Effects of Ground Corn Particle Size on Ethanol Yield and Thin Stillage Soluble Solids

The effects of ground corn particle size on ethanol yield and soluble solids in thin stillage was evaluated using a 2‐L laboratory dry‐grind procedure. The procedure was optimized for grinding, liquefaction, sacchari‐fication, and fermentation parameters. The optimized procedure was reproducible with a coefficient of variation of 3.6% in ethanol yield. Five particle size distributions of ground corn were obtained using a cross‐beater mill equipped with five screens (0.5, 2, 3, 4, and 5 mm). Particle size had an effect on ethanol yield and on soluble solids concentration in thin stillage. The highest ethanol yield of 12.6 mL/100 mL of beer was achieved using a 0.5‐mm screen in the cross‐beater mill. Treatment using the 0.5‐mm mill screen resulted in soluble solids concentration of 25.1 g/L and was higher than soluble solids concentrations obtained with other screens. No differences in soluble solid concentrations were observed in samples of thin stillage obtained from 2, 3, 4, and 5‐mm screens which had a mean yield of 16.2 g/L. By optimizing particle size for maximum ethanol yield and minimum solids in thin stillage, dry‐grind corn plants could realize reduced capital and operating costs.     

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.     

Rye and Triticale as Feedstock for Fuel Ethanol Production

Normal gravity rye and triticale mashes, containing 20–21 g of dissolved solids per 100 mL of mash liquid, were fermented with active dry yeast at 27°C. Fermentations were completed within 48 hr for rye, and within 72 hr for triticale. Supplementation of mashes with urea at a concentration of 8 mM accelerated rates of sugar consumption and fermentation, and reduced fermentation time from 48 to 36 hr for rye, and from 72 to 48 hr for triticale. Rye fermented faster than triticale, due to its higher level of free amino nitrogen. Ethanol yields were 356–363 L/tonne of 14% moisture rye grain, and 362–367 L/tonne of 14% moisture triticale. Fermentation efficiencies, which were 90–91% for triticale, and 91–93% for rye, and ethanol yields were comparable to those obtained from wheat and were not affected significantly by urea supplementation. The replacement of wheats by less expensive crops such as rye and triticale would provide good economic opportunities and alternatives for the fuel alcohol industry.     

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%).     

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.     

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.     

Enzyme production by wood-rot and soft-rot fungi cultivated on corn fiber followed by simultaneous saccharification and fermentation

This research aims at developing a biorefinery platform to convert lignocellulosic corn fiber into fermentable sugars at a moderate temperature (37 °C) with minimal use of chemicals. White-rot (Phanerochaete chrysosporium), brown-rot (Gloeophyllum trabeum), and soft-rot (Trichoderma reesei) fungi were used for in situ enzyme production to hydrolyze cellulosic and hemicellulosic components of corn fiber into fermentable sugars. Solid-substrate fermentation of corn fiber by either white- or brown-rot fungi followed by simultaneous saccharification and fermentation (SSF) with coculture of Saccharomyces cerevisiae has shown a possibility of enhancing wood rot saccharification of corn fiber for ethanol fermentation. The laboratory-scale fungal saccharification and fermentation process incorporated in situ cellulolytic enzyme induction, which enhanced overall enzymatic hydrolysis of hemi/cellulose components of corn fiber into simple sugars (mono-, di-, and trisaccharides). The yeast fermentation of the hydrolyzate yielded 7.8, 8.6, and 4.9 g ethanol per 100 g corn fiber when saccharified with the white-, brown-, and soft-rot fungi, respectively. The highest ethanol yield (8.6 g ethanol per 100 g initial corn fiber) is equivalent to 35% of the theoretical ethanol yield from starch and cellulose in corn fiber. This research has significant commercial potential to increase net ethanol production per bushel of corn through the utilization of corn fiber. There is also a great research opportunity to evaluate the remaining biomass residue (enriched with fungal protein) as animal feed.     

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.     

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.     

Solid-substrate fermentation of corn fiber by Phanerochaete chrysosporium and subsequent fermentation of hydrolysate into ethanol

The goal of this study was to develop a fungal process for ethanol production from corn fiber. Laboratory-scale solid-substrate fermentation was performed using the white-rot fungus Phanerochaete chrysosporium in 1 L polypropylene bottles as reactors via incubation at 37 degrees C for up to 3 days. Extracellular enzymes produced in situ by P. chrysosporium degraded lignin and enhanced saccharification of polysaccharides in corn fiber. The percentage biomass weight loss and Klason lignin reduction were 34 and 41%, respectively. Anaerobic incubation at 37 degrees C following 2 day incubation reduced the fungal sugar consumption and enhanced the in situ cellulolytic enzyme activities. Two days of aerobic solid-substrate fermentation of corn fiber with P. chrysosporium, followed by anaerobic static submerged-culture fermentation resulted in 1.7 g of ethanol/100 g of corn fiber in 6 days, whereas yeast ( Saccharomyces cerevisiae) cocultured with P. chrysosporium demonstrated enhanced ethanol production of 3 g of ethanol/100 g of corn fiber. Specific enzyme activity assays suggested starch and hemi/cellulose contribution of fermentable sugar.     

Optimization of Fermentation Temperature and Mash Specific Gravity for Fuel Alcohol Production

The effects of fermentation temperature and dissolved solids concentration adjusted by changing mashing water-to-grain ratios on wheat fermentation efficiencies, fermentation times, final ethanol concentrations, and ethanol production rates were studied by using response surface methodology. Final ethanol concentrations in fermentors depended primarily on mash specific gravities. Predictably, increases in fermentation temperatures dramatically reduced fermentation times and thereby shortened fermentation cycles. The highest ethanol production rates were achieved with a high fermentation temperature of 30°C and a low water-to-grain ratio of 2.0. At these settings, an ethanol concentration of 13.6% (v/v) was attained with a fermentation time of 54 hr and an ethanol production rate of 2.45 mL of ethanol/L/hr. Optimization of operating conditions suggested in the current study will provide existing fuel alcohol plants with increased productivity without alteration of plant equipment or process flow.     

固定化酵母粒子强化及其对甜高粱茎秆汁液乙醇发酵的影响

采用正交试验设计开展了三亚乙基四胺(TTA)和戊二醛(GLU)的浓度和处理时间对海藻酸钙固定化酵母粒子的化学强度影响的试验研究，并以甜高粱茎秆汁液为原料，在5 L的反应器中进行乙醇发酵试验，考察强化后的固定化酵母粒子对乙醇发酵的影响。结果表明，最优的固定酵母粒子强化处理的方案为：TTA浓度为0.5%，处理时间为120 min；GLU浓度为0.5%，处理时间为8 min。连续8批次的甜高粱茎秆汁液乙醇发酵试验结果表明，最优组合强化后固定化酵母粒子用于乙醇发酵时，平均乙醇得率和变异系数(CV%)分别为84.78%和8.08%，而未强化的固定化酵母籽子为84.32%和9.68%，可见，最优组合强化后的固定化酵母粒子的发酵性能略优于未强化的固定化酵母籽子。该文为固定化酵母发酵甜高粱茎秆汁液制取生物乙醇技术的研究提供了参考。     

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.     

Recovery of Fiber in the Corn Dry-Grind Ethanol Process: A Feedstock for Valuable Coproducts

A new process was developed to recover corn fiber from the mash before fermentation in dry-grind ethanol production. In this process, corn is soaked in water (no chemicals) for a short period of time and then degermed using conventional degermination mills. In the remaining slurry, corn coarse fiber is floated by increasing the density of the slurry and then separated using density differences. The fiber recovered is called quick fiber to distinguish it from the conventional wet-milled fiber. This study evaluated the percent of quick fiber recovery for a normal yellow dent and high oil corn hybrid. The quick fiber was analyzed for levels of corn fiber oil, levels of ferulate phytosterol esters (FPE) and other valuable phytosterol components in the oil and compared with conventional wet-milled corn coarse and fine fiber samples. Fiber samples were also analyzed and compared for yields of potentially valuable corn fiber gum (CFG, hemicellulose B). Comparisons were made between the quick fiber samples obtained with and without chemicals in the soakwater. An average quick fiber yield of 6–7% was recovered from the two hybrids and represented 46–60% of the total fiber (fine and coarse) that could be recovered by wet-milling these hybrids. Adding steep chemicals (SO₂ and lactic acid) to the soakwater increased the quick fiber yields, percent of FPE recoveries, and total percent of phytosterol components to levels either comparable to (for the dent corn hybrid) or higher than (for the high oil corn hybrid) those recovered from the total conventional wet-milled fiber samples. CFG yields in the quick fiber samples were comparable to those from the wet-milled fiber samples. CFG yields in the quick fiber samples were not significantly affected by the addition of chemicals (SO₂ and lactic acid) to the soakwater.     

固定化酵母粒子强化及其对甜高梁茎秆汁液乙醇发酵的影响

采用正交试验设计开展了三亚乙基四胺(TTA)和戊二醛(GLU)的浓度和处理时间对海藻酸钙固定化酵母粒子的化学强度影响的试验研究,并以甜高粱茎秆汁液为原料,在5 L的反应器中进行乙醇发酵试验,考察强化后的固定化酵母粒子对乙醇发酵的影响.结果表明,最优的固定酵母粒子强化处理的方案为TTA浓度为0.5%,处理时间为120 min;GLU浓度为0.5%,处理时间为8 min.连续8批次的甜高粱茎秆汁液乙醇发酵试验结果表明,最优组合强化后固定化酵母粒子用于乙醇发酵时,平均乙醇得率和变异系数(CV%)分别为84.78%和8.08%,而未强化的固定化酵母籽子为84.32%和9.68%,可见,最优组合强化后的固定化酵母粒子的发酵性能略优于未强化的固定化酵母籽子.该文为固定化酵母发酵甜高粱茎秆汁液制取生物乙醇技术的研究提供了参考.     

Corn Endosperm Fermentation Using Endogenous Amino Nitrogen Generated by a Fungal Protease

Fractionating the corn kernel to separate endosperm from germ and pericarp improves corn ethanol processing by increasing fermentation throughput and generating salable coproducts. One fractionation technology, dry fractionation (DF), suffers from loss of germ‐derived nutrients and amino acids, resulting in poor fermentation performance. Such deficiencies may be addressed by increasing nitrogen and other nutritional supplementation. As an alternative to exogenous nitrogen source, we investigated the use of a fungal protease to generate free amino nitrogen (FAN) from corn endosperm. Incubation of endosperm with protease did not affect subsequent liquefaction and saccharification. FAN supplementation through proteolysis resulted in fermentation being 99% complete in 48 hr, compared to 93% maximum with urea supplementation. Viable cell growth rates were similar in FAN and urea‐supplemented fermentations. Urea and FAN addition resulted in similar fermentation characteristics and similar FAN consumption rates as with FAN alone, which was indicative that FAN was assimilated preferentially. Increased amounts of maltose remaining after fermentation were correlated with initial FAN concentrations in mash. This observed trend was implicated in ethanol yield reduction of 2 g/L at high protease loading (generating 1.6 mg of FAN/g of glucose substrate) compared to a urea control. Using a glucose and maltose solution, we confirmed higher residual maltose in fermentations supplemented with high FAN concentrations. Use of protease to generate optimal FAN concentration in mash (1.2 mg of FAN/g of glucose substrate) could improve economics of dry fractionated corn ethanol production by increasing fermentation rates and, consequently, reducing fermentation time.     

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

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

高压脉冲电场对酿酒酵母发酵能力的影响

为探究高压脉冲电场(PEF)在酒精发酵工业上的实际应用,本研究以酿酒酵母为试验材料,在设定脉宽、频率、作用时间等参数不变的条件下,以电场强度为唯一变量,分别采用电场强度为1、6 kV·cm^-1的PEF对酵母进行预处理。通过检测发酵底液中酵母生长量、葡萄糖消耗量和乙醇产出量的变化,探究PEF对酿酒酵母发酵能力的影响。结果表明,经12 h发酵后,在电场强度为1 kV·cm^-1的PEF刺激作用下,酿酒酵母葡萄糖消耗量提高了10.18%,乙醇产出量提高了11.05%;当电场强度为6 kV·cm^-1时,葡萄糖消耗量和乙醇产出量均降低。本研究结果为提高酿酒酵母的发酵能力提供了一种新方法。     

Economics of Germ Preseparation for Dry-Grind Ethanol Facilities

A detailed economic analysis of a 914 tonnes/day (36,000 bu/day) “Quick Germ” ethanol process was performed. The Quick Germ ethanol process is a combination of a dry-grind and a wet-milling ethanol process. The Quick Germ ethanol process increases the coproduct value in the dry-grind ethanol process by recovering germ before fermentation. Germ is recovered using the conventional wet-milling degermination process. Economic assessment of the Quick Germ process proved profitable. The savings achieved by recovering germ as a coproduct and by increasing the fermentor capacity due to removal of nonfermentables from the corn mash will reduce the manufacturing cost of ethanol by 2.69 ¢/L (10.19 ¢/gal or $0.265/bu) when compared to the conventional dry-grind ethanol process.     

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.