Influence of Stenocarpella maydis Infected Corn on the Composition of Corn Kernel and Its Conversion into Ethanol

Widespread epidemics of Stenocarpella ear rot (formerly Diplodia ear rot) have occurred throughout the central U.S. Corn Belt in recent years, but the influence of S. maydis infected grain on corn ethanol production is unknown. In this study, S. maydis infected ears of variety Heritage 4646 were hand‐harvested in 2010 from a production field in central Illinois and segregated into one of five levels of ear rot severity based upon visual symptoms. The concentration of ergosterol, a sterol produced by fungi but not plants, was observed to increase with the severity of ear rot (127–306.5 μg/g), and none was detected in the control corn. Corn test weight declined with progression of the disease and was 42.6% lower for the most severely rotted grain from ears infected early in their development. Accompanying changes in composition were also apparent. Crude fat and oil contents decreased (from 4.7 to 1.5%) and fiber increased (from 6.6 to 9.6%), but starch content remained largely invariant. Oil composition also varied among the infected samples. Control and infected corn samples were subjected to ethanol fermentation with a laboratory‐scale corn dry‐grind ethanol process. Ethanol yields for control and infected samples were similar on an equivalent weight basis (2.77–2.85 gal/bu). In comparison with the control, S. maydis infection altered the distillers dried grains with solubles (DDGS) properties, wherein the crude protein was significantly higher and oil significantly reduced, and ash, fiber, and yield per ton were not significantly different. Based upon these results, we conclude that Stenocarpella ear rot has the potential to affect DDGS composition but not ethanol yield on an equivalent weight basis.     

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.     

Comparison of Modified Dry‐Grind Corn Processes for Fermentation Characteristics and DDGS Composition

Three different modified dry‐grind corn processes, quick germ (QG), quick germ and quick fiber (QGQF), and enzymatic milling (E‐Mill) were compared with the conventional dry‐grind corn process for fermentation characteristics and distillers dried grains with solubles (DDGS) composition. Significant effects were observed on fermentation characteristics and DDGS composition with these modified dry‐grind processes. The QG, QGQF, and E‐Mill processes increased ethanol concentration by 8–27% relative to the conventional dry‐grind process. These process modifications reduced the fiber content of DDGS from 11 to 2% and increased the protein content of DDGS from 28 to 58%.     

Comparison of Enzymatic (E‐Mill) and Conventional Dry‐Grind Corn Processes Using a Granular Starch Hydrolyzing Enzyme

A new low temperature liquefaction and saccharification enzyme STARGEN 001 (Genencor International, Palo Alto, CA) with high granular starch hydrolyzing activity was used in enzymatic dry‐grind corn process to improve recovery of germ and pericarp fiber before fermentation. Enzymatic dry‐grind corn process was compared with conventional dry‐grind corn process using STARGEN 001 with same process parameters of dry solid content, pH, temperature, enzyme and yeast usage, and time. Sugar, ethanol, glycerol and organic acid profiles, fermentation rate, ethanol and coproducts yields were investigated. Final ethanol concentration of enzymatic dry‐grind corn process was 15.5 ± 0.2% (v/v), which was 9.2% higher than conventional process. Fermentation rate was also higher for enzymatic dry‐grind corn process. Ethanol yields of enzymatic and conventional dry‐grind corn processes were 0.395 ± 0.006 and 0.417 ± 0.002 L/kg (2.65 ± 0.04 and 2.80 ± 0.01 gal/bu), respectively. Three additional coproducts, germ 8.0 ± 0.4% (db), pericarp fiber 7.7 ± 0.4% (db), and endosperm fiber 5.2 ± 0.6% (db) were produced in addition to DDGS with enzymatic dry‐grind corn process. DDGS generated from enzymatic dry‐grind corn process was 66% less than conventional process.     

Use of Pigmented Maize in Both Conventional Dry‐Grind and Modified Processes Using Granular Starch Hydrolyzing Enzyme

In the dry‐grind ethanol process, distillers dried grains with solubles (DDGS) is the main coproduct, which is primarily used as an ingredient in ruminant animal diets. Increasing the value of DDGS will improve the profitability of the dry‐grind ethanol process. One way to increase DDGS value is to use pigmented maize as the feedstock for ethanol production. Pigmented maize is rich in anthocyanin content, and the anthocyanin imparts red, blue, and purple color to the grain. It is reported that anthocyanin would be absorbed by yeast cell walls during the fermentation process. The effects of anthocyanin on fermentation characteristics in the dry‐grind process are not known. In this study, the effects of anthocyanin in conventional (conventional starch hydrolyzing enzymes) and modified (granular starch hydrolyzing enzymes [GSHE]) dry‐grind processes were evaluated. The modified process using GSHE replaced high‐temperature liquefaction. The ethanol conversion efficiencies of pigmented maize were comparable to that of yellow dent corn in both conventional (78.4 ± 0.5% for blue maize, 74.3 ± 0.4% for red maize, 81.2 ± 1.0% for purple maize, and 75.1 ± 0.2% for yellow dent corn) and modified dry‐grind processes using GSHE (83.8 ± 0.8% for blue maize, 81.1 ± 0.3% for red maize, 93.5 ± 0.8% for purple maize, and 85.6 ± 0.1% for yellow dent corn). Total anthocyanin content in DDGS from the modified process was 1.4, 1.9, and 2.4 times of that from the conventional process for purple, red, and blue maize samples, respectively. These results indicated that pigmented maize rich in anthocyanin did not negatively affect the fermentation characteristics of the dry‐grind process and that there was a potential to use pigmented maize in the dry‐grind process, especially when using GSHE.     

Pericarp Fiber Separation from Corn Flour Using Sieving and Air Classification

In the dry‐grind process, starch in ground corn (flour) is converted to ethanol, and the remaining corn components (protein, fat, fiber, and ash) form a coproduct called distillers dried grains with solubles (DDGS). Fiber separation from corn flour would produce fiber as an additional coproduct that could be used as combustion fuel, cattle feed, and as feedstock for producing valuable products such as “cellulosic” ethanol, corn fiber gum, oligosaccharides, phytosterols, and polyols. Fiber is not fermented in the dry‐grind corn process. Its separation before fermentation would increase ethanol productivity in the fermenter. Recently, we showed that the elusieve process, a combination of sieving and elutriation (air flow), was effective in fiber separation from DDGS. In this study, we evaluated the elusieve process for separating pericarp fiber from corn flour. Corn flour remaining after fiber separation was termed “enhanced corn flour”. Of the total weight of corn flour, 3.8% was obtained as fiber and 96.2% was obtained as enhanced corn flour. Neutral detergent fiber (NDF) of corn flour, fiber, and enhanced corn flour (dry basis) were 9.0, 61.5, and 5.7%, respectively. Starch content of corn flour, fiber, and enhanced corn flour (dry basis) were 68.8, 23.5, and 71.3%, respectively. Final ethanol concentration from enhanced corn flour (14.12% v/v) was marginally higher than corn flour (13.72% v/v). No difference in ethanol yields from corn flour and enhanced corn flour was observed. The combination of sieving and air classification can be used to separate pericarp fiber from corn flour. The economics of fiber separation from corn flour using the elusieve process would be governed by the production of valuable products from fiber and the revenues generated from the valuable products.     

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.     

Comparison of Raw Starch Hydrolyzing Enzyme with Conventional Liquefaction and Saccharification Enzymes in Dry‐Grind Corn Processing

In a conventional dry‐grind corn process, starch is converted into dextrins using liquefaction enzymes at high temperatures (90–120°C) during a liquefaction step. Dextrins are hydrolyzed into sugars using saccharification enzymes during a simultaneous saccharification and fermentation (SSF) step. Recently, a raw starch hydrolyzing enzyme (RSH), Stargen 001, was developed that converts starch into dextrins at low temperatures (<48°C) and hydrolyzes dextrins into sugars during SSF. In this study, a dry‐grind corn process using RSH enzyme was compared with two combinations (DG1 and DG2) of commercial liquefaction and saccharification enzymes. Dry‐grind corn processes for all enzyme treatments were performed at the same process conditions except for the liquefaction step. For RSH and DG1 and DG2 treatments, ethanol concentrations at 72 hr of fermentation were 14.1–14.2% (v/v). All three enzyme treatments resulted in comparable ethanol conversion efficiencies, ethanol yields, and DDGS yields. Sugar profiles for the RSH treatment were different from DG1 and DG2 treatments, especially for glucose. During SSF, the highest glucose concentration for RSH treatment was 7% (w/v), whereas for DG1 and DG2 treatments, glucose concentrations had maximum of 19% (w/v). Glycerol concentrations were 0.5% (w/v) for RSH treatment and 0.8% (w/v) for DG1 and DG2 treatments.     

Dry‐Grind Processing of Corn with Endogenous Liquefaction Enzymes

An amylase corn has been developed that produces an α‐amylase enzyme that is activated in the presence of water at elevated temperatures (>70°C). Amylase corn in the dry‐grind process was evaluated and compared with the performance of exogenous amylases used in dry‐grind processing. Amylase corn (1–10% by weight) was added to dent corn (of the same genetic background as the amylase corn) as treatments and resulting samples were evaluated for dry‐grind ethanol fermentation using 150‐g and 3‐kg laboratory procedures. Ethanol concentrations during fermentation were compared with the control treatment (0% amylase corn addition or 100% dent corn) which was processed with a conventional amount of exogenous α‐amylase enzymes used in the dry‐grind corn process. The 1% amylase corn treatment (adding 1% amylase corn to dent corn) was sufficient to liquefy starch into dextrins. Following fermentation, ethanol concentrations from the 1% amylase corn treatment were similar to that of the control. Peak and breakdown viscosities of liquefied slurries for all amylase corn treatments were significantly higher than the control treatment. In contrast, final viscosities of liquefied slurries for all amylase corn treatments were lower than those of the control. Protein, fat, ash, and crude fiber contents of DDGS samples from the 3% amylase corn treatment and control were similar.     

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.     

Separation of Fiber from Distillers Dried Grains with Solubles (DDGS) Using Sieving and Elutriation

A process was developed to separate fiber from distillers dried grains with solubles (DDGS) in a dry‐grind corn process. Separation of fiber from DDGS would provide two valuable coproducts: 1) DDGS with reduced fiber, increased fat, and increased protein contents; and 2) fiber. The process, called elusieve process, used two separation methods, sieving and elutriation, to separate the fiber. Material carried by air to the top of the elutriation column was called the lighter fraction and material that settled to the bottom of the column was called the heavier fraction. We evaluated the compositions of fractions produced from sieving and elutriation. Two commercial samples of DDGS were obtained from two dry‐grind corn plants. Sieving over four screens (869, 582, 447, and 234 μm openings) created five size categories. The two smallest size categories contained >40% (w/w) of the original DDGS and had reduced fiber and increased protein and fat contents relative to the original DDGS. Elutriation of the remaining three size categories increased protein and fat contents and reduced fiber contents in the heavier fractions. Elutriation at air velocities of 1.59–5.24 m/sec increased the protein content of the heavier fraction by 13–41% and increased the fat content of the heavier fraction by 4–127% compared with the bulk fractions of each size category. This process was effective in separating fiber from both DDGS samples evaluated. Elusieve process does not require changes in the existing dry‐grind process and can be implemented at the end of the dry‐grind process.     

Analysis of Heat Transfer Fouling by Dry‐Grind Maize Thin Stillage Using an Annular Fouling Apparatus

In dry‐grind processing to produce ethanol from corn, unfermented solids are removed from ethanol by distillation and dried to produce distillers dried grains with solubles (DDGS), an animal food. Fouling of thin stillage evaporators has been identified as an important energy consumption issue in dry‐grind facilities. Using an annular fouling apparatus, four batches of thin stillage were analyzed to determine repeatability of fouling rate and induction period measurements. Dry solids, protein and ash concentrations, and pH were correlated to fouling rate and induction period to determine how variation in thin stillage from the same dry‐grind facility affects these fouling parameters. Effects of increasing Reynolds number (Re) in the laminar region on fouling rate, induction period, and fouling deposit protein and ash concentrations were also determined. Repeatability of fouling rate measurements was similar to other studies (CV < 7.0%) but repeatability of induction period measurements was high relative to other studies (CV < 88.7%). Fouling rate increased with increasing dry solids concentration. Thin stillage at Re = 440 had shorter induction periods and greater fouling rates than at Re = 880. Fouling deposits collected from Re = 440 tests had similar protein concentrations and lower ash concentrations compared with deposits from Re = 880 tests.     

Fate of Bt Protein and Influence of Corn Hybrid on Ethanol Production

Corn hybrids were compared to determine the fate of recombinant Bt protein (CRY1Ab from Bacillus thuringiensis) in coproducts from dry grind and wet‐milled corn during production of fuel ethanol. Two pairs of Bt and non‐Bt hybrids were wet milled, and each fraction was examined for the presence of the Bt protein. Bt protein was found in the germ, gluten, and fiber fractions of Bt hybrids. In addition, one set of Bt and non‐Bt hybrids were treated by the dry‐grind ethanol process and Bt protein was monitored during each step of the process. The Bt protein was not detected after liquefaction. Subsequent experiments determined that the Bt protein is rapidly denatured at liquefaction temperatures. Finally, five hybrids were compared for ethanol yield after dry grinding. Analysis of fermentation data with an F‐test revealed the percent of total starch available for conversion into ethanol varied significantly among the hybrids (P < 0.002), indicating ethanol yield is not exclusively dependent on starch content. No difference, however, was observed between Bt and non‐Bt corn hybrids for either ethanol productivity or yield.     

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.     

Effects of Selected Harvest Moistures and Frozen Storage Times on Selected Yellow Dent Corn: Wet‐Milling Yields and Starch Pasting Properties

The effects of harvest moistures and frozen storage times on corn wet‐milling yields and the pasting properties of the resulting starch were studied. Pioneer hybrid P‐0916‐XR harvested at three moisture contents (49, 35, and 21% wb) were stored frozen for three days or for five months, followed by wet‐milling. The pasting properties of the resulting starch were evaluated with a Rapid Visco Analyzer. The yields of starch and germ increased by 1.2 and 1.9 percentage points, respectively, when harvest moisture decreased from 49 to 21% wb, whereas the yields of steep water solids, total fiber, and gluten decreased by 2.1, 0.7, and 0.6 percentage points, respectively. The frozen corn had lower coarse fiber yields but higher cellular fiber yields. The starch pasting properties showed that peak and breakdown viscosities decreased by 8% (3,824 ± 36 versus 3,520 ± 38 cP) and 13% (2,336 ± 47 versus 2,029 ± 60 cP), respectively, when harvest moisture decreased from 49 to 21% wb, whereas peak time increased by 5% (6.32 ± 0.06 versus 6.62 ± 0.07 min). The setback and final viscosities of starch from long‐term frozen storage (five months) were 14% (1,574 ± 65 versus 1,828 ± 79 cP) and 8% (3,063 ± 27 versus 3,317 ± 101 cP) lower, respectively, than that from control (unfrozen) corn.     

Methods for Characterization of Residual Starch in Distiller's Dried Grains with Solubles (DDGS)

The objective of this study was to establish methods for determining the content and components of residual starch in distiller's dried grains with solubles (DDGS), a coproduct from dry‐grind corn ethanol production. Four DDGS prepared in our laboratory and one DDGS obtained from a commercial ethanol manufacturer were used for the study. Quantitative analysis of total residual sugar (TRS) in DDGS was performed by determining d ‐glucose produced by enzymatic hydrolysis of oligosaccharides and residual starch remaining in hexane‐defatted DDGS after being dispersed in 90% DMSO. The TRS consisted of free glucose, oligosaccharides, and residual starch. The commercial manufacturer's DDGS contained more TRS (15.8%, w/w db) than the laboratory‐processed DDGS (2.4–2.9%, w/w db). The content of residual starch remaining in the commercial DDGS (5.5% w/w db) was also larger than the laboratory‐processed DDGS (1.9–2.5% w/w db). Analyses of molecular weight distribution showed that the residual starch in DDGS consisted of short‐chain amylose and amylopectin, respectively, as the major and minor components. The short‐chain amylose molecules constituted 86.5–94.1% of the residual starch. The major population of the short‐chain amyloses had an average degree of polymerization (DP) of 85, closely resembling the length of enzyme‐resistant fragments of amylose‐lipid complexes.     

Effect of Single‐Screw Extruder Die Temperature,Amount of Distillers' Dried Grains With Solubles (DDGS), and Initial Moisture Content on Extrudates

Corn distillers' dried grains with solubles (DDGS) was extruded with corn meal in a pilot plant single‐screw extruder at different extruder die temperatures (100, 120, and 150°C), levels of DDGS (0, 10, 20, and 30%) and initial moisture contents (11, 15, and 20% wb). In general, there was a decrease in water absorption index (WAI), water solubility index (WSI), radial expansion, and L* value with an increase in DDGS level, whereas a* value and bulk density increased. Increase in extruder die temperature resulted in an increase in WSI and WAI but a decrease in L* and bulk density. Peak load was highest at 30% DDGS as compared with 0, 10, and 20% DDGS extrudates. Die temperature of 120°C and initial moisture content of 20% resulted in least peak load. The a* value remained unaffected by changes in extruder die temperature. Radial expansion was highest at extruder die temperature of 120°C. Maximum WAI, WSI, radial expansion, and L* value were obtained at 15% initial moisture content. An increase in initial moisture content, in general, decreased L* value and bulk density but increased a* value of extrudates.     

Effects of Protease and Urea on a Granular Starch Hydrolyzing Process for Corn Ethanol Production

The dry grind process using granular starch hydrolyzing enzymes (GSHE) saves energy. The amount of GSHE used is an important factor affecting dry grind process economics. Proteases can weaken protein matrix to aid starch release and may reduce GSHE doses. Two specific proteases, an exoprotease and an endoprotease, were evaluated in the dry grind process using GSHE (GSH process). The effect of protease and urea addition on GSH process was also evaluated. Addition of these proteases resulted in higher ethanol concentrations (mean increase of 0.3–1.8 v/v) and lower distillers' dried grains with solubles (DDGS) yields (mean decrease of 1.3–8.0% db) compared with the control (no protease addition). As protease levels and GSHE increased, ethanol concentrations increased and DDGS yields decreased. Protease addition reduced the required GSHE dose. Final mean ethanol concentrations without urea (15.2% v/v) were higher than with urea (15.0% v/v) in GSH process across all protease treatments.     

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.     

Bulk Density of Long- and Medium-Grain Rice Varieties as Affected by Harvest and Conditioned Moisture Contents

The effect of harvest and conditioned moisture contents (MC) on bulk density (BD) of rough, brown, and white rice was determined for several long- and medium-grain rice varieties harvested from different Arkansas locations. The results indicated that harvest MC (HMC) significantly affected the BD of freshly harvested and conditioned rough rice. Higher HMC levels resulted in lower BD levels for rough rice over the conditioned MC range from 11 to 29% (wb). There was a strong linear relationship between rough rice BD and conditioned MC for a given HMC. The harvest location had a more pronounced effect on rough rice BD of long-grain varieties than of medium-grain varieties. The BD of brown and white rice was less dependent on harvest location and conditioned MC than the BD of rough rice. Rice harvested at low MC level (≈13%) gave a higher BD for both brown and white rice than that harvested at higher MC levels. Medium-grain varieties showed a higher BD for brown and white rice than did long-grain varieties.