Evaluation of Waxy Grain Sorghum for Ethanol Production

The objective of this research was to investigate the fermentation performance of waxy grain sorghum for ethanol production. Twenty‐five waxy grain sorghum varieties were evaluated with a laboratory dry‐grind procedure. Total starch and amylose contents were measured following colorimetric procedures. Total starch and amylose contents ranged from 65.4 to 76.3% and from 5.5 to 7.3%, respectively. Fermentation efficiencies were in the range of 86.0–92.2%, corresponding to ethanol yields of 2.61–3.03 gallons/bushel. The advantages of using waxy sorghums for ethanol production include easier gelatinization and low viscosity during liquefaction, higher starch and protein digestibility, higher free amino nitrogen (FAN) content, and shorter fermentation times. The results showed a strong linear relationship between FAN content and fermentation rate. Fermentation rate increased as FAN content increased, especially during the first 30 hr of fermentation (R² = 0.90). Total starch content in distillers dried grains with solubles (DDGS) was less than 1% for all waxy varieties.     

Impact of Deficit Irrigation on Maize Physical and Chemical Properties and Ethanol Yield

The objective of this research was to study the effect of irrigation levels (five levels from 102 to 457 mm of water) on the physical and chemical properties and ethanol fermentation performance of maize. Twenty maize samples with two crop rotation systems, grain sorghum–maize and maize–maize, were harvested in 2011 and evaluated at the Kansas State University Southwest Research‐Extension Center near Garden City, Kansas, under a semiarid climate. Results showed that maize kernel weight, density, and breakage susceptibility decreased as irrigation level decreased. Starch contents of maize samples grown under a low irrigation level were approximately 3.0% lower than those under a high irrigation level. Protein contents ranged from 9.24 to 11.30% and increased as irrigation level decreased. Maize flour thermal and rheological properties were analyzed by differential scanning calorimetry and the Micro Visco‐Amylo‐Graph‐U device. Starch gelatinization temperature increased significantly as irrigation level decreased, whereas starch pasting viscosity decreased as irrigation level decreased. Free amino nitrogen (FAN) was significantly affected by irrigation level: it increased as irrigation decreased. Ethanol fermentation efficiency ranged from 90.96 to 92.48% and was positively correlated with FAN during the first 32 h of fermentation (r² = 0.645). Deficit irrigation had a negative impact on ethanol yield. The maize with lower irrigation yielded about 4.0% less ethanol (44.14 mL/100 g of maize) than maize with high irrigation (45.92 mL/100 g of maize). Residual starch contents in the distillers dried grains with solubles were in a range of 0.80–1.02%. In conclusion, deficit irrigation had a significant effect on physical properties, chemical composition, ethanol yield, and fermentation efficiency of maize.     

A Rapid Protein Digestibility Assay for Identifying Highly Digestible Sorghum Lines

Protein digestibility in sorghum (Sorghum bicolor (L.) Moench) lines was determined using two standard procedures (pepsin digestibility and pH‐stat) and compared with a newly developed, rapid electrophoresis‐based screening assay. The new assay was based on the rate of α‐kafirin disappearance after pepsin digestion. α‐Kafirin, the major sorghum storage protein, makes up ≈60–70% of the total protein in the grain. In the new assay, samples were first digested with pepsin for 1 hr, and undigested proteins were then analyzed by SDS‐PAGE. The intensitizes of the undigested α‐kafirin bands were measured. Higher band intensity indicated lower protein digestibility. The new assay was significantly correlated with the standard pepsin digestibility assay (r = −0.96, n = 16) after which it was patterned. The same was true of the pH‐stat procedure (r = −0.85, n = 16). This implies that the new assay is comparable to existing procedures and can be used for screening sorghum lines for protein digestibility. Two groups consisting of high‐protein digestibility and wild‐type sorghum lines were identified when the new assay was tested on 48 sorghum lines derived from crosses of wild‐type and mutant high protein digestibility lines, indicating that the new assay was efficient in differentiating between the two groups. Advantages of the new assay over the standard procedures include considerable reduction in analysis time and sample size required for the analysis. For example, analysis time was reduced by 20% and sample size by 10% when the new assay was used as compared with the pH‐stat procedure. We estimate that ≈60 sorghum lines can be screened in a day by a single operator using the new assay.     

Evaluation of Nebraska Waxy Sorghum Hybrids for Ethanol Production

To evaluate the ethanol production performance of waxy sorghum hybrids and the effects of location and harvest year on ethanol yield, samples of four waxy sorghum hybrids collected from two Nebraska locations (Mead and Lincoln) in both 2009 and 2010 were tested for ethanol production in a dry‐grind process. No significant difference (P = 0.216) in starch contents was observed among the four hybrids, but starch contents of the hybrids were significantly affected by growth location (P = 0.0001) and harvest year (P = 0.0258). Location, hybrid, and harvest year all had significant effects on ethanol fermentation efficiency in the dry‐grind process. Lincoln sorghum samples showed higher (P = 0.022) ethanol fermentation efficiency (90.4%) than did Mead sorghum samples (90.0%). Sorghums harvested in 2010 had higher (P < 0.001) ethanol fermentation efficiency (91.1%) than those harvested in 2009 (89.3%). The 2009 sorghum flours had more amylose‐lipid complexes than the 2010 samples did, and amylose‐lipid complexes as previously reported had adverse effects on ethanol fermentation. Residual starch contents in distillers dried grains with solubles (DDGS) were significantly affected by hybrid and harvest year (P < 0.0001), but we observed no difference in protein content in DDGS from the four hybrids.     

Genetic Analysis of Kafirins and Their Phenotypic Correlations with Feed Quality Traits,In Vitro Digestibility,and Seed Weight in Grain Sorghum

Twenty‐three entries of grain sorghum (Sorghum bicolor (L.) Moench), including eight inbred lines (five males and three females) and 15 hybrids, were evaluated to determine the proportion of γ, αII, and β‐αI‐kafirins and their association with contents of crude protein, fat, and starch; protein digestibility; in vitro dry matter disappearance; and seed weight. The male lines included three normal‐seeded lines (TX2737, TX435, and P954063) and two large‐seeded lines (Eastin1 and PL‐1). Female lines consisted of three common U.S. seed parent lines (Wheatland, Redlan, and SA3042). The lines and their hybrids were grown under dryland conditions at two locations in Kansas using a randomized complete block design. The effects of genotype, location, and males were significant for all kafirins. Wide variations in composition and general combining ability (GCA) for kafirin content were noted among parent lines and hybrids, with TX2737, Eastin1, and PL1 having the largest GCA values for γ (1.37), αII (1.99), and β‐αI (2.57), respectively. Correlations among kafirins ranged from ‐0.89 to 0, whereas those of kafirins with feed quality traits, digestibility, and seed weight ranged from ‐0.45 to 0.48.     

Transgenic sorghum with altered kafirin synthesis: kafirin solubility, polymerization, and protein digestion

Transgenic sorghum (TG) lines with altered kafirin synthesis, particularly suppression of γ-kafirin synthesis, and improved protein quality have been developed. The proportion of kafirin extracted with 60% tert-butyl alcohol alone was greatly increased in the TG lines. However, the total amount of kafirin remained unchanged. Further, in the TG lines, the kafirin was much less polymerized by disulfide bonding. There was also evidence of compensatory synthesis of other kafirin proteins. Cooked protein digestibility was increased in the TG form, even after removal of interfering starch. The TG protein bodies were intermediate in appearance between the normal type and the invaginated high digestibility mutants. Hence, the increased protein digestibility of these TG lines is probably related to their lower levels of disulfide-bonded kafirin polymerization, allowing better access of proteases. This work appears to confirm that disulfide bond formation in kafirin is responsible for the reduced protein digestibility of cooked sorghum.     

Germination‐Improved Ethanol Fermentation Performance of High‐Tannin Sorghum in a Laboratory Dry‐Grind Process

A high‐tannin sorghum cultivar with 3.96% tannin content was used to study the effects of germination on its ethanol fermentation performance in a laboratory dry‐grind process. High‐tannin sorghum sample was germinated for 3 and 4 days. Original and germinated samples were analyzed for tannin, starch, protein, free amino nitrogen (FAN), and glucose content. Endosperm structures and flour pasting properties of germinated and nongerminated sorghum samples were examined using a scanning electron microscope (SEM) and rapid visco analyzer (RVA). Germination reduced tannin content from 3.96% to negligible levels. The free fermentable sugars (glucose, maltose, and maltotriose) in the germinated samples were significantly higher than those in the nongerminated control. Judged by the starch (starch plus dextrin) and free amino nitrogen contents in the mashed samples, germination improved degree of hydrolysis for starch by 13–20% and for protein by 5‐ to 10‐fold during mashing. Germination significantly shortened the required fermentation time for ethanol production by 24–36 hr, increased ethanol fermentation efficiency by 2.6–4.0%, and reduced the residual starch content in the distillers dried grain with solubles (DDGS) compared to the nongerminated control. Ethanol yield for the 3‐day germinated samples was 2.75 gallons/bushel, which was 3.1% higher than the 2.67 gallons for the nongerminated control. Ethanol yield for the 4‐day germinated sorghum was 2.63 gallons/bushel due to excessive loss of starch during germination.     

Ethanol‐Production Performance of Ozone‐Treated Tannin Grain Sorghum Flour

Ozone has been reported as being able to degrade macromolecules such as cellulose, starch, lignins, and tannins in the textile, pulping, and water‐treatment industries. Thus, we hypothesized that ozone treatment may also inactivate tannin activity and increase fermentation efficiency of tannin sorghum lines. The objective of this research was to study the physicochemical properties of ozone‐treated whole tannin grain sorghum flour and its fermentation performance in ethanol production. Results showed that the ethanol yields from ozone‐treated tannin grain sorghums were significantly higher than yields from the untreated flour. The fermentation efficiency of ozone‐treated tannin grain sorghum was approximately 90%, which was 8–14% higher than that of untreated samples at the 36th hr of fermentation. At the end of 72 hr of fermentation, the efficiencies of ozone‐treated sorghum flour were 2–5% higher than those of untreated samples. Measured tannin levels of ozone‐treated samples decreased significantly from 3.8 to 2.7%. Gel‐permeation chromatographic results indicated that both degradation and polymerization processes might have happened to starch molecules during ozone treatment. Rapid Visco Analyzer data showed that the setback of viscosity of ozone‐treated flour was lower than that of untreated flours. Distillers dried grains with solubles made from ozone‐treated sorghum were low in residual starch (<1%) and high in crude protein (≈35%). Therefore, ozonation could be a novel and useful method to improve ethanol yield and fermentation efficiency of tannin grain sorghum.     

Impact of mashing on sorghum proteins and its relationship to ethanol fermentation

Nine grain sorghum cultivars with a broad range of ethanol fermentation efficiencies were selected to characterize the changes in sorghum protein in digestibility, solubility, and microstructure during mashing and to relate those changes to ethanol fermentation quality of sorghum. Mashing reduced in vitro protein digestibility considerably, and a large amount of polymers cross-linked by disulfide bonds were developed during mashing. As a marker of cross-linking, protein digestibility of the original samples was highly related to conversion efficiency. gamma-Kafirin (%) neither correlated to ethanol yield nor conversion efficiency significantly. Solubility of proteins in an alkaline borate buffer in conjunction with SDS decreased substantially after mashing. Solubility and the SE-HPLC area of proteins extracted from mashed samples were highly correlated with ethanol fermentation. Ethanol yield increased and conversion efficiency improved notably with the increase of extracted proteins from mashed samples. SE-HPLC total area could be used as an indicator to predict ethanol fermentation. CFLSM images proved that sorghum proteins tended to form highly extended, strong web-like microstructures during mashing. The degree of protein cross-linking differed among samples, and more open microstructures were observed in samples with higher conversion efficiencies. The web-like protein matrix was found to hold not only starch granules but also some oligosaccharides or polysaccharides inside. The formation of web-like microstructures because of cross-linking reduced conversion efficiency.     

Maize Proximate Composition and Physical Properties Correlations to Dry‐Grind Ethanol Concentrations

Dry‐grind ethanol plants incur economic losses because of seasonal variations in ethanol yields. One possible cause associated with ethanol yield variability is incoming grain quality. There is little published information on factors causing variation in dry‐grind ethanol concentrations. The objective of this study was to determine relationships between rapidly measurable corn quality attributes (physical parameters and chemical composition) and dry‐grind ethanol concentrations. Corn samples obtained from a Midwestern ethanol plant were analyzed for physical quality parameters (test weight, kernel weight, true density, percent stress cracks, and moisture content) and composition (starch, protein, oil, and soluble sugars contents) and then processed with a laboratory‐scale dry‐grind procedure. There were significant (P < 0.05) variations in corn quality parameters and ethanol concentrations. Correlation coefficients were significant (P < 0.05) but low (–0.50 < r < 0.50) between starch content and final ethanol concentrations (72 h) and total soluble sugar content and ethanol concentrations at 72 and 48 h. Ethanol concentrations (at 24, 48, and 72 h) were predicted as a function of a combination of grain quality factors using multiple regression methods; however, the R² values obtained were low. Variations in ethanol concentrations were not related to physical and chemical composition quality factors. Other factors, such as structural and physiologic attributes of corn grain, need to be evaluated.     

A Novel Modified Endosperm Texture in a Mutant High‐Protein Digestibility/High‐Lysine Grain Sorghum (Sorghum bicolor (L.) Moench)

Development of high‐protein digestibility (HPD)/high‐lysine (hl) sorghum mutant germplasm with good grain quality (i.e., hard endosperm texture) has been a major research objective at Purdue University. Progress toward achieving this objective, however, has been slow due to challenges posed by a combination of genetic and environmental factors. In this article, we report on the identification of a sorghum grain phenotype with a unique modified endosperm texture that has near‐normal hardness and possesses superior nutritional quality traits of high digestibility and enhanced lysine content. These modified endosperm lines were identified among F₆ families developed from crosses between hard endosperm, normal nutritional quality sorghum lines, and improved HPD/hl sorghum mutant P721Q‐derived lines. A novel vitreous endosperm formation originated in the central portion of the kernel endosperm with opaque portions appearing both centrally and peripherally surrounding the vitreous portion. Kernels exhibiting modification showed a range of vitreous content from a slight interior section to one that filled out to the kernel periphery. Microstructure of the vitreous endosperm fraction was dramatically different from that of vitreous normal kernels in sorghum and in other cereals, in that polygonal starch granules were densely packed but without the typically associated continuous protein matrix. We speculate that, due to the lack of protein matrix, such vitreous endosperm may have more available starch for animal nutrition, and possibly have improved wet‐milling and dry‐grind ethanol processing properties. The new modified endosperm selections produce a range that approaches the density of the vitreous parent, and have lysine content and protein digestibility comparable to the HPD/hl opaque mutant parent.     

Extraction of β‐Glucan from Oat Bran in Laboratory Scale

Effects of various enzymes and extraction conditions on yield and molecular weight of β‐glucans extracted from two batches of commercial oat bran produced in Sweden are reported. Hot‐water extraction with a thermostable α‐amylase resulted in an extraction yield of ≈76% of the β‐glucans, while the high peak molecular weight was maintained (1.6 × 10⁶). A subsequent protein hydrolysis significantly reduced the peak molecular weight of β‐glucans (by pancreatin to 908 × 10³ and by papain to 56 × 10³). These results suggest that the protein hydrolyzing enzymes may not be pure enough for purifying β‐glucans. The isolation scheme consisted of removal of lipids with ethanol extraction, enzymatic digestion of starch with α‐amylase, enzymatic digestion of protein using protease, centrifugation to remove insoluble material, removal of low molecular weight components using dialysis, precipitation of β‐glucans with ethanol, and air‐drying.     

Ethanol Production from Pearl Millet Using Saccharomyces cerevisiae

Four pearl millet genotypes were tested for their potential as raw material for fuel ethanol production in this study. Ethanol fermentation was performed both in flasks on a rotary shaker and in a 5‐L bioreactor using Saccharomyces cerevisiae (ATCC 24860). For rotary‐shaker fermentation, the final ethanol yields were 8.7–16.8% (v/v) at dry mass concentrations of 20–35%, and the ethanol fermentation efficiencies were 90.0–95.6%. Ethanol fermentation efficiency at 30% dry mass on a 5‐L bioreactor reached 94.2%, which was greater than that from fermentation in the rotary shaker (92.9%). Results showed that the fermentation efficiencies of pearl millets, on a starch basis, were comparable to those of corn and grain sorghum. Because pearl millets have greater protein and lipid contents, distillers dried grains with solubles (DDGS) from pearl millets also had greater protein content and energy levels than did DDGS from corn and grain sorghum. Therefore, pearl millets could be a potential feedstock for fuel ethanol production in areas too dry to grow corn and grain sorghum.     

Extraction and Film Properties of Kafirin from Coarse Sorghum and Sorghum DDGS by Percolation

The high cost of kafirin and zein restricts their use for bioplastic and food applications. Effective, simple, and rapid kafirin/zein isolation processes are required. Here a percolation‐type aqueous ethanol solvent extraction process from coarse meals (grits) and coarse sorghum distillers dried grains and solubles (DDGS) for kafirin and zein isolation employing a low ratio of extractant to meal (2.5:1) was investigated, which is potentially applicable in the grain bioethanol industry. Postextraction filtration times were more than twice as fast using coarse meals compared with fine flours. Washing the meals prior to extraction to remove starch improved protein preparation purity to 73–85% compared with 68–72% for unwashed meals. Hence, no subsequent filtration or centrifugation step is required to clean up the kafirin/zein solution prior to solvent evaporation. With a single extraction step, kafirin/zein yields were 48% (protein basis) for DDGS and 53–70% for washed sorghum/maize meals. Cast films were used as a model bioplastic system to evaluate extracted kafirin/zein functional properties. DDGS kafirin films had rough surfaces but had the lowest water uptake and in vitro digestibility, owing to heat‐induced disulfide crosslinking during DDGS processing. Extraction by percolation using coarse meal/DDGS has potential to improve kafirin/zein viability.     

Size Distribution and Properties of Wheat Starch Granules in Relation to Crumb Grain Score of Pup‐Loaf Bread

Twelve hard winter wheat flours with protein contents of 11.8–13.6% (14% mb) were selected to investigate starch properties associated with the crumb grain score of experimentally baked pup‐loaf bread. The 12 flours were classified in four groups depending on the crumb grain scores, which ranged from 1 (questionable‐unsatisfactory) to 4 (satisfactory). Flours in groups 1, 2, 3, and 4 produced breads with pup‐loaf volumes of 910–1,035, 1,000–1,005, 950–1,025, and 955–1,010 cm³, respectively. Starches were isolated by a dough handwashing method and purified by washing to give 75–79% combined yield (dry flour basis) of prime (62–71%) and tailing (7–16%) starches. The prime starch was fractionated further into large A‐granules and small B‐granules by repeated sedimentation in aqueous slurry. All starches were assayed for weight percentage of B‐granules, swelling power (92.5°C), amylose content, and granular size distribution by quantitative digital image analysis. A positive linear correlation was found between the crumb grain scores and the A‐granule sizes (r = 0.65, P < 0.05), and a polynomial relationship (R² = 0.45, P < 0.05) occurred between the score and the weight percentage of B‐granule starch. The best crumb grain score was obtained when a flour had a weight percentage of B‐granules of 19.8–22.5%, shown by varietal effects.     

Differential responses of grain yield,grain protein,and their associated traits to nitrogen supply in soft red winter wheat

Increased application of nitrogen fertilizers has significantly raised grain yield and protein concentration in wheat. However, only 30–50% of applied fertilizer nitrogen are usually utilized by the plant. In this study, four soft red winter wheat genotypes (Triticum aestivum L., IL07‐4415, MD05W10208‐11‐8, OH06‐150‐57 and Sisson) were grown under three different nitrogen regimes (high, medium, and low) in a greenhouse, and grain yield, grain protein concentration, nitrogen use efficiency (NUE) and their associated traits were evaluated. Among the four genotypes, a high‐yielding cultivar, Sisson, exhibited superior performance in terms of grain weight plant⁻¹ and NUE for yield (NUEY) at low nitrogen due to maintained grain number spike⁻¹ and harvest index. Significant yield losses due to nitrogen limitation were attributable to reduced spike number plant⁻¹ and grain number spike⁻¹ in the other genotypes. Interestingly, a linear relationship between NUEY and NUE for grain protein (NUEP) was detected at high (R ² = 0.67) and low (R ² = 0.42) nitrogen; both of these traits were positively correlated with grain number spike⁻¹, 1000‐seed weight, and harvest index under nitrogen‐limited conditions (R ² = 0.35–0.48). These results suggest that simultaneous improvement of NUEY and NUEP could be achieved through the selection of the three yield components (grain number spike⁻¹, 1000‐seed weight, and harvest index) at low nitrogen.     

Relationship Between Wheat (Triticum aestivum L.) Grain Hardness and Wet‐Milling Quality

Grain hardness variation has large effects on many different end‐use properties of wheat (Triticum aestivum). The Hardness (Ha) locus consisting of the Puroindoline a and b genes (Pina and Pinb) controls the majority of grain hardness variation. Starch production is a growing end‐use of wheat. The objective of this study was to estimate the differences in starch yield due to natural and transgenically conditioned grain hardness differences. To accomplish this goal, a small‐scale wet‐milling protocol was used to characterize the wet‐milling properties of two independent groups of isogenic materials varying in grain hardness and in Pin expression level. The first group of lines consisted of hard/soft near‐isogenic lines created in cultivars Falcon or Gamenya in which lines carried either the Pina‐D1a (functional) or the Pina‐D1b (null) alleles of Pina. The second group of lines consisted of Pina, Pinb, or Pina and Pinb overexpressing lines created in Hi‐Line, a hard red spring wheat. Soft near‐isogenic lines had higher starch extractability than the hard Pina null counterparts. This difference in starch extractability was more pronounced between Hi‐Line and its transgenic isolines, with highest levels of extractable starch observed in the transgenic isoline with intermediate grain texture. The results demonstrate that the Ha locus and puroindoline expression are both linked to wet‐milling starch yield and that selection for increased Ha function increases starch yield through the enhanced separation of starch granules and the protein matrix during wet milling.     

Distribution of β‐Glucan in the Grain of Hull‐less Barley

Nine hull‐less barley (HB) containing waxy (0–7% amylose), normal (≈25% amylose), or high amylose (≈42% amylose) starch with normal or fractured granule make‐up and 4–9% (1→3)(1→4)‐β‐d ‐glucans (β‐glucan) were pearled to remove 70% of the original grain weight in 10% intervals. The pearled fractions were analyzed for β‐glucan distribution within HB grain. Protein content of the pearled fractions indicated that the three outermost fractions contained pericarp and testa, aleurone, and subaleurone tissues, respectively. For all HB, β‐glucan and acid‐extract viscosity were very low in the outermost 20% of the kernel. For low β‐glucan HB, β‐glucan content was the greatest in the subaleurone region and declined slightly toward inner layers. For high β‐glucan HB, however, more than 80% of grain β‐glucan was distributed more evenly throughout the endosperm. Acid extract viscosity was significantly (P < 0.01) correlated with total (r = 0.75) and soluble (r = 0.87) β‐glucan content throughout the kernel of all HB. Growing conditions, location and year, had significant effects on the concentration of protein, starch and β‐glucan. However, protein, starch, and β‐glucan distribution patterns were not affected by growing conditions. The difference in β‐glucan distribution between low and high β‐glucan HB may explain the difference in milling performance of HB with low or high β‐glucan.     

Factors Impacting Ethanol Production from Grain Sorghum in the Dry-Grind Process

The goal of this research is to understand the key factors affecting ethanol production from grain sorghum. Seventy genotypes and elite hybrids with a range of chemical compositions and physical properties selected from ≈1,200 sorghum lines were evaluated for ethanol production and were used to study the relationships of composition, grain structure, and physical features that affect ethanol yield and fermentation efficiency. Variations of 22% in ethanol yield and 9% in fermentation efficiency were observed among the 70 sorghum samples. Genotypes with high and low conversion efficiencies were associated with attributes that may be manipulated to improve fermentation efficiency. Major characteristics of the elite sorghum genotypes for ethanol production by the dry-grind method include high starch content, rapid liquefaction, low viscosity during liquefaction, high fermentation speed, and high fermentation efficiency. Major factors adversely affecting the bioconversion process are tannin content, low protein digestibility, high mash viscosity, and an elevated concentration of amylose-lipid complex in the mash.     

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.