Cross‐Linked Resistant Starch: Preparation and Properties

Resistant starches (RS) were prepared by phosphorylation of wheat, waxy wheat, corn, waxy corn, high‐amylose corn, oat, rice, tapioca, mung bean, banana, and potato starches in aqueous slurry (≈33% starch solids, w/w) with 1–19% (starch basis) of a 99:1 (w/w) mixture of sodium trimetaphosphate (STMP) and sodium tripolyphosphate (STPP) at pH 10.5–12.3 and 25–70°C for 0.5–24 hr with sodium sulfate or sodium chloride at 0–20% (starch basis). The RS₄ products contain ≤100% dietary fiber when assayed with the total dietary fiber method of the Association of Official Analytical Chemists (AOAC). In vitro digestion of four RS₄ wheat starches showed they contained 13–22% slowly digestible starch (SDS) and 36–66% RS. However after gelatinization, RS levels fell by 7–25% of ungelatinized levels, while SDS levels remained nearly the same. The cross‐linked RS₄ starches were distinguished from native starches by elevated phosphorus levels, low swelling powers (≈3g/g) at 95°C, insolubilities (<1%) in 1M potassium hydroxide or 95% dimethyl sulfoxide, and increased temperatures and decreased enthalpies of gelatinization measured by differential scanning calorimetry.     

White Pan Bread and Sugar‐Snap Cookies Containing Wheat Starch Phosphate,A Cross‐Linked Resistant Starch

Pup‐loaf bread was made with 10, 30, and 50% substitution of flour with wheat starch phosphate, a cross‐linked resistant starch (XL‐RS4), while maintaining flour protein level at 11.0% (14% mb) by adding vital wheat gluten. Bread with 30% replacement of flour with laboratory‐prepared XL‐RS4 gave a specific volume of 5.9 cm³/g compared with 6.3 g/cm³ for negative control bread (no added wheat starch), and its crumb was 53% more firm than the control bread after 1 day at 25°C, but 13% more firm after 7 days. Total dietary fiber (TDF) in one‐day‐old bread made with commercial XL‐RS4 at 30% flour substitution increased 3–4% (db) in the control to 19.2% (db) in the test bread, while the sum of slowly digestible starch (SDS) plus resistant starch (RS), determined by a modified Englyst method, increased from 24.3 to 41.8% (db). The reference amount (50 g, as‐is) of that test bread would provide 5.5 g of dietary fiber with 10% fewer calories than control bread. Sugar‐snap cookies were made at 30 and 50% flour replacement with laboratory‐prepared XL‐RS4, potato starch, high‐amylose (70%) corn starch, and commercial heat‐moisture‐treated high‐amylose (70%) corn starch. The shape of cookies was affected by the added starches except for XL‐RS4. The reference amount (30 g, as‐is) of cookies made with commercial XL‐RS4 at 30% flour replacement contained 4.3 g (db) TDF and 3.4 g (db) RS, whereas the negative control contained 0.4 g TDF and 0.6 g RS. The retention of TDF in the baked foods containing added XL‐RS4 was calculated to be >80% for bread and 100% for cookies, while the retention of RS was 35–54% for bread and 106–113% for cookies.     

Influence of Particle Size on the Twin-Screw Extrusion of Corn Meal

Effects of particle size (50–1,622 μm), screw speed (200–400 rpm), and feed moisture content (19–22%) on twin-screw extrusion of corn meal were investigated using a full-factorial design. Torque, specific mechanical energy, and product temperature generally showed no change within the commonly used particle-size range (100–1,000 μm), but each value dropped significantly as the particle size increased >1,000 μm. Die pressure was influenced by the three-way interaction of particle size, screw speed, and feed moisture content. The highest moisture level (22%), largest particle size (1,622 μm), and two lowest screw speeds (200 and 300 rpm) were the only conditions where the starch was <97.5% of transformation (gelatinization). Consequently, these two conditions also showed the least expansion and hardest product.     

Effect of DDGS,Moisture Content,and Screw Speed on Physical Properties of Extrudates in Single‐Screw Extrusion

Three isocaloric (3.5 kcal/g) ingredient blends containing 20, 30, and 40% (wb) distillers dried grains with solubles (DDGS) along with soy flour, corn flour, fish meal, and mineral and vitamin mix, with net protein adjusted to 28% (wb) for all blends, were extruded in a single‐screw laboratory‐scale extruder at screw speeds of 100, 130, and 160 rpm, and 15, 20, and 25% (wb) moisture content. Increasing DDGS content from 20 to 40% resulted in a 37.1, 3.1, and 8.4% decrease in extrudate durability, specific gravity, and porosity, respectively, but a 7.5% increase in bulk density. Increasing screw speed from 100 to 160 rpm resulted in a 20.3 and 8.8% increase in durability and porosity, respectively, but a 12.9% decrease in bulk density. On the other hand, increasing the moisture content from 15 to 25% (wb) resulted in a 28.2% increase in durability, but an 8.3 and 8.5% decrease in specific gravity and porosity, respectively. Furthermore, increasing the screw speed and moisture content of the blends, respectively, resulted in an increase of 29.9 and 16.6% in extruder throughput. The extrudates containing 40% DDGS had 8.7% lower brightness, as well as 20.9 and 16.9% higher redness and yellowness, compared with the extrudates containing only 20% DDGS. Increasing the DDGS content from 20 to 40% resulted in a 52.9 and 51.4% increase in fiber and fat content, respectively, and a 7.2% decrease in nitrogen free extract. As demonstrated in this study, ingredient moisture content and screw speed are critical considerations when producing extrudates with ingredient blends containing DDGS, as they are with any other ingredients.     

Metabolic Responses to Venezuelan Corn Meal and Rice Bran Supplemented Arepas (Breads)

The metabolic responses to South American foods remain to be determined. Using glycemic index (GI) and insulinemic index (II) values as references for therapeutic potential of foods, this study investigated the glucose responses to a typical Venezuelan corn bread (arepa) and to an arepa supplemented with rice bran. Adding rice bran to the bread increased the content of resistant starch and dietary fiber measured as total, soluble, and insoluble dietary fiber. It also increased the protein content of the arepa. Three meals, white wheat bread, 100% corn meal arepa, and an arepa supplemented with 20% rice bran, were administered within a one‐week period. Available starch in the foods was determined to provide 50 g of available carbohydrate per meal. To calculate the indices, bread was used as the reference. The GI and II of the two arepa meals were significantly smaller than the GI and II of white wheat bread, although the differences between the two types of arepas were not significant. It is concluded that Venezuelan arepas (corn meal bread) may have potential health benefits and that the presence of 20% rice bran in the arepa meal did not produce a significant improvement in the glucose response. Due to the presence of antioxidant elements in the supplemented arepa and its higher protein, dietary fiber, and resistant starch content, it may have a potential preventive effect against the development of other pathologies.     

Evaluation of gamma-irradiation in cocoa husk

gamma-Irradiation was investigated as a technique to improve the hygienic quality of cocoa husk. Cocoa husk is a byproduct of cocoa bean processing industry. It contains approximately 57.5% (w/w) dietary fiber (nonstarch polysaccharides plus lignin), 15% (w/w) crude protein, 10.7% (w/w) mineral elements, 2.32% (w/w) cocoa butter, and 2.8% (w/w) carbohydrates (free sugars plus starch). The effect of irradiation on the growth rates of microorganisms are reported. Total counts, enterobacteriaceae, coliforms, Staphylococcus aureus, Streptococcus "D" of Lancefield, and yeast and mold counts before and after irradiation at 5, 8, and 10 kGy were determined. Cocoa husk was irradiated in open containers. An irradiation dose of 5 kGy was already sufficient to decrease the microbial counts to a very low level. No alteration in dietary fiber was measured in the irradiated product and no significant differences were detected between irradiated and nonirradiated cocoa husk.     

Protease Treatment to Improve Ethanol Fermentation in Modified Dry Grind Corn Processes

To improve fractionation efficiency in modified dry grind corn processes, we evaluated the effectiveness of protease treatment in reducing residual starch in endosperm fiber. Three schemes of protease treatment were conducted in three processes: 1) enzymatic milling or E‐Mill, 2) dry fractionation with raw starch fermentation or dry RS, and 3) dry fractionation with conventional fermentation or dry conv. Kinetics of free amino nitrogen production were similar in both dry and wet fractionation (E‐Mill), indicating that proteolysis was effective in all three schemes. At the end of fermentation, endosperm fiber was recovered and its residual starch measured. Using protease treatment, residual starch in the endosperm fiber was reduced by 1.9% w/w (22% relative reduction) in dry conv and 1.7% w/w (8% relative reduction) in dry RS, while no reduction was observed in the E‐Mill process. Protease treatment increased ethanol production rates early in fermentation (≤24 hr) but final ethanol concentrations were unaffected in both dry RS and E‐Mill. In dry conv, the addition of protease resulted in a decline in final ethanol concentration by 0.3% v/v, as well as a higher variability in liquefaction product concentration (higher standard deviations in the glucose and maltose yields). Protease treatment can be used effectively to enhance modified dry grind processes.     

Characterization and In Vivo Hydrolysis of Amylose–Stearic Acid Complex

Objectives of this study were to compare thermal properties, swelling power, and enzymatic hydrolysis of a type 5 resistant starch (RS5) with that of normal corn starch (NCS) and high‐amylose corn starch (HA7). The RS5 was prepared by complexing debranched HA7 with stearic acid (SA). Because of amylose‐helical‐complex formation with SA, the RS5 starch granules showed restricted swelling at 95°C. The RS5 displayed a larger RS content (67.8%) than the HA7 (33.5%) and NCS (0.8%), analyzed following AOAC method 991.43 (AACC International Approved Method 32‐07.01). When the cooked RS5, HA7, and NCS were used to prepare diets for rats with 55% (w/w) starch content, RS contents of the diets were 33.7, 15.8, and 2.6%, respectively. After the diet was fed to the rats in week 1, ≈16% of the starch in the RS5 diet was found in the feces, substantially greater than that of the HA7 diet (≈6%) and NCS diet (0.1%). The percentage of starch not being utilized in the RS5 diet decreased to ≈5% in week 9, which could be partially attributed to fermentation of RS5 by gut microflora. Large proportions (68–99%) of the SA in the RS5 diet were unabsorbed and discharged in the rat feces. The results suggest that the interactions between starch and SA can be used to enhance resistance of starch to in vitro and in vivo digestion.     

Selected Functional Properties of Extruded Starch Acetate and Natural Fibers Foams

Response surface methodology was employed to study the functional properties of starch acetate foams blended with 0, 7.5, and 15% wood, oat, and cellulose fibers. The blends were extruded with 14, 17, and 20% ethanol as a blowing agent, using a twin‐screw extruder with 160°C barrel temperature and 225 rpm screw speed. Physical characteristics of the extrudates including radial expansion ratio, unit density, and bulk density; and mechanical properties including unit spring index and bulk spring index were determined. Scanning electron micrographs were taken to observe foam cell textures. Higher fiber content resulted in lower radial expansion. Ethanol content had a positive effect on foam expansion. Higher expansion was obtained in starch acetate‐cellulose foams because better starch acetate‐fiber matrix was formed. Mechanical properties increased with higher fiber and ethanol contents. Micrographs showed that uniform cell structures were associated with better mechanical properties.     

Effects of Thermomechanical Extrusion and Particle Size Reduction on Bioconversion Rate of Corn Fiber for Ethanol Production

The objective of this research was to evaluate the effect of thermomechanical extrusion and particle size (PS) reduction on the bioconversion rate of corn fiber for ethanol production. Extrusion was conducted at a screw speed of 300 rpm, feed rate of 120 g/min, feed moisture content of 30%, melt temperature of 140°C, and die diameter of 3 mm. Raw and extruded corn fiber were separated into three different PSs (1 > PS ≥ 0.5, 0.5 > PS ≥ 0.3, and 0.3 > PS ≥ 0.15 mm) with a wire sieve. Extrusion pretreatment and PS reduction resulted in a significant (P < 0.05) difference in physical properties and color values of extruded corn fiber as a result of accelerated degradation of corn fiber structure. Significant increase in water solubility index of extruded corn fiber at 0.3 > PS ≥ 0.15 mm was an indication of high degradation of starch during extrusion for higher release of polysaccharides. Moreover, extruded corn fiber at PS reduction 0.3 > PS ≥ 0.15 mm also significantly increased (P < 0.05) ethanol yield (69.11 g/L) and conversion (68.18%) by increasing protein digestibility and free amino nitrogen, which are essential for higher fermentation efficiency.     

Exotic Corn Lines with Increased Resistant Starch and Impact on Starch Thermal Characteristics

Ten parent corn lines, including four mutants (dull sugary2, amylose‐extender sugary2, amylose‐extender dull, and an amylose‐extender with introgressed Guatemalen germplasm [GUAT ae]) and six lines with introgressed exotic germplasm backgrounds, were crossed with each other to create 20 progeny crosses to increase resistant starch (RS) as a dietary fiber in corn starch and to provide materials for thermal evaluation. The resistant starch 2 (RS2) values from the 10 parent lines were 18.3–52.2% and the values from the 20 progeny crosses were 16.6–34.0%. The %RS2 of parents was not additive in the offspring but greater RS2 in parents was correlated to greater RS2 in the progeny crosses (r = 0.63). Differential scanning calorimetry (DSC) measured starch thermal characteristics, revealing positive correlations of peak gelatinization temperature and change in enthalpy with %RS2 (r = 0.65 and r = 0.67, P ≤ 0.05); however, % retrogradation (a measure of RS3) and retrogradation parameters did not correlate with %RS2. The %RS2 and onset temperature increased with the addition of the ae gene, likely because RS delays gelatinization.     

Production of Boiling‐Stable Granular Resistant Starch by Partial Acid Hydrolysis and Hydrothermal Treatments of High‐Amylose Maize Starch

The purpose of the present work was to examine whether partial acid hydrolysis (PAH) of a high‐amylose maize starch (ae‐VII) would enhance the effects of hydrothermal treatments to produce granular resistant starch (RS) that is stable to further heat treatment at atmospheric pressure. PAH ae‐VII starches were prepared by heating 35% (w/v) suspensions with 1% (w/w) HCl at 25°C for 6, 30, and 78 hr. Native and PAH starches were then treated by annealing (ANN) or heat‐moisture treatment (HMT). ANN was done at 70% moisture at 50, 60, or 70°C for 24 hr, and HMT was done at 30% moisture at 100, 120, or 140°C for 80 min. RS that survives boiling during analysis was determined by a modification of the AOAC method for determining total dietary fiber. RS was also determined by the Englyst method. Little change in the gelatinization enthalpy was found for ae‐VII starch after PAH, ANN, or HMT as individual treatments. After PAH, either ANN or HMT led to decreased gelatinization enthalpy. HMT and ANN alone increased boiling‐stable RS but decreased total RS. After PAH of ae‐VII, either ANN or HMT tended to increase the yield of boiling‐stable granular RS, with the greatest yield (≤63.2%) observed for HMT.     

Water Solubility and Macromolecular Properties of Corn Meal Extrudates as Affected by Epichlorohydrin

Degermed corn meal adjusted to 18% moisture content (db) with epichlorohydrin (ECH) content at 0, 0.5, 1, or 2% (w/w) were extruded with a twin-screw laboratory extruder at a screw speed of 140 rpm. Compression and metering barrel zones were set at 100, 120, or 140°C. Water solubility (WS) of ground extrudates ranged from 7.6 ± 1.1% to 14.3 ± 1.3%. ECH content had a significant (P < 0.01) negative effect on WS, while barrel temperature and the interaction between barrel temperature and ECH content were not significant (P > 0.05). Presumably, ECH reduced WS of extrudates through cross-linking between hydroxyl groups on starch and protein molecules. Gel-permeation chromatography patterns for both 100 and 140°C barrel temperatures showed that high molecular weight carbohydrates in the extrudates decreased with increasing ECH content without a simultaneous increase in low molecular weight carbohydrates. This suggested that the decrease in high molecular weight fractions was due to insolubilization by cross-linking rather than degradation. SDS-PAGE revealed that two protein bands of ≈29 and 17.5 kDa disappeared, and a new band appeared at 45 kDa with increasing ECH content. This indicated that, most likely, ECH reacted with protein in addition to reacting with starch. However, glycoprotein and starch-protein complexes were not identified with electrophoresis.     

Effect of RS4 Resistant Starch on Extruded Ready‐to‐Eat (RTE) Breakfast Cereal Quality

A phosphorylated cross‐linked type 4 resistant wheat starch (RS4) containing 85.5% total dietary fiber (TDF) replaced 5–20% of the whole corn flour in an extruded ring‐shaped ready‐to‐eat breakfast cereal formulation. TDF content of the dry ingredient blend increased by roughly 3.6% for every 5% of added RS4. TDF loss during extrusion processing increased as RS4 level increased; however, a high percentage (78–89%) of the TDF content was retained in the final product. Product density increased as level of RS4 increased, but no effect on the specific mechanical energy was observed. X‐ray microtomography showed that RS4 addition did not affect internal air‐cell wall thickness, air‐cell size, or porosity. Moreover, addition of 5 or 10% RS4 did not affect expansion, physical appearance, initial crispness, or bowl life of the cereal rings. High levels of RS4 (15 and 20%) decreased cereal ring diameter but increased initial (dry) product crispness and extended bowl life. In general, RS4 addition level did not affect moisture content or moisture uptake of cereal rings during soaking in milk. Furthermore, moisture content and moisture uptake did not appear to influence the crispness of milk‐soaked cereal rings.     

Effects of Calcium Hydroxide and Processing Conditions on Corn Meal Extrudates

The effects of added calcium hydroxide (0.0, 0.15, 0.25, and 0.35%) and processing conditions, feed moisture content (mc) (16, 18, and 20%) and barrel temperature (130 and 150°C) on characteristics of corn meal extrudates were studied. Extruder screw speed was maintained at 130 rpm. Corn meal was extruded with a single-screw extruder (Brabender model GNF 1014/2) with a screw compression ratio of 3:1. The highest values (P < 0.05) for radial expansion and the lowest values for density and breaking force of extrudates were found for the treatment with 0.00% calcium hydroxide extruded at 16% feed mc and 130°C barrel temperature. This treatment was statistically different from the other treatments. Best values for radial expansion of samples extruded with added calcium hydroxide were for the samples with 0.15% calcium hydroxide at 18% feed mc and 130°C barrel temperature, followed by the sample with 0.35% calcium hydroxide at 16% feed mc and 130°C barrel temperature. Water absorption index and water solubility index were affected by calcium hydroxide and extrusion conditions evaluated. Extrudates had large numbers of flattened and sheared granules. Increases in calcium hydroxide increased extrudate yellowness. The combined action of calcium hydroxide and extrusion conditions completely modified the organized structure of the starch and suggest the formation of a starch-calcium complex (crystalline region). The texture of the extruded products was crispy after puffing.     

Effect of Bonding Forces on Corn Starch Isolation

Chemical treatments with a number of low‐toxicity or nontoxic reagents were applied to corn slurry to investigate the disruption or weakening of common bonding forces between corn starch and proteins, such as hydrogen bonds, disulfide bonds, electrostatic interactions, or combinations thereof, to improve the corn starch isolation process. Starch and proteins could be easily separated by disrupting disulfide bonds with 1% l ‐cysteine (w/v). The most effective reagents for hydrogen bonds and electrostatic interactions were 3M urea and pH 7.5 separately. The sequence treatment of hydrogen bonds, disulfide bonds, and electrostatic interactions (namely, sequence treatment of 1M urea, 1% l ‐cysteine, and pH 7.5) led to the highest amount of starch in corn slurry and facilitated the corn starch isolation.     

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 genotype and processing on the in vitro rate of starch hydrolysis and resistant starch formation in peas (Pisum sativum L.).

The formation of resistant starch (RS) and the rate of starch hydrolysis were evaluated in vitro in a wild type of green-seeded pea genotype RRRbRb BC3 (33-Am) with 32.7% amylose content and in two mutants RRrbrb BC3 (23-Am) and rrRbRb BC3 (65-Am) with amylose contents of 23.3 and 65.1%, respectively. Pea samples were intact or homogenized and subjected either to autoclaving or to boiling at atmospheric pressure. The amount of RS (total starch basis) varied from 6.2 to 12.9% in the 23-Am products and from 31.2 to 33.4% in the 65-Am products. The RS level of the 33-Am product with a regular amylose content was 11.0%. Both the 23-Am and the 65-Am products were abundant sources of dietary fiber (39 and 34%, dry matter basis, respectively) versus 23% in the regular pea product. The amylose/amylopectin ratio was an important determinant of the rate of starch hydrolysis. The hydrolysis indices (HI) and predicted glycemic indices were lowest in the 65-Am peas (HI range = 42-59) as compared to the 23-Am peas (HI range = 53-84). It is concluded that the pea genotypes covered a wide range in starch availability, which is likely to affect nutritional parameters such as glycemic responses and colonic delivery of starch.     

Effect of Complexation Temperature on Thermal Properties and Enzymatic Hydrolysis of Starch–Oleic Acid Complexes

The effect of complexation temperature (30, 60, and 90°C) on the gelatinization properties, glass transition, enzymatic hydrolysis, and crystalline structure of high‐amylose corn starch–oleic acid (HACS‐OA) complexes created by a dimethyl sulfoxide (DMSO)‐based complexation method and of normal corn starch–oleic acid (NCS‐OA) complexes created by an alkaline‐based complexation method were investigated by using differential scanning calorimetry, thermogravimetric analysis, and X‐ray diffractometry. The results indicated that the highest complex indices were found in the complexes created at 30 and 60°C with the DMSO‐based complexation method and alkaline‐based complexation method, respectively. The X‐ray diffraction patterns of both HACS‐OA complexes and NCS‐OA complexes created at different complexation temperatures were the V‐type pattern. For the complexes created by the two methods, both the melting temperature and the glass transition temperature increased obviously with increasing complexation temperature. Complexation temperature also influenced the enzymatic hydrolysis rate of starch‐OA complexes.     

Resistant Starch Formation in Tortillas from an Ecological Nixtamalization Process

The objective of this work was to study the formation of resistant starch (RS) in tortillas from an ecological nixtamalization process compared with the traditional nixtamalization process. The RS increased through all the steps of tortilla production. It was found that the increase of the RS corresponds mainly to the formation of RS5 (V‐amylose‐lipid complex), but in tortillas two major types of RS coexist: RS5 and RS3 (retrograded starch). In general, tortillas from the ecological nixtamalization process gave higher values of protein, lipids, total dietary fiber, insoluble fiber, soluble fiber, and RS compared with tortillas from the traditional nixtamalization process and commercial flour. The highest glycemic index (GI) occurred in the tortillas from commercial flour, whereas tortillas from 0.4% CaCO₃ and 0.6% CaSO₄ were classified as medium‐GI (GI 50–70). Tortillas from 0.6% CaCl₂ had the lowest value of GI. The ecological nixtamalization processes caused significant differences in quality and nutritional properties of tortillas.