Low α-Amylase Starch Digestibility of Cooked Sorghum Flours and the Effect of Protein

The comparably low starch digestibility of cooked sorghum flours was studied with reference to normal maize. Four sorghum cultivars that represent different types of endosperm were used. Starch digestibilities of 4% cooked sorghum flour suspensions, measured as reducing sugars liberated following α-amylase digestion, were 15–25% lower than for cooked maize flour, but there were no differences among the cooked pure starches. After the flours were predigested with pepsin to remove some proteins, the starch digestibility of cooked sorghum flours increased 7–14%, while there was only 2% increase in normal maize; however, there was no effect of pepsin treatment on starch digestibility if the flours were first cooked and then digested. After cooking with reducing agent, 100 mM sodium metabisulfite, starch digestibility of sorghum flours increased significantly while no significant effect was observed for maize. Also, starch solubility of sorghum flours at 85 and 100°C was lower than in maize, and sodium metabisulfite increased solubility much more in sorghum than in maize. Differential scanning calorimetry results of the flour residue after α-amylase digestion did not show any peaks over a temperature range of 20–120°C, indicating that sorghum starches had all undergone gelatinization. These findings indicate that the protein in cooked sorghum flour pastes plays an important role in making a slowly digesting starch.     

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.     

Detection of Proteins in Starch Granule Channels

Proteins were detected in channels of commercial starches of normal maize, waxy maize, sorghum, and wheat through labeling with a protein‐specific dye and examination using confocal laser scanning microscopy (CLSM). The dye, specifically 3‐(4‐carboxybenzoyl)quinoline‐2‐carboxaldehyde (CBQCA), fluoresces only after it reacts with primary amines in proteins, and CLSM detects fluorescence‐labeled protein distribution in an optical section of a starch granule while it is still in an intact state. Starch granules in thin sections of maize kernels also had channel proteins, indicating that proteins are native to the channels and not artifacts of isolation. Incubation of maize starch with protease (thermolysin) removed channel proteins, showing that channels are open to the external environment. SDS‐PAGE analysis of total protein from gelatinized commercial waxy maize starch revealed two major proteins of about M_r 38,000 and 40,000, both of which disappeared after thermolysin digestion of raw starch. Commercial waxy maize starch granule surface and channel proteins were extracted by SDS‐PAGE sample buffer without gelatinization of the granules. The major M_r 40,000 band was identified by MALDI‐TOF‐MS and N‐terminal sequence analysis as brittle‐1 (bt1) protein.     

Genetic Diversity in Properties of Starch from Zimbabwean Sorghum Landraces

Starch was isolated from 95 sorghum landraces from Zimbabwe using an alkali steep and wet‐milling procedure. The physicochemical properties of sorghum starch were examined for potential use in Southern Africa. All the landraces evaluated had a normal endosperm indicated by the amylose content of the starches. Starch properties were not correlated to most of the physical grain quality traits evaluated. Grain hardness was weakly correlated to starch gel adhesiveness (r = 0.36) and amylose content (r = 0.38) (P < 0.001). The mean peak viscosity (PV) of the sorghum starches was 324 Rapid Visco Analyser units (RVU) compared with 238 RVU in a commercial corn starch sample; PV was 244–377 RVU. Some landraces had low shear‐thinning starches, implying good paste stability under hot conditions. Pasting properties were highly correlated among the sorghum starches. The starch gel hardness showed considerable variation (44–71 g) among the landraces. Gelatinization peak temperatures were 66–70°C. The thermal properties of starches were not correlated with starch swelling and pasting properties. Genotype grouping by highest and lowest values in each category would allow selection of sorghums based on a specific attribute depending on the desired end use.     

Interaction Between Sorghum Protein Extraction and Precipitation Conditions on Yield,Purity, and Composition of Purified Protein Fractions

Sorghum proteins have the potential to be used as a bio‐industrial renewable resource for applications such as biodegradable films and packaging. This project was designed to evaluate the effect of interactions between sorghum protein extraction and precipitation conditions on the yield, purity, and composition of sorghum protein fractions. Proteins were extracted with 70% ethanol under nonreducing conditions, with ultrasound, or under reducing conditions using either sodium metabisulfite or glutathione as the reducing agent. Several conditions were used to isolate the extracted proteins through precipitation, including lowering ethanol concentrations alone or in combination with lowering to pH 2.5, or by adding 1M NaCl to the extract. Combinations of these conditions were also tested. All precipitation conditions effectively precipitated proteins and lowering the pH and adding 1M NaCl to the extracts enhanced precipitation in some cases. However, the conditions that precipitated the maxium amount of protein or highest purity of protein varied according to how the proteins were initially extracted. Precipitated proteins were characterized by RP‐HPLC, SEC, HPCE, and SDS‐PAGE to compare the protein fractions composition. Nonreduced and sonicated samples had a much wider M_w distribution than reduced extracts. Thus, extraction and precipitation conditions influenced the isolated proteins yield, purity, and composition. Because the extraction and purification processes influenced the composition, purity, and biochemical properties, it may be possible to prepare protein fractions with unique functionalities for specific end‐uses.     

Application of High‐Intensity Ultrasound and Surfactants in Rice Starch Isolation

High‐intensity ultrasound was evaluated as an alternative method to isolate rice starch without the use of chemicals as in the traditional alkaline steeping method. Surfactants, including sodium dodecyl sulfate (SDS), sodium stearoyl lactylate (SSL), and Tween 80, at 0.1, 0.3, or 0.5% combined with high‐intensity ultrasound were also investigated for rice starch isolation. A rice flour slurry (33%) was subjected to sonication for 15, 30, or 60 min at an amplitude of 25, 50, or 75% and at 40 or 50°C. The starch yield was not significantly affected by the treatment temperature and ranged from 46.7 to 76.2% (starch dry basis) after the sonication treatment; the protein and damaged starch contents of the isolated starches were 0.9–1.7% and 3.1–3.5% (dry basis), respectively. The combination of 0.5% SDS and high‐intensity ultrasound improved the starch yield to 84.9% with low residual protein, however, little improvement was observed with SSL or Tween 80. The pasting properties of isolated starch as measured by a Rapid Visco‐Analyser were affected by the treatment temperature and by the amount of residual protein and damaged starch. The thermal properties of the isolated starch were not changed by sonication and the amylose content remained unchanged. The surface of the isolated starch was not damaged by sonication as shown by scanning electron microscopy. High‐intensity ultrasound, alone or combined with SDS, showed a great potential for rice starch isolation in a short period of time without generating alkaline effluent.     

Laboratory Procedure to Wet‐Mill 100 g of Grain Sorghum into Six Fractions

A small‐scale (100 g of grain) procedure was developed to wet‐mill grain sorghum into six fractions by modifying the procedure of Eckhoff et al (1996). The wet‐milling process was repeated five times on commercial grain sorghum, and the mean yield (69.4%) of starch (≤0.3% protein) varied by 0.3%, whereas the yields of fiber, gluten, and germ plus bran fractions varied by 5–6%. The starch fraction accounted for ≈95% of that in the grain, while the total solids recovered was 99.0%. Four other samples of grain sorghum gave 92–95% recoveries of starches and 98.2–99.8% recoveries of total solids. All grain sorghum starches had lightness (L*) values and pasting curves nearly equal to those of a commercial maize starch.     

Effects of Amylose,Corn Protein,and Corn Fiber Contents on Production of Ethanol from Starch‐Rich Media

The effects of amylose, protein, and fiber contents on ethanol yields were evaluated using artificially formulated media made from commercial corn starches with different contents of amylose, corn protein, and corn fiber, as well as media made from different cereal sources including corn, sorghum, and wheat with different amylose contents. Second‐order response‐surface regression models were used to study the effects and interactions of amylose, protein, and fiber contents on ethanol yield and conversion efficiency. The results showed that the amylose content of starches had a significant (P < 0.001) effect on ethanol conversion efficiency. No significant effect of protein content on ethanol production was observed. Fiber did not show a significant effect on ethanol fermentation either. Conversion efficiencies increased as the amylose content decreased, especially when the amylose content was >35%. The reduced quadratic model fits the conversion efficiency data better than the full quadratic model does. Fermentation tests on mashes made from corn, sorghum, and wheat samples with different amylose contents confirmed the adverse effect of amylose content on fermentation efficiency. High‐temperature cooking with agitation significantly increased the conversion efficiencies on mashes made from high‐amylose (35–70%) ground corn and starches. A cooking temperature of ≥160°C was needed on high‐amylose corn and starches to obtain a conversion efficiency equal to that of normal corn and starch.     

Noodle Quality as Related to Sorghum Starch Properties

Starch was extracted from 10 sorghum genotypes and physicochemical properties (amylose content and pasting, textural, and thermal properties) were evaluated. The amylose content was 24–30%. DC‐75 starch had the highest peak viscosity (380 Rapid Visco Analyser units). Gelatinization peak temperature occurred over a narrow range (67–69°C). Genotypes Kasvikisire and SV2 produced white starches. Starches from other genotypes were different shades of pink. The starch noodles prepared were, accordingly, either white or pink. Cooking enhanced the pink coloration of noodles. Cooking loss, noodle rehydration, and elasticity were evaluated. Cooking loss was low (mean 2.4%). Noodle elasticity was highly correlated with starch pasting properties of hot paste viscosity (HPV) (r = 0.81, P < 0.01) and cold paste viscosity (CPV) (r = 0.75, P < 0.01). Noodle rehydration was significantly correlated to the initial swelling temperature of starch (T_i) (r = ‐0.91, P < 0.001) and gelatinization peak temperature (T_p) (r = 0.69, P < 0.05). The findings suggest a potential area of food application for sorghum genotypes of different grain colors. Evaluation of starch properties could be a good starting point for selecting sorghum genotypes with superior noodle‐making properties.     

Physical Properties and Enzymatic Digestibility of Phosphorylated ae,wx, and Normal Maize Starch Prepared at Different pH Levels

Phosphorylated starches were prepared with sodium tripolyphosphate (STPP) at pH 6, 8, and 10 from waxy (wx, 3.3% amylose), normal (22.4% amylose), and two high-amylose (ae, 47 and 66% amylose) maize starches. After phosphorylation, the gelatinization peak temperature (T_p) decreased and pasting peak viscosity (PV) increased for all the starches except wx, which showed a slight increase in gelatinization temperature. There was a substantial effect of phosphorylation pH on paste viscosity. More crosslinking was found in ae starches with phosphorylation at pH 10. Sodium ions apparently decreased PV of all the phosphorylated starches while only slightly affecting PV of native starches. Phosphorylation increased swelling power of some of the starches, with maximum swelling power at phosphorylation pH 8 and minimum at pH 10. Maximum swelling power for wx starch after preparation was at pH 8 and minimum at pH 6. After phosphorylation, the clarity and freeze-thaw stability of all the starches was greatly increased compared with the native starches. Phosphorylation increased digestibility of ae starches but had little effect on wx and normal starches. After phosphorylation, the adhesiveness, springiness, and cohesiveness of all starch gels generally increased, the hardness of 47% ae and wx starches increased, and that of normal starches decreased. Enthalpy of gelatinization decreased after phosphorylation with the greatest decrease observed for ae starches. When the phosphorylation pH increased from 6 to 10, the brightness (L*) of all the phosphorylated starches decreased, while a* and b* of all the phosphorylated starch increased. Scanning electron micrographs showed some erosion on the surface of starch granules after phosphorylation.     

General Application of Raman Spectroscopy for the Determination of Level of Acetylation in Modified Starches

A method using Raman spectroscopy was recently developed for the determination of the degree of acetylation in modified wheat starch. In this article, we show that the method can be generalized to a wide range of starches of different botanical origin and amylose content. Calibration sets were used to develop regression equations for 11 types of acetylated starches, including cereal (rice, maize, wheat) and noncereal (potato and sweetpotato) sources. The calibration lines were then used to predict the level of acetylation of starch samples with unknown level of acetylation using their Raman spectra. In each case, R² > 0.98 for linear regression of Raman vs. titrimetric determination of acetylation. The Raman-based calibration curves allow fast and nondestructive determination of the degree of acetylation for different types of starches.     

Starch Molecular Mass and Size by Size-Exclusion Chromatography in DMSO-LiBr Coupled with Multiple Angle Laser Light Scattering

The weight average molar mass (M_w) and root mean square radii of starches from waxy maize (Amioca), waxy rice flour, cassava, Hylon V, Hylon VII, and potato amylose were determined by size-exclusion chromatography (SEC) and multiple-angle laser light scattering (MALLS). Dimethylsulfoxide (DMSO) containing 50 mM LiBr was used to dissolve the starches and also served as the mobile phase. SEC with large particle size polystyrene divinylbenzene packing materials and MALLS instrumentation were evaluated for the ability to separate and determine molar mass (MM) of starch polymers, respectively. The determination of M_w by MALLS is necessary because the M_w of many cereal starches exceeds the available molecular standards by one or two orders of magnitude. The M_w depends on the method of calculation. The M_w (Berry method) of starch from waxy corn was 2.27 × 10⁸ Da, waxy rice 8.9 × 10⁷ Da, cassava 5.7 × 10⁷ Da, Hylon V 2.7 × 10⁷ Da, Hylon VII 4.8 × 10⁶ Da, and potato amylose 1.9 × 10⁵ Da. Recovery dropped dramatically for molecules with root mean square radii >200 nm.     

Extrusion of Cross‐Linked Hydroxypropylated Corn Starches I. Pasting Properties

A series of cross‐linked (0, 0.014, 0.018, 0.024, and 0.028% POCl₃, dry starch basis) hydroxypropylated (8%) corn starches were extruded using a Leistritz micro‐18 co‐rotating extruder. Process variables included moisture, barrel temperature, and screw design. Differential scanning calorimetry and X‐ray diffraction studies showed the level of starch crystallinity decreased with increasing severity of extrusion conditions. Pasting properties of the extruded starches were examined using a Rapid Visco Analyser. Pasting profiles of starches extruded at different conditions displayed different hot paste viscosity and final viscosity. Increasing starch moisture content during extrusion and level of cross‐linking increased starch viscosity (P < 0.0001), whereas increasing extrusion temperature and shear decreased starch viscosity (P < 0.0001). Interactions were found between level of cross‐linking and screw design and between extrusion temperature and starch moisture content (P < 0.0001).     

Effect of Steeping Treatment on Pasting and Thermal Properties of Sorghum Starches

Chemical treatments in wet milling could improve the physico‐chemical properties of starch isolated from high‐tannin sorghums. Sorghums Chirimaugute (medium‐tannin), DC‐75 (high‐tannin), and SV2 (tannin‐free) were steeped in water, dilute HCl (0.9%, v/v), formaldehyde (0.05%, v/v), and NaOH (0.3%, w/v) solutions before wet milling and starch separation. Pasting, textural, and thermal properties of starch were determined. Steeping in NaOH resulted in starches with higher peak viscosity (PV), cool paste viscosity (CPV), and setback than when water, HCl, and formaldehyde were used. The time to PV (P_time) and PV temperature (P_temp) were markedly reduced by treatment with NaOH. NaOH could have caused a degree of pregelatinization. HCl treatment gave starches with higher P_temp and P _time, presumably due to delayed granule swelling. Gel hardness was largely determined by the starch amylase content. The low hardness of DC‐75 starch gels was slightly improved in NaOH‐treated grains. Gelatinization temperatures of sorghum starches were generally low, regardless of steeping treatment. Starch from NaOH‐treated grain generally had slightly higher gelatinization temperatures than when water, HCl, or HCHO was used. Chemical treatments during steeping of sorghum grains greatly affected starch properties. Dilute alkali steeping during wet milling could be used to improve properties of starch isolated from tannin‐containing sorghums.     

Thermal Properties of Corn Starch Extraction Intermediates by Differential Scanning Calorimetry

Thermal properties of corn starch extraction intermediates from four types of corn were studied using differential scanning calorimetry. Starch at four different stages of extraction, including a standard single-kernel starch isolation procedure and three starch extraction intermediates, was isolated from mature corn kernels of B73 and Oh43 inbreds and the mutants of waxy (wx) and amylose extender (ae) in an Oh43 background. Differences in thermal properties and moisture and protein contents of starch from the extraction stages were statistically analyzed. Most thermal properties (gelatinization and retrogradation onset temperatures, gelatinization and retrogradation ranges, gelatinization and retrogradation peak temperatures, gelatinization and retrogradation enthalpies, peak height index, and percentage of retrogradation) of starches extracted at stage 3 intermediate (a procedure that did not include a final washing step) were similar to those of starch extracted by the standard single-kernel isolation procedure. Values for gelatinization peak temperature, gelatinization enthalpy, and peak height index were different between the standard and the stage 3 intermediate. The values obtained from starches extracted at stage 3, however, were consistent and predictable, suggesting that this extraction intermediate might be used in screening programs in which many starch samples are evaluated. By using the stage 3 extraction, samples could be evaluated in three rather than four days and the procedure saved ≈0.5 hr of labor time. The other two starch extraction intermediates, which excluded filtering and washing or filtering, washing, and steeping, produced starch with thermal properties generally significantly different from starch extracted by the standard single-kernel isolation procedure.     

Optimum Steeping Process for Wet Milling of Sorghum

Pioneer 8500, a red hard sorghum hybrid, was steeped batchwise using three steeping solutions at 50°C: SO₂ solution; SO₂ solution containing 1.25% (w/w) of a commercial multiple‐enzyme preparation (Novo SP249); and SO₂ solution with the addition of 0.5% (w/w) lactic acid. Novo SP249 contained pectolytic, cellulolytic, hemicellulolytic, and proteolytic activities and small amounts of saccharolytic activities. Three SO₂ concentrations (0.1, 0.2, and 0.3% w/v) prepared by dissolving sodium bisulfite in distilled water and three steeping times (24, 36, and 48 hr) were used. Incorporation of multiple enzymes into the SO₂ resulted in an increase in starch yield with reduced protein content compared with the SO₂ solution alone. The best wet‐milling performance for sorghum resulted from the SO₂ solution containing 0.5% lactic acid; it produced the whitest starch with the highest yield and the lowest protein content. Both higher SO₂ concentration of the steeping solution and longer steeping time led to higher starch yield, lower protein content in starch, and whiter starch. However, no significant differences in starch yield, protein content in starch, and starch color occurred between SO₂ concentrations of 0.2 and 0.3% for all three steeping solutions. The optimum steeping process for wet milling of sorghum was using a 0.2% SO₂ solution with 0.5% lactic acid for 36 hr at 50°C. Under these conditions, the starch yield, protein content in starch, and L value of starch color were 60.2% (db), 0.49% (db), and 92.7, respectively, which were not significantly different from the best values from the 48‐hr steeping using the solution with 0.3% SO₂ and 0.5% lactic acid.     

Slowly Digestible Starch from Debranched Waxy Sorghum Starch: Preparation and Properties

Effects of debranching time, storage time, and storage temperature on production and structural properties of slowly digestible starch (SDS) were investigated. Waxy sorghum starch was hydrolyzed by isoamylase for various times (0–24 hr), and the variously debranched products were stored at ‐30, 1, and 30°C for 1–6 days. Optimal conditions for SDS production were isoamylase treatment for 8 hr and storage at 1°C for three days, resulting in SDS content of 27.0% in the optimum product. Microscopic observation revealed that rapidly digestible starch (RDS) and SDS were removed from the edges and surfaces of the optimum product by α‐amylase digestion. Digestion conditions that removed RDS and SDS resulted in a residue with a higher transition temperature and enthalpy than raw starch on a differential scanning calorimetric thermogram. Removal of RDS alone did not cause distinct decrements of peak temperature (T_p) and enthalpy (ΔH) compared with stored starch. The optimum SDS product showed an amorphous type of X‐ray diffractogram. Digestive removal of RDS from the optimum product gave a residue with X‐ray peaks similar to B type, which supports that it is partly crystalline. Removal of RDS and SDS gave broader peaks in the X‐ray pattern.     

Hot‐Water Solubilities and Water Sorptions of Resistant Starches at 25°C

Resistant starches (RS) were prepared from wheat starch and lintnerized wheat starch by autoclaving and cooling and by cross‐linking. Heat‐moisture treatment also was used on one sample to increase RS. The experimental resistant starches made from wheat starch contained 10–73% RS measured as Prosky dietary fiber, whereas two commercial resistant starches, Novelose 240 and 330, produced from high‐amylose maize starch, contained 58 and 40%, respectively. At 25°C in excess water, the experimental RS starches, except for the cross‐linked wheat starch, gained 3–6 times more water than the commercial RS starches, and at 95°C gained 2–4 times more. Cross‐linked RS4 wheat starch and Novelose 240 showed 95°C swelling powers and solubilities of 2 g/g and 1%, and 3 g/g and 2%, respectively. All starches showed similar water vapor sorption and desorption isotherms at 25°C and water activities (a_w) < 0.8. At a_w 0.84–0.97, the resistant starches made from wheat starch, except the cross‐linked wheat starch, showed ≈10% higher water sorption than the commercial resistant starches.     

Physicochemical Properties of Pueraria Root Starches and Their Effect on the Improvement of Buckwheat Noodle Quality

The physicochemical properties of starches from cultivated Pueraria thomsonii Benth were examined and compared with those of P. lobata (Willd.) Ohwi and other root starches, and the effect of pueraria root starches on the improvement of buckwheat noodle quality was investigated. The total content of isoflavones in P. thomsonii root starches was higher than in P. lobata root starches, and the size and uniformity of those particles displayed a significant difference. The gel stabilities of pueraria root starches were similar and more favorable than those of potato starch and sweet potato starch. For the amylose molecular properties of pueraria root starches, the λ_max and blue value index were higher than those of the potato starch and the sweet potato starch, whereas the amylose content and degree of polymerization were much lower in comparison. However, amylopectin branch lengths of pueraria root starches were shorter. Thus, pueraria root starches could improve the quality of buckwheat noodles and enhance their nutritional function. Therefore, pueraria root starches may be regarded as raw materials that influence the quality of buckwheat noodles.     

Physicochemical Properties of Common and Tartary Buckwheat Starch

Physicochemical properties of starch of three common (Fagopyrum esculentum) and three tartary (F. tataricum) buckwheat varieties from Shanxi Province, China, were compared. Starch color, especially b*, differed greatly between tartary (7.99–9.57) and common (1.97–2.42) buckwheat, indicating that removal of yellow pigments from tartary buckwheat flour may be problematic during starch isolation. Starch swelling volume in water of reference wheat starch (2.8% solids and 92.5°C) was 20.1 mL; for the three common buckwheat starches it was 27.4–28.0 mL; and for the three tartary buckwheat starches it was 26.5–30.8 mL. Peak gelatinization temperature (T_p) in water was 63.7°C for wheat starch, 66.3–68.8°C for common buckwheat and 68.8–70.8°C for tartary buckwheat. T_p of all samples was similarly delayed (by 4.0–4.8°C) by 1% NaCl. Enthalpy of gelatinization (ΔH) was higher for all six buckwheat starches than it was for wheat starch. However, one common buckwheat sample had significantly lower ΔH than the others. Starch pasting profiles, measured by a Rapid Visco-Analyzer, were characteristic and similar for all six buckwheat starches, and very different from the reference wheat starch. A comparison of pasting characteristics of common and tartary buckwheat starches to wheat starch indicated similar peak viscosity, higher hot paste viscosity, higher cool paste viscosity, smaller effect of NaCl on peak viscosity, and higher resistance to shear thinning. Texture profile analysis of starch gels showed significantly greater hardness for all buckwheat samples when compared to wheat starch.