RU2803496C2 - Method for obtaining monoacylglyceride oils and food products containing monoacylglyceride oils - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: food product (variants), processed oil, and a method for producing processed oils with a high content of monoacylglycerides (MAG). The food product contains processed oil and has a total caloric content of 25 kcal to 1000 kcal, characterized in that from 5% to 75% of the total calorie content is supplied by said processed oil, where the processed oil is obtained from an oil source and contains less than 10 wt.% triacylglycerides (TAG) and more than 50 wt.% MAG based on the total weight of the processed oil, and where the processed oil additionally contains non-oily ingredients derived from a natural source of oil and present in this oil source, and non-oily ingredients are not added to the processed oil.

EFFECT: method for producing food, enzyme-modified oil, practically free of TAG and food products based on oil for individuals with impaired digestive system.

62 cl, 8 dwg, 5 tbl, 10 ex

Description

Cross reference to related applications

[001] This application claims priority to US Provisional Application No. 62/571910, filed October 13, 2017, which is incorporated herein by reference in its entirety.

Prerequisites for creating the invention

[002] Chronic deficiency in the secretion of digestive enzymes by the pancreas is called exocrine pancreatic insufficiency (EPI). In the absence of these digestive enzymes, in patients suffering from EPI, nutrients from food cannot be properly digested, and these patients may suffer from malnutrition and abdominal disorders. EPI most often occurs in individuals with chronic pancreatitis and a number of other chronic gastrointestinal disorders. EPI is also observed in patients suffering from cystic fibrosis. The effects of EPI can be reduced by pancreatic enzyme replacement therapy (PERT), in which the individual is given enzyme capsules with each meal. Typically, PERT therapy involves the administration of pancreatic enzymes extracted from the porcine pancreas.

[003] Lipids are energy-dense compounds that are a source of essential long-chain fatty acids. Consumed lipids are digested by lipases secreted by the pancreas to form free fatty acids (FFA) and monoacylglyceride (MAG). Blocking the release of lipase from the pancreas results in very poor absorption of fats and oils. In patients suffering from EPI, this can lead to significant malnutrition as calories, essential fatty acids, and fat-soluble nutrients become trapped in undigested lipid particles and are lost from the body.

[004] There is currently a clinical need for alternative nutritional sources that can be absorbed by individuals with EPI and do not require additional PERT.

[005] Partially hydrolyzed fats and oils in the form of MAG are easily digestible by individuals with EPI and do not require PERT. MAG oil products have been clinically evaluated as capsule-based dietary supplements, however, these capsules have been used to eliminate unpleasant taste. As for the usual sources MAG oils, the feedstock oil was processed chemically or enzymatically to produce MAG, which was then extracted with solvents and distilled to fractionate the MAG from other components of the feedstock oils. These MAG products are commercially available as relatively pure products containing only small amounts of FFA, DAG and TAG impurities and virtually no other compounds. Thus, standard sources of MAG oils often do not contain other natural compounds found in the oil, such as tocopherol.

[006] There is a clinical need for very high energy calorie foods that can be consumed by individuals with an ineffective or impaired digestive system. In addition to individuals with pancreatic pathologies (eg, patients with cystic fibrosis, pancreatitis, and pancreatic cancer), other patients with diagnosed or undiagnosed exocrine pancreatic insufficiency (EPI) will experience benefit from consuming such foods. Additionally, individuals with bile duct dysfunction (cholestasis) may benefit from “pre-digested” fats that do not require bile acids for emulsification. High-calorie “energy bars” and drinks are available for sale. But these foods are not suitable for individuals who cannot digest (hydrolyze) the fats present in these foods. To date, lipids have not been prepared in liquid form (shakes) or solid forms (bars) suitable for use without PERT.

[007] Accordingly, there is a need for high-calorie foods that can be consumed by individuals with an ineffective or impaired digestive system. This application describes a process for producing an edible enzyme modified oil (EMO) that is substantially free of triacylglycerides (TAG).

The essence of the invention

[008] The present invention relates to a product containing processed oil obtained from an oil source. In one embodiment of the invention, the processed oil contains MAG in an amount equal to or greater than 40 wt.% of the total weight of the processed oil, where the processed oil does not contain TAG or contains TAG in an amount equal to or less than 5 wt.% of the total weight of the processed oil, and where the processed oil contains non-oil ingredients derived from a natural oil source and normally present in that oil source, and therefore the non-oil ingredients are not added to the processed oil.

[009] In some embodiments of the invention, the source of the oil of the specified product comes from plants, animals or fish.

[0010] In some embodiments, the non-oil ingredients of the product are selected from antioxidants, vitamins, and mixtures thereof.

[0011] In some embodiments of the invention, the specified product contains more than 1 wt.% MAG based on the total weight of the product.

[0012] In some embodiments of the invention, the specified product contains more than 50 wt.% MAG based on the total weight of the product.

[0013] The present invention also relates to a food product. In one embodiment of the invention, the food product contains oil and has a caloric content of from about 1 kcal/gram to about 5 kcal/gram, wherein from about 20% to about 50% of the calories are derived from said oil.

[0014] In some embodiments of the invention, the oil of the specified food product is a processed oil obtained from an oil source, where the processed oil contains MAG in an amount equal to or greater than 40 wt.% by weight of the total weight of the processed oil, where the processed oil does not contain TAG or contains TAG in an amount that is equal to or less than about 5 wt.% by weight of the total weight of the processed oil, and where the processed oil contains non-oil ingredients derived from a natural oil source and normally present in that oil source, and therefore no non-oil ingredients are added into processed oil.

[0015] In some embodiments of the invention, the source of oil of the specified food product comes from plants, animals or fish.

[0016] In some embodiments, the non-oily ingredients of the food product are selected from antioxidants, vitamins, and mixtures thereof.

[0017] In some embodiments of the invention, the specified product contains more than 1 wt.% MAG based on the total weight of the product.

[0018] In some embodiments of the invention, the specified product contains more than 50 wt.% MAG based on the total weight of the product.

[0019] In some embodiments, the food product has a total weight of from about 25 grams to about 500 grams.

[0020] In some embodiments, the food product has a total calorie content of from about 1 kcal to about 5 kcal per gram.

[0021] In some embodiments of the invention, said food product may further comprise a source of carbohydrates.

[0022] In some embodiments of the invention, said food product may further comprise a source of protein.

[0023] In some embodiments, said oil provides 5 to 95% of the total calories in the food product.

[0024] The present invention also relates to a method for producing monoacylglycerol enriched oil. In one embodiment of the invention, the method includes mixing a starting oil containing triacylglycerols (TAGs), a buffer solution and a first enzyme capable of hydrolyzing said TAGs to free fatty acids (FFAs) to form a first reaction mixture; reacting said reaction mixture under conditions sufficient to hydrolyze said TAGs with said first enzyme for a first period of time to produce an aqueous phase and a lipid reaction product (free fatty acid); inactivating said first enzyme in said reaction product; collecting said lipid reaction product; mixing said lipid reaction product and edible glycerin and a second enzyme capable of esterifying FFA to form a second reaction mixture; reacting said second reaction mixture for a second period of time to produce a reaction product of a lipid oil phase and a glycerol phase; inactivating said second enzyme in said reaction product; adding salt to the reaction mixture and separating the lipid oil phase from said glycerol phase; and collecting said lipid oil phase.

[0025] In some embodiments of the invention, the specified source oil is an oil isolated from plants, animals or fish.

[0026] In some embodiments, the first enzyme is an AY lipase.

[0027] In some embodiments, said first period of time is a period of time sufficient to hydrolyze at least 94% of the TAG in said feed oil.

[0028] In some embodiments of the invention, said first period of time is from approximately 14 hours to 24 hours.

[0029] In some embodiments, said step of reacting said reaction mixture under conditions sufficient to hydrolyze said TAG with said first enzyme is carried out at a temperature of from about 30°C to about 35°C.

[0030] In some embodiments of the invention, said steps of mixing a feed oil containing triacylglycerols (TAGs), a buffer solution, and a first enzyme capable of hydrolyzing said TAGs to free fatty acids (FFAs), and reacting said reaction mixture under conditions sufficient to hydrolysis of said TAGs with said first enzyme to form FFA is carried out under a nitrogen atmosphere.

[0031] In some embodiments, said second enzyme is lipase G.

[0032] In some embodiments of the invention, the second period of time is a period of time sufficient to enrich the MAG in the lipid oil phase from approximately 60% to 95%.

[0033] In some embodiments of the invention, the second period of time is from about 24 hours to about 72 hours.

[0034] In some embodiments, said step of reacting said second reaction mixture for a second period of time to produce a lipid oil phase and a glycerol phase is carried out at a temperature of from about 17°C to about 23°C.

[0035] In some embodiments, the method further includes drying said reaction product by applying a vacuum for a third period of time sufficient to remove at least a portion of the water from the reaction product.

[0036] In some embodiments of the invention, the specified stage of drying the specified reaction product is carried out at a temperature of 20°C - 30°C.

[0037] In some embodiments of the invention, said drying step is carried out during a second period of time.

[0038] In some embodiments of the invention, said step of inactivating said second enzyme is accomplished by heating said reaction product.

[0039] In some embodiments of the invention, said heating is carried out at a temperature of at least 70°C for at least 1 hour.

[0040] In some embodiments of the invention, said step of separating said lipid oil phase from said glycerol phase comprises adding sodium chloride to said reaction product.

[0041] In some embodiments of the invention, the final concentration of sodium chloride is up to 0.3 wt.% sodium chloride.

[0042] In some embodiments of the invention, the method, prior to mixing said lipid reaction product and edible glycerol and a second enzyme capable of esterifying FFA and glycerol, further comprises restoring a nitrogen atmosphere over said lipid reaction product.

[0043] In some embodiments of the invention, said steps of removing at least a portion of said aqueous phase and replacing said at least a portion of said aqueous phase with an approximately equivalent volume of water and incubating for a second period of time are repeated before performing said step of collecting said lipid reaction product .

[0044] In some embodiments of the invention, the method further includes adding tocopherol to the specified lipid oil phase after collecting the specified lipid oil phase.

Brief description of drawings

[0045] Figure 1 shows the TLC separation of the components of the original vegetable oil, intermediate FFA and final MAG oil.

[0046] Figure 2 shows the TLC separation of the components of the original vegetable oil, intermediate FFA and final MAG oil.

[0047] Figure 3 shows the distribution of FFA, MAG, DAG and TAG in the "base nutrition shake" and enzyme-modified oil product of the invention ("GBFS") method.

[0048] Figure 4 is a flowchart in the form of a diagram illustrating a method for preparing enzyme modified oil.

[0049] Figure 5 shows the distribution of amino acids and peptides in ready-to-drink nutritional drinks and GBFS hydrolyzed pea protein.

[0050] Figure 6A shows the NMR spectrum of authentic TAG (trysterol).

[0051] Figure 6B shows the NMR spectrum of an enzyme-modified oil obtained from almond oil.

[0052] Figure 7 shows the increase in serum triglyceride levels following administration of a ready-to-drink EMO cocktail.

[0053] Figure 8 shows the increase in serum triglyceride levels after administration of MAG-based RTDS without PERT.

Detailed Description of the Disclosed Subject Matter of the Invention

[0054] It should be noted that the foregoing general description and the following detailed description are provided for purposes of illustration and explanation only and do not limit the scope of the claimed invention. In this application, the use of words in the singular includes words in the plural, the articles “a” or “an” means “at least one”, and the conjunction “or” means “and/or” unless otherwise stated. Moreover, as used herein, the term “including”, as well as other forms such as “includes” and “included”, are not limiting. In addition, terms such as “element” or “component” cover elements or components containing a single object and elements or components containing more than one object, unless otherwise specified.

[0055] The section headings presented here are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited in this application, including, but not limited to, patents, patent applications, articles, books and treatises, are expressly and fully incorporated herein by reference for all purposes. In the event that one or more of the included documents and similar materials has a definition of a term that conflicts with the definition of that term given in this application, then the definition given in this application shall prevail.

[0056] The term "enriched" in the context of the present invention means an amount greater than the amount in the starting material. For example, a MAG-enriched oil is an oil in which the MAG content is higher than the original MAG content before the enrichment process, or the original oil has a higher percentage of MAG than the oil before the enrichment process. The enrichment process can be carried out by converting TAG to MAG and thereby increasing the content and percentage of MAG and decreasing the content or percentage of TAG.

[0057] Triacylglycerol (“TAG”), also known as a triglyceride, is a glyceride consisting of three fatty acid chains covalently linked to a glycerol molecule via ester bonds. TAGs can also be classified as long or medium chain molecules. Long chain TAGs contain fatty acids with 14 or more carbon atoms, and medium long chain TAGs contain fatty acids with 6-12 carbon atoms. Long-chain TAGs may include omega-3 and omega-6 fatty acids. Medium chain TAGs contain saturated fatty acids and therefore do not contain omega-6 or omega-3 fatty acids. Energy sources include long chain TAG (LCT) and medium chain triglycerides (MCT).

[0058] Diacylglycerol (“DAG”), also known as a diglyceride, is a glyceride consisting of two fatty acid chains covalently linked to a glycerol molecule via ester bonds.

[0059] Monoacylglycerol (“MAG”), also known as a monoglyceride, is a glyceride consisting of a single fatty acid chain covalently linked to a glycerol molecule via an ester bond.

[0060] As used herein, the term "refined oil" means a non-natural oil composition that is substantially free of triacylglycerols ("TAGs") or has a reduced amount of TAG compared to pre-modified or pre-processed oil.

[0061] As used herein, the terms “enzyme modified oil” or “EMO” refer to processed oil in which TAGs have been enzymatically converted to MAGs, for example, through the enzymatic conversion of the invention.

[0062] As used herein, the term "food product" means a manufactured or non-naturally occurring food product. It should be noted that the food product mentioned herein, even if it is industrially manufactured and generally not found in nature, may contain various combinations of natural ingredients, where said combinations either do not occur in nature or they exist in nature but are not present in the relative quantities in which they are used in a given food product.

[0063] As used herein, the term “non-oil ingredient” means an ingredient present in a natural oil source that is not a MAG, DAG, TAG, FFA, or lipid.

[0064] In some embodiments, source oils may include, for example, but are not limited to, oils derived from plants, such as olive oil, almond oil, flaxseed oil, sunflower oil, corn oil, canola oil, palm oil, soybean oil or oil of animal origin such as fish oil, sardine oil or anchovy oil or algae oil or mixtures thereof. In one aspect of the invention, the starting oil includes a mixture of olive oil, sunflower oil and linseed oil, wherein from about 50 wt.% to about 80 wt.% of the total mass of the original oil is olive oil; from approximately 10 wt.% to approximately 30 wt.% of the total mass of the original oil is sunflower oil, and from approximately 5 wt.% to approximately 20 wt.% of the total mass of the original oil is linseed oil. In another aspect of the invention, from about 50 wt.% to about 80 wt.% of the total mass of the original oil is olive oil; from approximately 10 wt.% to approximately 30 wt.% of the total mass of the original oil is linseed oil; and from approximately 5 wt.% to approximately 20 wt.% of the total mass of the original oil is sunflower oil.

[0065] In some embodiments, a method for producing a MAG-enriched product includes a first step of hydrolyzing TAG. As a non-limiting example, hydrolysis of TAG can be accomplished using a lipase such as lipase AY (Amano Enzymes, USA Elgin IL, USA), or any non-regiospecific lipase that cuts the chain at the sn-1, sn-2 and sn-3 positions .

[0066] In some embodiments of the invention, the first stage of TAG hydrolysis can be carried out at a temperature of from about 30°C to 35°C. As a non-limiting example, the first stage of TAG hydrolysis can be carried out at a temperature of 30°C - 35°C, 31°C - 35°C, 32°C - 35°C, 33°C - 35°C, 34°C - 35°C, 30°C - 34°C, 31°C - 34°C, 32°C - 34°C, 33°C - 34°C, 30°C - 33°C, 31°C - 33° C, 32°C - 33°C, 30°C - 32°C, 31°C - 32°C, 30°C - 31°C or 30°C, 31°C, 32°C, 33°C, 33°C or 35°C.

[0067] In some embodiments of the invention, the first stage of TAG hydrolysis can be carried out over a period of time from approximately 14 hours to 24 hours. As a non-limiting example, the first stage of TAG hydrolysis can be carried out for a period of time from 14 hours to 20 hours, from 14 hours to 16 hours, from 18 hours to 24 hours, from 22 hours to 24 hours, from 18 hours to 20 hours , or approximately within 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours or 24 hours.

[0068] In some embodiments, the first TAG hydrolysis step results in the hydrolysis of substantially all of the TAG. As a non-limiting example, the first TAG hydrolysis step results in TAG hydrolysis of 94% - 100%, 95% - 100%, 96% - 100%, 97% - 100%, 98% - 100%, 99% - 100%, 94% - 99%, 95% - 99%, 96% - 99%, 97% - 99%, 98% - 99%, 94% - 98%, 95% - 98%, 96% - 98%, From 97 % - 98%, 94% - 97%, 95% - 97%, 96% - 97%, 94% - 96%, 95% - 96% or 94% - 95%, or at least 94%, 95 %, 96%, 97%, 98%, 99% or 100%.

[0069] In some embodiments, the method of producing a MAG-enriched product includes a second glycerol esterification step to increase the MAG oil content. As a non-limiting example, this second esterification step can be carried out using a lipase such as lipase G (Amano Enzymes, USA Elgin IL, USA), or any regiospecific lipase that catalyzes esterification at the sn-1 position but does not efficiently catalyze the formation a second or third glycerol ester (to produce DAG and TAG).

[0070] In some embodiments, a second glycerol esterification step to increase the MAG oil content results in an enrichment of MAG in the product of approximately 70-95%. As a non-limiting example, the MAG oil content can be increased by 70% - 95%, 75% - 95%, 80% - 95%, 85% - 95%, 90% - 95%, 70% - 90%, 75% - 90%, 80% - 90%, 85% - 90%, 70% - 85%, 75% - 85%, 80% - 85%, 70% - 80%, 75% - 80%, 70% - 75 % or 70%, 75%, 80%, 85%, 90% or 95%.

[0071] In some embodiments of the invention, the second stage of glycerol esterification can be carried out at a temperature of from about 17°C to 23°C. As a non-limiting example, glycerol esterification can be carried out at a temperature of 17°C - 23°C, 18°C - 23°C, 19°C - 23°C, 20°C - 23°C, 21°C - 23°C C, 22°C - 23°C, 17°C - 22°C, 18°C - 22°C, 19°C - 22°C, 20°C - 22°C, 21°C - 22°C, 17°C - 21°C, 18°C - 21°C, 19°C - 21°C, 20°C - 21°C, 17°C - 20°C, 18°C - 20°C, 19° C - 20°C, 17°C - 19°C, 18°C - 19°C, 17°C - 18°C or 17°C, 18°C, 19°C, 20°C, 21°C, 22°C or 23°C.

[0072] In some embodiments of the invention, the second stage of glycerol esterification can be carried out over a period of about 24 hours to 72 hours. By way of non-limiting example, the second glycerol esterification step may be carried out for a period of 24 hours to 72 hours, 36 hours to 72 hours, 48 hours to 72 hours, 60 hours to 72 hours, 24 hours to 60 hours , from 36 hours to 60 hours, from 48 hours to 60 hours, from 24 hours to 48 hours, from 36 hours to 48 hours, from 24 hours up to 36 hours or within 24 hours, 30 hours, 36 hours, 42 hours, 48 hours, 54 hours, 60 hours, 66 hours or 72 hours.

[0073] In some embodiments, the method for producing a MAG-enriched product includes a third step of lipase inactivation and phase separation.

[0074] The resulting product of the above embodiments of the invention is a processed oil having a MAG content equal to or greater than 40 wt.% of the total weight of the processed oil. In some aspects of the invention, the MAG content is between about 40 wt.% to approximately 99 wt.% of the total weight of the processed oil. In some aspects of the invention, the MAG content is between about 50 wt.% to approximately 99 wt.% of the total weight of the processed oil. In some aspects of the invention, the MAG content is between about 60 wt.% to approximately 99 wt.% of the total weight of the processed oil. In some aspects of the invention, the MAG content is between about 70 wt.% to approximately 99 wt.% of the total weight of the processed oil. In some aspects of the invention, the MAG content is between about 80 wt.% to approximately 99 wt.% of the total weight of the processed oil. In some aspects of the invention, the MAG content is between about 50 wt.% to approximately 80 wt.% of the total weight of the processed oil. In any of the above aspects, the TAG content is equal to or less than 5 wt.% of the total mass of the processed oil. In any of the above aspects, the TAG content is equal to or less than 4 wt.%, equal to or less than 3 wt.%, equal to or less than 2 wt.%, equal to or less than 1 wt.% of the total mass of processed oil.

[0075] In some embodiments of the invention, the product contains a processed oil according to the invention.

[0076] In some embodiments of the invention, the processed oil has a MAG content equal to or greater than 40 wt.% of the total weight of the processed oil. As a non-limiting example, recycled oil has a MAG content of approximately 40% - 95%, 50% - 95%, 60% - 95%, 70% - 95%, 80% - 95%, 90% - 95%, 40% - 90%, 50% - 90%, 60% - 90%, 70% - 90%, 80% - 90%, 40% - 80%, 50% - 80%, 60% - 80% 70% - 80%, 40% - 70%, 50% - 70%, 60% - 70%, 40% - 60%, 50% - 60%, 40% - 50% or approximately 40%, 45%, 50%, 55%, 60 %, 65%, 70%, 75%, 80%, 85%, 90% or 95% by total weight of oil processed.

[0077] In some embodiments, the recycled oil either contains no TAG or has a TAG content that is equal to or less than 5% by weight of the total processed oil. As a non-limiting example, processed oil has a TAG content of approximately 0% - 5%, 1% - 5%, 2% - 5%, 3% - 5%, 4% - 5%, 0% - 4%, 1 % - 4%, 2% - 4%, 3% - 4%, 0% - 3%, 1% - 3%, 2% - 3%, 0% - 2%, 1% - 2%, 0% - 1% or 0%, 1%, 2%, 3%, 4% or 5% by total weight of processed oil. As another non-limiting example, the TAG content may be less than 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%.

[0078] In some embodiments, the processed oil contains non-oil ingredients derived from and present in the natural oil source, and therefore the non-oil ingredients are not added to the processed oil. By way of non-limiting example, such non-oily ingredients may include antioxidants such as tocopherols, which include alpha-tocopherol, beta-tocopherol, delta-tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, delta-tocotrienol or gamma-tocotrienol and other vitamins such as vitamin K and structurally similar 2-methyl-1,4-naphthoquinone derivatives. In some embodiments, the antioxidant is selected from natural antioxidants (eg, mixed tocopherols or ascorbic acid) and synthetic antioxidants (eg, butylated hydroxyanisole or butylated hydroxytoluene).

[0079] In some embodiments, the product or food product may contain at least 1% MAG. By way of non-limiting example, the food product may contain at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13% , 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more MAG by weight of the product or food product and any amount between these values. In another non-limiting example, the product or food product may contain from 5 wt.% to 15 wt.% MAG based on the total weight of the product or food product.

[0080] In some embodiments of the invention, the food product may further contain a source of carbohydrates. By way of non-limiting example, such a carbohydrate source may contain simple sugars such as glucose and fructose derived from carbohydrate sources such as fruit syrups and agave syrups. Other sources of carbohydrates include other plant sugar syrups, starches, and a number of sugar alcohols.

[0081] In some embodiments of the invention, the food product may further comprise a source of protein. In some aspects, the protein source may be hydrolyzed or partially hydrolyzed. By way of non-limiting example, such a protein source may include milk protein (casein and whey) and other plant proteins including soy protein, rice and rice bran protein, lentils, chickpeas, peanuts, almonds, kelp (helical), quinoa seed protein, mycoprotein , chia seed protein and hemp seed protein. Hydrolyzed protein can be completely hydrolyzed, and pea protein is enriched with peptides ranging from 1 to 10 amino acids in length. In some embodiments, the protein is enriched with peptides ranging from 1 to 10 amino acids in length by approximately 25-75% compared to commercially available partially hydrolyzed protein and other whey hydrolysate products such as Peptamen and Crucial. As a non-limiting example, the protein is enriched in peptides 1 to 10 amino acids in length by at least 25% - 75%, 35% - 75%, 45% - 75%, 55% - 75%, 65% - 75%, 25% - 65%, 35% - 65%, 45% - 65%, 55% - 65%, 25% - 55%, 35% - 55%, 45% - 55%, 25% - 45%, 35% - 45 %, 25% - 35% or by 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75%.

[0082] In some embodiments of the invention, the food product may be liquid, semi-solid, or solid. By way of non-limiting example, the semi-solid may include a pudding, mousse, popsicle or ice cream-like product. By way of non-limiting example, the liquid may be a cocktail or other beverage. As a non-limiting example, the solid may be a bar or other solid food product.

[0083] In some embodiments of the invention, the product or food product further comprises a viscosity modifying agent. The viscosity changing agent may be, for example, but is not limited to, xanthan gum or acacia gum. In some embodiments, the product or food product may further comprise a structure-enhancing or stability-enhancing component such as, for example, but not limited to, gum arabic, sunflower lecithin, and xanthan gum. In some embodiments of the invention, the product or food product further comprises a source of fiber, such as, for example, but limited to, oligosaccharides. In some embodiments of the invention, the product or food product may further contain a food preservative, such as, for example, but limited to, sodium benzoate or potassium sorbate.

[0084] In some embodiments, the product or food product further comprises a flavoring agent, masking agent, or blocking agent. By way of non-limiting example, flavors may include chocolate, vanilla, strawberry or other flavors.

[0085] In some embodiments, the food product has a total weight of from about 25 grams to about 500 grams. By way of non-limiting example, a food product may have a weight of 25 grams to 500 grams, 50 grams to 500 grams, 100 grams to 500 grams, 250 grams to 500 grams, 25 grams to 250 grams, 50 grams to 250 grams. grams, from 100 grams to 250 grams, from 25 grams to 100 grams, from 50 grams to 100 grams, from 25 grams to 50 grams or a total mass that is less than or equal to 25 grams, 50 grams, 100 grams, 150 grams, 200 grams, 250 grams, 300 grams, 350 grams, 400 grams, 450 grams or 500 grams.

[0086] In some embodiments, the food product has a total caloric content of from about 200 kcal to about 1000 kcal. By way of non-limiting example, a food product may have a caloric value of from about 200 kcal to 1000 kcal, from 400 kcal to 1000 kcal, from 600 kcal to 1000 kcal, from 800 kcal to 1000 kcal, from 200 kcal to 800 kcal, 400 kcal to 800 kcal. kcal, from 600 kcal to 800 kcal, from 200 kcal to 600 kcal, from 400 kcal to 600 kcal, from 200 kcal to 400 kcal or a number of calories less than or equal to 200 kcal, 300 kcal, 400 kcal, 500 kcal , 600 kcal 700 kcal, 800 kcal, 900 kcal or 1000 kcal.

[0087] In some embodiments, between 20% and 75% of the calories in the food product come from oil or fat. In one aspect of the invention, the oil or fat is a processed oil. In yet another aspect of the invention, the processed oil has a MAG content equal to or greater than 40 wt.% of the total weight of the processed oil. In other aspects of the invention, the processed oil has a MAG content of from about 40% to about 99% of the total weight of the processed oil. In yet another aspect of the invention, the processed oil has a TAG content of less than 5 wt.% of the total weight of the processed oil. As a non-limiting example, 20% - 50%, 30% - 50%, 40% - 50%, 20% - 40%, 30% - 40%, 20% - 30%, 20% - 75%, 30% - 75%, 40% - 75%, 50% - 75%, 60% - 75%, 70% - 75%, 20% - 70%, 30% - 70%, 40% - 70%, 50% - 70% , 60% - 70%, 20% - 60%, 30% - 60%, 40% - 60%, 50% - 60% or approximately 20%, 21%, 22%, 23%, 24% 25%, 26 %, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% , 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74% or 75% calories in food product comes from any of the above-described processed oils.

[0088] In some embodiments, approximately 20-50% of the calories in the food product come from a carbohydrate source. As a non-limiting example, 20% - 50%, 30% - 50%, 40% - 50%, 20% - 40%, 30% - 40%, 20% - 30% or approximately 20%, 21%, 22% , 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39 %, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% of the calories in a food come from a carbohydrate source.

[0089] In some embodiments, approximately 10% to 50% of the calories in the food product come from a protein source. As a non-limiting example, 10% - 50%, 20% - 50%, 30% - 50%, 40% - 50%, 10% - 40%, 20% - 40%, 30% - 40%, 10% - 30%, 20% - 30%, 10% - 20% or approximately 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21 %, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36% 37%, 38 %, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% of the calories in a food come from a protein source.

[0090] It should be noted that the food product embodiments disclosed above may be combined.

[0091] In some embodiments of the invention, EMO-based products are consumed by individuals with a poorly functioning digestive system, such as, but limited to, individuals suffering from EPI, or individuals who eat while undergoing PERT. In some embodiments, EMO-based products are consumed by individuals who require faster or more complete conversion of lipids to serum triglycerides. Thus, the present invention relates to a method for developing a nutritional regimen for a human or animal with a poorly functioning digestive system. This method includes administering to said patient a food product described in any of the above embodiments or combinations of such embodiments. In one aspect of the invention, a human or animal suffers from an acute digestive disorder (eg, childhood EPI) or a chronic digestive disorder. Such groups of individuals are groups of individuals suffering from cystic fibrosis, pancreatitis, pancreatic cancer or cholestasis, gastrointestinal cancer, celiac disease, diabetes mellitus, Crohn's disease, short bowel syndrome, Solinger-Ellison syndrome; individuals following pancreatic and intestinal surgery, and individuals with other conditions in which digestion is impaired or ineffective. In other aspects of the invention, a person or animal may experience the benefit of faster and more efficient digestion even in the absence of any specific disease (eg, older adults and athletes).

Examples

Example 1: Method for producing a MAG-enriched product compared to a TAG-enriched parent oil

[0092] The process for producing a MAG-enriched product from a TAG-enriched parent oil involves 3 key steps: (1) a mild enzyme-catalyzed triglyceride (TAG) hydrolysis reaction, sequentially converting neutral oils into specific combinations of FFA, MAG, DAG, and Low residue TAG; (2) esterification with glycerol to produce predominantly high amounts of MAG having low concentrations of FFA; and (3) isolation of the modified lipid product, which is achieved by phase separation with or without the use of a centrifuge.

[0093] Step 1: Enzymatic conversion of triacylglycerols

[0094] Prepare a buffer solution . Sodium citrate solution (100 mM, pH 5.8) was prepared in a stirred tank reactor. 11.1 L of deionized (DI) water was placed in a mixing vessel, the stirrer was set to 200 rpm and 0.213 kg of citric acid (anhydrous) was added. After dissolving the powder, the pH was adjusted to pH=5.8 by adding sodium hydroxide solution (approximately 0.121 kg). 220 ml was removed for subsequent preparation of the enzyme solution.

[0095] Preparation of enzyme solution . An enzyme solution was prepared in a separate 250 ml bottle with gentle stirring: 200 ml of citrate buffer solution was placed in a mixing vessel. 10 g of AY AMANO lipase was then added and the bottle was shaken until the enzyme dissolved.

[0096] Conversion of a mixture of triglyceride oils . Three vegetable oils: olive oil, linseed oil and sunflower oil were added to the vessel to obtain a total of 10 kg of vegetable oil mixture. Vacuum was then applied to reduce the pressure to approximately 20 mmHg and the product was degassed (specifically, dissolved oxygen was removed). Stirring was set to 200 rpm and the mixture was heated to 33°C and stirred for approximately 15 minutes to remove any dissolved gas. The vacuum was then replaced with an atmosphere of nitrogen gas. Once the mixture was sufficiently dispersed, 220 ml of the enzyme preparation in buffer solution was added. Stirring was continued and the reaction was monitored by TLC analysis for 24 hours until conversion to FFA was complete.

[0097] The reactor temperature was increased to 70°C and stirring was again carried out for 1 hour to inactivate the enzyme.

[0098] Stirring was stopped and the phases were separated after approximately 60 minutes.

[0099] The aqueous (bottom) phase was removed along with a small amount of the oil phase to ensure removal of residual protein at the interface.

[00100] Step 2: Esterification with glycerol. Typically, transesterification of FFA with glycerol produces a mixture of MAG, DAG and TAG. We have found that by significantly lowering the temperature (below 25°C) and removing water (by evaporation) as it forms in the reaction mixture, the amount of MAG in the product can be increased significantly (by at least 60%, but not above 95%). This result was unexpected.

[00101] In this reaction, the reaction product of Step 1 (approximately 10 L) was cooled to approximately 30°C and stirred at 300 rpm. 10 kg of food grade glycerol was added to the lipid mixture and the temperature was maintained at ~30°C. The mixture was stirred to obtain a dispersion of oil and glycerin. To dry the reaction mixture, a vacuum was applied: first, a vacuum was applied (25 mmHg, Torr), and then a receiver was installed to collect water. After the residual water had evaporated, 20 g of Amano lipase G dissolved in water (50 ml) was added to the reactor. The temperature was reduced to 23°C, and the vacuum was adjusted to 5 mmHg. using an oil diaphragm pump and a cold trap to collect water. The mixture was stirred at 300 rpm at 23°C under vacuum for 72 hours, after which the vacuum was removed and the mixture was treated under nitrogen gas. The mixture was analyzed after 72 hours by TLC to assess conversion to MAG as shown in Figure 1.

[00102] Step 3: Lipase inactivation and phase separation . After completion of the reaction, the lipase was inactivated by heating the mixture (under nitrogen gas) to 70°C for 1 hour. At this time, MAG oil and glycerin were thoroughly mixed, since they are very difficult to separate using traditional gravity or centrifugal methods. After much experimentation, the authors discovered that the lipid could be separated from excess glycerol by adding 0.3 wt% salt (NaCl) to the reaction mixture with stirring. The product mixture was then allowed to cool to approximately 60°C and left without stirring for approximately 1 hour.

[00103] The lipid oil phase was separated from the remaining heavier glycerol phase. The glycerol phase was removed: it contained some salt, residual water and dissolved inactivated enzyme that was present at the visible interface. The glycerol phase can be reused after membrane filtration and should be stored for reprocessing. Tocopherol (vitamin E) was added to a concentration of 200 ppm. (0.02 wt.%) oil product. The hydrolyzed oil (approximately 10 kg) was ready for use and could be stored under nitrogen.

[00104] Product storage. The final hydrolyzed products were transferred to food containers with nitrogen gas for storage and transportation.

Example 2: Characteristics of the oil obtained in Example 1.

[00105] The reaction products and overall procedure can be evaluated using thin layer chromatography and gas chromatography.

[00106] Thin layer chromatography testing. The components of the oil samples were separated using TLC plates (Analtech Uniplate GHL silica gel containing inorganic binder, 20x20 cm, 250 μm). The solvent was a solution of hexane:diethyl ether:acetic acid (70:30:1). Typical sample sizes were 3 μL. Once the solvent front approached the top of the plate (~1 cm), the plates were removed from the TLC tank and the solvent was evaporated in a fume hood. The components were visualized using iodine vapor (at room temperature) in a TLC tank and relative intensities were assessed using colorimetric imaging (Amersham 600 imager). After 15 minutes in the tank, the plates were removed and photographed. The intensity of the spots decreased after 30-60 minutes.

[00107] Determination of physical properties was carried out to evaluate product consistency, color, water content, fats and oils (mixed).

[00108] Figure 1 shows the final results of steps 1-3 in preparing a MAG-enriched product compared to the original TAG-enriched oil as described in Example 1. Figure 2 shows that the tocopherol originally present in olive oil was maintained after the steps described in Example 1. Figure 2 shows that the tocopherol spot can be traced above the TAG spot in all three lanes.

[00109] Fatty acid profile testing using gas chromatography . Lipid components including C10:0 capric acid, C12:0 lauric acid, C14:0 myristic acid, C16:0 palmitic acid, C18:0 stearic acid, C18:1 oleic acid, C18:2 linoleic acid and alpha-linolenic acid C18:3 was analyzed after derivatization as fatty acid methyl esters and compared with standards. For derivatization, the sample (500 μl) was added to a 5 ml reaction tube containing 2 ml boron trifluoride solution (12% in methanol), 20 μl dimethoxypropane and 100 μl tridecanoic acid solution as internal standard (10 mg/ml). The reaction tube was shaken and incubated in a heating block at 60°C for 30 minutes.

[00110] The reaction tube was removed from the heating block and allowed to cool for 15 minutes. Then 1 ml of distilled water was added to quench the reaction, and then 1 ml of hexane was added. The reaction tube was shaken for 60 seconds and the phases were allowed to separate for 3 minutes. The upper (hydrophobic) phase was transferred to a 1.5 ml tube containing approximately 50 mg sodium sulfate (anhydrous). After shaking for 60 seconds, the 1.5 mL tube was centrifuged and ~500 μL of the clarified, dried hydrophobic phase was transferred to the sample vessel for gas chromatography.

[00111] Samples were analyzed using an Agilent 7890A gas chromatograph with a flame ionization detector and the Agilent Openlab CDS Chemstation computer program. The GC column is an Omegawax 100 column (15 × 0.1 mm × 0.1 µm). The results were converted to wt% using the internal standard.

[00112] Figure 3 shows the distribution of FFA, MAG, DAG and TAG during the process of preparing the ENSURE product and the GBFS product according to the invention. The percentages of FFA, MAG, DAG and TAG in the oils were determined using thin layer chromatography.

Example 3: Another example of a method for preparing a MAG-enriched product compared to a TAG-enriched original oil.

[00113] The following procedure is represented by the flowchart in Figure 4.

[00114] Vegetable oil was added to citric acid and sodium hydroxide (caustic soda) in deionized water and heated to 33°C ± 2°C. Low vacuum was applied for degassing to remove oxygen. After this, lipase AY was added. Hydrolysis of the mixture was carried out for 14-24 hours in a nitrogen atmosphere at a temperature of 33°C ± 2°C.

[00115] Lipase was inactivated at 70°C ± 2°C for 1 hour. The aqueous phase with the inactivated enzyme was poured off. Then glycerin was added. The reaction mixture was then cooled to 22°C ± 2°C. Lipase G was then added and the water was evaporated under moderate vacuum.

[00116] Transesterification was carried out for 72 hours under high vacuum (approximately 720 mmHg) at a temperature of 20°C ± 2°C. Lipase G was inactivated at 70°C ± 2°C for one hour, and salt was added to the reaction mixture.

[00117] Enzyme inactivation and phase separation were carried out under nitrogen atmosphere for 1 hour at 70°C ± 2°C. The aqueous phase with the inactivated enzyme, glycerol and salt was poured off. The reaction mixture was cooled to 60°C ± 2°C. After this, an antioxidant was added. The final product was stored at 4°C ± 2°C under nitrogen atmosphere.

Example 4: Ready-to-use composition comprising MAG and hydrolyzed protein

[00118] The product of the invention was formulated as a conventional "milkshake" that included a source of fats, proteins, carbohydrates, vitamins and fibers in addition to the traditional surfactants and stabilizing agents typically found in these products. An individual serving was 250 ml. The Ingredients label listed the following ingredients: Water, Organic Agave Syrup, Hydrolyzed Pea Protein, Hydrolyzed Oil Blend, Gum Arabic, Sunflower Lecithin, Xanthan Gum, Oligosaccharides, Potassium Sorbate, Sodium Benzoate, Instant Coffee, Natural Organic Vanilla Flavor, Vitamin C , vitamin E succinate, vitamin A palmitate, niacinamide, calcium D-pantotinate, pyrodoxine-HCl, thiamine-HCl, riboflavin, vitamin D3, folic acid, cyanocobalamin, vitamin K2.

[00119] Carbohydrates were provided in the form of simple sugars (glucose and fructose) from fruit and agave syrups.

[00120] The protein in the product was partially hydrolyzed pea protein (PURIS Pea Protein 870H, World Food Processing LLC, Turtle Lake, WI 54889).

[00121] Autoami also produced fully hydrolyzed pea protein (EHP) with peptides in a size range that were biologically more suitable for transport across the intestinal wall. EHP was prepared by subsequent enzymatic hydrolysis. For example, partially hydrolyzed pea protein (Puris 870H pea protein described above) was dissolved in 100 mM phosphate buffer to a concentration of 25 mg/ml. The enzyme was then added and the reaction mixture was incubated at 50°C overnight. Three different commercially available GRAS enzymes were evaluated: alkalase, thermoase, and flavororzyme.

[00122] The average size and distribution of peptides in protein samples were assessed using size exclusion chromatography. Samples were dissolved in 100 mM phosphate buffer, pH 6.8 to a concentration of 25 mg/ml, and analyzed on a Shimadzu HPLC with UV detector (214 nm) using a Phenomenex Yarra 3 μm SEC-2000 column (elution with 100 mM sodium phosphate buffer (pH 6.8), flow rate 0.8 ml/min at room temperature). Samples were compared to a molecular weight standard (Phenomenex SEC Standard Part ALO-3042). Dimensions were estimated using a calibration curve constructed from known molecular weight standards. The average size of non-hydrolyzed pea protein was ~200 amino acids. The average size of Puris Pea 870H partially hydrolyzed pea protein was 34 amino acids with a significant number of amino acids in the range of 2-40, similar to the number of amino acids in other hydrolyzed protein products (whey based).

[00123] Figure 5 shows the distribution of amino acids and peptides in ready-to-drink nutritional drinks as described by E. Phillips et al., 2005 Peptide-Based Formulas: the Nutraceuticals of Enteral Feeding? EPCN October: 40-45 compared to fully hydrolyzed GBFS pea protein described above. Figure 5 shows the percentage of amino acids (Y-axis) of size 1 amino acid, 2-4 amino acids, 9-10 amino acids, 10-40 amino acids or more than 40 amino acids in the Oeotamen, Perative, Cricual, Pivot and GBFS products according to the invention (X-axis) . The average size of GBFS EHPs was 3–4 amino acids, with most being 1–7 amino acids.

[00124] The fat was obtained as a restructured lipid MAG and was prepared from a mixture of olive oil (70%), sunflower oil (21%) and flaxseed oil (9%) to provide the energy value and other benefits of polyunsaturated fatty acids (PUFAs), Omega-6 PUFAs and omega-3 PUFAs. The omega-6/omega-3 ratio was ~4/1.

[00125] Gum arabic, sunflower lecithin and xanthan gum are common GRAS food ingredients used to improve the structure and stability of the beverage. Oligosaccharides provide indigestible fiber. Potassium sorbate and sodium benzoate are common food preservatives. Instant coffee and vanilla add flavor.

[00126] A vitamin pack containing fat-soluble and water-soluble vitamins has been included.

[00127] A prototype product labeled "Nutritional" is shown in Table 1. In Table 1, ** indicates percent daily values (%DV) based on a 2000 calorie diet. † - daily value (DV) not established.

Additional facts
Serving size 250 ml Amount per serving %DV** Calories 379 kcal Total fat 19 G 24% Cholesterol - mg 0% Total carbohydrates 32 G 12% Dietary fiber 2 G 7% Sahara thirty G Protein 20 G 40% Vitamin A 320 mcg 35% Vitamin C 480 mg 35% Vitamin D 7 mcg 35% Vitamin E 5 mg 35% Vitamin K 42 mcg 35% Thiamine 400 mcg 35% Riboflavin 450 mcg 35% Niacin 6 mg 35% Vitamin B6 600 mcg 35% Folate 132 mcg 35% Vitamin B12 1 mcg 35% Biotin 10 mcg 35% Pantothenic acid 2 mg 35% Calcium 480 mg 35% Iron 6 mg 35% Phosphorus 450 mg 35% Chloride - mg † Sodium 175 mg 7% Potassium 1600 mg 35%

[00128] Sensory assessment. FFAs derived from a vegetable oil mixture were rated as unpalatable on a taste panel. Surprisingly, the MAG oil produced from this mixture was palatable and resembled the original triglyceride oil in taste and consistency. When MAG oil was incorporated into the RTD product described above, the aroma was acceptable and did not differ from similar commercial products with intact (undigested) lipids and proteins.

Example 5: Receiving multiple EMO batches

[00129] The time taken to prepare EMO as described in Example 1 was evaluated, resulting in an oil that had a MAG content of greater than 70%; MAG+FFA and TAG more than >85%; and TAG 5% or less.

[00130] Table 2 illustrates TLC plate scans indicating the concentrations of MAG and FFA in an enzyme modified oil (EMO) prepared from a mixture of olive oil/linseed oil/sunflower oil (7/2/1 ratio). In these experiments, stage 2 was extended to 84 hours to establish upper time and temperature limits to avoid TAG formation.

Experiment TAG DAG MAG FFA FFA+MAG 1 4 26 56 13 70 2 4 eleven 73 13 85 3 5 10 76 9 85

The reaction can be monitored substantially in real time by TLC analysis and terminated at any time in step 2 to obtain the desired amounts of MAG, DAG, TAG and FFA. A reaction time of 72 hours for Stage 2 was found to be a practical and productive termination point, as shown in Example 6.

Example 6: Evaluation of Enzyme Modified TAG Free Almond Oil by NMR Analysis.

[00131] TAG-free EMO was prepared using almond oil. Reaction conditions were as described in Example 1. To minimize TAG formation, step 2 was carried out for 72 hours.

[00132] Figures 6A-6B illustrate ¹³ C-NMR analysis of EMO obtained from almond oil. Figure 6A shows that ^{the 13} C-NMR signal is associated with authentic TAG (trysterol) and characteristic peaks at 62.173 ppm. and 68.921 ppm Figure 6B illustrates ^{the 13} C-NMR signal for EMO. This means that no noticeable TAG signals were observed. Integration of the actual signals indicates an acylglycerol distribution between MAG:DAG=88%:12%. The samples were analyzed on a JEOL ECA600II NMR spectrometer operating with proton radiation at a frequency of 600 MHz and carbon radiation at a frequency of 150 MHz. Samples were prepared in 5 mm tubes and blocked with CDCl ₃ at ambient temperature.

Example 7: Clinical trials of a ready-to-drink cocktail based on EMO

[00133] This study was designed to show that patients with EPI can consume foods prepared with TAG-free oils, which were prepared by the authors, without adding any enzyme supplements, and these patients will metabolize lipids with serum TAG production, but will not have the symptoms (flatulence, cramps and steatorrhea) associated with lipid intake without PERT.

[00134] This clinical trial is a single-center, randomized, double-blind, crossover study evaluating an EMO-based ready-to-drink shake (RTDS) for blood lipid levels to evaluate safety, tolerability, and palatability compared to a standard nutritional supplement. used in conjunction with Pancreatic Enzyme Replacement Therapy (“PERT”) capsules.

[00135] Patients came to the clinic after an overnight fast and an evening meal of a standardized meal. Group 1 (10 patients) received RTDS plus placebo for PERT, and Group 2 (10 patients) received a standard nutritional supplement for PERT. Serial blood samples collected over 6 hours (0, 1, 2, 3, 4, 5, 6 hours) were obtained from patients in both experimental groups without repeated administration of RTDS or standard dietary supplementation. Water could be consumed throughout the study.

[00136] Patients returned to the clinic for treatment 2 (crossover treatment) after fasting overnight and eating a standardized meal in the evening. Group 1 patients were given a standard PERT nutritional supplement, and Group 2 patients were given RTDS along with a PERT placebo. Serial blood samples collected over 6 hours (0, 1, 2, 3, 4, 5, 6 hours) were obtained from patients in both experimental groups without repeated administration of RTDS or standard dietary supplementation.

[00137] The lipid dose was 0.5 g/kg body weight. The lipid in the standard dietary supplement was a mixture of TAGs isolated from canola oil, high oleic sunflower oil, and corn oil. In the health drink, MAG was obtained from almond oil. The lipid dosage for each patient was based on well-known “lipid tolerance tests,” which are similar in protocol and scope to known glucose tolerance tests. The recommended dose for these tests is 0.5-1.0 g/kg administered over 20-30 minutes. Samples were taken at the beginning of the test and hourly for 6-8 hours. Serum triglyceride levels were measured using standard laboratory methods.

[00138] The PERT dose used in the study was that recommended by the manufacturers and was 2500 IU of lipase activity per gram of fat consumed. This is equivalent to 3-4 capsules administered during the crossover phase of the study.

[00139] Figure 7 shows the increase in serum triglyceride levels after administration of a ready-to-drink EMO cocktail to two patients. In Figure 7, the dotted lines show patients who took 0.5 g of enzyme-modified almond oil per kg body weight, and the solid lines show patients who took 0.5 g canola oil, high oleic sunflower oil and corn oil per kg body weight where such oil was included in the RTDS consumed within half an hour. Figure 7 shows that the enzyme-modified oil in RTDS was absorbed by patients and converted into serum triglycerides. This treatment was well tolerated by patients.

[00140] Figure 8 illustrates an increase in serum triglyceride levels in another patient following consumption of test beverages. In Figure 8, the dotted lines show serum triglycerides after MAG-based RTDS without PERT, and the solid line shows standard RTDS treatment plus PERT. Patients received 0.5 g of enzyme-modified almond oil per kg body weight or 0.5 g of corn oil per kg body weight, where such oil was included in the RTDS, consumed within half an hour. This treatment was well tolerated by patients.

[00141] In this patient, lipid digestion and conversion to serum triglycerides occurred significantly more rapidly after administration of a MAG-based RTDS without PERT than after administration of a TAG-based product (as standard of care) with PERT. This suggests that the patient, in addition to EPI, suffered from cholestasis (impaired flow of bile from the liver), and not simply an enzyme deficiency, and therefore could not emulsify properly in canola oil, high oleic sunflower oil, and corn oil when taking a standard drink. Emulsification of the oil into very small droplets is necessary to create the surface area required for pancreatic lipase to hydrolyze the oil into MAG and FFA, which are transported into the enterocytes. Liver bile acids are required to achieve this activity. The TAG in a standard medicinal drink cannot be emulsified into the microemulsions required for optimal lipase activity in the small intestine, and thus serum triglyceride levels did not increase as quickly as they do with the MAG formulation, which does not require additional emulsification or activity lipases.

Example 8: High Calorie Ready-to-Drink Shake Without PERT

[00142] High-calorie RTDS can be prepared as follows: deionized water (~60% of final volume) is added to the main mixing vessel and the water is heated to ~60°C. Then it is mixed with hydrolyzed protein and agave syrup is added. Use a hand mixer for mixing. Warm (60°C) EMO and lecithin are mixed in a separate container. After the lecithin has dissolved, EMO/lecithin is added to the aqueous phase and mixed. Water is then added again to achieve the final mass (volume). It is then emulsified using a high shear blender. To make very high calorie RTDS, higher levels of components are added.

[00143] Once the beverage base is prepared, various flavorings, masking agents, and blockers such as chocolate, vanilla, strawberry, and the like can be added to create unique products.

[00144] This method can also be used to prepare high-calorie foods in semi-solid form, such as pudding-like products, mousses, popsicles, and ice creams, using a beverage base formulation with added viscosity changing agents such as xanthan gum and acacia gum.

[00145] Primary ingredients and nutritional values are listed in Table 3 for high-calorie (1.5 kcal/ml) RDTS.

High-calorie components Mass. % kcal/g kcal/325 ml drink kcal/ml EMO 7.5 9 219 0.7 Agave sugar 14 4 182 0.6 Hydrolyzed protein 5 4 65 0.2 Pea fat 0.7 9 21 0.1 Sunflower lecithin 0.5 9 15 0.0 Total 28 502 1.5

[00146] Primary ingredients and nutritional values are shown in Table 4 for very high calorie (2.5 kcal/ml) RDTS.

High-calorie components Mass. % kcal/g kcal/325 ml drink kcal/ml EMO 12 9 351 1.1 Agave sugar 20 4 260 0.8 Hydrolyzed protein 10 4 130 0.4 Pea fat 1.4 9 42 0.1 Sunflower lecithin 0.5 9 15 0.0 Total 44 797 2.5

Example 9: High Calorie Bar Without PERT

[00147] The high-calorie bar can be prepared as follows: deionized water (~60% of final volume) is added to the main mixing vessel and the water is heated to ~60°C. Then it is mixed with hydrolyzed protein and agave syrup is added. In a separate container, mix warm (60°C) EMO and lecithin using a hand mixer. After the lecithin has dissolved, EMO/lecithin is added to the aqueous phase and mixed. Water is then added again to achieve the final mass (volume).

[00148] Primary ingredients and nutritional values are shown in Table 5 for the PERT-free energy bar.

High-calorie components Mass. % in product kcal/g kcal/60 g bar EMO 9 9 81 Agave sugar 15 4 60 Hydrolyzed protein 19 4 76 Pea fat 2.7 9 24 Sunflower lecithin 0.5 9 15 Total 46 256

[00149] After preparing the bar base, various flavorings, masking agents, and blockers such as chocolate, vanilla, and the like can be added to create unique products.

Example 10. Making a ready-to-drink cocktail.

[00150] Deionized water is added to the main mixing vessel and the water is heated to ~60°C ± 2°C. Once the water reaches 60°C, add sugar syrup (such as agave syrup or date syrup) slowly, stirring lightly. The solution is then mixed. The individual solids (vitamin/micronutrient mixture and hydrolyzed protein) are weighed and combined in a separate container. Dry ingredients are mixed thoroughly. Then slowly add the mixed dry ingredients directly into the main mixing vessel with gentle shaking. After this, the EMO is weighed in a separate container. EMO is heated to 60°C ± 2°C. The sunflower lecithin is then slowly added to the warm EMO with moderate stirring (if necessary) until the sunflower lecithin is completely mixed into the solution. After this, the EMO/sunflower lecithin mixture is slowly added to the main mixing vessel. Distilled water is then added to increase the volume to 95% by weight of the total liquid, and left at 70°C ± 2°C. Fragrances and color are slowly added to the main mixing vessel. Next, a stabilizer (such as acacia gum) is slowly added to the main mixing vessel. The solution is mixed for 20 minutes (to increase viscosity). The solution temperature is maintained at 70°C ± 2°C. The solution is brought to the final volume by adding a sufficient amount of distilled water. The product is passed through a homogenizer(s) to reduce the pressure to obtain stable emulsions. The product is pasteurized or sterilized. The product is then cooled to room temperature. After this, the product is poured into packaging.

[00151] Therefore, the present invention is well adapted to achieve the above-mentioned objects and advantages, as well as its inherent advantages. Although various changes may be made by one skilled in the art, such changes are intended to be within the scope of the present invention, as illustrated generally by the appended claims.

[00152] The above description of specific embodiments of the invention has been presented for purposes of illustration and description thereof. Representative embodiments of the invention have been selected and described to better understand the principles of the disclosure of the invention and its practical application, and to enable others skilled in the art to best use the subject matter of the invention and the various embodiments thereof with various modifications suitable for the particular application contemplated. Various features and disclosures of various embodiments of the invention may be combined and are included within the scope of the disclosure of the present invention.

Claims (72)

1. A food product containing processed oil and having a total calorie content of from about 25 kcal to about 1000 kcal, wherein from about 5% to about 75% of the total calorie content comes from said processed oil, wherein the processed oil is derived from an oil source, wherein the processed oil oil contains less than 10 wt. % triacylglycerides (TAG) by weight of the total oil, where the processed oil contains more than 50 wt. % monoacylglyceride (MAG) of total processed oil and wherein the processed oil further contains non-oil ingredients derived from a natural oil source and present in that oil source, and therefore the non-oil ingredients are not added to the processed oil. 2. Food product according to claim 1, where the processed oil contains more than 60 wt. % MAG of total processed oil. 3. Food product according to claim 1, where the processed oil contains more than 70 wt. % MAG of total processed oil. 4. Food product according to claim 1, where the processed oil contains more than 80 wt. % MAG of total processed oil. 5. Food product according to claim 1, where the processed oil contains more than 90 wt. % MAG of total processed oil. 6. Food product according to any one of paragraphs. 1-5, wherein from about 10% to about 60% of the total caloric content comes from said processed oil. 7. Food product according to any one of paragraphs. 1-5, wherein from about 20% to about 50% of the total caloric content comes from said processed oil. 8. Food product according to any one of paragraphs. 1-5, wherein from about 25% to about 45% of the total caloric content comes from said processed oil. 9. Food product according to any one of paragraphs. 1-5, wherein from about 30% to about 40% of the total caloric content comes from said processed oil. 10. Food product according to any one of paragraphs. 1-5, additionally containing a source of carbohydrates. 11. The food product according to claim 10, wherein said carbohydrate source is fruit syrup or agave syrup. 12. The food product according to claim 10, wherein said carbohydrate source contains simple sugars. 13. The food product of claim 10, wherein about 20% to about 50% of the calories come from a carbohydrate source. 14. Food product according to claim 10, additionally containing a source of protein. 15. The food product of claim 14, wherein about 10% to about 50% of the calories come from a protein source. 16. The food product according to claim 14, wherein said protein source contains hydrolyzed or partially hydrolyzed protein. 17. The food product according to claim 16, wherein said hydrolyzed protein is selected from hydrolyzed pea protein and a whey hydrolyzed product. 18. Food product according to any one of paragraphs. 1-5, additionally containing a source of protein. 19. The food product of claim 18, wherein about 10% to about 50% of the calories come from a protein source. 20. The food product according to claim 18, wherein said protein source contains hydrolyzed or partially hydrolyzed protein. 21. The food product according to claim 20, wherein said hydrolyzed protein is selected from hydrolyzed pea protein and a whey hydrolyzed product. 22. The food product according to claim 1, wherein the non-oil ingredients are selected from antioxidants, vitamins and mixtures thereof. 23. The food product according to claim 22, wherein said antioxidant is tocopherol. 24. The food product according to claim 23, wherein said tocopherol is selected from α-tocopherol, β-tocopherol, δ-tocopherol, γ-tocopherol, α-tocotrienol, β-tocotrienol, δ-tocotrienol and γ-tocotrienol. 25. A food product containing processed oil, a source of carbohydrate and a source of protein and having a total weight of from about 25 grams to about 500 grams with a caloric content of from about 1 kcal per gram to about 5 kcal per gram, wherein the processed oil contains from about 10% to approximately 50% of the total calorie content, where the processed oil has a MAG content exceeding more than 50 wt. % of the total weight of the processed oil, and a TAG content equal to or less than 10 wt. % of the total weight of the processed oil, where the processed oil is derived from an oil source and where the processed oil additionally contains non-oil ingredients derived from a natural oil source and present in that oil source, and therefore the non-oil ingredients are not added to the processed oil. 26. Refined oil obtained from an oil source, where the processed oil has a MAG content equal to or greater than 40 wt. % of the total weight of the processed oil, where the processed oil does not contain TAG or has a TAG content equal to or less than 10 wt. % of the total weight of the processed oil, and where the processed oil contains non-oil ingredients derived from a natural oil source and present in that oil source, and therefore the non-oil ingredients are not added to the processed oil. 27. The refined oil of claim 26, wherein said oil source comes from a source selected from plants, animals, algae or fish. 28. Refined oil according to claim 26, wherein said oil source is of vegetable origin. 29. The refined oil of claim 26, wherein said oil source is selected from the group consisting of olive oil, sunflower oil, corn oil, almond oil, canola oil, palm oil, soybean oil, flaxseed oil, and mixtures thereof. 30. Recycled oil according to any one of paragraphs. 26-29, wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof. 31. The processed oil of claim 30, wherein said antioxidant is tocopherol. 32. The processed oil of claim 31, wherein said tocopherol is selected from the group consisting of α-tocopherol, β-tocopherol, δ-tocopherol, γ-tocopherol, α-tocotrienol, β-tocotrienol, δ-tocotrienol and γ-tocotrienol. 33. Recycled oil according to any one of paragraphs. 26-29, wherein said processed oil has a MAG content of approximately 50 wt. % to approximately 95 wt. % of the total mass of processed oil. 34. The refined oil of claim 33, wherein said refined oil has a TAG content of approximately 5 wt. % to approximately 0.5 wt. % of the total mass of processed oil. 35. Recycled oil according to any one of paragraphs. 26-29, wherein said processed oil has a TAG content of approximately 5 wt. % to approximately 0.5 wt. % of the total mass of processed oil. 36. A method for producing processed oil according to any one of paragraphs. 1-35, including: mixing a source oil containing triacylglycerols (TAG), where TAG is present in an amount of more than 50 wt. % of the total mass of the original oil; a buffer solution and a first enzyme capable of hydrolyzing said TAGs to free fatty acids (FFAs) to obtain a first reaction mixture; reacting the first reaction mixture under conditions sufficient to hydrolyze said TAGs with said first enzyme for a first period of time to form an aqueous phase and a first lipid reaction product containing FFA; inactivating said first enzyme in said first lipid reaction product; collecting said first lipid reaction product by removing it from the aqueous phase; mixing said first lipid reaction product with food grade glycerol and a second enzyme capable of esterifying FFA to form a second reaction mixture; reacting said second reaction mixture for a second period of time to form a second lipid reaction product comprising a lipid oil phase and a glycerol phase; inactivating said second enzyme in said second lipid reaction product; adding salt to the reaction product and separating the lipid oil phase from said glycerol phase; And collecting said lipid oil phase. 37. The method according to claim 36, wherein said starting oil is an oil obtained from plants, animals or fish. 38. The method according to claim 36, wherein said starting oil is a vegetable oil or a mixture of vegetable oils. 39. The method of claim 36, wherein said starting oil is a vegetable oil selected from the group consisting of olive oil, sunflower oil, corn oil, almond oil, canola oil, palm oil, soybean oil, linseed oil and mixtures thereof. 40. The method according to claim 36, wherein said first enzyme is a lipase. 41. The method according to claim 40, wherein said first enzyme is lipase AY. 42. The method according to claim 36, wherein said buffer solution is a sodium citrate solution. 43. The method of claim 36, wherein said first period of time is a period of time sufficient to hydrolyze at least 94% of the TAG in said feed oil. 44. The method of claim 36, wherein said first period of time is from about 14 to about 24 hours. 45. The method of claim 36, wherein said step of reacting said reaction mixture under conditions sufficient to hydrolyze said TAGs with said first enzyme is carried out at a temperature of from about 30°C to about 35°C. 46. The method of claim 36, wherein said steps of mixing a starting oil containing triacylglycerols (TAGs), a buffer solution and a first enzyme capable of hydrolyzing said TAGs to free fatty acids (FFAs), and reacting said reaction mixture under conditions sufficient to cause hydrolysis the specified TAG with the specified first enzyme to FFA is carried out under a nitrogen atmosphere. 47. The method of claim 36, wherein said second enzyme is a lipase. 48. The method of claim 36, wherein said second enzyme is lipase G. 49. The method of claim 36, wherein said second period of time is a period of time sufficient to enrich the MAG in the lipid oil phase to approximately 60-95%. 50. The method of claim 36, wherein said second period of time is from about 24 hours to about 72 hours. 51. The method of claim 36, wherein said step of reacting said second reaction mixture for a second period of time to form a lipid oil phase and a glycerol phase is carried out at a temperature of about 17 to 23°C. 52. Method according to any one of paragraphs. 36-51, further comprising drying said second lipid reaction product by applying a vacuum for a third period of time sufficient to remove at least a portion of the water from the second lipid reaction product. 53. The method according to claim 52, where the specified stage of drying the specified second lipid reaction product is carried out at a temperature of 20-30°C. 54. The method according to claim 52, wherein said drying step is carried out during a second period of time. 55. Method according to any one of paragraphs. 36-51, wherein said step of inactivating said second enzyme is carried out by heating said second lipid reaction product. 56. The method according to claim 55, wherein said heating is carried out at a temperature of at least 70°C for at least 1 hour. 57. Method according to any one of paragraphs. 36-51, wherein said step of separating said lipid oil phase from said glycerol phase comprises adding sodium chloride to said second lipid reaction product. 58. The method according to claim 57, where the final concentration of sodium chloride is up to 0.3 wt. % sodium chloride. 59. Method according to any one of paragraphs. 36-51, which, before mixing said first lipid reaction product and edible glycerol and a second enzyme capable of esterifying FFA and glycerol, further includes re-applying a nitrogen atmosphere over said first lipid reaction product. 60. Method according to any one of paragraphs. 36-51, further comprising adding tocopherol to said lipid oil phase after collecting said lipid oil phase. 61. Method according to any one of paragraphs. 37-51, where the lipid oil phase contains MAG in an amount of approximately 40 wt. % to approximately 99 wt. % of the total weight of the lipid oil phase, and where the lipid oil phase either does not contain TAG or contains TAG in an amount of about 0.1 wt. % to approximately 10 wt. % of the total mass of the lipid oil phase. 62. Method according to any one of paragraphs. 36-61, where water is removed from the second reaction mixture over a second period of time.