Liquid crystalline phase compositions and methods of making them

Abstract

The present invention relates to enzymatic methods for producing a stable lipid-based liquid crystalline phase of Fd3m or Hll space group, compositions comprising such phases, and uses thereof, wherein the starting mixture comprises 30-65 wt% polar lipids and 35-70 wt % oil which are well dispersed and triacylglycerol lipase is added to produce oil hydrolysate which in presence of aqueous solvent results in liquid crystal formation.

Description

[0001]P92248PC 1 COMPOSITIONS AND METHODS OF MAKING THEM Technical field of the invention The present invention relates to enzymatic methods for producing a stable lipid- based liquid crystalline phase, compositions comprising such phases, and uses thereof. Background of the invention Liquid crystalline phases are structures formed by the self-assembly of amphiphilic molecules, such as lipids, in an aqueous environment. These phases are known for their highly organized and unique molecular arrangements. Unlike emulsions, which are mixtures of immiscible liquids stabilized by emulsifiers, liquid crystalline phases exhibit specific geometrical patterns and are characterized by their unique physical properties. The Fd3m and HII liquid crystalline phases represent two distinct arrangements. The Fd3m phase is a discontinuous cubic structure, notable for its intricate three- dimensional network containing reversed micelles of two different sizes. The HII phase, also known as the inverted hexagonal phase, is characterized by its hexagonally packed tubular structure. These phases have significant potential across various applications, including drug delivery systems, food technology, and cosmetic formulations. However, the production of these specific liquid crystalline phases can be challenging, particularly when aiming for cost-effectiveness and alignment with clean label standards. Traditional methods often involve complex processing and may require the use of synthetic or potentially health-impacting additives. Therefore, there is a need for innovative, low-cost, and effective methods to produce these specialized liquid crystalline phases, aligning with the growing consumer demand for products that are both effective and composed of natural, health-conscious ingredients. P92248PC 2 Summary of the invention The invention relates to compositions comprising a stable liquid crystalline phase comprising lipids organized in the Fd3m or HII space group, and methods of producing them. The invention relates in a first aspect to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids, and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Adding an aqueous solvent to the starting mixture, and mixing; and c. Adding at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase, and mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. The invention relates in a second aspect to a liquid crystalline phase comprising lipids organized in the Fd3m or HII space group, said liquid crystalline phase being obtained or obtainable by the methods of the invention. The invention relates in another aspect to a liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said liquid crystalline phase comprising a phase mixture comprising: i. in the range from 30 to 65 wt% polar lipids; ii. in the range from 35 to 70 wt% oil; and iii. an aqueous solvent; wherein said liquid crystalline phase has a lipid structure characterized by a small- angle X-ray scattering (SAXS) profile defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44 for the Fd3m space group and / or 1, ¥3, 2, ¥7, 3, ¥12 for the HII space group. P92248PC 3 In a third aspect, the invention relates to uses of the liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group obtained or obtainable by the methods of the invention. In another aspect, the invention relates to a hydrolysate mixture comprising: i. in the range from 30- 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and iii. less than 15 wt% other components. In a fourth aspect, the invention relates to the use of triacylglycerol lipases to produce the liquid crystalline phases organized in Fd3m or HII space group according to the invention. In a further aspect, the invention relates to a product selected from the group consisting of a food, a feed, a cosmetic, and a nutraceutical, wherein said product comprises a liquid crystalline phase as described herein. A still further aspect of the invention relates to the liquid crystalline phase as described herein for use as a medicament. Brief description of the figures Fig 1- SAXS diagram on liquid crystalline phase formed in Example 2. This diagram shows well-defined liquid crystalline phase with Fd3m with patterns with peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44. Fig 2- SAXS diagram on liquid crystalline phase formed in Example 7. This diagram shows SAXS diagram characteristic for less defined liquid crystalline phases. Seen as patterns of peaks that could be partially or fully overlapped, in SAXS measurements where Fd3m is defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44. Fig 3 - Light microscopy images of the hydrolysate mixture before addition of water (a), and 5-20 seconds after addition of water (b). On Fig 3a, a boundary is visible, which separates air (to the left) from the hydrolysate mixture (to the right). On Fig 3b, the LCP structure is seen in the middle. P92248PC 4 Detailed description of the invention The invention is based on the surprising discovery that when oil hydrolysate is mixed with polar lipids in the presence of water, the components present in the mix will assemble into liquid crystalline phases comprising lipids organized in Fd3m and / or HII space group. These liquid crystalline phases (LCP) are useful in many applications, such as in food, feed, cosmetic, nutraceutical and pharmaceutical applications. The presence of Fd3m and / or HII space group confers particular stability. In this invention, the liquid crystalline phase formation of the invention can be achieved using oil entirely originating from natural oils, without any synthetic components, such as synthetic lipids or emulsifiers. The method of the invention provides a means to avoid the use of chemicals or compounds that may be viewed as undesirable or unhealthy by the consumer, such as organic solvents. Further, the methods of the invention provide robust methods by which LCP organized in the Fd3m and / or HII space groups may be produced in significant quantities. The method is particularly effective at converting the starting mixture to liquid crystalline phase. Previously, LCP organized in Fd3m space groups has only been known to be formed in a narrow range of conditions. In contrast, the present invention allows use of several types of lipids and LCPs of the invention are formed under a wide range of conditions. The methods may be implemented using plant-based oils thereby avoiding components less desirable due to their impact on plant, animal, or human health. The methods of the invention furthermore provide methods for producing stable compositions comprising bioactive molecules. Definitions MAG: Monoacylglycerol; DAG: diacylglycerol; TAG: triacylglycerol; FFA: free fatty acids The term hydrolysis used in the context of hydrolysis of triacylglycerols, means that at least one fatty acid chain has been removed from the glycerol backbone by lipase. As used herein and in the appended claims, the singular forms “a,” “or,” and “the” include plural referents unless the context clearly dictates otherwise. P92248PC 5 It is understood that the embodiments described herein include “consisting” and / or “consisting essentially of” embodiments. As used herein, except where the context requires otherwise due to express language or necessary implication, the word “comprise” or variations such as “comprises” or “comprising” is used in an inclusive sense, i.e. to specify the presence of the stated features but not to preclude the presence or addition of further features in various embodiments. Overview of methods of the invention The hydrolysis products of oil are combined with an aqueous solvent, such as water, and polar lipids to assemble to the liquid crystalline phases organized in the Fd3m and / or HII space groups. The inventors have surprisingly identified the ratio of hydrolysis products to polar lipids needed, as well as the parameters required, for formation of these liquid crystalline phases. Hydration of the components is important. Thus, a certain amount of water must be present, and the components must be allowed to hydrate for the LCPs to form. It will be appreciated that there are many ways of generating hydrolysis products, of providing polar lipids, of ensuring the hydration of these components, and of mixing the components. The present invention encompasses all suitable means of each of these, if the identified ratio of hydrolysis products to polar lipids is fulfilled, resulting in the formation of liquid crystalline phases. Thus, the invention relates in a first aspect to the following methods for producing LCP comprising lipids organized in Fd3m and / or HII space group. Note that the disclosure regarding parameters in relation to the methods of the invention, is relevant also to all other aspects of the invention. Liquid crystalline phases (LCPs) Liquid crystals have properties between those of liquids and solid crystals. They are classified in different types, including lyotropic, thermotropic and metallotropic liquid crystals. The present invention concerns lyotropic liquid crystals, which means that they are built up of amphiphilic molecules and a solvent, which can be water. The invention relates to method of producing liquid crystalline phases comprising lipids organized in Fd3m and / or HII space group. As used herein, the term liquid crystalline phase refers to the lipid-containing phase which is formed when the oil P92248PC 6 hydrolysis products and the polar lipids organize into specific space groups. This LCP organizes and separates from the residual aqueous phase. The LCP according to the invention organizes the lipid components in specific space groups which are identifiable using SAXS, see further descriptions below, as wells as FIG.1 and 2. The Fd3m and HII space groups are particularly stable arrangement of molecules. The inventors have identified under what circumstances Fd3m or HII phases are formed, either in large amounts (well defined) or in smaller amounts sometimes together with other phases (less defined). The classification is based on the very typical pattern of peaks seen in SAXS for these phases, see below. When no significant amounts of Fd3m or HII are formed, the structure is referred to herein as disordered (when we cannot identify any typical LC phase using SAXS) or "lamellar". Lamellar phase is for example seen with the low amounts of polar lipids (PL4-PL25). This space group is not associated with stability. Viscoelastic properties of the liquid crystalline phases, such as Fd3m, have a storage modulus typically exceeding 10000 Pa further indicating their stability. Well-defined liquid crystalline phase: Products containing polar lipid-based liquid crystalline phase with the Fd3m, or HII nonlamellar mesomorphic space group. Seen as characteristic patterns of peak ratios in SAXS measurements where Fd3m is defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44, and HII at the peak positions at the q values in the ratios of 1, ¥3, 2, ¥7, 3, ¥12. See exemplary SAXS diagram in Fig 1. Less defined liquid crystalline phases. Products containing polar lipid-based liquid crystalline phase with the Fd3m, and / or HII space group. Seen as patterns of peaks that could be partially or fully overlapped, in SAXS measurements where Fd3m is defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44 and HII at the peak positions at the q values in the ratios of 1, ¥3, 2, ¥7, 3, ¥12. See exemplary SAXS diagram in Fig 2. Lamellar or lipid micellar aggregates. Formed especially at polar lipid contents of 25 % or below. In SAXS measurements the lamellar phase is defined by peak positions at the q values in the ratios of 1, 2, 3, 4, 5, 6. P92248PC 7 Further, some embodiments relate to the method according to the invention, wherein at least 50 wt% of the lipids from the starting mixture are incorporated in the lipid-based liquid crystalline phase; for example, at least 60%, 70, 80, 90, 95, 96, 97%; or for example in the range of 50 to 98 wt%. Thus, some embodiments relate to the method according to the invention, wherein at least 50 wt% of the lipids present in the stable liquid crystalline phase are organized in the Fd3m or HII space group, for example at least 60wt%, 70, 80, 90, 95, 96, 97 wt%; or for example 50 to 98 wt%. Methods for producing LCP The present invention relates in a first aspect to methods for producing stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group. Thus, the invention in one embodiment relates to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: A. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids; and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; B. Conducting step (B1) and (B2) in any order or simultaneously: B1. Adding an aqueous solvent to the starting mixture, and mixing; and B2. Contacting the mixture of step (A) or (B1) with at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase; and C. Subjecting the mixture of step (B) to further mixing; whereby a liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. In an embodiment of the invention, step (B1) precedes step (B2). P92248PC 8 In another embodiment of the invention, step (B2) precedes step (B1). Optionally, the lipase is removed prior to step (C). For example, if step (B2) precedes step (B1), the lipase may be removed in between step (B2) and (B1). In some embodiments, the aqueous solvent is added in a ratio of at least 0.01:1, such as at least 0.2:1, relative to the volume of the starting mixture. In a related embodiment, the invention relates to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids; and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Adding an aqueous solvent to the starting mixture, and mixing; and c. Adding at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase, and mixing; whereby a liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. In some embodiments, the aqueous solvent is added in a ratio of at least 0.01:1, such as at least 0.2:1, relative to the volume of the starting mixture. An embodiment of the invention relates to the method as described herein, wherein step a precedes steps (b) and (c), and wherein step (b) and step (c) can proceed in any order or simultaneously. Another embodiment of the invention relates to the method as described herein, wherein step (b) precedes step (c). A further embodiment of the invention relates to the method as described herein, wherein step (c) precedes step (b).In a further embodiment, the invention relates to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Mixing oil and polar lipids, to form a starting mixture consisting of: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil; and P92248PC 9 iii. less than 15 wt% other components wherein the oil and polar lipids are well-dispersed in each other; b. Adding water in a ratio at least 0.2:1 relative to the volume of starting mixture, and mixing; c. Contacting the mixture of step (b) with at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase; and d. Subjecting the mixture of step (c) to further mixing; whereby a liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. Without wishing to be bound by theory, it appears that hydrolysis products from lipase reactions organize together with polar lipids to form the liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group. It is preferred that a relatively large fraction of the lipids are incorporated into the LCP. Therefore, an embodiment of the invention relates to the method as described herein, wherein at least 50 wt% of the lipids from the starting mixture are incorporated in the liquid crystalline phase. Another embodiment of the invention relates to the method as described herein, wherein at least 50 wt% of the lipids in the liquid crystalline phase are organized in the Fd3m or HII space group. Oils In some embodiments of the method of the invention, oil and polar lipids are mixed to form a starting mixture. The starting mixture may also provide as a pre-mixed mixture. Therefore, an embodiment of the invention relates to the method as described herein, wherein said oil and said polar lipids of the starting mixture have been pre- mixed The term oil as used herein refers to a lipid composition comprising mainly triglycerides, and which is liquid at 40 τC. Typically, oil comprises in the range from 94 to 99 wt% triacylglycerol. In particular embodiments, the oil may comprise from P92248PC 10 95- 99 wt% TAG, or for example at least 97 wt% TAG, or at least 98 wt% TAG at least 99.5 wt% TAG, or consist essentially of TAG. The oil according to the invention may comprise oils from several sources or may consist of oil from a single source. Particular embodiments relate to where the oil is selected from the group consisting of plant oil, microbial oil, marine oil, animal oil and any combination thereof. Plant oil as used herein refers to any oil which is derived from a plant, such as from seeds, leaves, or any other part of a plant. Microbial oil as used herein refers to any oil which is derived from a microorganism. Examples of microbial oil are for example algal oil. Marine oil as used herein refers to oil which is derived from any marine-dwelling organism, such as one or more of algae, seaweed, fish, or plankton. Fish oil as used herein refers to oil which is derived from any fish. Fish oils are an example of animal oil. The selection of the oil to be used in the method of the invention may be based on particular diet or customer requirements. For example, vegan products may be made using one or more of plant oils and / or microbial oils. Compositions high in omega-3 fatty acids may be made using one or more marine oils. Particular embodiments of the invention relate to where the oil comprises or consists of a plant oil and / or marine oil. In particular embodiments, for example when relating to food, feed, and cosmetic applications, the oil may be one or more plant oil, such as one or more oil selected from the group consisting of oil from soybean, grape seed, cocoa, olive, palm, rice bran, rapeseed, canola, sunflower, safflower, peanut, cottonseed, almond, coconut, palm kernel, shea, corn, hazelnut, linseed, wheat germ and sesame oils; or one or more marine oil, such as one or more selected from the group consisting of fish oil, algal oil and seaweed oil. In particular embodiments, the invention relates to where the oil is a food grade oil, in particular a food grade plant oil. While animal fats are typically solid at 40 τC, they may be processed to become liquid at room temperature, for example as with ghee. Alternatively, the method of the invention may be performed at temperatures where the animal fat is liquid. The method of the invention relates to where the lipids are in liquid form. Thus, particular embodiments relate to oils and polar lipids which are in liquid form at 40oC. However, in principle any fat may be used as oil and / or source of polar lipids, P92248PC 11 as long as the method is performed under parameters where the lipid is in liquid form. For example, the method may be performed at high temperature such that lipids are melted, or at a combination of temperature and pressure where lipids are in liquid form. Polar lipids As used herein, the term polar lipids refers to lipids which have both hydrophobic and hydrophilic parts. In some embodiments the polar lipids comprise one or more phospholipids. In further embodiments, the polar lipids comprise one or more glycolipids. Further embodiments of the invention relate to where the polar lipid comprise one or more phospholipids, and / or one or more glycolipids. Phospholipids may be selected from the group comprising phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidic acid, and combinations thereof. Glycolipids may be selected from the group consisting of digalactosyldiglyceride (DGDG), Monogalactosyldiglyceride (MGDG), Digalactosylmonoacylglyceride (DGMG), Monogalactosylmonoacylglyceride and combinations thereof (MGMG). The polar lipids may be from any suitable source, for example being endogenous to the oil, being added exogenously to the oil, or any combination thereof. A particular embodiment relates to where the polar lipids comprise exogenously added polar lipids. The term lecithin as used herein refers to a fraction of oils which has high amount of polar lipids. Lecithins are readily available sources of polar lipids. In addition to polar lipids, lecithins may contain neutral fats, in particular TAGs. In some embodiments of the invention, the polar lipids comprise or consists of one or more lecithins, for example at one or more lecithins selected from the group consisting of plant lecithins, microbial lecithins, marine lecithins, animal lecithins and combinations thereof. Particular embodiments of the invention relate to where the polar lipid comprises or consists of one or more plant lecithins. Any plant lecithin may be used in the method of the invention. Examples of plant lecithin include lecithin from sunflower, soybean, oat, rapeseed, and cottonseed. P92248PC 12 Some embodiments of the invention relate to where the polar lipid comprises or consists of one or more animal lecithins, such as for example one or more of egg yolk and milk. Oat oil is particularly high in polar lipids, up to 20-40% wt, with digalactosyldiglyceride (DGDG) and phosphatidylcholine being the most abundant ones. In particular embodiments of the methods of the invention, the oil provided is oat oil and the polar lipids are provided by oat oil lecithin. Other components The starting mixture comprises less than 15% wt of other components, besides polar lipids, and oil. These may comprise or consist for example of sterols, carbohydrates or proteins present in the oil and / or polar lipids. Other components may also comprise one or more additives as described below. Therefore, an embodiment of the invention relates to the method as described herein, wherein the starting mixture comprises less than 15 wt% other components. In some embodiments the starting mixture comprises less than 12%, less than 10%, less than 9%, less than 8%, less than 7%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% of other components, (all % are by wt). In other embodiments, the starting mixture comprises in the range from 0.5 % to 15%, 0.5 to 10%, 0.5 to 8% , 0.5 to 6%, 0.5 to 5%, 0.5 to 4% , 0.6 to 3% or for example in the range from 0.05% to 6%, 0.05 to 4% by weight of other components. Starting mixture In one method of the invention, oil and polar lipids are mixed to form a starting mixture comprising in the range from 30 to 65 wt% polar lipids and in the range from 35 to 70 wt% oil. In one embodiment, the starting mixture consists of: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil; and iii. less than 15 wt% other components. P92248PC 13 This ratio of polar lipids to oil is important for the formation of the crystalline phase, see Example 5. The starting mixture may be formed by combining one or more oils with a further composition comprising one or more polar lipids. For example, an oil may be combined with a polar lipid, such as a lecithin, to reach a mixture within the range specified by the method of the invention. Any oil may be combined with any polar lipid. In some embodiments, an oil may be selected which already innately has the amount of polar lipids according to the invention, and addition of further polar lipids is not needed. Thus, one embodiment of the method of the invention relates to where no exogenous polar lipids are added. In alternative embodiments, method of the invention comprises a step (a) of mixing at least one lecithin and at least one oil, to form a starting mixture consisting of in the range of from 30 to 65 wt% lecithin and in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other. Mixing The method of the invention comprises mixing the starting mixture. This ensures that the polar lipids are dispersed in the oil. The mixing may be by any suitable means, and the person of skill will be able to select such means. In some embodiments this mixing takes place prior to contacting with lipase. In some cases, an oil may already have intrinsically the amount of polar lipids specified by the invention. In this case, the oil may still be subjected to mixing in order to ensure that the polar lipids are dispersed throughout the oil. In some embodiments, the mixing may be for example for at least 20 mins, such as at least 30, 40, 60, 90 or 120 mins. In other examples, the mixing may be in the range from 20 mins to 10 hours, or for example 1 hr to 10 hrs. Water Without wishing to be bound by theory, water is thought to have two functions in the method of the invention. Water is needed for enzymatic activity of step (c). Addition of water also initiates the formation of the LCP, and some water becomes P92248PC 14 a constituent of the LCP comprising lipids organized in Fd3m and / or HII space group that is formed in the method of the invention. Thus, in some embodiments, the method of the invention comprises a step of addition of an aqueous solvent, such as water, to the starting mixture in order to reach an amount which is at least 0.2 parts aqueous solvent to 1 part starting mixture, based on the volume of each part. The upper limit is not critical as excess water is excluded from the formed LCP into aqueous phase. Thus, the volume ratio may be for example in the range of from 0.2 to 10:1, such as from 0.3:1 to 6:1; or for example 0:2.1 to 5:1, or for example 1.2:1 to 4:1. Accordingly, an embodiment of the invention relates to the method as described herein, wherein the volume ratio of aqueous solvent to starting mixture is in the range of 0.2:1 to 10:1, such as from 0.3:1 to 6:1.In some variants of the methodology, only very little aqueous solvent is needed to initiate formation of the LCP, e.g. when adding the aqueous solvent to a hydrolysate mixture. Thus, an embodiment of the invention relates to the method as described herein, wherein an aqueous solvent is added to reach a ratio of at least 0.01:1 relative to the volume of hydrolysate mixture, such as at least 0.02:1, such as at least 0.05:1, such as at least 0.1:1, such as at least 0.2:1 relative to the volume of hydrolysate mixture. Thus, in some embodiments, the aqueous solvent is added in a ratio of at least 0.01:1, such as at least 0.2:1, relative to the volume of the starting mixture. Another embodiment of the invention relates to the method as described herein, wherein the aqueous solvent comprises at least 70 vol% water, such as at least 80 vol% water, such as at least 90 vol% water, such as at least 95 vol% water, such as at least 99 vol% water. A further embodiment of the invention relates to the method as described herein, wherein the aqueous solvent consists of water. Mixing in the presence of water ensures that the components of the LCP can assemble in an efficient way. The transport of water to the reversed micelles of the Fd3m structure is a potentially slow step, that is facilitated by mixing. Further, this mixing contributes to efficient enzyme hydrolysis. The water may be in any suitable form. For example, the at least one lipase of step (c) may be provided in step (b) in aqueous solution. Said aqueous solution of lipase P92248PC 15 may form part or all of the water that is added. Thus, particular embodiments relate to where lipase is provided in aqueous solution. Accordingly, an embodiment of the invention relates to the method as described herein, wherein the at least one lipase is comprised within the aqueous solvent. Aqueous solvent The water may be provided as an aqueous solvent comprising primarily water or consisting entirely of water. Accordingly, the aqueous solvent may comprise an additional polar solvent. The additional polar solvent may be, but is not limited to, methanol, ethanol, acetone, acetonitrile, ethylene glycol, preferably, methanol or ethanol. The volume ratio between the water and the additional polar solvent may be from 100:0 to 80:20, such as from 100:00 to 90:10, such as from 100:00 to 95:5. A too high concentration of the additional polar solvent interferes with enzyme functionality and is not desired. Lipase activity The inventors have identified that the generation of diacylglycerols, monoacylglycerols and free fatty acids in the hydrolysis of triacylglycerols contribute to the formation of the liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group. The at least one lipase according to the invention may be any lipase which will hydrolyse triacylglycerol. Thus, one embodiment relates to the method according to the invention where, under the conditions of step (c) or step (a1), the main activity of said at least one lipase is hydrolysis of triacylglycerols. Some embodiments of the invention relate to where the mixture of oil, polar lipids and aqueous solvent, such as water, is contacted with at least one lipase under conditions that allow lipase activity, such that at least 1% of the triacylglycerol in the oil is hydrolysed by said at least one lipase. In particular embodiments, the at least one lipase comprises or consists of a triacylglycerol lipase [EC 3.1.1.3]. P92248PC 16 In further particular embodiments, the at least one lipase comprises or consists of lipase having sn 1,3 positional specificity, such lipases preferentially cleave at positions 1 and 3 of the glycerol backbone. The sn 1,3 lipase may be any lipase with this positional specificity. For example, the sn 1,3 lipase may be selected from the group consisting of Rhizomucor miehie lipase (e.g. RM lipase from Novozymes), Thermomyces lanuginosus Lipase (e.g. Lipozyme TL lipase), and Rhizomucor Arrhizus lipase (e.g. Palatase from Novozymes), as well as any variants or modifications of these, such as heat stable variants, or the like. Some embodiments of the method of the invention relate to where, under the conditions of step (c) or (a1), said lipase has little or no phospholipase activity. In some embodiments, the at least one lipase is not classified as a phospholipase. The conditions for activity of enzyme may be selected by the skilled person. The parameters include temperature, amount of water, mixing speed, incubation time and amount of enzyme. The skilled person will be able to select the conditions under which the enzyme will achieve the hydrolysis of the triacylglycerol. The lipase may be provided in any formulation compatible with hydrolysis of the oil, and the skilled person will be able to select such formulations. Typically, the lipase is provided in aqueous suspension in the method according to the invention and incubated with the oil under mixing. Alternatively, the lipase may be provided as immobilized lipase, see further below. Any combination of the various formulations, such as aqueous and / or immobilized, may be used; in any sequence including use in parallel, use at the same time and / or sequential use. Lipase - immobilized Some embodiments of the invention relate to wherein the at least one lipase comprises or consists of immobilized lipase. In some embodiments of the invention, at least one lipase may be provided as immobilized lipase. For example, the lipase may be any of the lipases described above, which is immobilized. Immobilized lipase means that the at least one lipase is attached covalently or noncovalently to solid particles, such as beads. The immobilized enzyme particles can be used in various types of reactors, including stirred tank, rotating bed, and packed bed reactors. Immobilized lipases have the advantage of being able to be P92248PC 17 re-used. Further, it provides a means for control over the degree of hydrolysis of the triacylglycerol. The reaction can be stopped by separating the immobilized lipase from the reaction mixture (hydrolysate mixture). An even further advantage is that removal of the immobilized lipase ensures an enzyme-free product. In one embodiment, the at least one lipase comprises or consists of a triacylglycerol lipase, such as a sn-1,3 specific lipase, which is immobilized, for example immobilized TL lipase. Degree of hydrolysis The reaction products of lipase hydrolysis of triacylglycerol are important for the formation of the stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group of the invention, see Example 7. Thus, some embodiments of the method of the invention comprise a step (c) of contacting the mixture of step (b) with at least one lipase under conditions that allow lipase activity, such that at least 1% wt of the triacylglycerol in the oil is hydrolysed by said at least one lipase. Hydrolysed here means removal of at least one fatty acid chain from glycerol backbone. In other embodiments, the amount of triacylglycerol that is hydrolysed is at least 5 wt%, such as at least 30, at least 40 at least 50 at least 55, at least 65, at least 70, at least 75 at least 80, at least 85 at least 90, at least 92, at least 94 at least 95, at least 96, at least 97, at least 98, at least 99, or at least 99% wt. Alternatively, the invention relates in some embodiments to where the LCP comprises no more than 60 wt% remaining triacylglycerol, for example in no more than 50, 45, 40, 38, 35, 32, 30, 27, 25, 20, 18, 15, 12, 10, 8, 5, 4%, or no more than 3 wt% triacylglycerol remaining calculated as wt % based on the amount of triacylglycerol in the starting mixture comprising oil, polar lipids and possibly other components (see step (a)). In particular embodiments, the LCP comprises no triacylglycerol, i.e., that all triacylglycerol subject to lipase has been hydrolysed. In some embodiments, the LCP comprises in the range of at least 35% FFA, no more than 20% DAG and no more than 10% TAG; or for example at least 40% FFA, no more than 15% DAG, and no more than 10% TAG. The amount may be calculated as peak area percentage in the TLC data (see Example 6). P92248PC 18 Without wishing to be bound by hypothesis, it appears that the level of TAG and FFA are more important to the formation of the LCP than the levels of DAG and MAG. Thus, particular embodiments relate to where the LCP of the invention comprise in the range of at least 35% FFA and no more than 10% TAG; for example at least 40% FFA and no more than 10% TAG; such as at least 50% FFA and no more than 10% TAG; or for example at least 60% FFA and no more than 5% TAG, or at least 60% FFA and no more than 3% TAG, or at least 60% FFA and essentially no TAG. Low amounts (such as below 10%) of TAG seem to be particularly useful when aiming to form an LCP comprising lipids organized in the Fd3m space group. Typically, LCPs comprising lipids organized in the HII space group contains higher amounts of TAG compared to LCPs comprising lipids organized in the Fd3m space group. For example, LCPs comprising lipids organized in the HII space may comprise TAG in the range of from about 15% to 60%, such as from 25% to 58%, for example from 35% to 55%. Further mixing The methods of the invention comprise a first mixing, done to ensure the dispersion of polar lipids in oil, see above. The methods of the invention may further comprise a mixing whereby lipase is contacted with the mixture. The mixing to ensure lipase contact with substrate and / or water, may be done by any suitable means. Some embodiments of the invention relate to where the formation of LCP is allowed to continue for at least 12 hrs, such as at least 14 hrs, at least 20 hrs, or for example in the range from 12 to 100 hrs, or for example in the range from 16 to 100 hrs. Another embodiment of the invention relates to the method as described herein, wherein the liquid crystalline phase formation is allowed to continue for at least 12 hours, such as at least 14 hours, preferably at least 16 hours. The long formation time is needed to ensure sufficient lipase activity. The LCP formation happens rapidly as soon as sufficient degrees of hydrolysis has occurred. Sufficient hydrolysis has occurred when TAG levels are within the ranges as described herein. P92248PC 19 Therefore, in embodiments wherein a hydrolysate mixture is formed, then it is the formation of the hydrolysate mixture that takes time. Therefore, if a hydrolysate mixture is stored for at least the above-mentioned period, then the LCP formation is instantaneous upon addition of water. By instantaneous is meant no more than 1 minute, for example no more than 50 sec, such as no more than 40 sec, for example no more than 30 sec, such as no more than 20 sec, for example no more than 10 sec, such as no more than 5 sec. Additives The inventors have shown that liquid crystalline phases of the invention can be made to comprise additives. The additive may also be referred to as an ingredient. Thus, some embodiments relate to the method according to the invention, wherein at least one additive is added, resulting in a stable liquid crystalline phase comprising said additive. These one or more further ingredients may be added to the aqueous composition if water-soluble, or if lipophilic, may be mixed with the oil and / or polar lipids of the process, and is incorporated into the LCP. Accordingly, an embodiment of the invention relates to the method as described herein, wherein the at least one additive is added to the starting mixture or to the hydrolysate mixture if the additive is lipophilic or the at least one additive is added with the aqueous solvent if the additive is water-soluble. The further ingredient may be any ingredient. They may for example be bioactive molecules, such as vitamins, pharmaceutically active ingredient, or the like. In other embodiments, alternatively or additionally, the additive may comprise one or more of colourants, fragrances, and flavouring agents. The additives may be selected based on the needs and aims of the product and are not particularly limited. An embodiment of the invention relates to the method as described herein, wherein said at least one additive is selected from the group consisting of bioactive molecules, pharmaceutically active ingredients, colourants, fragrances, and flavouring agents. P92248PC 20 Further steps The method of the invention may comprise one or more further steps, whereby the LCP is processed further. For example, the method of the invention may include a step of spray-drying the LCP. Spray drying may be done for example by dispersing the LCP, followed by e.g., sonication to get particles dispersed in solution. This solution is then spray-dried to achieve a final product in powder form. Thus, one embodiment of the invention relates to the spray-dried LCP of the invention. Further methods for producing LCP The LCPs are formed when oil hydrolysates, polar lipids and an aqueous solvent, such as water, are combined in amounts according to the invention. In some embodiments, hydrolysis of oil is performed after oil and polar lipids are mixed, in the presence of water. In these embodiments, the generated hydrolysis products (e.g. FFA, MAG, and / or DAG) and the polar lipids assemble to LCP as the hydrolysis progresses. This principle may be referred to as “one pot production”. In some embodiments, LCP is produced in a two-step method, where the hydrolysis of oil and the formation of LCP are performed in separated reactions. Thus, in a first step, hydrolysis of oils is performed, yielding hydrolysis products (a hydrolysate). In a separate, second step, the hydrolysis products are combined with water and polar lipids in amounts according to the invention, and the LCP are formed. The two steps may be separated in time or may be physically separated. Examples of separation in time are for example where hydrolysis of oil is performed first; after which the at least one enzyme is inactivated or removed; and thereafter the hydrolysate is combined with polar lipids and water in amounts according to the invention, thereby forming LCP. Example of physical separation is for example where hydrolysis of oil is performed in a reactor bed using immobilized lipase; after which the hydrolysis reaction mixture is separated from the immobilized lipase by for example transferring the hydrolysis reaction mixture containing the hydrolysis products to another vessel, leaving the immobilized lipase behind; and thereafter combining the hydrolysis products with water and polar lipids in amounts according to the invention in said vessel, thereby forming LCP. Thus, the enzymatic hydrolysis of oil to yield hydrolysis product may be done before, concurrently with, or after mixing with polar lipids. P92248PC 21 Thus, the invention relates in some embodiments to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a1. Contacting an oil with at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase to obtain an oil hydrolysate; b1. Optionally removing the at least one lipase from the oil hydrolysate, c1. Mixing the oil hydrolysate from step a1 or b1 with polar lipids, to form a hydrolysate mixture comprising: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and wherein the oil hydrolysate and polar lipids are well-dispersed in each other; d1. Adding an aqueous solvent to the hydrolysate mixture, and further mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. In some embodiments, the aqueous solvent is added to reach a ratio of at least 0.01:1, such as at least 0.2:1, relative to the volume of hydrolysate mixture. Another embodiment of the invention relates to the method as described herein, wherein said method comprises the steps of: a1. Contacting an oil with at least one lipase under conditions that allow lipase activity, such that at least 1% wt of the triacylglycerol in the oil is hydrolysed by said at least one lipase; b1. Optionally removing lipase from reaction mixture, c1. Mixing the oil hydrolysate from step a1 or b1 with polar lipids, to form a mixture consisting of: i. in the range from 30- 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and iii. less than 15 wt% other components wherein the oil hydrolysate and polar lipids are well-dispersed in each other; P92248PC 22 d1. Adding water to reach a ratio in the range of at least 0.2:1 relative to the starting mixture and mixing; whereby a stable lipid-based liquid crystalline phase is formed. The above methods may be stopped after step c1, so as to obtain a hydrolysate mixture. Said hydrolysate mixture is storage stable. Then, a method of forming a liquid crystalline phase comprises the steps of: i. providing a hydrolysate mixture; ii. adding aqueous solvent to the volume of the hydrolysate mixture and mixing; whereby a stable lipid-based liquid crystalline phase is formed. In some embodiments, the aqueous solvent is added to reach a ratio of at least 0.01:1, such as at least 0.2:1, relative to the volume of hydrolysate mixture, and further mixing. When aqueous solvent, such as water, is added to the hydrolysate mixture, then the LCP formation is typically instantaneous. By instantaneous is meant no more than 1 minute, for example no more than 50 sec, such as no more than 40 sec, for example no more than 30 sec, such as no more than 20 sec, for example no more than 10 sec, such as no more than 5 sec. Hydrolysate mixture The disclosure also relates to the hydrolysate mixture as such. The hydrolysate mixture has been found to be surprisingly stable. Additives may be added to the hydrolysate mixture after prolonged storage followed by mixing with an aqueous solvent to form a desired LCP. If the additives are fragile it may be preferred to delay incorporation into the system and for some applications it may therefore be advantageous to delay addition of additive and aqueous solvent. Thus, an aspect of the present invention relates to a hydrolysate mixture comprising: i. in the range from 30- 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and iii. optionally, less than 15 wt% other components. P92248PC 23 An embodiment of the invention relates to a method of forming the hydrolysate mixture comprising the steps of: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids, and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Adding at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase, and mixing. Another embodiment of the present invention relates to a method of forming a hydrolysate mixture comprising the steps of: a1. Contacting an oil with at least one lipase under conditions that allow lipase activity, such that at least 1% wt of the triacylglycerol in the oil is hydrolysed by said at least one lipase; b1. Optionally removing lipase from reaction mixture, c1. Mixing the oil hydrolysate from step a1 or b1 with polar lipids, to form a hydrolysate mixture comprising: i. in the range from 30- 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and iii. less than 15 wt% other components wherein the oil hydrolysate and polar lipids are well-dispersed in each other. A related embodiment of the invention relates to a method of forming a hydrolysate mixture comprising the steps of: a1. Contacting an oil with at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase to obtain an oil hydrolysate; b1. Optionally removing the at least one lipase from the oil hydrolysate, c1. Mixing the oil hydrolysate from step a1 or b1 with polar lipids, to form a hydrolysate mixture comprising: P92248PC 24 i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and wherein the oil hydrolysate and polar lipids are well-dispersed in each other. The hydrolysate mixture can be produced with or without additives and stored for later use. Thus, at a later time, an aqueous solvent may be added to the hydrolysate mixture to form a desired LCP. The hydrolysate mixture may comprise the lipase. Alternatively, when the lipase is removed in step b1, then the hydrolysate mixture comprises no or almost lipase. LCP achieved by the methods of the invention The invention relates in a second aspect to a liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said liquid crystalline phase being obtained or obtainable by the methods of the invention. Upon the final step of mixing in the methods of the invention, the oil hydrolysate, polar lipids, and some of the water come together to form the liquid crystalline phase where the lipids are organized in specific space groups. As the LCP is formed, it separates from a residual aqueous phase. The liquid crystalline phase comprises or consists of oil hydrolysis products, polar lipids and an aqueous solvent, such as water, and any further ingredients, such as additives. The additives may be described as being encapsulated or enclosed in the LCP of the invention. Previous methods of making liquid crystalline phases only show transient LCPs with Fd3m. In contrast, the present invention relates to the formation of stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group. The LCP comprising lipids organized in Fd3m and / or HII space group according to the invention are stable. Stability correlates with the extent of formation of Fd3m and / or HII. Thus, the invention relates in some embodiments to where the LCP has well-defined crystalline phase. This can be determined by the SAXS diagram, but also by visual inspection, and stability tests (see Example 4). P92248PC 25 Thus, in some embodiments, the LCP is stable for at least 4 days at room temperature (25°C), such as for 5, 6. 7 days; or for example for 1, 2, 3, or 4 weeks at 25°C, or for example for 1, 2 or 3 months at 25°C. Particular embodiments relate to where the LCP is stable for at least 2 months at 25°C, such as is stable for at least 3 months, at least 4 months at 25 °C. Additionally, or alternatively, in some embodiments, the LCP according to the invention is stable for at least 4 weeks at 4°C such as for at least 5, at least 6 or at least 7 weeks; or for example for at least 1, 2, 3, or 4 months at 4°C, or for example for at least 1, 2 or 3 years at 4°C. Particular embodiments relate to where the LCP is stable for at least 6 months at 4°C, such as is stable for at least 12 months or at least 18 months at 4°C. In an embodiment, a liquid crystalline phase is obtained or obtainable by the method of the disclosure, said liquid crystalline phase comprising in the range from 30 to 65 wt% polar lipids, and having a lipid structure characterized by a small- angle X-ray scattering (SAXS) profile defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44 for the Fd3m space group or 1, ¥3, 2, ¥7, 3, ¥12 for the HII space group. An aspect of the invention relates to a liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said liquid crystalline phase comprising a phase mixture comprising: i. in the range from 30 to 65 wt% polar lipids; ii. in the range from 35 to 70 wt% oil; and iii. an aqueous solvent; wherein said liquid crystalline phase has a lipid structure characterized by a small- angle X-ray scattering (SAXS) profile defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44 for the Fd3m space group and / or 1, ¥3, 2, ¥7, 3, ¥12 for the HII space group. In the context of the present application, a phase mixture is a composition comprising polar lipids, oil and aqueous solvent, for example, the phase mixture may comprise from in the range from 30 to 65 wt% polar lipids; in the range from 35 to 70 wt% oil; and an aqueous solvent. P92248PC 26 An embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the volume ratio between the aqueous solvent and the phase mixture is at least 0.01:1, such as at least 0.2:1. It has been found that particular contents of triacylglycerol (TAG) and free fatty acids (FFA) leads to well-defined and very stable liquid crystalline phases with lipids organized in the Fd3m space group. Accordingly, an embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the lipids are organized in the Fd3m space group. The ratio between TAG and FFA may be given by the peak intensity of the peaks corresponding to TAG and FFA, respectively, as determined by TLC. In brief, the area under each peak can be determined and the ratio be calculated as peak area of TAG to peak area of FFA. Alternatively, the ratio may be given as the wt% of TAG to wt% of FFA. An embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the ratio between triacylglycerol (TAG) and free fatty acids (FFA) is equal to or less than 1:5. Another embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the ratio between TAG and FFA is equal to or less than 1:10, such as equal to or less than 1:20, such as equal to or less than 1:30, such as equal to or less than 1:40, such as equal to or less than 1:50, such as equal to or less than 1:60, such as equal to or less than 1:70, such as equal to or less than 1:80, such as equal to or less than 1:90, such as equal to or less than 1:95, such as equal to or less than 1:99. A further embodiment of the invention related to the liquid crystalline phase as described herein, wherein the liquid crystalline phase comprises substantially no TAG. For some applications it may be preferred to produce liquid crystalline phases as described herein, wherein the lipids are organized in the HII space group. As demonstrated herein, these LCPs are formed at higher amounts of TAG, i.e. when less hydrolyzation has occurred. In particular, the content of TAG may be higher than FFA. Accordingly, an embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the lipids are organized in the HII space group. P92248PC 27 Another embodiment of the present invention relates to the to the liquid crystalline phase as described herein, wherein the content of TAG is higher or equal to the content of FFA. Such LCPs can be in the HII phase. A further embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the ratio between triacylglycerol (TAG) and free fatty acids (FFA) is in the range of 1:1 to 50:1, such as 2:1 to 25:1, such as 3:1 to 20:1, such as 5:1 to 15:1. Yet another embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the ratio between TAG and FFA is in the range of 12:1 to 6:1. A still further embodiment of the invention relates to the liquid crystalline phase as described herein, wherein the liquid crystalline phase comprises less than 15 wt% other components. The liquid crystalline phase may be loaded with additives to yield functionalities beneficial in a broad variety of products. The additives may be components of importance to the development and preparation of food compositions or nutraceuticals for which high stability is a premium property, or could be active pharmaceutical ingredients (API) that requires sustained release and / or stability. Thus, an embodiment of the invention relates to the liquid crystalline phase as described herein, further comprising one or more additives. Another embodiment of the invention relates to the liquid crystalline phase as described herein, wherein said one or more additives are selected from the group consisting of bioactive molecules, pharmaceutically active ingredients, colourants, fragrances, and flavouring agents. The LCP of the disclosure typically comprise from 10 to 35 wt% water, such as from 15 to 25 wt% water. An aspect of the invention relates to a product selected from the group consisting of a food, a feed, a cosmetic, and a nutraceutical, wherein said product comprises a liquid crystalline phase as described herein. P92248PC 28 Uses In a third aspect, the invention relates to uses of the compositions obtained or obtainable by the methods of the invention. An aspect of the invention relates to the use of a liquid crystalline phase as described herein in the preparation of a food, a feed, a cosmetic, or a nutraceutical. Another aspect of the invention relates to the liquid crystalline phase as described herein for use as a food, feed, ingredient, cosmetic, nutraceutical or excipient. Alternatively, the liquid crystalline phase may comprise pharmaceutical active ingredient (API). In that case, the liquid crystalline phase may be used within medicine for delivery of the API. Thus, an aspect of the invention relates to the liquid crystalline phase as described herein for use as a medicament. An embodiment of the invention relates to the liquid crystalline phase as described herein for use in medicine, for example as nanocarrier of active pharmaceutical ingredients. Lipase for use in producing LCP In a fourth aspect, the invention relates to the use of triacylglycerol lipases in the methods of the invention. Thus, the invention relates to a triacylglycerol lipase, in particular having sn 1,3 positional specificity, for use to produce the LCP of the invention. In particular embodiments, this use is in any method according to the present invention. Specific embodiments The invention relates in one specific embodiment to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Mixing oil and polar lipids, to form a starting mixture consisting of: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil; and iii. less than 5 wt% other components P92248PC 29 wherein the oil and polar lipids are well-dispersed in each other; b. Adding water to reach a ratio in the range of at least 0.2:1 relative to the starting mixture, and mixing; c. Contacting the mixture of step (b) with at least one triacylglycerol lipase having sn 1, 3 specificity, under conditions that allow lipase activity, such that at least 50 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase; and d. Subjecting the mixture of step (c) to further mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. The invention relates in a related embodiment to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Adding an aqueous solvent to the starting mixture, and mixing; c. Contacting the mixture of step (b) with at least one triacylglycerol lipase having sn 1, 3 specificity, under conditions that allow lipase activity, such that at least 50 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase; and d. Subjecting the mixture of step (c) to further mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. In some embodiments, the aqueous solvent is added to reach a ratio of at least 0.01:1, such as at least 0.2:1, relative to the volume of starting mixture, and further mixing. The invention relates in a related embodiment to a method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: P92248PC 30 a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids; and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Contacting the mixture of step (a) with at least one triacylglycerol lipase having sn 1, 3 specificity, under conditions that allow lipase activity, such that at least 50 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase; c. Adding an aqueous solvent to the starting mixture; and d. Subjecting the mixture of step (c) to mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed. In some embodiments, the aqueous solvent is added to reach a ratio of at least 0.01:1 for example at least 0.2:1 relative to the volume of starting mixture, and further mixing. The listing or discussion of an apparently prior published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge. Preferences, options and embodiments for a given aspect, feature or parameter of the invention should, unless the context indicates otherwise, be regarded as having been disclosed in combination with any and all preferences, options and embodiments for all other aspects, features and parameters of the invention. This is especially true for the description of the method for producing the stable crystalline phase and all its features, which may readily be part of the stable crystalline phase per se or obtained by the method or its use as described herein. Embodiments and features of the present invention are also outlined in the following items. The invention will now be described in further details in the following non-limiting examples. EXAMPLES P92248PC 31 Example 1- Lipases Various lipases were tested for usefulness in producing LCPs. The lipases tested are described in Table 1 below. They are commercially available enzymes from Novozymes, and the activity is defined by Novozymes as: The 1LU is the amount of enzyme activity which liberates 1 Njmol of titratable butyric acid from the substrate glycerol tributyrate per minute under defined standard conditions. 1LU is equal to 1IUN. 1 PLU is the amount of enzyme activity which generates 1 Njmol of propyl laurate per minute under defined standard conditions. The experiments were done using oat oil that has been enriched with polar lipid fraction from oat. Thus, 300 mg polar lipid-enriched oat oil, containing 40 wt% polar lipids and 1.2 ml aqueous buffer were added to 4.5 mL glass sample tubes. 10 μl of lipase solution was added, and the samples incubated on a thermomixer (700 RPM) at 40 °C degrees for 16 hrs. After incubation, the samples were removed from incubator and the formation of liquid crystalline phase assessed by Small Angle X-ray Scattering (SAXS), see below and the results are presented Table 1 and Figure 1. Phase identification using SAXS. Most SAXS-experiments were carried out at the SAXS laboratory at the Division of Physical Chemistry, Dept of Chemistry, Lund university. A few experiments were carried out at the CoSAXS beamline of the MAX IV laboratory, Lund University. Phases were identified based on the pattern of peaks appearing in the SAXS experiments. Fd3m is characterized by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44, HII at the peak positions at the q values in the ratios of 1, ¥3, 2, ¥7, 3, ¥12. Table 1 Sample Lipase name, source, Comments Results and specificity; activity A No lipase 0.1M pH 7 phosphate Disordered buffer BLecitase Ultra0.1M pH 7 phosphate Disordered (Novozymes); PLA1 sn-1 buffer directed CLipozyme RM,0.1M pH 7 phosphate Fd3m Rhizomucor miehei, buffer (Novozymes); sn1,3 directed; 275 IUN / g P92248PC 32 DLipozyme RM,Tap water Fd3m Rhizomucor miehei, (Novozymes) ;sn1,3 directed, 275 I UN / g ELipozyme TL,0.1M pH 7 phosphate Fd3m Thermomyces buffer lanuginosus, (Novozymes), sn1,3 directed, 100 KLU / g FPalatase (Novozymes),0.1M pH 7 phosphate Fd3m 20000L sn1,3 directed, buffer GLipozyme CALB L,0.1M pH 7 phosphate Fd3m Candida Antarctica buffer (Novozymes), Non- specific / sn-2 directed; 5000 LU / g HLipozyme TL IM,Tap WaterFd3mThermomyces lanuginosus, (Novozymes), 250 IUN / g ILipozyme RM IM,Tap WaterFd3mRhizomucor miehei, (Novozymes), 150 IUN / g *K=Kilo, LU = Lipase unit, PLU = Propyl Laurate Unit, IUN = Interesterification Unit. 1LU is the amount of enzyme activity which liberates 1 Njmol of titratable butyric acid per minute under defined standard conditions. 1LU is equal to 1IUN. 1 PLU is the amount of enzyme activity which generates 1 Njmol of propyl laurate per minute under defined standard conditions, as defined by Novozymes. Thus, the formation of the LCPs according to the invention require the hydrolysis of the triacylglycerols of an oil. Buffer and tap water work equally well as water phase. Example 2- Time course The method was carried out according to the method in Example 1. The lipase used was Lipozyme TL. The formation of the LCP is allowed to continue for differing amounts of time. Table 2 Time, h Outcome 6 Disordered 12 Disordered P92248PC 33 16 Fd3m 24 Fd3m 48 Fd3m 96 Fd3m The results show that development of LCP occurs over time. The LCP formation causes a phase separation with essentially all of the lipids forming the LCP phase and the excess water forming a separate phase. It is seen that even after 96 hrs the developed LCP remain, thus indicating good stability of the LCP. Example 3 – Temperature The method was carried out according to the method in Example 1, but incubation time was 24 h. Samples were then transferred to various temperatures and their structures determined by SAXS. Thus, the Fd3m phase can exist in a wide range of temperatures. Table 3 Oat Polar Water, Time, Temp, Mixing, Outcome oil, lipids ml h C RPM mg (wt%) 300 40 1.2 24 25 700 Fd3m 300 40 1.2 24 35 700 Fd3m 300 40 1.2 24 45 700 Fd3m 300 40 1.2 24 55 700 Fd3m 300 40 1.2 24 65 700 Fd3m Example 4 - Stability LCPs were made according to the method in Example 1 using oat oil with 40% polar lipids and Lipozyme TL. The LCPs were stored as described, and the stability checked visually as well as with SAXS equipment. Phase separation in the context of this experiment refers to where the LCP breaks down into an oil and a water phase and the oil phase no longer has any identifiable organization. For example, the SAXS profiles associated with LCP (see definitions) P92248PC 34 are no longer identified in the LCP. The table indicates how long the LCP is intact, until it breaks down (phase separation). Thus, lamellar mesophase structures within a few days. LCP comprising lipids organized in Fd3m and / or HII space group display greater stability. Table 4 System created Storage Stability- duration of LCP Lamellar mesophase 25°C 2 days before phase separation (See Example 5) Lamellar mesophase 4°C 1-2 weeks before phase separation (see Example 5) Liquid crystalline phase with 25°C At least 2 months before the Fd3m space group phase separation Liquid crystalline phase with 4°C More than a year before phase the Fd3m space group separation or other apparent negative effect on stability Liquid crystalline phase with 4°C More than year before phase the HII space group separation or other apparent negative effect on stability Example 5 -Amount of polar lipids Oat oils containing various amounts of polar lipids from oat (named PL4 to PL60, having from 4 to 60 %wt polar lipids based on total wt) were used in the method described in Example 1. 10 μl of Lipozyme TL (Novozymes) lipase was used. The results are shown in Table 5. Table 5 Oat oil Water, Time, Temp, Mixing, Outcome with ml h C RPM Polar lipid content (wt%); 4 1.2 16 40 700 Lamellar 15 1.2 16 40 700 Lamellar 25 1.2 16 40 700 Lamellar 30 1.2 16 40 700 Fd3m 40 1.2 16 40 700 Fd3m 50 1.2 16 40 700 Fd3m P92248PC 35 60 1.2 16 40 700 HII Thus, in the case of oat oil, the formation of the Fd3m phase requires the presence of at least 30 % polar lipids. At polar lipid content of 60 %, inverse hexagonal phase is formed instead. Example 6 – Effect of dispersion The method was carried out according to the general method in Example 1 using 300 mg oat oil which contained 40% polar lipids by weight (PL40), 1.2 ml water and 10 μl lipase (Lipozyme TL, Novozymes). After incubation for 16 h, the LCP was collected, and dispersed in water by sonication. This made it possible to pipette the dispersion, which was analysed by SAXS at the CoSAXS beamline of MAX IV, Lund University. The analysis showed that the dispersed product contained HII phase. Thus, dispersion by sonication under the conditions used causes the conversion of Fd3m phase to HII phase. The HII phase can be of interest because its water domains are more connected than those of Fd3m, leading to facilitated diffusion of hydrophilic molecules. Furthermore, a liquid dispersion can be handled by pumping, pipetting etc., which cannot be done with a stiff bulk LCP. Example 7 - Oil hydrolysate products form liquid crystalline phases Sunflower oil was hydrolysed using immobilized TL IM lipase from Novozymes, using the general method of Example 12. Samples were taken at different time points and analysed with the TLC method that is sensitive for TAGs, MAGs, DAGs and FFAs. The samples containing hydrolysis products were combined with lecithin, water (2 -step method, see Example 12) to produce liquid crystalline phases. The LCPs were analysed with SAXS, see Table 6 for results. Two control samples were run using the general method of Example 1. These samples were incubated for 16 hours to ensure full hydrolysis of TAG. The LCP that was formed was analysed by TLC Analysis, see Table 6. TLC Analysis The lipid hydrolysis products were extracted according to Bligh and Dyer (Canadian Journal of Biochemistry and Physiology, 1959. 37(8): p. 911-917) P92248PC 36 and analysed using thin layer chromatography (TLC). The lipids were separated on TLC silica 60 plates (Merck) using heptane: diethyl ether:acetic acid (70:30:1) as mobile phase and visualized under UV light after spraying with primulin solution. The TLC plates were analysed using a scanner (CAMAG) and the peak intensities were recorded. The sensitivity of the method is not equal for all sample components, however the results are still useful for demonstrating the ongoing conversion of TAGs to DAGs, MAGs and FFAs. Data is generated using the software visionCATS, where approximate peak areas are generated. The results of TLC and SAXS analysis are shown in Table 6. The numbers for each of “PL, MAG”, “FFA”, “DAG”, and “TAG” corresponds to peak intensities as measured in the TLC analysis. It could be seen that the amount of free fatty acids and diacylglycerol increased significantly after enzymatic hydrolysis. The increase was greater with longer incubation times and was accompanied by a decrease in triacylglycerol. The formation of Fd3m structure correlates with the removal (hydrolysis) of TAG and generation of FFA. In contrast, the concentration of MAG and DAG respectively do not change as drastically as that of FFA and TAG. In the well-defined Fd3m phase, most of the components are FFAs (60% peak area approximately). TAG and PL combined make up approximately 40% peak area. It can be observed that in the well-defined Fd3m there is no TAG. Table 6 Time course PL, MAG FFA DAG TAG LCP (minutes) 0 16,73 0,72 4,61 59,33 micelles 15 20,24 5,12 7,71 57,9 HII 30 15,07 7,48 12,2 57,69 HII 60 15,16 8,64 9,7 55,92 HII 90 26,59 40,6 12,5 9,9 Fd3m, less defined 120 30,41 39,35 8,25 8,76 Fd3m, less defined 330 25,35 45,91 10,6 9,28 Fd3m, less defined End point PL, MAG FFA DAG TAG LCP 16 hours 13,54 66,27 12,96 0 Fd3m, well defined 16 hours 22,71 63,5 6,16 0 Fd3m, well defined P92248PC 37 PL=polar lipids; MAG=monoacylglycerol; DAG=diacylglycerol; TAG=triacylglycerol; FFA=free fatty acids. Example 8 - Dispersion of polar lipids in the oil is important for LCP formation. Samples were prepared according to the method of Example 1. Peanut oil was mixed with polar lipids in the form of sunflower lecithin (see Example 9). In one case, the mixture was thoroughly mixed for 2h to obtain a visually homogeneous dispersion before addition of water and enzyme. In the other case, water and enzyme were added directly and incubation started. Both mixtures were incubated 16 h at 700 rpm and the products were examined (see Table 7). Thus, dispersion of the polar lipids in the oil is important for the later formation of Fd3m phase. Table 7 Peanut Lecithin, Water, Pre- Incubation Temp, Mixing Outcome oil, mg mg ml mixing time, h °C during time incubation, H, T°C R and RPM 300 300 1.2 0 h 16 40 700 Disordered 300 300 1.2 2 h, 16 40 700 Fd3m 40- 60°C, 700 Example 9 - LCP can be formed with various oils Samples were prepared according to the method of Example 1, using the following oils: peanut oil, flaxseed oil, rapeseed oil, and coconut oil. The oils were food grade oil used for cooking and were commercially available in grocery shop. Peanut oil has a high smoke temperature. Flaxseed oil has a high content of omega-3 fatty acids, in particular ALA. The beneficial qualities of linseed oil can be reduced if used at high temperatures. Coconut oil is solid below 25°C. See Table 8 for composition of the oils. P92248PC 38 Sunflower lecithin was purchased from health shop and had the following composition per 100 g: Phospholipids - 99 g; where saturated fat was 12 g; monounsaturated fat was 23 g and polyunsaturated fat was 64 g. The composition contained 0 g carbohydrates, 0 g protein and 0 g salt. Table 8 Contents per 100 Sunflower Peanut oil, Flaxseed oil, Coconut oil, ml oil Per 100 g Energy 3404 kJ, 3382 kJ, 823 3441 kJ, 837 884 kcal 828 kcal kcal kcal Fat (lipids) 92 g 91.4 93 g 100 g - Saturated fat 11 g 16.4 9.3 g 86.5 g - Monounsaturated 25 g 52.1 18.6 g 5.8 g fat - Polyunsaturated 56 22.9 65.1 g 1.8 fat Carbohydrates 0 0 0 0 Protein 0 0 0 0 Salt 0 0 0 0 300 mg of the oil was mixed well with various amounts of sunflower lecithin (see above) to obtain a visually homogeneous dispersion. 1.2 ml water and 10 μl lipase (TL) solution was added followed by incubation for 16h at 40°C under mixing at 700 rpm. The results are shown in Table 9. Thus, Fd3m phase can be made by combining vegetable oils from one source with polar lipids from other plant species. Further, the examples show that the method is suitable for producing Fd3m phases with various oils. Table 9 Oil, Sunflower Water, Outcome 300 mg lecithin, mg ml Sunflower 0 1.2 Disordered Sunflower 150 1.2 Disordered Sunflower 200 1.2 Disordered Sunflower 300 1.2 Fd3m Rapeseed 200 1.2 Disordered Rapeseed 300 1.2 Fd3m P92248PC 39 Rapeseed 500 1.2 Disordered Coconut 0 1.2 Disordered Coconut 200 1.2 Disordered Coconut 400 1.2 Fd3m Coconut 500 1.2 Fd3m Peanut 150 1.2 Disordered Peanut 200 1.2 Disordered Peanut 300 1.2 Fd3m Flaxseed 0 1.2 Disordered Flaxseed 100 1.2 Disordered Flaxseed 200 1.2 Disordered Flaxseed 300 1.2 Fd3m Example 10 - LCP can be made to comprise additives LCP were prepared according to the method of Example 1. 300 mg of oat oil containing 40 % polar lipids (PL40), was mixed with 1.2 ml water and 10 μl lipase solution (TL lipase) and incubated for 16 h at 40τC under mixing at 700 RPM with thermomixer. The following additives were tested as inclusions: Curcumin from Curcuma longa (Turmeric), powder (CAS: 458-37-7), Sigma-Aldrich; Vitamin D (Ergocalciferol, CAS: 50-14-6), >98%, Sigma-Aldrich; Iron(II) sulphate heptahydrate: ACS reagent, ^99.0%, (CAS: 7782-63-0) Sigma-Aldrich; Fish Oil: Capsules, ICA AB (the capsules were cut open and the fish oil recovered and used in the experiments). The water-soluble additives (curcumin, iron sulphate) were dissolved in the water before the water was mixed with oil. The fat-soluble additive (fish oil, Vitamin D) was dissolved in oil before the oil was mixed with water. For the trials with 5-40% fish oil, the amount of PL40 was reduced correspondingly. The results are shown in Table 10. The results show that additives can be included and become encapsulated in the LCP. Thus, the LCPs can be made to comprise additives such as bioactive molecules. Table 10 Additive Amount Outcome P92248PC 40 Curcumin 30 mg / g oil Fd3m Vitamin D 10 ug / ml oil Fd3m Fish oil 5 wt% Fd3m Fish oil 10 wt% Fd3m Iron Sulphate 83 mg / ml oil Fd3m Octyl glucoside 1 % Fd3m 2% Fd3m 5% Fd3m, less defined 1-Methyl-3- 16.67-333.3 μg / ml Fd3m phenylpropylamine oil Benzyl acetone 16.67-333.3 μg / ml Fd3m oil Phenylethylamine 16.67-166.7 μg / ml Fd3m oil Example 11 - Immobilized enzyme – Two-step process LCPs were produced by on a larger scale in a method where the hydrolysis employed immobilized lipase and a rotating bed reactor (RBR, SpinChem RBR S2). Approximately 8 grams of immobilized TL IM (Novozymes) were added to the RBR. In the first step, sunflower oil (see Example 9) was hydrolysed. 120 mL of oil was poured into RBR, and 15 mL of water added, the temperature was set to 60°C, and the bed was allowed to rotate at 600 RPM for 2 hours. After the hydrolysis, TL IM was removed from the reactor. The second step consisted of adding approximately 120g of sunflower lecithin (see Example 9) and leaving the reactor on slow RPMs (around 50) until lecithin was well mixed with the oil. Visual inspection showed no remaining cloudiness, and no visible particles in the mixture, indicating it was well-mixed. Finally, approximately 200 mL of tap water was added to the reactor and the temperature reduced to 40°C, the mixing speed lowered to 50 RPM and the reaction allowed to run for 24 hours. About 300-400g LCP was formed. When analysed with SAXS, the LCP was confirmed to have Fd3m structure. The experiment was repeated with rapeseed oil (see Example 9), with same results. P92248PC 41 Example 12 - Immobilized enzyme LCP is prepared using the general method: 300 mg oat oil containing 40 % polar lipids (PL-40) is mixed with 1.2 ml water. In this case the enzyme is added in immobilized form (the commercial preparation TL IM, Novozymes). After incubation for 16 h at 40 °C with 700 RPM, the enzyme is removed, and the product examined and is found to contain Fd3m phase. In this case the product is completely free of enzyme, which can be desired in certain applications. Furthermore, the removal of enzyme makes sure that the enzymatic hydrolysis does not continue thereafter. Example 13 – LCP formation with hydrolysate mixture Hydrolysate mixture is prepared according to the disclosure. Then water is added so as form LCP. Light microscopy was used to visualize the hydrolysate mixture before addition of water, and 5-20 seconds after addition of water, see Fig 3. As is evident LCP formation is seen within the first 20 seconds after water is added to the hydrolysate mixture. Example 14 – LCP formation with coconut oil Coconut oil (bought in ICA, food grade) was hydrolysed with TL IM immobilized lipase in the rotating bed reactor until approximately 85% of TAG were hydrolysed, as analysed with GC-MS and GC-FID methods. Hydrolysed coconut oil was mixed with different amounts (wt%) of sunflower lecithin (ICA) so as to form hydrolysate mixtures. Then 2 mL of water was added to 300 mg of hydrolysate mixture. Additionally, a reference sample without water was prepared. Then, SAXS measurements were performed after 12 hours of reaction time, so as to determine the structure of the formed LCP (see table 11). Table 11. Polar lipid content (wt%) in the Phase hydrolysate mixture 27 Not clear, could be sponge phase P92248PC 42 33 Not clear, could be sponge phase 38 Fd3m 45 Fd3m 46 Fd3m 49 HII 52 HII 52 (REFERENCE: No water) No LCP structure Based on this, it is evident that a large range of polar lipids may be added, when aiming to form LCP structures. Lower amounts of polar lipids leads to formation of Fd3m phase, whereas higher amounts of polar lipids leads to formation of HII phase. Example 15 – LCP with additives The hydrolysate mixture comprising 52 wt% polar lipids of Example 14 was subjected to further treatment. Different levels of Haemoglobin were dispersed in the hydrolysate mixture. Then, the hydrolysate mixture including dispersed haemoglobin was added to the water, whereafter SAXS analyses was conducted so as to determine the LCP structure: 3 wt% haemoglobin in hydrolysate mixture: HII 11 wt% haemoglobin in hydrolysate mixture: HII 24 wt% haemoglobin in hydrolysate mixture: HII As is evident, the inclusion of from 3 wt% to 24 wt% additive directly to the hydrolysate mixture does not prevent subsequent LCP formation by addition of water. Example 16 – LCP formation with MCT oil MCT (medium chain triacylglycerol) oil (AAK, pharmaceutical grade) was hydrolysed with TL IM immobilized lipase in the rotating bed reactor until approximately 50-75% of TAG were hydrolysed, and around 50-25% were left – analysed with GC-MS and GC-FID methods. P92248PC 43 The hydrolysed MCT oil was mixed with 50 wt% of one of soybean lecithin NatraPhosTM Soya PC100-C Pharma (CRODA, pharmaceutical grade) and egg yolk lecithin NatraPhosTM PL95EN PHARMA (CRODA, pharmaceutical grade), so as to obtain two hydrolysate mixtures. After addition of water to each of the hydrolysate mixtures LCP having the HII structure was formed. Thus, hydrolysate mixtures can successfully be prepared from different polar lipids, and subsequently be turned into LCPs by addition of water.

Claims

P92248PC 44 CLAIMS 1. A method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids, and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Adding an aqueous solvent to the starting mixture, and mixing; and c. Adding at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase, and mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed.

2. The method according to claim 1, wherein step (a) precedes steps (b) and (c), and wherein step (b) and step (c) can proceed in any order or simultaneously.

3. The method according to any one of claims 1 or 2, wherein step (b) precedes step (c).

4. The method according to any one of claims 1 or 2, wherein step (c) precedes step (b); so as to form a hydrolysate mixture after step (c), which after addition of water in step (b) is converted into a stable liquid crystalline phase.

5. The method according to any one of claims 1 or 2, wherein the at least one lipase is comprised within the aqueous solvent.

6. The method according to any one of the preceding claims, wherein said oil and said polar lipids of the starting mixture have been pre-mixed.

7. A method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a1. Contacting an oil with at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase to obtain an oil hydrolysate;P92248PC 45 b1. Optionally removing the at least one lipase from the oil hydrolysate, c1. Mixing the oil hydrolysate from step a1 or b1 with polar lipids, to form a hydrolysate mixture consisting of: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and wherein the oil hydrolysate and polar lipids are well-dispersed in each other; d1. Adding an aqueous solvent to the hydrolysate mixture, and further mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed.

8. The method according to any one of the preceding claims, wherein the volume ratio of aqueous solvent to starting mixture is in the range of 0.01:1 to 10:1, such as from 0.2:1 to 10:1, such as from 0.3:1 to 6:

1.

9. The method according to any one of the preceding claims, wherein the aqueous solvent comprises at least 70 vol% water, such as at least 80 vol% water, such as at least 90 vol% water, such as at least 95 vol% water, such as at least 99 vol% water.

10. The method according to any one of the preceding claims, wherein the aqueous solvent consists of water.

11. The method according to any one of the preceding claims, wherein the starting mixture comprises less than 15 wt% other components.

12. The method of any one of the preceding claims, wherein the liquid crystalline phase formation is allowed to continue for at least 12 hours, such as at least 14 hours, preferably at least 16 hours.

13. The method according to any one of the preceding claims, wherein at least 50 wt% of the lipids from the starting mixture are incorporated in the liquid crystalline phase.P92248PC 46 14. The method according to any one of the preceding claims, wherein at least 50 wt% of the lipids in the liquid crystalline phase are organized in the Fd3m or HII space group.

15. The method according to any one of the preceding claims, wherein the stable liquid crystalline phase maintains its crystalline structure for at least 4 days at 25°C.

16. The method according to any one of the preceding claims, wherein the oil is selected from the group consisting of plant oil, microbial oil, marine oil, animal oil and any combination thereof.

17. The method according to any one of the preceding claims, wherein the polar lipids comprise one or more phospholipids.

18. The method according to any one of the preceding claims, wherein the polar lipids comprise one or one or more glycolipids.

19. The method according to any one of the preceding claims, wherein the polar lipids comprise or consist of one or more lecithins, for example one or more plant lecithins.

20. The method according to any one of the preceding claims, wherein under the conditions of step (c) or step (a1), the main activity of the at least one lipase is hydrolysis of triacylglycerols.

21. The method according to any one of the preceding claims, wherein the at least one lipase comprises or consists of triacylglycerol lipase.

22. The method according to claim 21, wherein the triacylglycerol lipase has sn1,3 positional specificity.

23. The method according to any one of the preceding claims, wherein the at least one lipase is selected from the group consisting of a lipase from R. Miehei, a lipase from R. Arrhizus, and a lipase from Thermomyces languniosus.P92248PC 47 24. The method according to any one of the preceding claims, wherein said at least one lipase comprises or consists of immobilized lipase.

25. The method according to any one of the preceding claims, wherein at least one additive is added, resulting in a liquid crystalline phase comprising said additive.

26. The method according to claim 25, wherein the at least one additive is added to the starting mixture or the hydrolysate mixture if the additive is lipohilic or the at least one additive is added with the aqueous solvent if the additive is water- soluble.

27. The method according to any one of claims 25 or 26, wherein said at least one additive is selected from the group consisting of bioactive molecules, pharmaceutically active ingredients, colourants, fragrances, and flavouring agents.

28. A liquid crystalline phase, obtained or obtainable by the method according to any one of the preceding claims.

29. A liquid crystalline phase, obtained or obtainable by the method according to any of the preceding claims, said liquid crystalline phase comprising in the range from 30 to 65 wt% polar lipids, and having a lipid structure characterized by a small-angle X-ray scattering (SAXS) profile defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44 for the Fd3m space group or 1, ¥3, 2, ¥7, 3, ¥12 for the HII space group.

30. A liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said liquid crystalline phase comprising a phase mixture comprising: i. in the range from 30 to 65 wt% polar lipids; ii. in the range from 35 to 70 wt% oil; and iii. an aqueous solvent; wherein said liquid crystalline phase has a lipid structure characterized by a small- angle X-ray scattering (SAXS) profile defined by peak positions at the q values in the ratios of ¥3, ¥8, ¥11, ¥12, ¥16, ¥19, ¥24, ¥27, ¥32, ¥44 for the Fd3m space group and / or 1, ¥3, 2, ¥7, 3, ¥12 for the HII space group.P92248PC 48 31. The liquid crystalline phase according to any one of claims 28-30, wherein the volume ratio between the aqueous solvent and the phase mixture is at least 0.01:1, such as at least 0.2:

1.

32. The liquid crystalline phase according to any one of claims 28-31, wherein the lipids are organized in the Fd3m space group.

33. The liquid crystalline phase according to any one of claims 28-32, wherein the ratio between triacylglycerol (TAG) and free fatty acids (FFA) is equal to or less than 1:

5.

34. The liquid crystalline phase according claim 33, wherein the ratio between TAG and FFA is equal to or less than 1:10, such as equal to or less than 1:20, such as equal to or less than 1:30, such as equal to or less than 1:40, such as equal to or less than 1:50, such as equal to or less than 1:60, such as equal to or less than 1:70, such as equal to or less than 1:80, such as equal to or less than 1:90, such as equal to or less than 1:95, such as equal to or less than 1:

99.

35. The liquid crystalline phase according to any one of claims 28-34, wherein the liquid crystalline phase comprises substantially no TAG.

36. The liquid crystalline phase according to any one of claims 28-35, wherein the liquid crystalline phase comprises less than 15 wt% other components.

37. The liquid crystalline phase according to any one of claims 28-36, further comprising one or more additives.

38. The liquid crystalline phase according to claim 37, wherein said one or more additives are selected from the group consisting of bioactive molecules, pharmaceutically active ingredients, colourants, fragrances, and flavouring agents.

39. A method of forming a hydrolysate mixture comprising the following steps: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids, and ii. in the range from 35 to 70 wt% oil;P92248PC 49 wherein the oil and polar lipids are well-dispersed in each other; b. Adding at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase, and mixing.

40. A method of forming a hydrolysate mixture comprising the steps of: a1. Contacting an oil with at least one lipase under conditions that allow lipase activity, such that at least 1 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase to obtain an oil hydrolysate; b1. Optionally removing the at least one lipase from the oil hydrolysate, c1. Mixing the oil hydrolysate from step a1 or b1 with polar lipids, to form a hydrolysate mixture comprising: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and wherein the oil hydrolysate and polar lipids are well-dispersed in each other.

41. A hydrolysate mixture as formed in any one of claims 4 and 39-40.

42. A hydrolysate mixture comprising: i. in the range from 30- 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil hydrolysate; and iii. less than 15 wt% other components.

43. A method of forming a liquid crystalline phase comprising the steps of: i. providing a hydrolysate mixture; ii. adding aqueous solvent to the hydrolysate mixture and mixing; whereby a stable lipid-based liquid crystalline phase is formed; 44. The method of claim 43, wherein the aqueous solvent is added in a ratio in the range of at least 0.01:1, such as at least 0.2:1, relative to the volume of the hydrolysate mixture.P92248PC 50 45. A liquid crystalline phase, obtained or obtainable by the method according to claims 43-44.

46. Use of a liquid crystalline phase according to any one of claims 28-38 in the preparation of a food, a feed, a cosmetic, or a nutraceutical.

47. The liquid crystalline phase according to claims 28-38 for use as a medicament.

48. A product selected from the group consisting of a food, a feed, a cosmetic, and a nutraceutical, wherein said product comprises a liquid crystalline phase according to any one of claims 28-38.

49. Use of a triacylglycerol lipase having sn1, 3 position specificity for producing the liquid crystalline phase according to any of claims 28-36.

50. The use according to claim 49, wherein the use is in a method according to any one of claims 1-27.

51. A method for producing a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group, said method comprising the steps of: a. Providing a starting mixture comprising: i. in the range from 30 to 65 wt% polar lipids and ii. in the range from 35 to 70 wt% oil; wherein the oil and polar lipids are well-dispersed in each other; b. Adding water to reach a ratio in the range of at least 0.2:1 relative to the volume of the starting mixture, and mixing; c. Contacting the mixture of step (b) with at least one triacylglycerol lipase having sn 1, 3 specificity, under conditions that allow lipase activity, such that at least 50 wt% of the triacylglycerol in the oil is hydrolysed by said at least one lipase; and d. Subjecting the mixture of step (c) to further mixing; whereby a stable liquid crystalline phase comprising lipids organized in Fd3m and / or HII space group is formed.

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