Reduction of glucosinolates in oilseed meal with high levels of antinutritionals

Abstract

A process of reducing the glucosinolate content in oilseeds that include sinigrin as a glucosinolate. The oilseeds are pennycress oilseeds. The oilseeds are conditioned to degrade some of the glucosinolates. The conditioned oilseeds are pressed to expel oil from the seeds. The press cake fraction of the oilseed is extracted with hexane, desolventized and toasted to reduce the glucosinolate content. Defatted meals with low glucosinolate content made by the process are also described.

Description

Docket No. PT-2457-WO-PCTREDUCTION OF GLUCOSINOLATES IN OILSEED MEAL WITH HIGH LEVELS OF ANTINUTRITIONALSCROSS-REFERENCE TO RELATED APPLICATIONS|0001| This application claims the benefit of U.S. Provisional Application No.63 / 924.956, filed November 25, 2025, and U.S. Provisional Application No. 63 / 736,770, filed December 20, 2024, which are incorporated by reference herein in its entirety.BACKGROUND

[0002] The development of new oilseed feedstock crop for biofuels (e.g. ethanol, biodiesel, bio-jet fuel), bio-industrial uses (e.g., bioplastics, lubricants) and specialty fatty acids (e.g., erucic acid) has been an area of interest. The members of the Brassicaceae family are of interest not only because of their potential for providing a high-quality feedstock oil but also because of their ability to be grown sustainably in many regions of the world. The dry oilseeds are crushed, pressed and oil is extracted, providing meals that can be used in animal feeds. The meals from oilseeds can be modified to alter the composition of some antinutritional factors.

[0003] Thlaspi arvense (Penny cress) is an annual overwintering flowering herb plant in the family Brassicaceae. It is native to Europe and areas of Central Asia. White, lavender or pink flowers with four petals develop between five to eight seeds. Penny cress is an emerging oil seed crop that is attracting growing interest because it can serve as a winter relay crop that can grow on marginal land. Pennycress products may potentially be used as animal feed, functional food, biodiesel, and for pharmaceuticals. Pennycress seed, oil, and defatted meal could potentially be widely used for different applications and can, therefore, be cntical crop commodities for many industries.

[0004] The pennycress seeds contain high concentrations of glucosinolates, specifically sinigrin, which makes the resulting meal (post oil-extraction) unattractive for inclusion in animal feed, due to studies demonstrating certain glucosinolates having anti-nutritive qualities for certain livestock. These various antinutritional factors present in pennycress meal can affect its use as livestock feed. The use of penny cress meal for animal feed is limited, for example, by the levels of sinigrin present in the meal. If the resulting defatted meal from pennycress oil process cannot be valorized, then the value chain is significantly reduced.Docket No. PT-2457-WO-PCTSUMMARY

[0005] The present disclosure provides a process for producing a meal from oilseeds wherein the process results in degrading the levels of glucosinolates and crude fiber in the defatted meal. The process includes conditioning oilseed material at a temperature of at least 60°C, or at least 70°C, or at least 80°C, or at least 90°C, or at least 100°C, or at least 110°C, or at least 120°C. The oilseed material comprises oilseeds and / or oilseed flakes from the Brassicctceae family. The conditioning inactivates the myrosinase enzy me in the oilseed material and degrades a portion of glucosinolates in the oilseed material. The process further comprises pressing the conditioned oilseed material to separate the oilseed material into an oil fraction and a cake fraction. The process includes further treating the cake fraction with a solvent comprising hexane to extract residual oil and generate a defatted cake. The process further comprises desolventizing the defatted cake to remove residual hexane and generate a defatted meal. The desolventizing step can further degrade the glucosinolates. The defatted meal comprises a total glucosinolate content that is less, preferably at least 10% less, or at least 20% less, or at least 30% less, or at least 40% less, or at least 50% less, or at least 60% less, or at least 70% less, or at least 80% less than the glucosinolate content of the starting seed material prior to conditioning. The defatted meal comprises a total sinigrin content that is less, preferably at least 10% less, or at least 20% less, or at least 30% less, or at least 40% less, or at least 50% less, or at least 60% less, or at least 70% less, or at least 80% less than the sinigrin content of the starting seed material prior to conditioning. The oilseeds may be Thlaspi arvense seeds.

[0006] The conditioning may further comprise adding moisture to the oilseed material. The conditioning may be conducted for a duration of 30 minutes and 120 minutes. The conditioning and pressing can reduce the glucosinolate content, wherein the glucosinolate content of the cake fraction may be reduced, preferably by at least about 20%, relative to the glucosinolate content in the oilseed material prior to conditioning. The oilseed material may be conditioned at a temperature in the range of 75°C to 120°C. The DT step may be performed at a temperature of at least 80°C. The DT step is performed at a temperature between 80°C to 130°C. The glucosinolate content of the defatted meal is less, preferably at least 10% less, relative to the glucosinolate content of the oilseeds. The level of hexane in the defatted meal is less than 200ppm. The process may further comprise flaking the oilseeds to produce oilseed flakes prior to conditioning. The process may further comprise drying / cooling the defatted meal to a moisture content of 12% or less.

[0007] The disclosure also provides a defatted meal of Pennycress oilseed.Docket No. PT-2457-WO-PCT

[0008] The disclosure also provides an animal feed additive comprising a defated meal produced from Pennycress oilseed.

[0009] The present disclosure provides a process for producing a meal from oilseeds with low crude fiber content. The process contains conditioning oilseed material at a temperature of at least 60°C, or at least 70°C, or at least 80°C, or at least 90°C, or at least 100°C. The oilseed material comprises oilseeds and / or oilseed flakes from the Brassicaceae family. The conditioning inactivates the myrosinase enzyme in the oilseed material and degrades a portion of crude fiber content in the oilseed material. The process further comprises pressing the conditioned oilseed material to separate the oilseed material into an oil fraction and a cake fraction. The process further treating the cake fraction with a solvent comprising hexane to extract residual oil and generate a defated cake. The process further comprises desolventizing-toasting (DT) the defated cake to remove residual hexane and generate a defated meal. The DT step further degrades the crude fiber content. The defated meal comprises a crude fiber content of less than 20wt% of the defated meal. The oilseeds may be from pennycress. The conditioning may further comprise adding moisture to the oilseed material. The conditioning may be conducted for a duration of 30 minutes and 120 minutes. The conditioning and pressing reduces the crude fiber content, wherein the crude fiber content of the cake fraction is reduced, preferably by at least about 10%, relative to the crude fiber content in the oilseed material prior to conditioning. The total crude fiber content of the defated meal is reduced, preferably by at least 20wt%, preferably by at least 30wt%, preferably by at least 40wt%, or preferably by at least 50wt%, or preferably be at least 60wt% after dry ing and cooling the defated meal relative to the content prior to drying and cooling. The moisture content of the defatted meal may be 15% or less. The oilseed material may be conditioned at a temperature in the range of 75° C. to 120° C. The DT step may be performed at a temperature of at least 60°C, or at least 70°C, or at least 80°C. The DT step is performed at a temperature between 60° C to 130° C. The DT reduces the crude fiber content by at least 10wt% relative to the crude fiber content in the defated cake prior to DT. The DT step comprises a period of greater than 40 minutes. The crude fiber content of the defatted meal is 10% less relative to the crude fiber content of the oilseeds. The level of hexane in the defated meal is less than 200ppm. The process may further comprise flaking the oilseeds to produce oilseed flakes prior to conditioning. The process may further comprise drying / cooling the defated meal to a moisture content of 12% or less, wherein the defated meal comprises a crude fiber content of less than 15% of the defated meal after drying and cooling.Docket No. PT-2457-WO-PCT

[0010] The disclosure also provides an animal feed additive comprising a defated meal produced from penny cress oilseed, wherein the defated meal comprises less than 15wt% of total crude fiber with a moisture content less than 12wt% and an oil content of less than 5wt%, dry weight, and a protein content of at least 30wt%, dry weight. The defated meal may comprise a crude fiber content of less than 12wt% of the defated meal after drying and cooling, with a moisture content less than 12wt% and an oil content of less than 5wt%, dry weight, and a protein content of less than 50wt%, dry weight.

[0011] The disclosure also provides for a feed ration comprising the animal feed additive. The feed ration may be suitable for ruminant livestock, monogastric livestock, or poultry livestock. The feed ration may be formulated to maximize weight gain and meat quality of beef cattle in the feed lot, or to maximize milk production of lactating dairy catle.BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The drawings illustrate generally, by way of example, but not by way of limitation, various aspects discussed herein.

[0013] FIG. 1 shows a schematic diagram of an exemplary process for generating the Pennycress meal with low glucosinolate levels.

[0014] FIG. 2 shows a schematic diagram of desolventizing / toasting apparatus.DETAILED DESCRIPTION

[0015] Reference will now be made in detail to certain aspects of the disclosed subject mater, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject mater will be descnbed in conjunction with the enumerated claims, it will be understood that the exemplified subject mater is not intended to limit the claims to the disclosed subject mater.

[0016] This disclosure relates to an animal feed additive comprising a defatted meal with low levels of total glucosinolates (GSL), or low levels of the aliphatic glucosinolates. or low levels of sinigrin. The defated meal is produced from oilseeds. The oilseeds can be from the Brassicaceae family. In one aspect, the defated meal is produced from pennycress. This disclosure also relates to a process for producing a defated meal from oilseeds, e.g. pennycress. In one aspect, the method for processing the oilseed material can reduce the GSL content and the crude fiber content in the defated meal. In one aspect, the defated meal is from oilseeds that may have been conditioned under '‘hot” conditions during the processing of the oilseeds. The methodDocket No. PT-2457-WO-PCT can include a step of conditioning under “hot” conditions and expelling oil from the seeds. The method may also include a step of extracting oilseed material with hexane and desolventizing- toasting (DT) the resulting oilseed material. In one aspect, the conditioning step and the DT step can reduce the GSL content and the crude fiber content in the defatted meal.

[0017] The term “animal feed” as used herein refers to processed formulations that are fed to livestock, e.g., cattle, poultry, fish and the like. Animal feeds may be formulated to provide optimum nutrition for applications, e.g., maximizing weight gain and meat quality of beef cattle in the feed lot, maximizing milk production of lactating dairy cattle and the like.

[0018] The term “oilseed” as used herein refers to any crop species where oil is extracted from the seeds of these grains for food or industrial purposes, and includes Brassicaceae oilseeds such as canola, and non-Brassicaceae oilseeds, such as flaxseed, soybean, safflower, and sunflower. Examples of crops that are used in the production of industrial feedstock oil are camelina and penny cress.

[0019] The term “oilseed material” as used herein refers to oilseeds and / or oilseed flakes.

[0020] The term “cake” or “pressed cake” or “expeller pressed cake” or “cake fraction” or“full-fat cake” as used herein refers to the remaining fraction of the seed content after it has gone through the flaking and cooking / conditioning stage and has been mechanically pressed to extrude the bulk of the oil. The term refers to the physical character of the meal at this stage, which has been compressed into a cake-like mass rich in protein and can still contain appreciable residual oil. For the present invention, the cake can be derived from any process that extracts oil from oilseeds, leaving the cake. These terms will be used interchangeably.

[0021] The term “defatted cake” as used herein refers to a solvent extracted full-fat cake. The full fat cake is solvent extracted, e.g., with hexane, to remove residual oil.

[0022] The term “meal” or “defatted meal” as used herein refers to defatted cake after it has been treated in the desolventizer / toaster. These terms will be used interchangeably.

[0023] The term “glucosinolates” or “GSL” as used herein refers to [3-thioglucoside N- hydroxysulfates with a variable side chain (R) and a sulfur-linked [3-d-glucopyranose moiety. They represent a large and heterogeneous family of naturally occurring compounds. Glucosinolates have the following chemical structure:Docket No. PT-2457-WO-PCT

[0025] The R groups are classified as aliphatic, aromatic, -methylthioalkyl, and heterocyclic (indole). Tissue compartmentalization has evolutionarily separated the chemically and thermally stable glucosinolates and their hydrolytic enzyme myrosinase, a beta thioglucosidase.

[0026] In pennycress, the glucosinolate profile has been examined, and the predominant glucosinolate is sinigrin. Sinigrin is an aliphatic glucosinolate that can break down into allyl isothiocyanate, which has a distinctive mustard-like pungency and biological activity. Other exemplary GSLs found in penny cress can include gluconasturtiin, progoitrin, gluconapin. Sinigrin can be found at levels up to 85-95% of total glucosinolates in penny cress seeds. The exact profile and levels of GSLs, e.g. sinigrin, may vary depending on variety, growing conditions, and harvest time.

[0027] Upon tissue disruption during mechanical processing, myrosinase can immediately react with glucosinolates. The hydrolytic degradants from glucosinolates are isothiocyanates (ITCs), which have the potential to induce anticarcinogenic effects by enhancing antioxidant potential and can ameliorate nonalcoholic fatty liver diseases by decreasing the level of lipid peroxidation in cells and tissues. Glucosinolates taste bitter and pungent to humans, chicken, fish, and pigs and can reduce feed intake and lead to growth retardation in finishing pigs when their content is high.

[0028] The term “crude fiber content” as used herein refers to the indigestible fiber present in the sample, fraction and / or defatted meal. The crude fiber content is the indigestible carbohydrate in the meal. The crude fiber content includes mainly cellulose and can also include hemicellulose and lignin.

[0029] The term “anti-nutritional factors” or “ANF” refers to a number of compounds found in oilseed meals that reduce the nutritional benefit of animal feed products in which the meal is used as an additive. Glucosinolates and isothiocyanates are classified as ANFs since they can impart a bitter and pungent taste to the feed ration when present in high enough concentrations resulting in reduced palatability and reduced intake of the meal. Crude fiber can also be an ANF in meals.

[0030] Method for processing oilseeds

[0031] In one aspect, the present description relates to a method for producing oilseed meal, e.g. defatted meal, with low levels of GSL and crude fiber. The defatted meal can be derived from a variety of oilseeds. The oilseeds can be from oilseed feedstock crop for biofuels (e.g. ethanol, biodiesel, bio-jet fuel), bio-industrial uses (e.g., bioplastics, lubricants) and specialty' fattyDocket No. PT-2457-WO-PCT acids (e.g., erucic acid). The oilseeds can be from crops from the members of the Brassicaceae family, for example, mustard seeds, rapeseeds and the like. The members of the Brassicaceae family can include, for example, members from the genus Brassica, Camelina, Crambe, Thlaspi, Sinapis, Raphanus and the like. The oilseeds used in the methods described herein can include oilseeds from, for example, Brassica carinata (Ethiopian mustard). Brassica napus (canola). Brassica juncea, Brassica nigra, Brassica oleracea, Camelina sativa (False flax), Thlaspi arvense (Pennycress), Crambe abyssmica (Crambe), Sinapis alba. Brassica rapa, Raphanus sativus (radish) and the like. Crops that provide a high-quality feedstock oil and have the ability to be grown sustainably may be used to produce the defatted meal described herein.

[0032] In one aspect, the oilseeds are Thlaspi arvense oilseeds, referred to herein as pennycress oilseeds. The present method will be described in the context of penny cress oilseeds and producing penny cress defatted meal, but it will be understood that other oilseeds can be used in the methods described herein to produce defatted meals with lower GSL content and lower crude fiber content relative to the oilseeds prior to conditioning. The methods can also degrade the GSL content and the crude fiber content of the oilseed materials during processing.

[0033] Pennycress oilseeds can be from different varieties of pennycress plants. The pennycress oilseeds can be oilseeds from a variety of GSL content and sinigrin content. The pennycress oilseeds can be from a variety with a high total GSL content or a variety with a low total GSL content. In one aspect, the pennycress oilseeds are from a variety with a low GSL. In one aspect, the low GSL variety can include total GSL at an amount of about 100 pmol / g of oilseeds or less, or about 90 pmol / g of oilseeds or less, or about 80 pmol / g of oilseeds or less, or about 70 pmol / g of oilseeds or less prior to conditioning. In one aspect, the low GSL variety can include total sinigrin at an amount of about 100 pmol / g of oilseeds or less prior to conditioning, or about 80 pmol / g of oilseeds or less, or about 60 pmol / g of oilseeds or less, or about 40 pmol / g of oilseeds or less, or about 20 pmol / g of oilseeds or less prior to conditioning.

[0034] In one aspect, the pennycress oilseeds can be from a variety with a high GSL variety. In one aspect, the high GSL variety can include total GSL at an amount of about 100 pmol / g of oilseeds or more, or about 120 pmol / g of oilseeds or more, or about 140 pmol / g of oilseeds or more, or about 160 pmol / g of oilseeds or more prior to conditioning. In one aspect, the high GSL variety can include total sinigrin at an amount of about 100 pmol / g of oilseeds or more, or about 120 pmol / g of oilseeds or more, or about 140 pmol / g of oilseeds or more, or about 160 pmol / g of oilseeds or more prior to conditioning.Docket No. PT-2457-WO-PCT

[0035] In one aspect, the method of processing the oilseeds may include cleaning the oilseeds. Generally, oilseeds are harvested from pennycress crops and cleaned. Cleaning may include separating weed seeds, grain seeds, leaves, dust, sand, earth pieces, stones, prods, sticks, metal, and other extraneous impurities from the oilseeds before storing. The oilseeds may enter the crushing plant with no more than 5wt%, or no more than 4wt%, or no more than 3wt%, or no more than 2wt%, or no more than 1 ,8wt% foreign materials in the oilseeds. The cleaning process may include removing dust and light material, for example, by aspiration, sieving and the like. The amounts of pods, fines, loose hulls can be minimized in the oilseeds.

[0036] In one aspect, the method comprises maximizing the removal of the pods from the oilseeds. The removal of pods from the oilseeds can be increased using finer screens to remove more of the pods. The pods contain high levels of crude fiber and minimizing the pods content in the oilseeds can reduce the crude fiber content in the oilseeds that are processed and thus, the defatted meal generated using the methods described herein. The oilseeds, preferably, enter the crushing plant with no more than 5wt%, no more than 4wt%, or no more than 3wt%, or no more than 2wt%, or no more than 1 ,8wt% of pods in the oilseeds.

[0037] In one aspect, the oilseed material may be preheated prior to other processing steps. Preheating may be performed to heat the oilseeds to between about 30°C and about 110°C, or between about 30°C and about 100°C, or between about 30°C and about 90°C, or between about 30°C and about 80°C, or between about 30°C and about 70°C, or between about 30°C and about 60°C, or between about 30°C and about 50°C, or between about 30°C and about 40°C. The preheating can be performed for less than 10 minutes, or less than 5 minutes, or less than 4 minutes, or less than 3 minutes, or less than 2 minutes. The preheated seeds are more easily pliable and thus less probable to shatter during the flaking and / or conditioning steps. Preheating may be conducted, for example, in bed heaters using warm air or direct steam. Externally heated rotary kilns may also be used for seed heating. Other methods of heating may be used and are also within the scope of this description.

[0038] In one aspect, the method may, optionally, include generating oilseed flakes from oilseeds in a flaker. The oilseeds may be flaked after preheating as described above. The oilseeds may be flaked prior to or after conditioning as described herein. The oilseeds may be flaked if the oilseeds are larger, e.g. greater than about 2mm. Oilseeds seeds larger than 2mm, or larger than 4mm, or larger than 5mm, may be flaked in a flaker. The method may include preheating the cleaned oilseeds prior to processing the oilseeds through a flaker to generate penny cress flakes.Docket No. PT-2457-WO-PCT

[0039] A variety of methods are known in the art for generating oilseed flakes from oilseeds and all are within the scope of this description. The method can comprise generating oilseed flakes by introducing the oilseeds into a flaking apparatus (flaker). The oilseeds may or may not be conditioned prior to flaking. Flaking can be applied to rupture the seed coat and to flatten the cotyledons. Flaking can enhance the oil output during processing. Flakers are known in the art. A process for flaking, crushing and refining oilseeds is described, for example, by Unger, E.H. “Processing” in Canola Chemistry, Production, Processing, and Utilization, p. 163- 188, 2011. The cleaned and optionally, conditioned, oilseeds can be crushed, for example, in a roller mill to generate flakes. The flakes can be. for example, between about 0.2 and 0.5mm in thickness, or between about 0.25 and 0.45, or between about 0.3-0.4mm, or between about 0.3 and 0.38 mm in thickness. The pressure to obtain the flakes can be, for example, between 500 and 750psi.

[0040] In one aspect, the oilseed material comprises fine oilseeds. Fine oilseeds may be used in the methods described herein without flaking. The oilseeds can be conditioned and further processed as described herein without being subject to a flaking step. If the cleaned seeds are fine oilseeds, the flaking step can be eliminated, and the cleaned fine oilseeds can be conditioned as described herein. Fine oilseeds are oilseeds that are less than or 3 mm, or less than 2 mm. or less than 1 mm may be conditioned as oilseeds. The method may not include a flaking step if the oilseeds are about 2mm or less. In one aspect, a flaking step may be eliminated prior to conditioning and / or pressing.

[0041] In one aspect, the amount of moisture in the incoming oilseed material, e.g., the oilseed entering the process described herein, can vary and may be based on plant configurations and the level of drying prior to processing. The seed moisture content may be at least 2wt%, or at least 5\\1%. or at least 7wt% by controlling the airflow. The seed moisture content can be less than 15wt%, or less than 12wt%, or less than 10wt% by controlling the airflow. The seed moisture content can be between about 2wt% and 15wt%, or between 5wt% and about 10wt%, or between about 7wt% and l lwt%, or between about 7wt% and 10wt% by controlling the airflow. In one aspect, the seed moisture can be in a range of 7.0 to 9.0 wt% when entering a flaking and / or conditioning step. Other moisture ranges are also within the scope of this description.

[0042] In one aspect, the method comprises conditioning the oilseed material. ■’Conditioning’' as used herein refers to treating the oilseed material to optimize expelling the oil from the oilseed material in the expeller. Conditioning the oilseed material can include, for example, heating the oilseed material to a selected temperature for a duration of time. The moistureDocket No. PT-2457-WO-PCT content of the oilseed material may be adjusted during conditionmg / heating. Conditioning the oilseed material can also include adding moisture to the oilseed material in the conditioner. Moisture can include the addition of water and / or steam.

[0043] In one aspect, the method can include conditioning the oilseed material in the presence of added moisture in the conditioner. The method can include adding water and / or steam to the oilseed material in the conditioner to a desired moisture. The moisture content in the conditioner can be at least 2wt%, or at least 5wt%, or at least 7wt% by weight of the oilseeds. The moisture content in the conditioner can be less than 15wt%, or less than 12wt%, or less than 10wt% by weight of the oilseeds. The moisture content in the conditioner can be between about 5wt% and 15wt%, or between 5wt% and about 10wt%, or between about 7wt% and 15wt%, or between about 7wt% and 10wt% by weight of the oilseeds. In one aspect, the moisture content in the conditioner can be in a range of 7.0 to 9.0 vA% when entering a flaking and / or conditioning step. Other moisture ranges are also within the scope of this description.

[0044] The method can further comprise conditioning the oilseed material by heating. The heating may use direct heating and / or indirect heating. Preferably, the oilseed material is indirectly heated in cookers. Indirect heating can include, for example, contacting the oilseed material with a hot surface. In one aspect, indirect heated can include the use of steam-heated tubes packed around the internal wall of the drum inside the conditioner as the conditioner heats the oilseed material.

[0045] In one aspect, the oilseed material can be conveyed to a heated drum where the oilseed material is conditioned. The oilseed material may be conditioned by heating to a temperature of, for example, at least 60°C, or at least 70°C, or at least 80°C, or at least about 90°C, at least 100 C. The oilseed material may be heated to no more than 150°C, or no more than 140°C, or no more than 130°C, or no more than 120°C. The oilseed material may be heated to between about 60°C and about 130°C, or between about 60°C and about 110°C, or between about 60°C and about 100°C, or between about 70°C and about 130°C. or between about 70°C and about 110°C, or between about 70°C and about 100°C, or between about 70°C and about 90°C, or between about 80°C and about 1 10°C, or between about 80°C and about 100°C, or between about 90°C and about 110°C. The oilseed material can be indirectly heated to reduce or control the moisture content of the flake or seeds to pressing. In one aspect, the oilseed material is heated to temperature greater than 50°C to inactivate the myrosinase activity. Preferably, the oilseed material is heated quickly through the temperature range between 40°C and 50°C, the active temperature range for myrosinase.Docket No. PT-2457-WO-PCT

[0046] The oilseed material may be conditioned for at least 15 min, or at least 20 minutes, or at least 30 min, or at least 45 min, or at least 60 minutes. The oilseed material may be conditioned for up to 180 minutes, or up to 150 minutes, or up to 120 minutes, or up to 60 min. The oilseed material may be conditioned for between 15 and 180 min, or between 20 and 150 minutes, or 20 and 120 minutes, or between 20 and 60 minutes, or between 30 and 180 minutes, or between 30 and 150 minutes, or between 30 and 120 minutes, or between 30 and 90min, or between 30 and about 60 minutes, or between 40 and 180 minutes, or between 40 and 150 minutes, or between 40 and 120 minutes, or between 40 and 60 minutes.

[0047] Without being bound by any theory, it is thought that heating the oilseed material in the conditioner can reduce oil viscosity bound to the seeds thereby promoting coalescence of oil droplets. This can increase the diffusion rate of hexane solvent into the pressed cake during the solvent extraction process. Heating can also contribute to the inactivation of enzymes such as myrosinase and lipase. The conditioning can include treating the oilseed material at the conditioning parameters, e.g., moisture, heat, time, as described herein to inactivate the enzymes, e.g., myrosinase.

[0048] In one aspect, the oilseed material is conditioned to substantially or completely inactivate the myrosinase enzyme activity. By substantially, it is meant that at least 70% of the myrosinase activity, or at least 80%, or at least 90%, or at least 95%, or at least 98%, or at least 99% of the myrosinase enzyme activity is eliminated compared to the myrosinase activity in the oilseed material prior to conditioning.

[0049] In one aspect, the oilseed material is conditioned to substantially or completely inactivate the lipase activity’. By substantially, it is meant that at least 70% of the lipase activity, or at least 80%, or at least 90%, or at least 95%, or at least 98%, or at least 99% of the lipase activity is eliminated compared to the lipase activity in the oilseed material prior to conditioning.

[0050] The GSL content in the penny cress oilseeds can vary depending on the variety, the growing conditions and the like. In one aspect, the GSLs in the pennycress oilseeds can be degraded during the conditioning step of the process for making the defatted oilseed meal. In one aspect, the aliphatic GSL can be degraded during the conditioning step. Aliphatic GSLs can include, for example, allylglucosinolate (sinigrin), benzylglucosinolate (glucotropaeolin), phenethylglucosinolate (gluconasturtiin), 2-hydroxy-3-butenyl glucosinolate (progoitrin), 3- butenyl glucosinolate (gluconapin). 4-pentenyl glucosinolate (glucobrassicanapin), 4- methylsulfmylbutyl glucosinolate (glucoraphanin), 4-methylsulfinyl-3-butenyl glucosinolate (glucoraphenin), 5-methylsulfinylpentyl glucosinolate (glucoberteroin), 9-methylsulfmylnonylDocket No. PT-2457-WO-PCT glucosinolate (glucocamelinin)and the like. In one aspect, sinignn, an aliphatic GSL, can be degraded during the conditioning step.

[0051] In one aspect, the conditioning of the oilseed material can reduce the total GSL content in the oilseed material. The GSL content of the oilseed material can be reduced by at least about 10%, or at least about 15%, or at least about 20%. or at least 25% relative to the GSL content in the oilseed material prior to conditioning. The GSL content of the oilseed material can be reduced by at most 80%, or at most 70%, or at most 60%, or at most 50%, or at most 40%, or at most 35% relative to the GSL content in the oilseed material prior to conditioning. The GSL content of the oilseed material can be reduced by between about 10% and about 80%, or between about 10% and about 70%, or between about 10% and about 60%, or between about 10% and about 50%, or between about 10% and about 40%, or between about 15% and about 70%, or between about 15% and about 60%, or between about 15% and about 50%, or between about 15% and about 40%, or between about 20% and about 70%. or between about 20% and about 60%, or between about 20% and about 50%, or between about 20% and about 45%, or between about 25% and about 70%, or between about 25% and about 60%, or between about 25% and about 50%, or between about 25% and about 40% relative to the GSL content in the oilseed material prior to conditioning.

[0052] In one aspect, the conditioning of the oilseed material can reduce the aliphatic GSL content in the oilseed material. The aliphatic content of the oilseed material can be reduced by at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least 25% relative to the aliphatic GSL content in the oilseed material prior to conditioning. The aliphatic GSL content of the oilseed material can be reduced by at most 80%, or at most 70%, or at most 60%, or at most 50%, or at most 40%, or at most 35% relative to the aliphatic GSL content in the oilseed material prior to conditioning. The aliphatic GSL content of the oilseed material can be reduced by between about 10% and about 80%, or between about 10% and about 70%, or between about 10% and about 60%, or between about 10% and about 50%, or between about 10% and about 40%, or between about 15% and about 70%. or between about 15% and about 60%, or between about 15% and about 50%, or between about 15% and about 40%, or between about 20% and about 70%, or between about 20% and about 60%, or between about 20% and about 50%, or between about 20% and about 45%, or between about 25% and about 70%, or between about 25% and about 60%. or between about 25% and about 50%. or between about 25% and about 40% relative to the aliphatic GSL content in the oilseed material prior to conditioning.Docket No. PT-2457-WO-PCT

[0053] In one aspect, the conditioning of the oilseed material can reduce the total simgrin content in the oilseed material. The sinigrin content of the oilseed material can be reduced by at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least 25% relative to the sinigrin content in the oilseed material prior to conditioning. The sinigrin content of the oilseed material can be reduced by at most 80%, or at most 70%. or at most 60%, or at most 50%, or at most 40%, or at most 35% relative to the sinigrin content in the oilseed material prior to conditioning. The sinigrin content of the oilseed material can be reduced by between about 10% and about 80%, or between about 10% and about 70%, or between about 10% and about 60%, or between about 10% and about 50%. or between about 10% and about 40%. or between about 15% and about 70%, or between about 15% and about 60%, or between about 15% and about 50%, or between about 15% and about 40%, or between about 20% and about 70%, or between about 20% and about 60%, or between about 20% and about 50%, or between about 20% and about 45%, or between about 25% and about 70%. or between about 25% and about 60%. or between about 25% and about 50%, or between about 25% and about 40% relative to the sinigrin content in the oilseed material prior to conditioning.

[0054] In one aspect, the conditioning of the oilseed material can reduce the crude fiber content in the oilseed material. The crude fiber content of the oilseed material can be reduced by at least about 5%. or at least about 10%, or at least about 15%, or at least about 20% relative to the crude fiber content in the oilseed material prior to conditioning. The crude fiber content of the oilseed material can be reduced by at most 40%, or at most 30%, or at most 20%, or at most 10% relative to the crude fiber content in the oilseed material prior to conditioning. The crude fiber content of the oilseed material can be reduced by between about 5% and about 40%, or between about 5% and about 30%, or between about 10% and about 40%, or between about 10% and about 30%, or between about 15% and about 40%, or between about 15% and about 30% relative to the crude fiber content in the oilseed material prior to conditioning.

[0055] In one aspect, the method further comprises pressing the conditioned oilseed material in an expeller. The conditioned oilseed material may be pressed in a large screw press expeller. In one aspect, the pressing can remove oil from the oilseed material to generate an oil fraction and a cake fraction. Cake fraction may also be referred to herein as pressed cake or fat cake or full fat cake. In one aspect, the pressing can remove at least about 30wt% of the oil, or at least about 40wt% of the oil, or at least about 50wt% of the oil, or at least about 60wt% of the oil in the oilseed material. The pressing may remove between about 30wt% to 80 wt%, or betweenDocket No. PT-2457-WO-PCT about 30wt% to 70 wt%, or between about 40wt% to 70 wt%, or between about 50wl% to 70wt% of the oil content in oilseed material.

[0056] In one aspect, the expeller / press may, for example, contain a screw that is enclosed in a cage formed by longitudinal bars, held by a heavy frame. The screw can squeeze the oilseed material across bars. The bars can be separated by metallic spacers to allow the oil to flow between the bars while the squeezed cake or pressed cake can be discharged to the extraction plant. Pressed oil will have some footing that may be extruded with the oil while pressing. These footings may be separated by setting and centrifuge. The heavy phase can be returned and mixed back with the feed of the press while the high phase (clarified pressed oil) can go to storage.

[0057] In one aspect, the cake fraction can include a reduced amount of glucosinolates relative to the glucosinolates in the oilseed material prior to the conditioning step. The cake fraction can include an amount of glucosinolates that are at least 5% less, or at least 10% less, or least 20% less, or at least 25% less, or at least 30% less, or at least 40% less relative to the glucosinolates in the oilseed material prior to the conditioning step.

[0058] In one aspect, the cake fraction can include a reduced amount of sinigrin relative to the sinigrin in the oilseed material prior to the conditioning step. The cake fraction can include an amount of sinigrin that is at least 5% less, or at least 10% less, or least 20% less, or at least 25% less, or at least 30% less, or at least 40% less relative to the sinigrin in the oilseed material prior to the conditioning step.

[0059] In one aspect, the cake fraction can include a reduced amount of crude fiber relative to the crude fiber in the oilseed material prior to the conditioning step. The cake fraction can include an amount of crude fiber that is at least 5% less, or at least 10% less, or least 20% less, or at least 25% less, or at least 30% less, or at least 40% less relative to the crude fiber in the oilseed material prior to the conditioning step.

[0060] In one aspect, the method further comprises treating the press cake fraction with hexane in a solvent extractor. The cake fraction can be treated with hexane as the solvent to extract residual oil from the cake fraction. A variety of solvent extractors are known in the art and can be used to treat the cake fraction with hexane. The extractor can be, for example, a continuous belt extractor that can move the cake fraction and the mixture (hexane plus oil) in opposite directions to achieve a continuous counter current extraction. Series of pumps move the mixture over the cake, so that the mixture with the highest concentration in oil is used to extract the entering cake fraction with the highest concentration in oil. At the opposite extremity of the extractor, pure hexane washes the cake fraction with the lowest concentration of oil.Docket No. PT-2457-WO-PCT

[0061] In one aspect, the cake fraction is extracted with solvent that can range between pure hexane and a mixture comprising hexane and oil. The mixture can comprise between about 10 and 40wt% of oil, or between about 20 to 30wt% of oil, or between about 20 and 25wt% of oil. Solvent / mixture percolates through the cake or submerges it, allowing the diffusion of the oil contained in the cake into the liquid phase. After leaving the solvent extractor, the resulting end cake is a defatted cake.

[0062] The defatted cake can include about 3wt% oil or less, 2wt% oil or less, or lwt% oil or less, 0.8wt% or less, or 0.6wt% oil or less, or 0.5wt% oil or less, or 0.4wt% oil or less. The defatted cake can include between about 0.5 and 3wt% oil, or between about 0.5wt% and 2.5 wt% oil, or between about 0.5wt% and 2wt% oil, or between about 1.0wt% and 3wt% oil, or between about l.Owt % and 2.5wt% oil, or between about l.Owt % and 2.0wt% oil.

[0063] The residual mixture (hexane / oil) separated from the defatted cake can include at least about 10wt% oil, or at least about 15wt% oil, or at least about 20wt% oil. or at least about 25wt% oil, or at least about 30wt% oil. The residual mixture (hexane / oil) separated from the defatted cake can include about 90wt% hexane or less, or 85 wt% hexane or less, or about 80wt% hexane or less, or about 75wt% hexane or less, or about 70wt% hexane or less. The residual mixture (hexane / oil) separated from the defatted cake can include 10-20wt% oil and 90-80wt% hexane, 15-25wt% oil and 85 to 75wt% hexane, or 20-30 wt% oil and 80 to 70 wt% hexane. The hexane can be removed by evaporation and stripping from the oil in the residual mixture to make extracted oil. This extracted oil can be mixed with the pressed oil to make a crude oil that is a combination of pressed and extracted oil.

[0064] In one aspect, the method further comprises desolventizing-toasting (DT) the defatted cake to remove the residual hexane and generate a defatted meal. The DT of the defatted cake can further degrade the GSLs and result in reducing and / or eliminating the GSL in the defatted meal. In one aspect, the DT step of the defatted cake can further degrade the crude fiber and result in reducing and / or eliminating the crude fiber content in the defatted material.

[0065] After the solvent extraction, the defatted cake can be transferred to a desolventizer- toaster, where it is heated. The DT step can remove remaining hexane, reduce GSL content and reduce the moisture content of the defatted cake. Most of the solvent is removed by heating the defatted cake on steam-heated plates. Removal of the final traces of solvent is carried out by injecting steam through the defatted cake. Injecting steam through the defatted cake can also toast the defatted cake.Docket No. PT-2457-WO-PCT

[0066] DT can be conducted in an apparatus having a variety of configurations and all are within the scope of this description. Desolventization in the DT can remove the solvent held in the extracted cake by means of evaporation and steam stripping. In one aspect, the DT apparatus can be a closed vessel with two or more zones. The zones can include a pre-desolventization zone, high desolventization zone(s). toasting zone, stripping zone, cooling zone and the like. Other zones may also be present in the DT apparatus that enhance the DT process.

[0067] In one aspect, the DT apparatus can include a vertical stack of cylindrical gas-tight pans that can be steam heated from the base. FIG. 2 is one exemplary DT apparatus configuration. Other DT apparatus with other configurations may also be used and are within the scope of this description. Starting from the top of the DT, the extracted defatted cake can be spread on tray one where it is equally distributed by means of a sweep arm. From there, the cake can be moved continuously from tray to tray through tray openings. Each of the toasting trays can have hollow punches for venting vapors from one tray to the next. Direct steam is sparged in the DT by the bottom tray and moved upward from tray-to-tray resulting in stripping hexane from the cake.

[0068] The main trays can be used for toasting the product. Main trays are perforated trays that can provide the steam. They are designed to provide both indirect heating and direct steam contact to remove the bulk of the solvent from the defatted extracted cake. The added steam can carry the evaporated hexane and get condensed in external condenser and returned to the hexane / water separator to separate the hexane from water. The hexane phase may be recycled and reused back in the extractor.

[0069] In one aspect, the defatted cake can be desolventized but not toasted. The defatted meal, without toasting, can have a moisture content between about 5wt% and about 15wt%, or between about 6wt% and about 13wt%, or between about 8wt% and about 12wt%, or between 9wt% and 1 lwt%. Moisture amounts in the defatted penny cress meal outside this range are also within the scope of this description.

[0070] Desolventizing and toasting generates a defatted meal with the hexane solvent reduced and / or eliminated. The DT step also lowers the moisture content in the defatted meal exiting the DT step. In one aspect, DT step can be conducted at a temperature of at least 80°C, or at least 90°C, or at least 100°C, or at least 105°C. The DT step can be conducted at a temperature between about 80°C and about 110°C, or between about 80°C and about 105°C, or between 80°C and 100°C, or between 85°C and 110°C, or between 85°C and 105°C. or between 85°C and 100°C, or between about 90°C and 110°C, or between 90°C and 105°C, or between 90°C and about 100°C,Docket No. PT-2457-WO-PCT or between 95°C and about 110°C, or between 95°C and about 105°C, or between 95°C and about 100°C, or between 100°C and about 110°C, or between about 100°C and 105°C.

[0071] The DT step can be conducted for at least 30 minutes, or at least 35 minutes, or at least 40 minutes, or at least 45 minutes, or at least 60 minutes. The DT step can be conducted for at most 120 minutes, or at most 90 minutes, or at most 60minutes, or at most 55 minutes, or at most 50 minutes, or at most 45 minutes. The DT step can be conducted for between about 30 minutes and about 120 minutes, or about 30 minutes and about 90 minutes, or between about 30 minutes and about 80 minutes, or between about 30 minutes and about 70 minutes, or about 30 minutes and 60 minutes, or between about 45 minutes and 90 minutes, or between about 45 minutes and about 80 minutes, or between about 45 minutes and 70 minutes, or between about 45 minutes and 65 minutes, or between about 45 minutes and 60 minutes, or between about 50 minutes and 90 minutes, or between about 50 minutes and 80 minutes, or between about 50 minutes and 70 minutes, between about 55 minutes and 65 minutes.

[0072] In one aspect, the DT may also include adding gum and / or soap stock out of an oil refining process to the extracted defatted cake prior to or in the DT. The soap stock may be added to add calories and / or to increase the nutritional value of the defatted meal to the health of the animal. In one aspect, the soap stock may be pennycress soap stock, a byproduct of refining Penny cress oil. The content of the gum (out of degumming process) and / or soap stock (out of the caustic refining process) can be about 50 % oil and about 50 % concentrated phospholipids or soap of fatty' acid. The composition of the soap stock can be, for example, 20 % oil, 20 % soap / gums, and 60 % water. Soap stock with other composition may also be used. The DT step can also dry the defatted cake to further remove the hexane and moisture form a defatted meal.

[0073] The DT step performed on the defatted cake can generate a defatted meal. The defatted meal can have a moisture content of less than 20wt% of moisture in the defatted meal, or less than 15wt%, or less than 13wt%, or less than 10wt%, or less than 81x4% of moisture in the defatted meal. The defatted meal can have a moisture content of between about 5wt% and about 25wt%, or between about 5wt% and about 20wt%, or between about 10wt% and about 25wt% or between about 10wt% and about 20wt%.

[0074] The defatted meal can have a hexane content of lOOOppm or less, or 800ppm or less, or 600ppm or less, 500ppm or less, 400ppm or less, 300ppm or less, 200 ppm or less, or lOOppm or less. The defatted meal can have a hexane content of between lOOppm and lOOOppm, or between lOOppm and 800ppm, or between lOOppm and 600ppm, or between lOOppm and 500ppm, or between 200ppm and lOOOppm, or between 200ppm and 800ppm, or between 200ppmDocket No. PT-2457-WO-PCT and 600ppm, or between 200ppm and 500ppm, or between 300ppm and lOOOppm, or between 300ppm and 800ppm, or between 300ppm and 600ppm, or between 300ppm and 500ppm, or between 400ppm and lOOOppm, or between 400ppm and 800ppm, or between 400ppm and 600ppm, or between 400ppm and 500ppm.

[0075] The GSL content and the crude fiber content in the defatted cake can be further degraded during the DT step to generate defatted meal with lower GSL content and lower crude fiber content. The DT step can reduce the GSL content in the defatted meal by at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50% relative to the GSL content in the whole oilseed. The DT step can reduce the GSL content in the defatted meal by between 20% and 80%, or between 20% and 70%, or between 20% and 60%, or between 30% and 80%, or between 30% and 70%, or between 30% and 60%, or between 40% and 80%, or between 40% and 70%, or between 40% and 60% relative to the GSL content in the whole oilseed.

[0076] The DT step can further reduce the crude fiber content in the defatted meal relative to the whole oilseed. The DT step can reduce the crude fiber content in the defatted meal by at least 10%, or at least 15%, or at least 20%, or at least 25% relative to the crude fiber content in the whole oilseed. The DT step can reduce the crude fiber content in the defatted meal by between 10% and 50%, or between 15% and 50%, or between 15% and 40%, or between 15% and 30%, or between 20% and 50%, or between 20% and 40%, or between 20% and 30%, relative to the crude fiber content in the whole oilseed.

[0077] The DT step may further comprise cooling the defatted meal. The temperature of the defatted meal exiting the DT can be about 80°C or lower, or about 70°C or lower, or about 65°C or lower. The temperature of the defatted meal exiting the DT can be between about 50°C and about 80°C, or between about 50°C and about 75°C, or between about 50°C and about 65°C, or between about 50°C and about 60°C, or between about 55°C and about 75°C, or between about 55°C and about 65°C, or between about 60°C and about 70°C.

[0078] The defatted meal exiting the DT can be further cooled and dried in a drier / cooler. The moisture content of the defatted meal can be reduced after the drying and cooling. The defatted meal after the drying and cooling may be cooled to temperature of less than 40°C, or less than 35°C, or less than 30°C. The defatted meal may be cooled to a temperature of between 20 and 40°C, or between 20 and 35°C, or between 20 and 30°C, or between 25 and 4-°C, or between 25 and 35°C after the drying and cooling. The moisture content of the defatted meal after drying and cooling can be less than 20wt%, or less than 18wt%, or less than 15wt%, or less than 12wt% based on the weight of the defatted meal. The moisture content of the defatted meal after dryingDocket No. PT-2457-WO-PCT and cooling can be between about 10wt% and 20wt%, or between about 10wt% and 18wt%, or between about 10wt% and 15wt%, or between about 10wt% and 12wt%, or between about 5wt% and 15\\ t%. or between about 5wt% and 12wt%, or between about 5wt% and 10wt% based on the weight of the defatted meal.

[0079] The defatted meal generated after the drying and cooling step can include a total GSL content of less than 150 pmol / gm (moisture and oil-free) of defatted meal, or less than 120 pmol / gm, or less than 100 pmol / gm, or less than 80 pmol / gm, or less than 60 pmol / gm of meal. The defatted meal generated after the DT step can include a total GSL content of between 20 and 200 pmol / gm of defatted meal, or between 20 and 150 pmol / gm, or between 20 and 100 pmol / gm, or between 20 and 80 pmol / gm, or between 5 and 100 pmol / gm, or between 5 and 80 pmol / gm, or between 5 and 60 pmol / gm.

[0080] The defatted meal generated after the drying and cooling step can include a crude fiber content of less than 25wt%, or less than 20wt%, or less than 17wt%, or less than 15wt%, or less than 12wt%. The defatted meal generated after the drying and cooling step can include a crude fiber content of between 10wt% and 25wt%, or between 10wt% and 20wt%, or between 10wt% and I 5wt%.

[0081] During the dry ing and cooling step, further residual hexane can be removed to lower the concentration of the hexane in the defatted meal. The concentration of the hexane can be less than 200ppm, or less than 150 ppm, or less than 100 ppm, or less than 50ppm, or less than 25ppm. The dried and cooled defatted meal can have a hexane content of between lOppm and 200ppm, or between lOppm and 150ppm, or between lOppm and lOOppm, or between lOppm and 75ppm, or between lOppm and 50 ppm, or between lOppm and 25ppm, or between 25ppm and 200ppm, or between 25ppm and lOOppm, or between 25ppm and 75ppm, or between 25ppm and 50ppm.

[0082] The moisture content in the dried meal can also be reduced to less than 20wt%, or less than 15wt%, or less than 12wt%, or less than 10wt%. The moisture content in the dried meal can be between 5wt% and 20wt%, or between 5wt% and 15wt%, or between 5wt% and 10wt%, or between 10wt% and 20wt%, or between 10wt% and 15wt%, or between 10wt% and 12wt%.

[0083] In one aspect, the methods described herein for generating the defatted meal do not include the use of exogenous myrosinase enzyme. The GSL content in the defatted meal is not significantly or at all reduced by the activity of the myrosinase enzyme.

[0084] The methods described herein reduce the GSL content during more than one step. In one aspect, the GSL content in the oilseeds is reduced during the conditioning step and the DTDocket No. PT-2457-WO-PCT step. The defatted meal generated by the process described herein can have reduced GSL content compared to the GSL content in the oilseeds. The degradation of the GSL may occur, for example, in the conditioning and / or the DT step. The method can reduce the GSL content in the defatted meal by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% compared to the GSL content in the oilseed material prior to conditioning. The method can reduce the GSL content in the defatted meal between 20% and 90%, or between 20% and 80%, or between 20% and 70%, or between 20% and 60%, or between 30% and 90%, or between 30% and 80%, or between 30% and 70%, or between 30% and 60%, or between 40% and 90%, or between 40% and 80%, or between 40% and 70%, or between 40% and 60%, or between 50% and 90%, or between 50% and 80%, or between 50% and 70%, or between 50% and 60%, or between 60% and 90%, or between 60% and 80%, or between 60% and 70% compared to the GSL content in the oilseed material prior to conditioning.

[0085] The methods described herein reduce the sinigrin content during more than one step. In one aspect, the sinigrin content in the oilseeds is reduced during the conditioning step and the DT step. The defatted meal generated by the process described herein can have reduced sinigrin content compared to the sinigrin content in the oilseeds. The degradation of the sinigrin may occur, for example, in the conditioning and / or the DT step. The method can reduce the sinigrin content in the defatted meal by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% compared to the sinigrin content in the oilseed material prior to conditioning. The method can reduce the sinigrin content in the defatted meal between 20% and 90%, or between 20% and 80%, or between 20% and 70%, or between 20% and 60%, or between 30% and 90%, or between 30% and 80%. or between 30% and 70%, or between 30% and 60%, or between 40% and 90%, or between 40% and 80%, or between 40% and 70%, or between 40% and 60%, or between 50% and 90%, or between 50% and 80%, or between 50% and 70%, or between 50% and 60%, or between 60% and 90%, or between 60% and 80%, or between 60% and 70% compared to the sinigrin content in the oilseed material prior to conditioning.

[0086] The defatted meal generated by the process described herein can have reduced crude fiber content compared to the crude fiber content in the oilseeds. The degradation of the crude fiber may occur, for example, in the conditioning and / or the DT step. The methods described herein reduce the crude fiber content during more than one step. In one aspect, the crude fiber content in the oilseeds is reduced during the conditioning step and the DT step. The defatted meal generated by methods described herein can have reduced crude fiber content compared to theDocket No. PT-2457-WO-PCT crude fiber content in the whole oilseeds due to the advantageous degradation of the crude fiber content in the conditioning and the DT step. The method can reduce the crude fiber content in the defatted meal by at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, compared to the crude fiber content in the oilseed material prior to conditioning. The method can reduce the crude fiber content in the defatted meal by at between 20% and 80%. or between 20% and 70%, or between 20% and 60%, or between 30% and 80%, or between 30% and 70%, or between 30% and 60%, or between 40% and 80%, or between 40% and 70%, or between 40% and 60%, or between 50% and 80%, or between 50% and 70%, or between 50% and 60%, or between 60% and 80%, or between 60% and 70% compared to the crude fiber content in the oilseed material prior to conditioning.

[0087] In one aspect, Fig. 1 shows a schematic flow diagram for a process of generating defatted meal with low total GSL levels and low crude fiber content. The process comprises cleaning the oilseeds. The oilseeds may be flaked in a flaker as described above. The oilseeds and / or the flakes may be conditioned. Conditioning can include injection of steam and / or water. Conditioning can include heating the oilseed material. The conditioned oilseed material can be transferred to an expeller. The expeller can separate the oil fraction from the cake fraction. The oil fraction can be further processed to separate any solids from the oil. The oil can be transferred to storage and any solids may be reintroduced into the expeller.

[0088] The cake fraction can be transferred to an extractor to remove the residual oil in the cake. Hexane can be introduced into the extractor and the cake can be permeated with the hexane as described above. The hexane / oil mixture can be removed from the extractor and further separated by distillation into hexane and oil. The extracted and now defatted cake can be transferred to a DT apparatus. Steam, and optionally gum or soap stock, can be added to the DT apparatus to reduce and / or remove the hexane in the defatted cake. The DT apparatus can also reduce the moisture content in the defatted cake to form a defatted meal. The defatted meal can be further processed by transferring to a dryer / cooler to further remove hexane traces and reduce the moisture content. The dried meal may be further ground and / or sifted prior to storage.

[0089] In one aspect, the present disclosure relates to a process for producing a meal of penny cress with reduced glucosinolate content comprising: a. conditioning oilseed material by cooking the oilseed material to a temperature of at least 60°C, wherein the oilseed material comprises oilseed flakes and fine oilseeds and wherein the conditioning inactivates the myrosinase enzyme;Docket No. PT-2457-WO-PCT b. pressing the oilseed material to remove oil from the oilseed material and producing an oil fraction and a cake fraction; wherein the oilseeds are from the Brassicaceae family and wherein the process degrades a portion of glucosinolates in the oilseed material relative to the glucosinolates in the oilseed material prior to conditioning.The method can further comprise the following steps: c. treating the cake fraction with a solvent comprising hexane to extract residual oil to generate a defatted cake fraction; and d. desolventizing-toasting (DT) the defatted cake fraction to remove residual hexane and further degrade the glucosinolates and generate a defatted meal.

[0090] In one aspect, the present disclosure relates to a process for producing a meal with a reduced crude fiber content from oilseeds comprising:

[0091] a. conditioning oilseed material at a temperature of at least 60°C. wherein the oilseed material comprises oilseeds and / or oilseed flakes;

[0092] b. pressing the conditioned oilseed material to separate the oilseed material into an oil fraction and a cake fraction; and wherein the oilseeds are from the Brassicaceae family and wherein the process inactivates the myrosinase enzy me in the oilseed material and degrades a portion of the crude fiber content in the oilseeds.

[0093] The method can further comprise the following steps:

[0094] c. treating the cake fraction with a solvent comprising hexane to extract residual oil and generate a defatted cake; and

[0095] d. desolventizing-toasting (DT) the defatted cake to remove residual hexane and generate a defatted meal, wherein the DT further degrades the crude fiber content and wherein the defatted meal comprises a total crude fiber content of less than 20% by weight;

[0096] In one aspect, the methods described herein do not include the addition of exogenous myrosinase enzyme. The degradation of the glucosinolates by myrosinase is minimized or eliminated in the methods described herein.

[0097] In one aspect, the methods described herein can degrade glucosinolates, preferably aliphatic glucosinolates. The glucosinolates can include, for example, sinigrin, glucotropaeolin, gluconasturtiin. and the like. In one aspect, the methods described herein can degrade sinigrin. The methods may degrade at least 20wt%, or at least 30wt%, or at least 50wt%, or at least 60wt%, or at least 70wt%, or at least 80wt%, of the glucosinolates present in the oilseeds prior toDocket No. PT-2457-WO-PCT conditioning. The methods may degrade at least 20wt%, or at least 30wt%, or at least 50wt%, or at least 60wt%, or at least 70wt%, or at least 80wt%, of the sinigrin present in the oilseeds prior to conditioning.Composition of Defatted Meal

[0098] In one aspect, the present description comprises an animal feed additive, wherein the animal feed additive comprises the defatted meal. The defatted meal can be derived from oilseeds.

[0099] Preferably, the defatted meal is from oilseeds from the members of the Brassicaceae family, for example, mustard seeds, rapeseeds and the like. The members of the Brassicaceae family can include, for example, members from the genus Brassica, Camelina, Crambe, Thlaspi, Sinapis, Raphanus and the like. The members of the Brassicaceae family can include, for example, Brassica carinata (Ethiopian mustard), Brassica napus (canola), Brassica juncea, Brassica nigra. Brassica oleracea, Camelina sativa (False flax), Thlaspi arvense (Pennycress), Crambe abyssinica (Crambe), Sinapis alba, Brassica rapa, Raphanus sativus (radish) and the like.

[0100] The GSL content in the oilseeds can vary and be dependent on the growing season, the variety, growth conditions and the like. In one aspect, the penny cress can be alow GSL variety'. In one aspect, the low GSL variety can include total GSL at an amount of about 100 pmol / g of oilseeds or less, or about 90 pmol / g of oilseeds or less, or about 80 pmol / g of oilseeds or less, or about 70 pmol / g of oilseeds or less prior to conditioning. In one aspect, the low GSL variety can include total sinigrin at an amount of about 100 pmol / g of oilseeds or less prior to conditioning, or about 80 pmol / g of oilseeds or less, or about 60 pmol / g of oilseeds or less, or about 40 pmol / g of oilseeds or less, or about 20 pmol / g of oilseeds or less prior to conditioning.

[0101] In one aspect, the pennycress can be a high GSL variety. In one aspect, the high GSL variety' can include total GSL at an amount of about 100 pmol / g of oilseeds or more, or about 120 pmol / g of oilseeds or more, or about 140 pmol / g of oilseeds or more, or about 160 pmol / g of oilseeds or more prior to conditioning. In one aspect, the high GSL variety can include total sinigrin at an amount of about 100 pmol / g of oilseeds or more, or about 120 pmol / g of oilseeds or more, or about 140 pmol / g of oilseeds or more, or about 160 pmol / g of oilseeds or more prior to conditioning.

[0102] The defatted meal, after drying and cooling, can have a moisture content of less than 20wt% of the meal, or less than 15wt% of the meal, or less than 10wt% of the meal, or less than 8wt% of the meal. The defatted meal has moisture content of between about 5wt% andDocket No. PT-2457-WO-PCT20wt%, or 6wt% and 15wt%, or 6wt% and 13wt% of the meal, or between 8wt% and 15wt%, or between 8wt% and 12wt%.

[0103] The defatted meal produced according to the methods described herein advantageously has a low GSL content. The defatted meal generated after the drying and cooling step can include a total GSL content of less than 150 pmol / gm (moisture and oil-free) of defatted meal, or less than 120 pmol / gm, or less than 100 pmol / gm, or less than 80 pmol / gm, or less than 60 pmol / gm of meal. The defatted meal generated after the DT step can include a total GSL content of between 20 and 200 pmol / gm of defatted meal, or between 20 and 150 pmol / gm, or between 20 and 100 pmol / gm, or between 20 and 80 pmol / gm, or between 5 and 100 pmol / gm, or between 5 and 80 pmol / gm, or between 5 and 60 pmol / gm.

[0104] The defatted meal, after drying and cooling, produced according to the method described herein comprises a crude fiber content of less than 25wt%, or less than 20wt%, or less than 17wt%, or less than 15wt%, or less than 12wt%. The defatted meal can include a crude fiber content of between 10wt% and 25wt%, or between 10wt% and 20wt%, or between 10wt% and 15wt%.

[0105] The defatted meal, after dry ing and cooling, produced according to the methods described herein comprises oil content of less than 5wt%, dry weight, or less than 4wt%, or less than 3.5wt%, or less than 3wt%, or less than 2wt%, dry weight. The defatted meal produced according to the method described herein comprise oil content of between about lwt% and about 5wt%, or between lwt% and about 4wt%, or between lwt% and 4wt%, or between 2wt% and 4wt%, or between about lwt% and about 3.5wt%, dry7weight.

[0106] The defatted meal, after drying and cooling, produced according to the method described herein comprises protein content of at least 30wt%, or at least 35wt%, or at least 40wt%, dry weight. The defatted meal produced according to the method described herein comprises protein content of between 30wt% and 50wt%, or between 35 \\t% and 45wt%, or between 40wt% and 43wt%.

[0107] In one aspect, the present description relates to a feed ration comprising an animal feed additive, wherein the animal feed additive includes the defatted meal described herein. The feed ration can be suitable for ruminant livestock, monogastric livestock, fish and / or poultry livestock. The ruminant livestock can be cattle. The feed ration can be formulated to maximize weight gain and meat quality of beef cattle in the feed lot and / or to maximize milk production of lactating dairy cattle. The monogastric livestock can be swine.Docket No. PT-2457-WO-PCT

[0108] In one aspect, the feed ration can further comprise the defatted meal. The feed ration can inlcude the defatted meal at an amount of at least lwt%, or at least 2wt%, or at least 3wt%, or at least 4wt%, or at least 5wt% of the feed ration, or at least 6wt%, or at least lwt%, or at least 8wt%, or at least 9wt%, or at least 10wt%.

[0109] Representative features of the present invention are set out in the following clauses, which stand alone or may be combined, in any combination, with one or more features disclosed in the text of the Specification.

[0110] The present invention is as set out in the following clauses:[OHl] Clause 1. A process for producing a meal from oilseeds comprising: a. conditioning oilseed material at a temperature of at least 60°C, wherein the oilseed material comprises oilseeds and / or oilseed flakes, wherein the conditioning inactivates the myrosinase enzyme in the oilseed material; b. pressing the conditioned oilseed material to separate the oilseed material into an oil fraction and a cake fraction; wherein the oilseeds comprise glucosinolates, wherein the glucosinolates comprise sinigrin, and wherein the process degrades at least a portion of the glucosinolates in the oilseed material and wherein the oilseeds are from the Brassicaceae family.

[0112] Clause 2: The process of clause 1 , wherein the process degrades at least about 10%, preferably at least about 20%, of the glucosinolates in the oilseed material relative to glucosinolates in the oilseed material prior to conditioning.

[0113] Clause 3. The process of any one of the preceding clauses, wherein the process degrades at least about 10%, preferably at least about 20%, of the sinigrin in the oilseed material relative to the sinigrin in the oilseed material prior to conditioning.

[0114] Clause 4. The process of any one of the preceding clauses, further comprising:

[0115] c. treating the cake fraction with a solvent comprising hexane to extract residual oil and generate a defatted cake; and

[0116] d. desolventizing / toasting (DT) the defatted cake to remove residual hexane and generate a defatted meal.

[0117] Clause 5. The process of any one of the preceding clauses, wherein the glucosinolates in the cake fraction are at least about 10% less, preferably at least about 20% less, than the starting oilseed material prior to conditioning.Docket No. PT-2457-WO-PCT

[0118] Clause 6. The process of any one of the preceding clauses, wherein the sinigrin in the cake fraction is at least about 10% less, preferably at least about 20% less, than the sinigrin in the starting oilseed material prior to conditioning.

[0119] Clause 7. The process of any one of the preceding clauses, wherein the conditioning is at a temperature of at least 100°C.

[0120] Clause 8. The process of any one of the preceding clauses, wherein the conditioning is at a temperature of at least 110°C.

[0121] Clause 9. The process of any one of the preceding clauses, wherein the conditioning degrades the glucosinolates in the oilseed material, preferably the conditioning degrades at least about 10%, preferably at least about 25%, of the glucosinolates relative to the glucosinolate content in the oilseed material prior to conditioning.

[0122] Clause 10. The process of any one of the preceding clauses, wherein the conditioning degrades the sinigrin in the oilseed material, preferably the conditioning degrades at least about 10%, preferably about 25%, of the sinigrin in the oilseed material relative to the sinigrin content in the oilseed material prior to conditioning.

[0123] Clause 11. The process of any one of the preceding clauses, wherein the DT step degrades the glucosinolates in the oilseed material.

[0124] Clause 12. The process of any one of the preceding clauses, wherein the DT step degrades the sinigrin in the oilseed material.

[0125] Clause 13. The process of any one of the preceding clauses, wherein the oilseeds are selected from group consisting of Brassica carinata (Ethiopian mustard), Brassica napus (canola), Brassica juncea, Brassica nigra, Brassica oleracea, Camelina saliva (False flax), Thlaspi arvense (pennycress), Crambe abyssinica (Crambe), Sinapis alba, Brassica rapa, Raphanus sativus (radish) and combinations thereof.

[0126] Clause 14. The process of any one of the preceding clauses, wherein the oilseeds are pennycress oilseeds.

[0127] Clause 15. A defatted meal from pennycress oilseeds produced by the process of clauses 1 -14.

[0128] Clause 16. An animal feed additive comprising a defatted meal produced from pennycress oilseeds, wherein the pennycress oilseed comprises about 140umol / g or less of glucosinolates.

[0129] Clause 17. An animal feed additive of clause 16, wherein the pennycress oilseed comprises about 50umol / g or less of glucosinolates.Docket No. PT-2457-WO-PCT

[0130] Clause 18. A feed ration comprising the animal feed additive of any one of clauses 16-17.

[0131] Clause 19. The feed ration of clause 18, wherein the feed ration is suitable for ruminant livestock, monogastric livestock, or poultry livestock.EXAMPLES

[0132] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[0133] Example 1. Reduction of glucosinolates through processing of penny cress meal at pilot scale.

[0134] Penny cress grain from two varieties (low GSL and high GSL) were used for a pilot crush and extraction. The pennycress was grown from September to July. A low glucosinolate (LG) variety and a high glucosinolate (HG) variety were processed and evaluated as described below.

[0135] Glucosinolate content and sinigrin content were determined using the method described in Meza et al. (2022), incorporated herein by reference. Pennycress glucosinolates in this example were calculated based on a moisture and oil free state to provide a more accurate comparison between stages of processing.

[0136] The control oilseeds were conditioned for 15 minutes at about 90°C. The oilseeds were also conditioned at "hot” conditions at 1 10°C or 120 C for 45 minutes as indicated in the Tables below. The conditioned seeds then entered the screw press. The oil and the full fat press cake were separated during this process. The full fat press cake containing all pennycress meal components was flushed with nitrogen and stored at -20°C overnight.

[0137] In the next step, the remaining oil from the full fat press cake was extracted using hexane extraction. Initial extraction occurred for 45 mins at 60°C followed by three additions of clean hexanes. Sparge steam was added to the extraction vessel to a temperature of 90°C-100°C and held for one hour to desolventize the cake. There was no control of moisture. Defatted meal (after extraction and desol ventizati on) was removed from the extractor and placed onto trays to further dry overnight.Docket No. PT-2457-WO-PCT

[0138] Table 1 shows the results of the analysis of the low GSL variety pennycress oilseeds after the indicated processing steps.Table 1

[0139] Table 2 shows the results of the analysis of the high GSL variety pennycress oilseeds after the indicated processing steps.Table 2Docket No. PT-2457-WO-PCT

[0140] Table 3 shows the results of the analysis of the low GSL variety pennycress oilseeds after the indicated processing steps comparing the control conditions and the “hot” conditions. The control conditions and the “hot” conditions are as listed in the Table.Table 3Docket No. PT-2457-WO-PCT

[0141] Table 4 shows the results of the analysis of the high GSL variety pennycress oilseeds after the indicated processing steps comparing the control conditions and the “hot” conditions. The control conditions and the “hot” conditions are as listed in the Table.Table 4

[0142] The GLS and the sinigrin content was reduced at conditioning step and the DT step. Conditioning at temperature 90°C and lower, resulted with less than 10 % GLS and Sinigrin degradation. While conditioning at higher temperature (110°C) resulted with greater than 25 % reduction / degradation of the GLS and Sinigrin. The higher temperature increased the extent of degradation.

[0143] Example 2. Reduction of crude fiber content through processing of pennycress meal at pilot plant

[0144] Pennycress seeds treated in Example Iwere analyzed for the crude fiber content. Table 5 shows the results of the crude fiber analysis of the low glucosinolate variety seeds treated as in Example 1.Table 5Docket No. PT-2457-WO-PCT

[0145] Table 6 shows the results of the crude fiber analysis of the high glucosinolate variety seeds treated as in Example 1.Table 6Docket No. PT-2457-WO-PCT

[0146] Table 7 shows the crude fiber content in the after the indicated step for the LG and HG varieties.Table 7Docket No. PT-2457-WO-PCT

[0147] The crude fiber content was reduced at conditioning step. The crude fiber was reduced by at least 25% in the pressed cake relative to the starting seed. The higher temperature increased the extent of degradation.

[0148] Conditioning at temperature above 90°C resulted in an expeller cake 0.3-0.7% lower in crude fiber compared to seed conditioned at 90°C and below.

[0149] In this document, the terms “a,” "an,” or '‘the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary' to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0150] Values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range were explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1. 1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0151] Unless expressly stated, ppm (parts per million), percentage, and ratios are on a by weight basis. Percentage on a by weight basis is also referred to as wt% or % (wt) below.

Claims

Docket No. PT-2457-WO-PCTCLAIMSWhat is claimed is:

1. A process for producing a meal from oilseeds comprising: a. conditioning oilseed material at a temperature of at least 60°C, wherein the oilseed material comprises oilseeds and / or oilseed flakes, wherein the conditioning inactivates the myrosinase enzyme in the oilseed material; b. pressing the conditioned oilseed material to separate the oilseed material into an oil fraction and a cake fraction; wherein the oilseeds comprise glucosinolates. wherein the glucosinolates comprise sinigrin, and wherein the process degrades at least a portion of the glucosinolates in the oilseed material and wherein the oilseeds are from the Brassicaceae family.

2. The process of claim 1, wherein the process degrades at least about 10%, preferably at least about 20%, of the glucosinolates in the oilseed material relative to glucosinolates in the oilseed material prior to conditioning.

3. The process of any one of the preceding claims, wherein the process degrades at least about 10%, preferably at least about 20%, of the sinigrin in the oilseed material relative to the sinigrin in the oilseed material prior to conditioning.

4. The process of any one of the preceding claims, further comprising: c. treating the cake fraction with a solvent comprising hexane to extract residual oil and generate a defatted cake; and d. desolventizing / toasting (DT) the defatted cake to remove residual hexane and generate a defatted meal.

5. The process of any one of the preceding claims, wherein the glucosinolates in the cake fraction are at least about 10% less, preferably at least about 20% less, than the starting oilseed material prior to conditioning,6. The process of any one of the preceding claims, wherein the sinigrin in the cake fraction is at least about 10% less, preferably at least about 20% less, than the sinigrin in the starting oilseed material prior to conditioning.Docket No. PT-2457-WO-PCT7. The process of any one of the preceding claims, wherein the conditioning is at a temperature of at least 100°C.

8. The process of any one of the preceding claims, wherein the conditioning is at a temperature of at least 110°C.

9. The process of any one of the preceding claims, wherein the conditioning degrades the glucosinolates in the oilseed material, preferably the conditioning degrades at least about 10%, preferably at least about 25%, of the glucosinolates relative to the glucosinolate content in the oilseed material prior to conditioning.

10. The process of any one of the preceding claims, wherein the conditioning degrades the sinigrin in the oilseed material, preferably the conditioning degrades at least about 10%. preferably about 25%, of the sinigrin in the oilseed material relative to the sinigrin content in the oilseed material prior to conditioning.

11. The process of any one of the preceding claims, wherein the DT step degrades the glucosinolates in the oilseed material.

12. The process of any one of the preceding claims, wherein the DT step degrades the sinigrin in the oilseed material.

13. The process of any one of the preceding claims, wherein the oilseeds are selected from group consisting of Brassica carinata (Ethiopian mustard), Brassica napus (canola), Brassica juncea, Brassica nigra, Brassica oleracea, Camelina saliva (False flax). Thlaspi arvense (penny cress). Crambe abyssinica (Crambe), Sinapis alba. Brassica rapa, Raphanus sativus (radish) and combinations thereof.

14. The process of any one of the preceding claims, wherein the oilseeds are penny cress oilseeds.Docket No. PT-2457-WO-PCT15. A defatted meal from penny cress oilseeds produced by the process of any one of claims 1-14.

16. An animal feed additive comprising a defatted meal produced from penny cress oilseeds, wherein the penny cress oilseed comprises about 140umol / g or less of glucosinolates.

17. An animal feed additive of claim 16, wherein the penny cress oilseed comprises about 50umol / g or less of glucosinolates.

18. A feed ration comprising the animal feed additive of any one of claims 16-17.

19. The feed ration of claim 18, wherein the feed ration is suitable for ruminant livestock, monogastric livestock, or poultry livestock.

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