Laboratory Yields and Process Stream Compositions from E‐Mill and Dry‐Grind Corn Processes Using a Granular Starch Hydrolyzing Enzyme

In dry‐grind corn processing, the whole kernel is fermented to produce ethanol and distillers dried grains with solubles (DDGS); the E‐Mill process was developed to generate coproducts in addition to DDGS. Compositions of thin stillage and wet grains obtained from the E‐Mill process will be different from the dry‐grind process. Knowledge of thin stillage compositions will provide information to improve coproducts from both processes. Laboratory dry‐grind and E‐Mill processes that used granular starch hydrolyzing enzymes (GSHE) were compared and process yields determined. Two methods, centrifugation and screening, were used to produce thin stillage and wet grains from the laboratory processes. Compositions of process streams were determined. In the dry‐grind process using GSHE, solids contents of beer, whole stillage, and wet grains were higher compared to the same fractions from the E‐Mill process using GSHE. Solids contents of mash for both processes were similar. Total solids, soluble solids, and ash contents of thin stillage were similar for the two processes. Fat content of thin stillage from E‐Mill was lower than that from the dry‐grind process; protein content of E‐Mill thin stillage was higher than that from dry‐grind thin stillage. Removal of germ and fiber before fermentation changed composition of thin stillage from the E‐Mill process. The screening method produced higher thin stillage and lower wet grains yields than using a centrifugation method. The screening method was less time consuming but resulted in limited wet grains material for additional analyses or processing. The centrifugation method of thin stillage separation removed more solids from thin stillage than the screening method.     

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.     

Effect of Corn Oil on Thin Stillage Evaporators

Removal of the germ at the front end of the dry-grind ethanol process using the Quick Germ process reduces the amount of oil in thin stillage. Thin stillage with 4–6% solids is dewatered to 25–30% solids by evaporation. Thin stillage evaporators in a dry-grind ethanol plant foul and have to be periodically taken down for maintenance and cleaning. Fouling caused by thin stillage containing different amounts of oil was studied using an annular fouling probe. It was determined that the rate of fouling in a drygrind ethanol plant is three times higher when compared with that in a wet-milling ethanol plant. The addition of oil to wet-milled thin stillage significantly affected the rate of fouling. Fouling resistance increased with an increase in oil concentration for wet-milled thin stillage up to a concentration of 1.41%. At a concentration of 1.47%, the rate of fouling decreased. As the concentration of oil increased in dry-grind ethanol thin stillage, the rate of fouling decreased. These results suggest that the Quick Germ process will reduce the rate of heat transfer equipment fouling in a drygrind ethanol plant, which will decease capital costs and maintenance costs.     

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

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

Effect of pH on Fouling Characteristics and Deposit Compositions in Dry‐Grind Thin Stillage

Dry‐grind corn processing facilities produce ethanol, carbon dioxide, and distillers dried grains with solubles (DDGS). To produce DDGS, dry‐grind corn processors concentrate thin stillage in multieffect evaporators. Concentration of thin stillage uses large amounts of energy, and efficient operation is important for long‐term economic stability of the industry. Little data are available on fouling of evaporators during thin stillage concentration. We evaluated how thin stillage pH and acid type used during pH adjustment affected fouling as measured by induction period, fouling rate, and deposit composition. Using an annular fouling apparatus, fouling tests were conducted at pH 3.5, 4.0, and 4.5. In a second experiment, we used two types of acid, HCl or H₂SO₄, to adjust thin stillage to pH 3.5. Induction periods were shorter at pH 3.5 than at pH 4.0 or 4.5. As pH increased, fouling deposit protein decreased and ash increased. Concentrations of most elements, including P, Ca, Mg, Mn, and K, increased with an increase in pH. Phosphorus was the most abundant mineral element in fouling deposits. Induction periods were similar for the two acids. Thin stillage pH has an influence on deposit concentration, fouling rate, and induction period.     

Effect of the corn breaking method on oil distribution between stillage phases of dry-grind corn ethanol production

The majority of fuel ethanol in the United States is produced by using the dry-grind corn ethanol process. The corn oil that is contained in the coproduct, distillers' dried grains with solubles (DDGS), can be recovered for use as a biodiesel feedstock. Oil removal will also improve the feed quality of DDGS. The most economical way to remove oil is considered to be at the centrifugation step for separating thin stillage (liquid) from coarse solids after distilling the ethanol. The more oil there is in the liquid, the more it can be recovered by centrifugation. Therefore, we studied the effects of corn preparation and grinding methods on oil distribution between liquid and solid phases. Grinding the corn to three different particle sizes, flaking, flaking and grinding, and flaking and extruding were used to break up the corn kernel before fermentation, and their effects on oil distribution between the liquid and solid phases were examined by simulating an industrial decanter centrifuge. Total oil contents were measured in the liquid and solids after centrifugation. Dry matter yield and oil partitioning in the thin stillage were highly positively correlated. Flaking slightly reduced bound fat. The flaked and then extruded corn meal released the highest amount of free oil, about 25% compared to 7% for the average of the other treatments. The freed oil from flaking, however, became nonextractable after the flaked corn was ground. Fine grinding alone had little effect on oil partitioning.     

Effect of Aflatoxin B1 on Dry‐Grind Ethanol Process

Aflatoxins, like all mycotoxins, are toxic fungal metabolites that can have adverse health effects on animals and human beings. Aflatoxins are a major concern for the dry‐grind corn processing industry as it is believed that aflatoxins affect yeast and reduce its efficacy in producing ethanol. In the present study, aflatoxin B1 (100, 200, 350, or 775 ppb) was added to mycotoxin‐free corn and laboratory‐scale fermentations were conducted. No effect of aflatoxin B1 was observed on the fermentation rates or final ethanol concentrations. Mean ethanol concentration in the fermenter was 14.01–14.51% (v/v) at 60 hr for all the treatments. In the dry‐grind ethanol process, 55% of aflatoxin B1 was detected in wet grains and 45% in thin stillage.     

Hydrolysis and Fermentation of Pericarp and Endosperm Fibers Recovered from Enzymatic Corn Dry‐Grind Process

A modified dry‐grind corn process has been developed that allows recovery of both pericarp and endosperm fibers as coproducts at the front end of the process before fermentation. The modified process is called enzymatic milling (E‐Mill) dry‐grind process. In a conventional dry‐grind corn process, only the starch component of the corn kernel is converted into ethanol. Additional ethanol can be produced from corn if the fiber component can also be converted into ethanol. In this study, pericarp and endosperm fibers recovered in the E‐Mill dry‐grind process were evaluated as a potential ethanol feedstock. Both fractions were tested for fermentability and potential ethanol yield. Total ethanol yield recovered from corn by fermenting starch, pericarp, and endosperm fibers was also determined. Results show that endosperm fiber produced 20.5% more ethanol than pericarp fiber on a g/100 g of fiber basis. Total ethanol yield obtained by fermenting starch and both fiber fractions was 0.370 L/kg compared with ethanol yield of 0.334 L/kg obtained by fermenting starch alone.     

Effects of Corn Sample,Mill Type,and Particle Size on Corn Curl and Pet Food Extrudates

The effects of corn sample, grinder type, and particle size of ground corn on the extrusion of corn curls and pet food were studied. Extrusion runs were conducted using a twin-screw extruder. Properties of corn curl and pet food extrudates were affected significantly by corn samples obtained from different parts of the country (Nebraska, Illinois, and Texas), even though grinding and extrusion parameters were held constant. The type of grinder used to grind the corn had an effect on extrusion properties. The volumetric expansion index (VEI) of extrudate from pin-milled samples was lower than that of extrudate from the same corn ground in a hammer mill or roller mill. Small particle size, obtained by grinding corn in a hammer mill with different screen sizes, produced extrudate with a significantly higher VEI than extrudate from coarse- or medium-sized particles.     

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.     

Development of an Ethanol Yield Procedure for Dry‐Grind Corn Processing

In 2008, the United States produced ethanol at a rate of 39.5 billion L/year; an additional 8.5 billion L/year capacity was under construction. Kernel composition and physical properties are not correlated with ethanol yield. A procedure that measured the potential of hybrids to produce ethanol would benefit corn seed companies, corn producers, and ethanol processors. The objective was to develop a laboratory procedure to measure ethanol yield from corn samples and evaluate the developed procedure for accuracy and precision. To determine parameters for routine analyses, effects of mill type, dry solids, and yeast addition were investigated separately followed by effects of fermentation time (T_f), glucoamylase dose, and yeast addition. Measurement of ethanol using HPLC and gravimetric (change in weight due to CO₂ loss) methods were compared. Using the procedure developed, ethanol yields for five diverse hybrids (dent, waxy, white, high oil, and high amylose) were measured. Effects of mill type, dry solids, T_f, glucoamylase dose, and yeast addition were significant (P < 0.05). The gravimetric method estimated higher yields (428 ± 10 L/tonne) than HPLC (405 ± 15 L/tonne) and had a higher level of precision. Both methods had coefficients of variations of <4% and gave similar conclusions. In the final procedure, we used corn (25 g/batch) liquefied with α‐amylase (60 min at 90°C) in 75 mL of distilled water. Simultaneous saccharification and fermentation was used (64 hr at 32°C) with glucoamylase and yeast. Gravimetric and HPLC methods measured differences in ethanol yield for the five hybrids (158–435 L/tonne). The method is suitable for routine testing of ethanol yield potential and as a reference method for verifying more rapid measurement techniques.     

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.     

Enzymatic Process for Corn Dry‐Grind High‐Solids Fermentation

A modified dry‐grind process that combined the use of conventional amylases (glucoamylase [GA]), phytase, and granular starch hydrolyzing enzymes (GSHE) to achieve low liquefaction viscosities and low glucose concentrations during simultaneous saccharification and fermentation (SSF) with a high slurry solids content (>33% w/w) was developed. Doses of GSHE and GA were optimized for the modified process. At 35% solids content, the modified process had 80% lower slurry viscosity, 24% lower peak glucose concentration, 7.5% higher final ethanol concentration, and 51% higher fermentation rate compared with the conventional dry‐grind process. At 40% solids content, the modified process had lower viscosities, lower peak and residual glucose concentrations, and higher ethanol concentrations than the conventional process; however, the results were in contrast to those for 35% solids content. At 40% solids content, SSF did not run to completion for conventional or modified processes, and more than 2.5% w/v of residual glucose was left in the fermentation broth. Final ethanol concentration achieved with the modified process at 40% solids content was 19.5% v/v, similar to the ethanol concentration achieved with the modified process at 35% solids content. At 35% slurry solids content, a GSHE level of 1.25 μL/g db of corn and a GA level of 0.25 μL/g db of corn were selected as optimum enzyme doses for the modified process.     

Effects of Alkali Debranning,Roller Mill Cracking and Gap Setting,and Alkali Steeping Conditions on Milling Yields from a Dent Corn Hybrid

Starch yield was significantly affected by all three main unit operations in alkali wet‐milling (debranning, roller milling, and steeping). The conditions for the three unit operations were studied using a single hybrid. Studies on debranning showed that optimal separation between pericarp and corn endosperm was obtained when corn was soaked in a 1.5–2% NaOH solution at 85°C for 5 min. Passing debranned corn through smooth roller mill once or twice did not affect the product yields, but passing the corn through the roller mill three times decreased the germ yield because of a large amount of broken germ. A 62% higher processing rate could be achieved when passing corn through the mill twice than by passing it through the mill once. The gap should be set at 2.0 mm when passing corn through the mill once, and it should be set at 3.5 mm for the first pass and 2.0 mm for the second pass when passing corn through the mill twice. Starch yield was more sensitive to NaOH concentration and steep temperature than to steep time. The highest starch yield was obtained when steeping corn in 0.5% NaOH for 1 hr at 45°C.     

Quantification of Physical and Chemical Properties,and Identification of Potentially Valuable Components from Fuel Ethanol Process Streams

Wider exploration of ethanol coproduct uses is necessary as the ethanol industry continues to face challenges. Currently, process streams such as thin stillage and condensed distillers solubles (CDS) are processed into distillers dried grains with solubles and used as animal feeds, but other higher value opportunities may exist. The objective of this study was to identify chemical components and quantify physical properties of CDS and thin stillage. Protein, organic acid, and sugar profiles were determined. Zein protein was identified, and glycerol was determined to have a concentration of 18.8 g/L in thin stillage and 63.2 g/L in CDS. Physical properties including density, thermal conductivity, thermal diffusivity, and rheological behaviors were also examined. Thermal conductivity of thin stillage and CDS was approximately 0.54 and 0.45 W/m°C, respectively. Quantification of the physical properties and identification of the chemical constituents pave the way for exploration of new value‐added uses for thin stillage and CDS.     

Solvent Extraction of Zein from Dry‐Milled Corn

Batch extraction of zein from dry‐milled whole corn with ethanol was optimum with 70% ethanol in water, an extraction time of 30–40 min, and temperature of 50°C. High yields (60% of the zein in corn) and high zein contents in the extracted solids (50%) were obtained at a solvent‐to‐solids ratio of 8 mL of 70% ethanol/g of corn. However, zein concentration in the extract was higher at lower ratios. Multiple extraction of the same corn with fresh ethanol resulted in a yield of 85% after four extractions, whereas multiple extractions of fresh corn with the same ethanol resulted in high (15 g/L) zein concentration in the extract. Optimum conditions for batch extraction of zein were 45°C, with 68% ethanol at a solvent‐to‐solids ratio of 7.8 mL/g for an extraction time of 55 min. Column extractions were also best at 50°C and 70% ethanol; a solvent ratio of 1 mL/g resulted in high zein concentrations in the extract (17 g/L) but yields were low (20%).     

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.     

Effect of Milling and Particle Size on Functionality and Physicochemical Properties of Cowpea Flour

Cowpeas (Vigna unguiculata) were milled through 0.5‐, 1.0‐, and 2.0‐ mm screens, and the flour was subsequently separated into different particle‐size ranges. Such procedures caused only minimal changes in moisture, fat, protein, ash, and total carbohydrate. The amount of extractable starch, however, varied from 34.5 to 52%. The effects of both mill screen and sieve mesh size were significant (P < 0.05). Differences in milling and separation procedures resulted in significant variations in water absorption (0.41–2.81 g of water/g of flour), solids lost (0.34–1.17 g/g of flour), and protein solubility (21.2–37.4%) (P < 0.05). Finely milled flours (91% moisture) had lower initial gelatinization temperatures (70–73°C), as measured by differential scanning calorimetry (DSC) (P < 0.01). Gelatinization peaks in high‐moisture flour were similar to that of pure starch. At lower moisture, a second peak was observed indicative of protein. Light‐scattering analysis showed that different conditions produced a bimodal particle‐size distribution when samples were suspended in water. The small size had relatively constant diameters (19–21 μm) and was associated with starch granules. The latter had a large size distribution and varying peak size and was associated with aggregated flour particles. These results indicate that changes in processing produces cowpea flours with differing chemical and physical properties.     

Quick Fiber Process: Effect of Mash Temperature,Dry Solids,and Residual Germ on Fiber Yield and Purity

Preliminary calculations showed that recovery of fiber before fermentation in the dry grind ethanol facilities known as the Quick Fiber process increases fermenter capacity and reduces ethanol production cost by as much as 4 ¢/gal. The objective of the current research was to evaluate the effect of mash temperature, dry solids, and residual germ on fiber yield and purity when using the quick fiber process. Fiber was recovered by flotation and skimming, while maintaining a specified temperature, dry solids, and residual germ in the mash. Varying temperature and dry solids in the mash resulted in a statistically significant effect on the fiber yield, neutral detergent fiber (NDF) content, and weight of NDF/100 g of dry corn. Varying residual germ in the mash resulted in statistically significant differences for NDF through dilution and the weight of NDF/100 g of dry corn. The highest fiber yield was 10.9% at 45°C, 23% dry solids, and 15% residual germ; the highest NDF was 50.9% at 30°C, 21% dry solids, and 0% residual germ. The highest weight of NDF/100 g of dry corn was observed at 45°C, 23% dry solids, and 0% residual germ.     

EFFECT OF SOIL AMENDMENT WITH THIN STILLAGE AND GLYCEROL ON PLANT GROWTH AND SOIL PROPERTIES

Controlled environment experiments were set up in 2007 and 2008 to evaluate the potential of using by-products of the biofuel industry as soil amendments to improve fertility and plant growth in Saskatchewan soils. Trials were run with thin stillage (a by-product of ethanol production) and glycerol (by-product of biodiesel production). Canola (B. napus L.) and wheat (T. aestivum) were grown as the test crop in amended pots. Plant yield, composition, and soil properties were measured after five weeks. The stillage was found to be an effective soil amendment for increasing plant biomass yield. Per unit of nitrogen (N) added, canola yields were less than that of urea when nitrogen was the only limitation, due to only a portion of the nitrogen in the thin stillage becoming available over the five week period. However, when nutrients other than nitrogen were limiting, canola dry matter yields with thin stillage amendment approached or exceeded that of urea, due to the ability of the amendments to supply other nutrients such as phosphorus in addition to nitrogen. Glycerol, an amendment that only contains carbon, hydrogen and oxygen, was effective in increasing soil organic carbon content, but required supplemental fertilizer to account for nutrient tie-up by microorganisms during decomposition in the soil. The amendments did not have any biologically significant effects on other soil chemical parameters measured, including soluble metals, pH or salinity.