Zein composition.

(54) Title: COMPOSITION OF ZEINA. (54) Title: ZEIN COMPOSITION.

(57) Summary

A method of producing bio-products from a raw material is described in a system configured to produce ethanol and distillery grains from a fermentation product. A system configured to process raw material to a fermentation product and bio-products including ethanol and food is described. A bioproduct produced from a fermentation product produced from a raw material in a bio-refining system is described.

(57) Abstract

A method of producing bioproducts from a feedstock in a system configured to produce ethanol and distillers grains from a fermented product is disclosed. A system configured to process feedstock into a fermented product and bioproducts including ethanol and meal is disclosed. A bioproduct produced from a fermented product produced from a feedstock in a biorefining system is disclosed.

ZEINA COMPOSITION

CROSS REFERENCE TO RELATED REQUESTS

The present invention claims the priority of and expressly incorporates for reference the following applications: (a) Provisional Patent Application No. 61/1 96,720 entitled “Extraction of Zein from Fermented Corn, which was filed on December 31, 2008 (as US Patent Application No. 12 / 347,566); (b) US Provisional Patent Application No. 61 / 207,868 entitled “Extraction of Zein from Fragmented Corn by Fermentation”, which was filed on December 31, 2008 (as US Patent Application No. 12 / 347,743); (c) Provisional Patent Application No. 61 / 161,313 entitled “Extraction of Zein from Fermented Corn”, which was filed on March 18, 2009; (d) US Provisional Patent Application No. 61 / 161,318 entitled “Extraction of Zein from Fractionated Corn by Fermentation”, which was presented on March 18, 2009; (e) US Provisional Patent Application No.

61 / 161,322 entitled "Extraction and Recovery of Zein", which was presented on March 18, 2009; and (f) US Provisional Patent Application No. 61 / 161,325 entitled "Extraction and Recovery of Zein", which was filed on March 18,

2009.

GOVERNMENT INTERESTS

The Government may have rights in an invention claimed in this application in accordance with Contract No. DE-FG3608G08033 with the Department of Energy.

TECHNICAL FIELD

The present application relates to a system for the extraction of protein from a fermentation product. The present application also relates to a system for the extraction of zein from fermented solids in the production of ethanol from corn. The present application further relates to an extracted zein composition comprising alpha-zein, beta-zein, and gamma-zein.

BACKGROUND

Zein is a group of plant proteins that can be extracted from corn or substrates containing corn protein, such as corn gluten meal. Zein is classified as GRAS (Generally Recognized As Safe) by the US Food and Drug Administration. and has a variety of commercial uses, including the manufacture of edible food packaging, edible films, biodegradable plastic resins, chewing gum base, tablet coating compounds, adhesives, paper cup coatings, soda lid liners , etc. Zein can also be processed into resins and other bio-plastic polymers, which can be extruded or rolled into a variety of plastic products. Zein has utility as a starting material for a variety of renewable and non-toxic polymer applications.

Zein belongs to a class of proteins called prolamins, soluble in alcohol. Zein comprises about forty to fifty percent of the total protein in corn, or about forty percent of the corn kernel. Zein has also been divided into four sub-classes: alphazein, beta-zein, gamma-zein, and delta-zein. Alpha-zein is the primary zein commercially used and represents about seventy percent of the zein in corn. Beta-zein represents about five percent of the zein in corn. Gamma-zein represents approximately twenty to twenty-five percent of the zein in corn, and delta-zein represents approximately one to five percent of the zein. Each type of zein (alpha, beta, gamma, and delta) has different amino acid profiles and exhibits slightly different properties. Zein can be extracted and recovered from corn or corn products from corn processing.

Corn gluten meal, a by-product of ethanol production through wet milling, is a typical starting material for the extraction of zein since its protein content is sixty percent or more. Sulfur dioxide or other chemicals that can be used during the preparation (for example, in a soaking procedure) of corn gluten meal can adversely affect the quality of zein.

Ethanol can be produced from raw materials (such as corn), cellulosic raw materials (such as stick grass or ears), or other plant material (such as sugar cane). Ethanol production. from corn it produces fermentation products (eg, co-products) that are suitable for use as starting materials for zein extraction.

In a wet milling procedure, ethanol is produced from corn by first soaking the corn kernels in water containing sulfur dioxide, and then separating the kernels into endosperm, fiber, and germ. The endosperm is further processed to produce corn starch and gluten, which can be dried into corn gluten meal. Corn gluten meal can comprise at least sixty percent protein, and is typically used as a starting material for zein extraction in commercial zein production. Sulfur dioxide or other chemicals that can be used during the preparation of corn gluten meal can adversely affect the quality of zein.

Ethanol can also be produced from corn using a dry milling procedure. In a dry milling process, a starch-containing material, such as corn, is ground into a flour and a slurry is formed with water and enzymes. The grout can be baked to liquefy the starch and to facilitate saccharification. Additional enzymes can be added to complete the saccharification to break the starch down into simple sugars (eg, glucose) that can be fermented using an ethanolgen (eg, yeast). Fermentation produces a fermentation product comprising a liquid portion or component and a solid portion or component. The liquid portion comprises ethanol and water and soluble components. The residual solids comprise, for example, proteins, fiber, oil, and other insoluble components.

The fermentation product comprising a liquid component and a solids component can be distilled to separate the ethanol and whole vinasse (eg, wet solids or fermentation solids). The entire stillage comprises residual solids and water, and can also be separated into a wet cake and thin stillage. The wet cake (wet solids) can be dried into flour such as dry distillery grains (DDG); The thin stillage can be reduced to syrup and added to the wet cake or flour during the drying procedure to produce dry distillers grains with solubles (DDGS). Flour, such as DDG and DDGS, can be used as an animal food product.

In accordance with an alternative procedure, for example, as described in the US Patent Application Publication

No. 2005/02391 81, the starch can be converted to sugars and can be fermented in a raw starch process without "baking" or liquefying. Heat damage to proteins and other components of the slurry can be avoided using the raw starch procedure.

A dry fractionation procedure that does not use sulfur dioxide can be used in place of dry milling to fractionate corn into endosperm, fiber, and germ. The amount of residual solids in the fermentation product can be reduced through fractionation and elimination of the fiber and germ, both low in starch, from the fermentation. The endosperm is primarily composed of starch and protein with small amounts of fiber and oil present. Zein is also concentrated in the endosperm; more than half of the endosperm protein may be composed of zein. When the endosperm is fermented, the residual solids comprise a high concentration of zein. The dry residual solids from endosperm fermentation are high in protein and result in a flour that is referred to as “high protein concentration dry distillery grains” (DDG HP).

Dried distillery grains (DDG) contain zein, but a high percentage or high quality of zein may not be recovered, if the product has been subjected to chemical or heat treatments.

It could be advantageous to provide a system for extracting protein from a fermentation product. It could also be advantageous to provide a method of producing bio-products from a raw material in a system configured to produce ethanol and distillery grains from a fermentation product. It could also be advantageous to provide a system configured to process raw material to a fermentation product and bioproducts including ethanol and flour. It could also be advantageous to provide a bio-product produced from a fermentation product produced from a raw material in a biorefining system.

BRIEF DESCRIPTION OF THE INVENTION

The present invention relates to a method of producing bioproducts from a raw material in a configured system or of producing ethanol and distillery grains from a fermentation product. The method comprises the steps of processing the raw material to a starch-containing component and creating a slurry comprising the starch-containing component. The method comprises the steps of preparing the starch containing component of the slurry to a fermentable component for fermentation and fermenting at least a portion of the fermentable component of the slurry to the fermentation product. The method comprises the steps of processing the fermentation product to bioproducts comprising ethanol and the distillery grains and producing a bio-product comprising a zein composition from the fermentation product.

The present invention relates to a system configured to process the raw material into a fermentation product and bioproducts including ethanol and flour. The system comprises a mill for processing the raw material to a starch-containing component so that a slurry comprising the starch-containing component can be formed. The system also comprises a container configured to contain the slurry and for the fermentation of a fermentable component from the starch-containing component to the fermentation product. The system comprises a distillation system for the recovery of ethanol from the fermentation product and the separation of wet solids from the fermentation product that are to be dried in the flour. The system comprises a system for extracting a bioproduct comprising a zein composition from the fermentation product, the fermentation product comprises a protein component, and the zein composition comprises at least a portion of the protein component from the fermentation product.

The present invention relates to the bio-product produced from a fermentation product from a raw material in a biorefining system. The bio-product comprises a zein composition comprising alpha-zein and beta-zein and gamma-zein. The zein composition has been extracted from a protein component of the fermentation product.

DESCRIPTION OF THE DRAWINGS

Figure 1 is a schematic diagram of an ethanol production facility.

Figures 2A to 2C are schematic block diagrams of an ethanol production facility.

Figures 3A to 3C are process flow diagrams of an ethanol production process.

Figure 4 is a schematic block diagram of the equipment used in an ethanol production facility, comprising a system for extracting zein from fermented solids.

Figures 5A through 5B are process flow diagrams of a system for extraction of zein from crude starch dry solids (DDG) and crude starch endosperm dry solids (DDG HP). Figure 5A is a procedural flow for zein extraction using a milling procedure. Figure 5B is a procedural flow for zein extraction using a fractionation procedure.

Figures 6A to 6B are process flow diagrams of a system for zein extraction. Figure 6A is a process flow for the extraction of zein from endosperm beer from raw starch. Figure 6B is a process flow for the extraction of zein from a wet cake.

Figures 7A through 7C are tables of the average composition of fermented solids. Figure 7A is a beer composition chart for conventional fermentation of raw starch and endosperm. Figure 7B is a chart of a wet cake composition for fermentation of raw starch. Figure 7C is a dry solids composition table (DDG) for conventional fermentation of raw starch and endosperm.

Figure 8 is a table for the average beer composition in conventional raw starch and endosperm fermentations.

Figure 9 is a table for the average protein content in starting materials for zein extraction.

Figure 10 is a table for operating conditions and parameters for a zein extraction vessel.

Figure 11 is a graphical representation of parameters and operating conditions for a zein extraction procedure.

Figure 12 in a table illustrating the extraction of zein produced from different starting materials.

Figure 13 is an illustrative chromatogram of the zein composition.

Figures 14A through 14C are tables for the zein composition that provide data from experimental extractions. Figure 14A is a table for the composition of zein extracted from beer on a laboratory scale. Figure 14B is a table for the zein composition extracted from high protein dry distillery grains on a laboratory scale. Figure 14C is a table for the zein composition extracted from high protein dry distillery grains on a pilot scale.

Figure 15 is a picture of the homogenization and temperature effect,

Figure 16 is a table for the zein gelation time at different storage conditions for zein solutions extracted from different starting materials.

Figures 17A through 17B are graphs of zein viscosity over time at different temperatures. Figure 17A is a graph of the zein viscosity that was extracted from high protein dry distillery grains. Figure 17B is a graph of the zein viscosity that was extracted from corn gluten meal.

DESCRIPTION OF THE MODALITIES

Figure 1 is a schematic block diagram of an ethanol plant. A method of producing bio-products from a raw material in a system configured to produce ethanol and distillery grains from a fermentation product can be used in the ethanol plant. The ethanol plant can comprise a system configured to process raw material into a fermentation product and bio-products including ethanol and flour.

The plant comprises a facility to produce corn-based ethanol and zein can be extracted from the fermentation solids, a component of the fermentation product. Fermentation solids can comprise beer, beer solids, wet solids, wet cake, or dry solids, flour distillery grains (eg DDG, DDGS, DDG HP). The ethanol plant can use various systems and methods, such as conventional starch liquefaction (for example, hydrolysis of baked starch or raw starch, among other procedures) to process corn (or other types of biomass).

Figures 2A to 2C are schematic block diagrams of an ethanol production facility. Figure 2A is a schematic block diagram of a system for an installation using a conventional "baked starch" fermentation procedure 200. In a "baked starch" ethanol plant that produces ethanol from corn, corn kernels are processed to separate the starch-containing material (eg, endosperm) from other matter (such as fiber and germ). The starch-containing material is then formed into a slurry with water and liquefied to facilitate saccharification, where the starch is converted to sugar (glucose) and fermentation, where the sugar is converted through ethanolgen (yeast) to ethanol . The product of fermentation (fermentation product) is beer comprising a liquid component and a solid component. A procedure typically used in a conventional ethanol plant, liquefaction of the starch-containing material is accomplished by “baking the slurry at a temperature at or above the gelatinization temperature of the starch (typically at or above 60-75 ° C).

Figure 2B is a schematic block diagram of a system for an installation using a raw starch fermentation procedure. ” In "raw starch" fermentation, the starch can be converted and fermented without "baking" or liquefying (as in the "baked starch" procedure). Figure 2C is a schematic block diagram of a system for an installation using a crude starch endosperm fermentation procedure 204. In the raw starch endosperm fermentation procedure, the corn kernel is fractionated into endosperm, germ, and fiber. The fractionation procedure aims to separate the starch-containing endosperm from the germ and fiber (which are low in starch). The endosperm is then supplied to the fermentation process of "raw starch.

Figures 3A to 3C are process flow diagrams of an ethanol production process. Figure 3A is a process flow diagram of a procedure for the conventional production of ethanol 300 and corresponds to Figure 2A where dry distillery grains (DDG) or dry distillery grains with solubles (DDGS) are the result of fermentation of baked starch. Figure 3B is a process flow diagram of a process for the production of ethanol comprising the hydrolysis step 302 of raw starch and corresponds to Figure 2B where DDG or DDGS are the result of the fermentation of raw starch. Figure 3C is a process flow diagram of a procedure for the production of crude starch endosperm ethanol 304 and corresponds to Figure 2C, where high protein distillery grains (DDG HP) is the result of Endosperm fermentation of raw starch.

The equipment used in an ethanol production facility for the extraction of zein from fermented solids (for example, DDG, HP, DDGS, DDG, beer, and wet cake) is illustrated in Figure 4. Substantially the same equipment is used. for each of the different fermentation procedures (conventional, raw starch, raw starch endosperm) and for each of the different starting materials (fermented solids, DDG, DDGS, DDG, beer, wet cake). According to an illustrative embodiment, the raw material used is DDG HP. According to another modality, the raw material used can be DDG.

In accordance with an illustrative embodiment, the zein extraction and recovery procedure may comprise three steps: extraction, refining (eg, purification), and recovery. The extraction step removes zein through solubilization; Zein is soluble in aqueous alcohol. The extraction equipment comprises a reactor vessel 402 where inlet fermentation solids are found (which can be processed through a hammer mill 404); the fermentation solids are the output of the various ethanol procedures shown in Figures 3A to 3C.

Other inlets to reactor vessel 402 comprise sodium hydroxide (NaOH, optional), alcohol, water, steam, and acid (optional). In accordance with certain embodiments, the alcohol can be selected from an alcohol composition comprising C1 to C7 alcohols (eg, methanol, ethanol, or propanol). According to certain modalities, alcohol is ethanol.

According to an illustrative embodiment, the aqueous alcohol (or aqueous ethanol) solution comprises an extraction agent, such as sodium hydroxide in an amount based on the dry solids of the starting material. According to an illustrative embodiment, up to 7.0 percent sodium hydroxide is used. According to a preferred embodiment, the sodium hydroxide concentration is 2.8-4.0 percent. According to the most preferred embodiment, the sodium hydroxide concentration is 3.2-3.8 percent.

In accordance with one embodiment, the contents of reactor vessel 402 are drained into a centrifuge, such as a disk nozzle centrifuge 406 (or basket centrifuge), for separation into a solids component and a liquid component comprising ethanol and zein. The zein can be refined through filtration and recovered from the liquid component through drying or precipitation.

As illustrated, the refining or purification step may comprise the use of multiple filters (or membranes) configured to remove matter with different sizes (eg, membrane filters having different pore sizes). The first filter 408 can have, for example, a pore size of one micrometer (for example, a micro filter). The first 408 filter is designed to protect the membrane from ultra-filtration by removing suspended solids that were removed in the separation step. The solids are sent to a 410 tank and the liquids are further processed to recover the zein. The liquids are then passed through a second filter 412, which may be a 10,000 molecular weight cut membrane filter intended to remove small molecular weight components and to concentrate the zein solution. The concentrate (for example, zein / ethanol solution) is sent to a maintenance tank 414 (mixing vessel or reactor) and the permeate is further processed for the recovery of ethanol (for example, through distillation).

The zein is recovered from the concentrate solution (for example from the second filter 412) by drying the solution (for example, with a 416 double drum vacuum dryer or a desolventizer). According to certain modalities, the zein solution can be precipitated by diluting the alcohol concentration to where the zein is no longer soluble (eg 50 percent (w / v) or less).

In one embodiment, zeolite can be used to remove impurities from the zein composition. A crude solution of a zein contained in an aqueous alcohol solvent is contacted with a zeolite adsorbent under conditions effective for adsorption of color and odor impurities in the zein solution in the zeolite. The treated solution can be separated from the adsorbent with the intention of recovering high quality zein dissolved in the aqueous alcohol solvent. Optionally, more impurities (eg, color or residual order) can be removed by contacting the treated solution with an activated carbon adsorbent or a mixture of activated carbon and zein adsorbents.

Figures 5A through 5B illustrate process flow diagrams for the extraction of zein from dry solids (DDG) and dry endosperm solids (DDG HP) from a crude starch fermentation procedure. Figure 5A illustrates a process flow 502 for zein extraction from a crude starch fermentation process (eg, without using fractionation). DDG or DDGS 504 are used for the extraction and recovery of zein 506 and may come from the ethanol production process 508 after drying step 510. Figure 5B illustrates a process flow 512 for extraction of zein with fractionation 514. During fractionation 514, the corn germ and fiber 516 are separated from endosperm 518 and removed, leaving endosperm 518 for fermentation 520. HP 522 DDG is used for extraction and recovery of 524 zein after 526 drying in the 528 ethanol production process.

Figures 6A through 6B illustrate process flow diagrams for zein extraction. Figure 6A illustrates a process flow 602 for extraction of zein from raw starch endosperm beer. In this case, corn 604 is fractionated 606 to remove the germ and fiber 608, leaving endosperm 610 for fermentation 612. A slurry 614 is created. Beer 614 is used for extraction and recovery of zein 616 after fermentation. Figure 6B illustrates a process flow 616 of zein extraction from a wet cake. As illustrated, corn kernel 618 undergoes fractionation 620 to remove germ and fiber 622 from endosperm 624. A slurry 626 is created. Endosperm 624 is saccharified and fermented 628 to a fermentation product, such as beer that separates 630 into a liquid component, shown as thin vinasse 632, and a solids component, shown compared to wet cake 634. Wet cake 634 it is used for the extraction of zein 636 and recovery 638.

Figures 7A to 7C are tables of the average composition (representative for commercial production) of fermented solids (DDG, DDG HP, DDGS, beer, and wet cake), intended to be used as starting materials for the extraction of zein.

Figure 7A is a beer composition chart and also visually depicted in Figure 8 which illustrates a chart for the average beer composition used in the examples in accordance with an illustrative embodiment. Data are presented for beer from three different types of procedures: conventional fermentation, raw starch fermentation, and endosperm fermentation. The numbers are in an "as such" condition. Beer composition and other information was gathered from six experiments.

The wet cake composition for raw starch fermentation is illustrated in Figure 7B. The average moisture content of the wet cake is 69.1 percent and the protein content is 31.0 percent dry matter.

Figure 7C illustrates a table for dry solids composition (DDG) for conventional fermentation, raw starch fermentation, and endosperm fermentation. The sulfur content and the protein content illustrated are the average composition as a percentage of dry matter.

A table for the average protein content 902 in starting materials for zein extraction is illustrated in Figure 9. The protein content for each starting material is divided into two subgroups, one for dry basis (left portion of the box) and one for the starting material in an “as such” condition (right side of the box). As illustrated, the starting material may be conventional beer, conventional DDGS, raw starch beer, wet raw starch cake, raw starch DDGS, endosperm beer, and / or endosperm DDGS. Concentration values ​​are represented as percentages.

Figure 10 illustrates a table for the operating conditions and parameters for a zein extraction. A typical scale, a preferred scale, and a highly preferred value or scale are provided for each of the ratio of solvent to solids, a ratio of solvent to solids for beer, a concentration of solvent ethanol, a concentration of sodium hydroxide, a temperature, and an extraction time. A graphical representation of these parameters and operating conditions is illustrated in Figure 11. The zein extraction parameter scales and operating conditions are shown using nested scales. The typical scale is represented by the identified external values, the preferred scale is identified by the identified internal values, and the highly preferred value or scale is identified within the dotted blocks.

Solvent to Solids Ratio 1102 is the weight of the solvent (ethanol and water combined) relative to the weight of the solids. A typical ratio of solvent to solids ratio is 4: 1 to 10: 1. A preferred ratio of solvent to solids is 4: 1 to 7: 1. A highly preferred ratio of solvent to solids is 5: 1.

For the extraction of zein from 1104 beer, a typical ratio of solvent to solids ratio is 6: 1 to 10: 1. A preferred ratio of solvent to solids is 7: 1 to 10: 1. A preferred ratio of solvent to solids is 7: 1 to 10: 1. A highly preferred ratio of solvent to solids is 7: 1 to 8: 1.

The concentration of solvent ethanol 1106 is the concentration by weight percentage of ethanol in the extraction solvent. A typical scale for the concentration of solvent ethanol is 40 to 90 percent. A preferred scale for the concentration of solvent ethanol is 50 to 80 percent. A highly preferred scale for the concentration of solvent ethanol is 60 to 70 percent.

The sodium hydroxide concentration 1108 is the weight of solids on a dry basis. A typical scale for sodium hydroxide concentration is 0 percent to 5 percent. A preferred scale is from 2.8 percent to 4.0 percent. A highly preferred scale is 3.4 percent to 3.6 percent.

The extraction temperature 1110 is the temperature of the slurry in the extraction vessel. A typical temperature scale for zein extraction is 20 to 78 ° C. A preferred temperature scale for zein extraction is 50 to 75 ° C. A highly preferred temperature scale for zein extraction is 68 to 70 ° C.

Extraction time 1112 is the length of time that the slurry is maintained at the extraction temperature. A typical extraction time is 20 to 120 minutes. A preferred extraction time is 25 to 60 minutes. A very preferred extraction time is 28 to 30 minutes.

EXAMPLES

Experiments and tests were conducted to evaluate zein compositions and yields of different starting materials and procedure. Figures 12 through 17 provide information related to the examples.

According to a series of example, the zein composition was extracted from different starting materials (beer, wet cake, and DDG) from different ethanol processes (baked starch fermentation, raw starch fermentation, and raw starch fermentation from endosperm) to compare zein yields. Zein was extracted using 70 percent ethanol in water with 3.5 percent sodium hydroxide (based on the dry solids of the starting material) at 70 ° C for 30 minutes. Figure 12 is a table showing illustrative zein extraction yields. Extraction from DDG (from raw starch fermentation) recovers more zein than extraction from DDG from a "baked starch" fermentation procedure (shown as "conventional fermentation). The combination of corn fractionation and raw starch fermentation provides better extraction efficiencies (as best partially explained by the higher protein content of DDG HP).

Figure 12 is an example of a laboratory scale extracted zein HPLC chromatogram for DDG HP. Alpha-zein (“zein), beta-zein (^p zein) and gamma-zein (^γ zein). The peaks for beta and gamma-zein were not completely separated using this technique, and the relative amounts of beta and gamma are reported as the sum total of both types of zein.

Experiments were conducted to study the yield and composition of zein extracted from different starting materials. Figures 14A through 14C are tables for the zein composition 10 detailing the results of the performance and composition experiments.

Zein was extracted from beer on a laboratory scale to study the performance and composition of the extracted zein. Figure 14A is a table of the zein composition extracted from the beer on a laboratory scale. Six examples are illustrated through experiments with two conducted experiments for each procedure (conventional fermentation, raw starch fermentation, and endosperm raw starch fermentation). The starting material for these experiments is illustrated in the table in Figure 7A. Data are from experimental extractions using solvent (ethanol) with ("YES) or without (" NO ") sodium hydroxide (NaOH). If sodium hydroxide is not used, the beta-zein and gamma-zein content of the extracted / recovered zein was both zero and below zero, as shown in Figure 14A. When sodium hydroxide is used, the beta-zein and gamma-zein content was much higher (up to around 25.9 percent in this example). The amount of zein recovered, in grams, dry basis was almost double the amount of zein recovered without the use of sodium hydroxide as shown in Figure 14A. The alpha, beta and gamma-zein composition of the zein was obtained through zein reverse phase liquid chromatography (HPLC) analysis.

A study was conducted to determine the yield and composition of zein extracted from high protein dry distillery grains on a laboratory scale. Data is from experimental extractions using 70 percent ethanol with 3.5 percent sodium hydroxide (based on the dry solids of the starting material). The alpha, beta, and gamma-zein composition of the zein was obtained through RPHPLC chromatogram analysis of the zein. The results of zein yield and composition from high protein dry distillery grains on a laboratory scale are shown in Figure 14B.

The bio-product comprises a zein composition comprising, on a weight basis, a protein component of at least 70 weight percent of the zein composition and a fat component of not more than 10 weight percent. The protein component comprises the zein composition.

Zein was extracted from high protein dry distillery grains on a pilot scale to study the performance and composition of the extracted zein. Data are from pilot scale extractions using 77 kg of solvent and 15.4 kg of dry HP DDG. In accordance with an illustrative embodiment for extracting zein from DDG, the equipment used for the extraction and recovery of zein comprises: an 80 liter reactor vessel; a 121.9 cm x 76.2 cm perforated centrifuge (commercially available from Sanborn Technologies of Walpole, Massachusetts) to separate suspended solids from the zein solution; a membrane filter containing a 20.06 cm 10,000 UF molecular weight cutoff membrane of polyethersulfone (commercially available from Parker Hannifin of Cleaveland, Ohio) to concentrate and remove impurities in the zein solution; and an 81.28 cm x 182.8 cm double vacuum drum dryer (available from Buflovak of Buffalo, New York) to dry the zein from the solution. The alpha, beta, and gamma-zein composition of the zein was obtained through analysis of zein RP-HPLC chromatograms. The performance and composition of the zein is shown in Figure 14C.

Zein was extracted from beer to study the homogenization and temperature effect of zein extraction and recovery. According to an illustrative embodiment, the zein extraction method comprises fermented beer with 50 grams (dry basis) of solids mixed with enough ethanol to produce 70 percent (w / w) of aqueous ethanol solution. The mixture was further diluted with an additional 70 percent (w / w) of aqueous ethanol to produce an extraction mixture with a solvent to solids ratio of 9: 1. The amount of ethanol extract and 70 percent aqueous ethanol required for extraction depends on the water content of the beer solids.

The extractions were carried out with the solvent, either containing sodium hydroxide or not containing sodium hydroxide. If sodium hydroxide was used, 2.3 ml of a 50 percent (w / w) solution was added to the solvent before heating. Solvent and beer solids were heated separately in closed containers at 70 ° C in a water bath. The hot solvent and beer solids were mixed and then homogenized for 3 minutes using a Polytron model PT-2100 homogenizer (available from Kinematica AG of Switzerland) in a 26 power apparatus.

After homogenizing, the mixtures were centrifuged using a HN-SII Model IEC centrifuge (available from International Equipment Co. of Needham Heights, MA) for 10 minutes at 5000 rpm. The component or solid fraction was air dried and retained for analysis. The liquid component or fraction (containing the zein) was neutralized to a pH of about 7 using 6N hydrochloric acid or 50 percent (w / w) sodium hydroxide. The zein contained in the liquid fraction was recovered by precipitating the solution in excess cold water (approximately 0 ° C). The zein was removed from the water by centrifuging for 10 minutes at 4600 rpm using a Beckman Model J-6B centrifuge (available from Beckman Coulter Inc. of Brea, CA). The liquid was discarded and the zein was air-dried and analyzed.

The results of the analysis are shown in Figure 15, which is a table showing the effect of the homogenization and temperature of the extraction and recovery of zein. According to one embodiment, the extraction time can be reduced by homogenizing the material at 70 ° C. Two recovery values ​​are illustrated, percent of theory for each conventional fermentation, raw starch fermentation, and endosperm raw starch fermentation. The first value for each is 30 minutes of stirring, 50 ° C, 70 percent ethanol, and 3.5 percent sodium hydroxide (based on the dry solids of the resulting material). The second value is for 3 minutes of homogenization, 70 ° C, 70 percent ethanol, and 3.5 percent sodium hydroxide (based on the dry solids of the starting material). Sample homogenization improves extraction efficiency and can also reduce the extraction time from 30 minutes to 3 minutes.

Zein gelation time was studied for different extraction starting materials. Figure 16 shows a table for the zein gelation time at different storage conditions of zein solutions extracted from different starting materials. Zein was extracted using 70 percent aqueous ethanol with 3.5 percent sodium hydroxide (based on the dry solids of the starting material) at 70 ° C for 20 minutes in a 50-liter reactor (available from Northland Stainless,

Tomahawk, Wl). The mixture was then centrifuged to remove solids in a 50.8 cm x 24.1 cm basket centrifuge (available from Sanborn Technologies of Walpole, MA) equipped with a polymeric filter cloth with 50 micron openings. The zein-containing liquid fraction was concentrated and purified by passing it through a 10,000 molecular weight cutoff ultrafiltration membrane (available from Parker-Hannifin of Cleveland, OH).

The zein solution was divided into three samples: one was stored at room temperature (approximately 22 ° C), one was stored at 5 ° C and one was stored at 40 ° C. The viscosity of the zein solutions was measured daily by emptying 300 milliliters of the sample into a 600 milliliter Griffen beaker, and viscosity measurements were taken with a Brookfield DV-1 8 digital viscosity meter available from Brookfield Engineering Laboratories of Middlenboro , MA). A spindle 2 to 100 revolutions per minute was used to test the viscosity of the solution. The viscosity of the samples was tested until a gel was formed.

Zein viscosity was studied over time at different temperatures, as shown in Figures 17A to 17B. Viscosity in centipoise (cP) is shown along the vertical axis and time, represented in days, is shown along the horizontal axis. Figure 17A shows the viscosity of zein that was extracted from high protein dry distillery grains and Figure 17B shows the viscosity of zein that was extracted from corn gluten meal. The ambient temperature was 22 ° C. The refrigerator temperature was 5 ° C and the oven temperature was 40 ° C.

Preliminary tests of viscosity in zein solutions showed that they were non-Newtonian and thixotropic. The zein solution extracted from corn gluten meal exhibited a lower viscosity (approximately 100 centipoises) than the zein solution extracted from high protein dry distillers grains (approximately 200 centipoises), but the extracted zein solution of corn gluten meal gelled faster and then reached a lower viscosity (17 days at room temperature and 10 days at 40 ° C, both at approximately 300 centipoises) compared to the zein solution extracted from high protein dry distillery grains, which obtained viscosities of more than 700 centipoises before gelling after 24 hours at room temperature and 12 days at 40 ° C . This difference in viscosity may be a result of the zein solution extracted from corn gluten harone containing lower molecular weight zein peptides due to the potential damage caused by sulfur dioxide used during wet milling. Both zein solutions took three to four months to gel when stored at 5 ° C.

The two zein solutions also appeared visually different. Although the zein solution extracted from the high-protein distillery dry grains contained a single phase, the zein solution extracted from the corn gluten meal contained two phases, which could be separated by laboratory centrifugation. When stored at room temperature, the lower phase gelled in less than a day, while the upper phase took approximately 38 days to gel.

In accordance with other alternative modalities, the production of the bio-product comprises extracting the zein composition by applying a solvent composition to the fermentation product and separating the fermentation product with the solvent composition into the zein composition and a solids component. The solvent composition comprises an agent. The agent can comprise sodium hydroxide. The agent can comprise potassium hydroxide. The agent can comprise an acid. The agent can comprise hydrochloric acid. The agent may comprise an extraction agent. The extraction agent can comprise an alkali hydroxide. The extraction agent can be sulfite. The extraction agent can comprise sodium metabisulfite. The extraction agent can comprise thiol. The extraction agent can comprise 2-mercaptoethanol.

In accordance with various modalities, the concentrations of the various proteins can be altered by processing to recover, within the available scale, a zein that purports to have a composition more specifically applicable to the intended commercial uses (such as increasing the percentage of beta and gamma-zein to allow more use in applications requiring resistance).

Conventional zein extraction methods use seventy percent aqueous ethanol to extract zein. The disclosed modalities provide methods where the ethanol concentration is increased after extraction, preferably to ninety percent. As a result of increasing the percentage of ethanol, more beta-zein and gamma-zein can precipitate from the zein solution.

The recovered zein yields depend on the time, temperature, particle size, and percentage of aqueous alcohol used for extraction. Sodium hydroxide can also be added to the solvent to increase zein yields. The use of sodium hydroxide (for example, as an extraction agent) in the solvent also changes the solubility of beta and gamma-zein and allows the use of beta and gamma-zein extraction. Other extracting agents can also be used, such as another alkaline hydroxide (for example, potassium hydroxide), an acid (hydrochloric acid or sulfuric acid), or a sulfite (for example, sodium sulfite, sodium bisulfate or sodium). The relative volumes of ethanol, extraction agent, and starting material will change based on the moisture content of the starting material and the target concentration of ethanol.

Zein recovery was improved by extracting at 50 ° C or higher and by adding 3.5 percent sodium hydroxide or other reducing agent to the aqueous alcohol. By increasing the pH of the extraction solvent using sodium hydroxide, the recovery of the zein is increased. Decreasing the pH also increases the amount of material recovered, but the recovered material has a lower protein content. By not adjusting the pH of the solvent, less zein was extracted.

The terms "dry distillery grains", "DDG", "dry distillery grains with solubles", #DDHS "," granulated material "," pelletized material ", or the like may refer to particulate matter. Although many types of biomass can be fermented in an alcohol plant producing various types of particulate products that will be transported to other locations, a corn-based ethanol plant that produces dry grains from distillery is discussed through this application for purposes Illustrative of material properties and operational aspects for this invention.

The word "illustrative" is used to refer to serving as an example, case, or illustration. Any aspect or design described as illustrative does not necessarily have to be constructed as preferred or advantageous over the other aspects or designs, nor does it mean excluding equivalent illustrative structures and techniques known to those skilled in the art. Rather, the use of the word illustrative is intended to present concepts in a concrete form, and the subject matter discussed is not limited by such examples.

The term “or” is intended to refer to an “or inclusive rather than an exclusive“ or ”. To the extent that the terms "comprise", "has", "contains", and other similar words are used in either the description or the claims, for the avoidance of doubt, such terms are intended to be inclusive in a manner similar to the term " It comprises as an open transition word without excluding any additional or other element.

In view of the illustrative apparatus and methods, the methodologies that can be implemented in accordance with the described subject matter will be best appreciated by referring to the flow diagrams of the various figures. Although for purposes of simplicity of explanation, the methodologies were shown and described as a series of blocks, it should be understood and appreciated that the claimed subject matter is not limited by the order of the blocks, as some blocks may occur in different orders and / or concurrently with other blocks than what is shown and described. Furthermore, not all of the illustrated blocks may be required to implement the methodologies.

It is important to note that the construction and arrangement of the elements of the subject matter described, as written in this application and as shown in the Drawings, is only illustrative. Although some embodiments have been described in detail in this description, those skilled in the art, who have reviewed this description, will readily appreciate that many modifications are possible (eg, variations in size, dimensions, structures, shapes, and proportions of the various elements, parameter heats, mounting arrangements, use of materials, colors, orientations, etc.) without materially departing from the novel teachings and advantages of the subject matter presented. For example, the elements shown as integrally formed, the operation of the interfaces may be inverted or otherwise varied, the length or width of the structures and / or members or connectors or other elements of the system may be varied, the nature or The number of adjustment positions provided between the elements can be varied. It should be understood that the elements and / or assemblies of the system can be constructed from any wide variety of materials that provide sufficient strength or durability, in any of a variety of colors, textures, and combinations. Accordingly, all such modifications are intended to be encompassed within the scope of the subject matter described. Other substitutions, modifications, changes and omissions can be made in the design, operating conditions and arrangement of the illustrative modalities without departing from the spirit of the present invention.