). The solubility of β‐glucan in the native and extruded Candle flour was substantially higher than that of β‐glucan in Phoenix. For both cultivars, β‐glucan in the extruded flours had solubility (primary solubility) values higher than in their native counterparts. The solubility of β‐glucan in the purified β‐glucan samples differed depending on the barley cultivar and the extrusion conditions employed. The glycosidic linkage profiles of purified soluble β‐glucan from native and extruded barley flours were determined in order to understand the changes in the primary structure of β‐glucan and the effect of extrusion on the β‐glucan structure‐solubility relationship.     

Viscoelastic properties of waxy and nonwaxy rice flours, their fat and protein-free starch, and the microstructure of their cooked kernels

Physicochemistry and structural studies of two types of japonica rice, low amylose Calmochi-101 (CM101) and intermediate amylose M-202 (M202), were conducted to determine similarities and differences between the rices perhaps attributable to amylose content differences. The rheological behavior of the gelation and pasting processes of flours and starches was determined with high accuracy and precision using a controlled stress rheometer. Fat and protein, although minor constituents of milled rice, were shown to have significant effects on the physicochemical and pasting properties of starches and flours. Removal of protein and lipids with aqueous alkaline or detergent solutions caused lower pasting temperatures and higher overall viscosity in both starches, compared with their respective flours. There was less viscosity difference between M202 flour and its starch when isolated by enzymatic hydrolysis of protein. The protease did not reduce internally bound lipids, suggesting that fats help to determine pasting properties of rice flours and their respective starches. Structural integrity differences in individual granules of waxy and nonwaxy rice flours, starches, and whole raw, soaked, and cooked milled grain were revealed by fracture analysis and scanning electron microscopy. Calmochi 101 and M202 did not differ in weight-averaged molar mass (Mw) and root-mean-square radii (Rz) between flours and starches, as determined by high-performance size exclusion chromatography (HPSEC) and multiple-angle laser light scattering (MALLS) (Park, I.; Ibanez, A. M.; Shoemaker, C. F. Starch 2007, 59, 69-77).     

Resistant Starch and Starch Thermal Characteristics in Exotic Corn Lines Grown in Temperate and Tropical Environments

Corn as a food that is heated and cooled to allow starch retrogradation has higher levels of resistant starch (RS). Increasing the amount of RS can make corn an even healthier food and may be accomplished by breeding and selection, especially by using exotic germplasm. Sixty breeding lines of introgressed exotic germplasm backgrounds, selected for high yield, were grown in three tropical and temperate locations and analyzed for starch thermal characteristics and RS levels. Although actual values for all starch characteristics were within normal levels, most characteristics had significant genotypic effects, and all had significant location effects. Thermal properties of retrograded starch were more influenced by the environment than the thermal properties of raw starch, making retrograded starch traits more heritable than raw starch traits. This suggests that a breeding strategy based on retrograded starch traits will have a better chance of success than a breeding strategy based on raw starch traits. A significant genotype effect for RS levels indicates that genotypic selection to raise the level of RS and increase the healthful aspects of corn food should be successful. Significant location effects indicate that breeders using winter nurseries to accelerate their breeding progress need to be careful when making selections using RS data collected on seed grown in the tropics. A small but highly significant correlation between RS and some thermal characteristics, especially percentage of retrogradation, indicates that we may be able to select promising genotypes for RS selection based on our extensive database of thermal characteristics collected on a wide number of diverse corn lines.     

Properties and Structures of Flours and Starches from Whole,Broken, and Yellowed Rice Kernels in a Model Study

The objective of this study was to compare the structure and properties of flours and starches from whole, broken, and yellowed rice kernels that were broken or discolored in the laboratory. Physicochemical properties including pasting, gelling, thermal properties, and X‐ray diffraction patterns were determined. Structure was elucidated using high‐performance size‐exclusion chromatography (HPSEC) and high‐performance anion‐exchange chromatography with pulsed amperometric detection (HPAEC‐PAD). The yellowed rice kernels contained a slightly higher protein content and produced a significantly lower starch yield than did the whole or broken rice kernels. Flour from the yellowed rice kernels had a significantly higher pasting temperature, higher Brabender viscosities, increased damaged starch content, reduced amylose content, and increased gelatinization temperature and enthalpy compared with flours from the whole or the broken rice kernels. However, all starches showed similar pasting, gelling, thermal properties, and X‐ray diffraction patterns, and no structural differences could be detected among different starches by HPSEC and HPAEC‐PAD. α‐Amylase may be responsible for the decreased amylopectin fraction, decreased apparent amylose content, and increased amounts of low molecular weight saccharides in the yellowed rice flour. The increased amount of reducing sugars from starch hydrolysis promoted the interaction between starch and protein. The alkaline‐soluble fraction during starch isolation is presumed to contribute to the difference in pasting, gelling, and thermal properties among whole, broken, and yellowed rice flours.     

Dual modification of starch via partial enzymatic hydrolysis in the granular state and subsequent hydroxypropylation

The effect of enzymatic pretreatment on the degree of corn and mung bean starch derivatization by propylene oxide was investigated. The starch was enzymatically treated in the granular state with a mixture of fungal alpha-amylase and glucoamylase at 35 degrees C for 16 h and then chemically modified to produce enzyme-hydrolyzed-hydroxypropyl (HP) starch. Partial enzyme hydrolysis of starch in the granular state appeared to enhance the subsequent hydroxypropylation, as judged from the significant increase in the molar substitution. A variable degree of granule modification was obtained after enzyme hydrolysis, and one of the determinants of the modification degree appeared to be the presence of natural pores in the granules. Enzyme-hydrolyzed-HP starch exhibited significantly different functional properties compared to hydroxypropyl starch prepared from untreated (native) starch. It is evident that the dual modification of starch using this approach provides a range of functional properties that can be customized for specific applications.     

Characteristics of pregelatinized ae mutant rice flours prepared by boiling after preroasting

As ae mutant rice, such as EM10, lacks the starch branching enzyme IIb, its amylopectin contains more long-chain glucans than that of ordinary Indica and Japonica rice grains. Although boiled grains of ae rice cultivars are too hard and nonsticky for table rice, they are promising in terms of biofunctionality, such as prevention of diabetes. The present paper investigates the characterization of a novel group of four ae mutant rice cultivars (EM72, EM145, EM174, and EM189). They were subjected to the evaluation for their main chemical components, physical properties, and enzyme activities at different grain conditions (raw milled rice, roasted rice, boiled rice, and rice boiled after preroasting). These mutant rice grains are characterized by high apparent amylose, high protein and high glucose contents, high pasting temperature, high α-amylase activities, high resistant starch, and low degree of gelatinization. A novel method was developed to maintain the high resistant starch contents of gelatinized rice grains. Rice boild after preroasting showed a higher ratio of resistant starch and a lower amount of glucose than ordinary boiled rice. It became possible to produce high-quality and biofunctional pregelatinized rice flours by boiling with frozen fruits, such as tomatoes, after rice grains had been preroasted. These ae mutants were found to be suitable materials for rice/fruit or rice/vegetable products to serve as palatable, low-glucose, and high resistant starch rice products.     

Evaluation of Liquid Nitrogen Freeze Drying and Ethanol Dehydration as Methods to Preserve Partially Cooked Starch and Masa Systems

Preservation of starch structure/properties, including structures formed during partial or complete cooking, are important when the impact of processing conditions is being studied. Two preservation techniques used to study changes in starch during thermal‐mechanical processing are commonly cited in the literature: 1) rapid freezing followed by lyophilization, and 2) a dehydration procedure using alcohols. A comparative determination on how these methods affect various starch structures has not been widely reported. Corn starch samples were collected from the Rapid Visco‐Analyser (RVA) at 3 min (swollen granules, 30°C), at the top of the pasting peak (gelatinized granules, 95°C), at the bottom of the trough (dispersed polymers, 95°C), and a completed RVA sample stored for 120 hr at 4°C (retrograded starch). Samples of masa were obtained by nixtamalizing corn. Differential scanning calorimetry (DSC) endotherms of starch and masa, and X‐ray diffraction (XRD) patterns of masa were evaluated after being preserved by alcohol‐ or freeze‐drying. No significant differences (P > 0.05) between methods were found for onset, end, and peak temperatures (°C), enthalpy (J/g) and % relative crystallinity in any of the samples analyzed. Liquid nitrogen freeze‐drying and ethanol dehydration are both effective methods of preserving various starch systems for structural changes detectible by DSC and XRD; freeze‐drying is generally less expensive and time‐consuming.