Mineral delivery formation

Abstract

This invention relates to a mineral delivery formulation comprising filaments, said filaments comprising protein, mineral and biopolymer. The invention also relates to methods for producing such formulations. The method also contemplates uses of the formulations.

Description

Mineral Delivery Formulation

[0001] This invention relates to a mineral delivery formulation comprising filaments, said filaments comprising protein, mineral and biopolymer. The invention also relates to methods for producing such formulations. The method also contemplates uses of the formulations.BACKGROUND

[0002] A balanced diet typically provides all the essential minerals required for normal bodily function in humans. However, certain individuals may require additional mineral supplementation, for example, due to the presence of intolerances or mineral deficiencies, to support a healthy lifestyle, to enhance the nutritional quality of diets for targeted populations with specific dietary needs, or for aligning with dietary preferences and trends, such as the growing transition from animal- to plant-based diets. For example, individuals having a dairy intolerance are likely to have a reduced intake of milk-based foods, potentially leading to a calcium deficiency. Alternatively, individuals who undertake vigorous physical exercise may benefit from additional mineral supplementation, for example, due to the loss of electrolytes via perspiration. Additionally, in the case of individuals who only consume plant-based proteins, supplementation is required due to most plant proteins not always naturally containing essential minerals such as calcium or iron.

[0003] Mineral deficiencies may include deficiencies of calcium, magnesium, iron, zinc, potassium, sodium, iodine, selenium, or any combination thereof. Further, individuals with such mineral deficiencies may suffer from one or more diseases or conditions selected from iron deficiency anaemia, osteoporosis, osteopenia, osteomalacia, hypocalcaemia, rickets, impaired immune function, muscle weakness, hypokalaemia, hypothyroidism, goitre, and cognitive impairments.

[0004] Supplementation with minerals has emerged as a viable solution to the above nutritional challenges (Farsi et al., 2022; Okeudo-Cogan et al., 2024; McClements and McClements, 2023). Calcium, an abundant mineral in the mammalian body, plays an important role in many fundamental physiological processes. The significance of calcium extends not only to individuals with specialised or plant-based diets, but to growing children who require sufficient calcium to build strong bones, and to those experiencing bone mass loss (osteoporosis) as part of the natural aging process, such as post-menopausal women and the elderly (Calvo and Whiting, 2022; Pointke and Pawelzik, 2022; Smolin and Grosvenor, 2019).

[0005] A number of different forms of mineral delivery systems have been described to attempt to circumvent the problems inherent with mineral delivery. Many of these aredesigned to be gellable proteins and / or polysaccharides associated with a mineral in a microgel particulate form.

[0006] One example is found in CN111529495, which relates to a high-gastric-stable protein emulsion microgel particles comprising pea protein isolate (PPI) functionalised with inulin by a Maillard reaction. The formulation contains calcium chloride to induce the emulsion to form a gel. Another example is described in EP2292102, which relates to a stable microcapsule comprising a protein-polysaccharide shell matrix and a core, said proteins including soy and pea, and a core comprising an emulsion of an aqueous dispersion or solution of at least one mineral salt in an edible oil. However, the formulations of CN111529495 and EP2292102 involve the use of oil, which raises nutritional concerns.

[0007] Other mineral delivery systems comprising the use of plant-based proteins have been described. For example, EP2731450 describes a food-grade encapsulate comprising gelled protein aggregates embedded in a continuous polysaccharide phase, with said gelled protein aggregates containing a mineral as a food-grade mineral or organic salts, or ions. EP2731462 relates to a food-grade microparticle encapsulate comprising gelled proteins, a colour component and metal ions, and optionally polysaccharide. WO2021 / 089972 relates to a gelled protein and / or gellable polysaccharide containing calcium, zinc, magnesium, iron, or transition metal cations. The primary focus of these documents is mineral delivery, but the texture and mouthfeel properties are equally important considerations in the production of orally administered mineral supplements, yet are not reported in these documents.

[0008] Filaments have emerged as potential systems for mineral delivery (Loveday et al., 2010; Loveday et al., 2011). In these approaches, it has been observed that the efficiency of filament formation was substantially enhanced by varying pH and NaCI or CaCl2 concentrations. Nevertheless, filament systems incorporating CaCl2 have shown worm-like structures with a size between 63 and 84 nm in length, giving rise to environmental concerns and animal / human health concerns as they are classified as nanomaterials, the use of which may induce toxic effects if incorporated in orally ingested systems.

[0009] Thus, there remains a need in the art to provide a food-grade, human safe mineral delivery system with optimized mouthfeel and bio-lubrication properties.

[0010] CN 113368130 discloses fibrous iron-carrying composites.

[0011] References:Calvo, M.S., and Whiting, S.J. (2022). “Perspective: School Meal Programs Require Higher Vitamin D Fortification Levels in Milk Products and Plant-Based Alternatives — Evidence from the National Health and Nutrition Examination Surveys (NHANES 2001-2018)”, Advances in Nutrition, 13 (5), pp. 1440-1449. https: / / doi . orq / 10.1093 adyances / nmacp68.Farsi, D. N., Uthumange, D., Munoz Munoz, J., & Commane, D. M. (2022). “The nutritional impact of replacing dietary meat with meat alternatives in the UK: A modelling analysis using nationally representative data”. British Journal of Nutrition, 127(11), pp. 1731-1741.Loveday, S. M.; Wang, X. L.; Rao, M. A.; Anema, S. G.; Creamer, L.; Singh, H., (2010). “Tuning the properties of p-lactoglobulin nanofibrils with pH, NaCI and CaCh”, International Dairy Journal, 20, pp. 571-579. hko ;. / / 4 r10 10 S / Hda:ry:.z0 Q 02 014.Loveday, S.M., Su, J., Rao, M.A., Anema, S.G., Singh, H. (2011). “ Effect of Calcium on the Morphology and Functionality of Whey Protein Nanofibrils’’, Biomacromolecules, 12(10), pp. 3780 - 3788. htps: / / doi.org / 10.1021 / bm201013b.McClements, I.F., McClements, D.J. (2023). Designing healthier plant-based foods: Fortification, digestion, and bioavailability, Food Research International, 169, 112853.Okeudo-Cogan, M.C., Yang, S., Murray, B.S., Ettelaie, R., Connell, S.D., Radford, S., Micklethwaite, S., Benitez-Alfonso, S., Yeshvekar, R., Sarkar, A. (2024). “Multivalent cations modulating microstructure and interactions of potato protein and fungal hyphae in a functional meat analogue”, Food Hydrocolloids, 149, 109569.Pointke, Marcel, and Elke Pawelzik. 2022. "Plant-Based Alternative Products: Are They Healthy Alternatives? Micro- and Macronutrients and Nutritional Scoring", Nutrients, 14 (3), 601 ■ htps: / / doi.org / 10.3390 / nu14030601.Smolin, L. A., and Grosvenor, M. B. (2019). Nutrition: Science and applications (4thed.). Wiley.BRIEF SUMMARY OF THE DISCLOSURE

[0012] The present invention relates to formulations useful in the delivery of minerals.

[0013] The formulations of the present invention are capable of being delivered without the need for additional fluids (e.g. water to assist swallowing a tablet). The formulations of the present invention also have good sensorial mouthfeel performance and bio-lubrication properties. For example, the formulations may have a low friction coefficient and / or a low viscosity. Additionally, the formulations may achieve the above advantages in the absence of oil.

[0014] Additionally, formulations of the present invention provide higher mineral concentrations compared to some natural and fortified food products without adverse effects on mouthfeel.

[0015] In a first aspect of the present invention, there is provided a formulation comprising one or more filaments, the one or more filaments comprising: a protein in an amount of from about 0.1% to less than about 5% by weight; a mineral in a concentration of from about 1 mM to less than about 150 mM; and a biopolymer in an amount of more than about 0.01 % to about 5% by weight; wherein the formulation has a pH of less than about 7.0.

[0016] In a second aspect of the present invention, there is provided a formulation comprising one or more filaments, the one or more filaments comprising: a protein in an amount of from about 0.1% to less than about 5% by weight; a mineral in a concentration of from about 1 mM to less than about 150 mM; and a biopolymer in an amount of more than about 0.01 % to about 5% by weight; wherein the weight ratio of biopolymer to protein in the formulation is in the range of from about x: 1 to about y: 1 , wherein x > 0.01 and y < 1 ; wherein the formulation has a pH of less than about 7.0; wherein the protein is ionically cross-linked by the mineral and / or the protein is crosslinked by the mineral via ion-dipole interactions.

[0017] In a third aspect of the present invention, there is provided a method of producing the formulation of the first aspect or second aspect, the method comprising: providing a solution comprising the protein and the mineral, the solution having a pH of less than about 7.0; heating the solution to form a protein-mineral intermediate; and combining the protein-mineral intermediate with the biopolymer to form the formulation of the first aspect or second aspect.

[0018] In a fourth aspect of the present invention, there is provided a formulation obtainable or obtained by the method of the third aspect.

[0019] In a fifth aspect of the present invention, there is provided a formulation of the first aspect or second aspect for use as a medicament.

[0020] In a sixth aspect of the present invention, there is provided a formulation of the first aspect or second aspect for use in treating a disease or condition caused by or associated with a mineral deficiency in a patient.

[0021] In a seventh aspect of the present invention, there is provided a use of a formulation of the first aspect or second aspect as a mineral supplement, wherein the use is not a method of treatment of the human or animal body.

[0022] In an eighth aspect of the present invention, there is provided a method for the treatment of a disease or condition caused by or associated with a mineral deficiency in a patient, the method comprising administering a formulation of the first aspect or second aspect to a patient in need thereof.BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Embodiments of the invention are further described hereinafter with reference to the accompanying drawings, in which:Figure 1 shows (A & B) light microscopy and (C & D) cryo-SEM images of exemplary potato protein filaments comprising xanthan gum and Ca2+ions manufactured at pH 2.0. (E) A confocal image depicting the distribution of fluorescent potato protein filaments. (F & G) EDX analysis depicting the distribution of Ca2+ions (fluorescing) within the plant protein-xanthan filaments.Figure 2 shows the results of friction coefficient as a function of entrainment speed of: (x) CaCh at a 100 mM concentration (Control 1), (+) potato protein filament(Po) + CaCh (100 mM) (Control 2), (o) xanthan gum + CaCh (100 mM) (Control 3), and (A) potato protein filaments + CaCh (100 mM) + xanthan gum. When present, the potato protein, xanthan gum and calcium are present in an amount of 1 wt%, 0.5 wt% and 100 mM respectively.Figure 3 shows the effect of 1 month storage time on the lubrication properties of the potato protein-xanthan gum-Ca filaments (Po + CaCh 100 mM + XG). The potato protein, xanthan gum and calcium are present in an amount of 1 wt%, 0.5 wt% and 100 mM respectively.Figure 4 shows the friction coefficient as a function of entrainment speed when varying CaCh concentration in the potato protein-xanthan gum-Ca filaments (Po + CaCh + XG). The potato protein and xanthan gum are present in an amount of 1 wt% and 0.5 wt%.Figure 5 shows the effect of protein type on the lubrication performance of the protein-xanthan gum-Ca filaments. The formulations include a protein concentration of 1wt%, CaCh concentration of 100 mM, and xanthan gum concentration of 0.5 wt%.Figure 6 shows (A) the effect of xanthan gum concentration (0.01 - 1wt%) on the viscosity of the potato protein-xanthan gum-Ca filaments, specifically with respect to the apparent viscosity and shear rate; and (B) the effect of xanthan gum concentration (0.01 - 1wt%) on the lubrication performance of the potato protein-xanthan gum-Ca filaments,specifically with respect to the friction coefficient and entrainment speed. Formulations include a potato protein concentration of 1wt% and CaCl2 concentration of 100 mM.Figure 7 shows (A) the effect of potato protein concentration (1 - 4wt%) on the lubrication properties of potato protein-xanthan gum-Ca filaments, specifically with respect to the friction coefficient and entrainment speed; and (B) the effect of potato protein concentration (1 - 4 wt%) on the viscosity of potato protein-xanthan gum-Ca filaments, specifically with respect to the apparent viscosity and shear rate. Formulations include a CaCh concentration of 100 mM, and xanthan gum concentration of 0.5wt %.Figure 8 shows the effect of pH (pH 2 - pH 9) on the lubrication performance of potato protein-xanthan gum-Ca filaments. (A) The effect of pH on the friction coefficient and entrainment speeds. (B) SEM images showing the structures of the potato protein-xanthan gum-Ca filaments at different pH. Formulations include a potato protein concentration of 1wt%, a CaCl2 concentration of 100 mM, and a xanthan gum concentration of 0.5 wt%.Figure 9 shows the effect of pH (pH 2 - pH 9) on the lubrication performance of lupin protein-xanthan gum-Ca filaments (Lu + CaCh + XG). (A) The effect of pH on the friction coefficient and entrainment speed. (B) SEM images showing the structures of the lupin protein-xanthan gum-Ca filaments at different pH. Formulations include a lupin protein concentration of 1wt%, a CaCl2 concentration of 100 mM, and a xanthan gum concentration of 0.5 wt%.Figure 10 shows the effect of mineral type on the lubrication performance of potato protein-xanthan gum filaments (Po + XG). Formulations include a potato protein concentration of 1wt%, a calcium chloride (CaCl2), magnesium citrate (CeHeMgO?), zinc citrate (C^H O Zns), or iron sulfate (FeSOs) concentration of 100 mM, and a xanthan gum concentration of 0.5wt%.DETAILED DESCRIPTION

[0024] The abbreviations used herein have their conventional meaning within the chemical and biological arts.

[0025] Throughout the description and claims of this specification, the words “comprise” and “contain” and variations of them mean “including but not limited to”, and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

[0026] Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and / or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive. The invention is not restricted to the details of any foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

[0027] The reader's attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.

[0028] For the avoidance of doubt, it is hereby stated that the information disclosed earlier in this specification under the heading “Background” is relevant to the invention and is to be read as part of the disclosure of the invention.Definitions

[0029] The term ‘biopolymer’ may refer to a biopolymeric hydrogel.

[0030] The term ‘protein’ includes the natural extract of the protein derived from plant or animal source and also to proteins that have been heat treated that results in partial unfolding I partial denaturation of the protein derived from plant or animal source.

[0031] The term ‘non-covalent interaction’ may refer to an ionic bond, a dipole-dipole interaction, an ion-dipole interaction, a Van der Waals interaction, a hydrogen bond, or a hydrophobic interaction.

[0032] The term ‘essential minerals for human biochemical processes’ may refer to any inorganic component that has an active role in a biochemical process within the human body, for example, in relation to growth, development, cognition, alertness, movement, energy production, utilization and / or maintenance of internal homeostasis.

[0033] The term ‘cross-linking’ may refer to the bonding of one protein component to another protein component via covalent and / or non-covalent interactions. The cross-linking may occur via a direct covalent and / or non-covalent interaction between protein components, or via non-covalent interactions of the protein components with the mineral component.

[0034] Without wishing to be bound by theory, the cross-linking of the protein components via the mineral component results in ‘nucleation’ of the protein components wherein a plurality of protein components are cross-linked to a single mineral component and / or wherein a plurality of mineral components are cross-linked to a single protein component. This nucleation results in a protein-mineral intermediate filament. The mineral acts as a ‘nucleator’ for the protein components.

[0035] The term ‘protein-mineral intermediate filament’ refers a single, thread-like structure comprising the protein and mineral components.

[0036] The term ‘filament’ in the context of the invention refers to a single, thread-like, flexible, cylindrical-shaped structure comprising the protein, mineral and biopolymer components.Formulation

[0037] The protein may be cross-linked by the mineral. For example, the protein may be ionically cross-linked by the mineral. Alternatively or additionally, the protein may be crosslinked by the mineral via ion-dipole interactions.

[0038] The protein may be associated with the biopolymer via covalent and / or non-covalent interactions. For example, the protein may be associated with the biopolymer via non- covalent interactions selected from one or more of: hydrogen bonding, hydrophobic interactions, dipole-dipole interactions, ion-dipole interactions, and ionic bonds. Alternatively or additionally, the protein may be associated with the biopolymer via covalent interactions, such as a covalent bond formed from a Maillard reaction (a covalent conjugation between amino groups of protein and carbonyl groups of reducing sugars in the biopolymers).

[0039] The protein may be associated with the biopolymer via non-covalent interactions. For example, the protein may be associated with the biopolymer via non-covalent interactions selected from one or more of: hydrogen bonding, hydrophobic interactions, Van der Waals interactions, dipole-dipole interactions, ion-dipole interactions, and ionic bonds. The protein may be associated with the biopolymer via non-covalent interactions selected from one or more of: hydrogen bonding, hydrophobic interactions, dipole-dipole interactions, ion-dipole interactions, and ionic bonds. The protein may be associated with the biopolymer via a combination of electrostatic interactions (e.g. dipole-dipole interactions, ion-dipole interactions, and / or ionic bonds) and hydrogen bonding.

[0040] Without wishing to be bound by theory, it is thought that the mineral assists in the nucleation of the protein components at acidic pH to form protein-mineral intermediate filaments, and the biopolymer acts as a ‘glue’ connecting adjacent protein-mineralintermediate filaments to one another, thereby facilitating the formation of a thread-like filament structure.

[0041] Given the above described interactions between the protein and the mineral, it may be that the biopolymer, which is introduced after the formation of the protein-mineral intermediate, is not associated with the mineral. Any interaction between the biopolymer and the mineral may be negligible relative to the above-described interactions between the protein and the mineral, and the biopolymer and the protein.

[0042] Alternatively, the biopolymer may be associated with the mineral via a non-covalent interaction. For example, the biopolymer may be associated with the mineral by a non- covalent interaction selected from the group consisting of a dipole-dipole interaction, an iondipole interaction, a Van der Waals interaction, a hydrogen bond, and / or a hydrophobic interaction. The biopolymer may be associated with the mineral via ion-dipole interactions.

[0043] Without wishing to be bound by theory, it is thought that the minerals are embedded in the protein uniformly or non-uniformly, mainly via ionic crosslinking with acidic amino acid groups and / or basic amino acid groups that are present on the surface of the protein. It is thought that these amino acid groups are available to interact with the minerals due to the unfolding of the protein via heat treatment. The heat treatment of the protein described herein results in partial unfolding of the protein rather than complete unfolding (and therefore complete denaturation) of the protein.

[0044] The formulation may be an oil-free formulation.

[0045] The weight ratio of biopolymer to protein in the formulation is in the range of from about x: 1 to about y: 1 , wherein x > 0.01 and y < 1.

[0046] The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.15: 1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.2:1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.25: 1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.3:1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.35: 1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.4:1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.45: 1 to about 1 :1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.5:1 to about 1 :1.

[0047] The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05: 1 to about 0.95: 1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.9:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.85:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05: 1 to about 0.8:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.75:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.7:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.65:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05: 1 to about 0.6: 1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.55:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.05:1 to about 0.5:1.

[0048] The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.9:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.85:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.8:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.75: 1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 : 1 to about 0.7:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.65:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.6: 1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.55: 1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.1 :1 to about 0.5:1.

[0049] The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.2:1 to about 0.8:1. The weight ratio of biopolymer to protein in the formulation may be in the range of from about 0.4: 1 to about 0.6: 1.

[0050] Preferably, the weight ratio of biopolymer to protein in the formulation is about 0.5:1 .

[0051] The protein may be selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein, duckweed protein, lactoferrin, lysozyme, gelatin, milk protein, bovine serum albumin, whey protein, casein, caseinate, egg protein, albumin, gluten, gelatin Type B, rice protein, soy protein, wheat protein, millet protein, legume, legumin, gluten, corn protein, zein, almond protein, peanut protein, pistachio protein, cashew nut protein, walnut protein, hazelnut protein, pine nut protein, coconut protein, insect protein, algal protein, microbial protein, spirulina, fungalprotein (mycoprotein), single cell protein, leaf protein, mung bean protein, red kidney bean protein, tomato seed protein, pumpkin seed protein, oat protein, flax seed protein, chickpea protein, hemp protein, rapeseed protein, quinoa protein, amaranth protein, barley protein, rye protein, sorghum protein and sesame protein in the form of isolates, concentrates or hydrolysates.

[0052] The protein may be selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein, lactoferrin, lysozyme, gelatin, milk protein, bovine serum albumin, whey protein, casein, caseinate, egg protein, albumin, gluten, gelatin Type B, rice protein, soy protein, wheat protein, millet protein, legume, legumin, gluten, corn protein, zein, almond protein, peanut protein, pistachio protein, cashew nut protein, walnut protein, hazelnut protein, pine nut protein, coconut protein, insect protein, algal protein, microbial protein, spirulina, fungal protein (mycoprotein), single cell protein, leaf protein, mung bean protein, red kidney bean protein, tomato seed protein, pumpkin seed protein, oat protein, flax seed protein, chickpea protein, hemp protein, rapeseed protein, quinoa protein, amaranth protein, barley protein, rye protein, sorghum protein and sesame protein in the form of isolates, concentrates or hydrolysates.

[0053] The protein may be selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein and duckweed protein.

[0054] The protein may be selected from the group comprising: potato protein, lupin protein, oat protein, sunflower protein, fava bean protein, and almond protein. The protein may be selected from potato protein and lupin protein. Preferably, the protein is potato protein.

[0055] The protein may be present in an amount in the range of from about 0.1% to about 4.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1% to about 4% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1% to about 3.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1% to about 3% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1% to about 2.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1% to about 2% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1 % to about 1.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.1% to about 1% by weight of the formulation.

[0056] The protein may be present in an amount in the range of from about 0.5% to about 4.5% by weight of the formulation. The protein may be present in an amount in the range offrom about 0.5% to about 4% by weight of the formulation. The protein may be present in an amount in the range of from about 0.5% to about 3.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.5% to about 3% by weight of the formulation. The protein may be present in an amount in the range of from about 0.5% to about 2.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.5% to about 2% by weight of the formulation. The protein may be present in an amount in the range of from about 0.5% to about 1.5% by weight of the formulation. The protein may be present in an amount in the range of from about 0.5% to about 1% by weight of the formulation.

[0057] The protein may be present in an amount in the range of from about 1 % to about 4.5% by weight of the formulation. The protein may be present in an amount in the range of from about 1 % to about 4% by weight of the formulation. The protein may be present in an amount in the range of from about 1% to about 3.5% by weight of the formulation. The protein may be present in an amount in the range of from about 1% to about 3% by weight of the formulation. The protein may be present in an amount in the range of from about 1% to about 2.5% by weight of the formulation. The protein may be present in an amount in the range of from about 1% to about 2% by weight of the formulation. The protein may be present in an amount in the range of from about 1 % to about 1.5% by weight of the formulation.

[0058] Preferably the protein is present in an amount of about 1% by weight of the formulation.

[0059] The mineral may comprise one or more essential minerals for human biochemical processes. The mineral may be selected from the group comprising calcium, magnesium, iron, zinc, potassium, sodium, iodine, selenium, or any combination thereof. The mineral may be selected from the group comprising calcium, magnesium, iron, zinc, potassium, sodium, or any combination thereof. The mineral may be selected from calcium, magnesium, zinc and iron. The mineral may be selected from calcium, magnesium, and zinc. The mineral may be selected from calcium and iron. The mineral may be calcium.

[0060] The mineral may be present in a concentration in the range of from about 1 mM to about 140 mM. The mineral may be present in a concentration in the range of from about 1 mM to about 130 mM. The mineral may be present in a concentration in the range of from about 1 mM to about 120 mM. The mineral may be present in a concentration in the range of from about 1 mM to about 110 mM. The mineral may be present in a concentration in the range of from about 1 mM to about 100 mM.

[0061] The mineral may be present in a concentration in the range of from about 10 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 20 mM to less than about 150 mM. The mineral may be present in a concentration inthe range of from about 30 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 40 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 50 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 60 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 70 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 80 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 90 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 100 mM to less than about 150 mM.

[0062] The mineral may be present in a concentration in the range of from about 40 mM to less than about 150 mM. The mineral may be present in a concentration in the range of from about 60 mM to about 140 mM. The mineral may be present in a concentration in the range of from about 75 mM to about 125 mM. The mineral may be present in a concentration in the range of from about 90 mM to about 110 mM. Preferably, the mineral is present in a concentration of about 100 mM.

[0063] The biopolymer may be a polysaccharide. The biopolymer may be selected from the group comprising: xanthan gum, K-carrageenan, i-carrageenan, A-carrageenan, agar, starch, maltodextrin, agarose, alginate, pectin, dextran sulphate, cellulose and modified cellulose (ethyl cellulose, microcrystalline cellulose, carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose), locust bean gum, gum Arabic, gum ghatti, konjac gum (glucomannan), Karaya gum, Cassia gum, gellan gum, guar gum, modified starches, chitosan, inulin, beta-glucans, gum tragacanth, and any negatively-charged polysaccharide. Preferably, the biopolymer is xanthan gum.

[0064] The biopolymer may be present in an amount in the range of more than about 0.01 % to about 5% by weight.

[0065] The biopolymer may be present in an amount in the range of more than about 0.01 % to about 4.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.01 % to about 4.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 4.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 4.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 4% by weight.

[0066] The biopolymer may be present in an amount in the range of more than about 0.01 % to about 3.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 3.6% by weight. The biopolymer may be present in an amount inthe range of more than about 0.01% to about 3.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 3.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 3% by weight.

[0067] The biopolymer may be present in an amount in the range of more than about 0.01 % to about 2.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.01 % to about 2.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 2.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 2.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 2% by weight.

[0068] The biopolymer may be present in an amount in the range of more than about 0.01 % to about 1.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 1.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 1.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 1.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.01% to about 1 % by weight.

[0069] The biopolymer may be present in an amount in the range of from more than about 0.01 % to about 0.95% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01 % to about 0.9% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01% to about 0.85% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01% to about 0.8% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01 % to about 0.75% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01% to about 0.7% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01% to about 0.65% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01 % to about 0.6% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01% to about 0.55% by weight. The biopolymer may be present in an amount in the range of from more than about 0.01% to about 0.5% by weight.

[0070] The biopolymer may be present in an amount in the range of more than about 0.1 % to about 5% by weight.

[0071] The biopolymer may be present in an amount in the range of more than about 0.1 % to about 4.8% by weight. The biopolymer may be present in an amount in the range of morethan about 0.1% to about 4.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 4.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 4.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 4% by weight.

[0072] The biopolymer may be present in an amount in the range of more than about 0.1 % to about 3.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 3.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 3.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 3.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 3% by weight.

[0073] The biopolymer may be present in an amount in the range of more than about 0.1 % to about 2.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 2.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 2.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 2.2% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 2% by weight.

[0074] The biopolymer may be present in an amount in the range of more than about 0.1 % to about 1.8% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 1.6% by weight. The biopolymer may be present in an amount in the range of more than about 0.1 % to about 1.4% by weight. The biopolymer may be present in an amount in the range of more than about 0.1% to about 1.2% by weight.

[0075] The biopolymer may be present in an amount in the range of from about 0.1% to about 1 % by weight. The biopolymer may be present in an amount in the range of from about 0.15% to about 1% by weight. The biopolymer may be present in an amount in the range of from about 0.2% to about 1% by weight. The biopolymer may be present in an amount in the range of from about 0.25% to about 1% by weight. The biopolymer may be present in an amount in the range of from about 0.3% to about 1% by weight. The biopolymer may be present in an amount in the range of from about 0.35% to about 1% by weight. The biopolymer may be present in an amount in the range of from about 0.4% to about 1% by weight. The biopolymer may be present in an amount in the range of from about 0.45% to about 1 % by weight. The biopolymer may be present in an amount in the range of from about 0.5% to about 1 % by weight.

[0076] The biopolymer may be present in an amount in the range of from about 0.1% to about 0.9% by weight. The biopolymer may be present in an amount in the range of from about 0.2% to about 0.8% by weight. The biopolymer may be present in an amount in therange of from about 0.4% to about 0.6% by weight. Preferably, the biopolymer may be present in an amount of about 0.5% by weight.

[0077] In some embodiments, the formulation may further comprise water.

[0078] The water may be present in an amount of at least about 90.0 % by weight. The water may be present in an amount of at least about 92.5 % by weight. The water may be present in an amount of at least about 95.0 % by weight. The water may be present in an amount of at least about 96.0 % by weight. The water may be present in an amount of at least about 97.0 % by weight. The water may be present in an amount of at least about 98.0 % by weight.

[0079] The water may be present in an amount in the range from about 90.0 % to about 99.0 % by weight. The water may be present in an amount in the range from about 92.5 % to about 99.0 % by weight. The water may be present in an amount in the range from about 95.0 % to about 99.0 % by weight. The water may be present in an amount in the range from about 96.0 % to about 99.0 % by weight. The water may be present in an amount in the range from about 97.0 % to about 99.0 % by weight. The water may be present in an amount in the range from about 96.0 % to about 98.0 % by weight. The water may be present in an amount in the range from about 96.0 % to about 97.0 % by weight.

[0080] The formulation may have a pH in of less than about 6.5. The formulation may have a pH in of less than about 6.0. The formulation may have a pH in of less than about 5.5. The formulation may have a pH in of less than about 5.0. The formulation may have a pH in of less than about 4.5.

[0081] The formulation may have a pH in the range of from about 2.0 to about 4.5. The formulation may have a pH in the range of from about 2.0 to about 4.0. The formulation may have a pH in the range of from about 2.0 to about 3.5. The formulation may have a pH in the range of from about 2.0 to about 3.0. The formulation may have a pH in the range of from about 3.0 to about 4.0.

[0082] Each of the one or more filaments may be at least about 1 pm in length. Each of the one or more filaments may be no more than about 100 mm in length.

[0083] Each of the one or more filaments may have a length in the range of from about 1 pm to about 100 mm. Each of the one or more filaments may have a length in the range of from about 50 pm to about 100 mm. Each of the one or more filaments may have a length in the range of from about 100 pm to about 100 mm. Each of the one or more filaments may have a length in the range of from about 500 pm to about 100 mm. Each of the one or more filaments may have a length in the range of from about 1 mm to about 100 mm.

[0084] Each of the one or more filaments may have a length in the range of from about 1 pm to about 50 mm. Each of the one or more filaments may have a length in the range of from about 1 pm to about 25 mm. Each of the one or more filaments may have a length in the range of from about 1 pm to about 10 mm. Each of the one or more filaments may have a length in the range of from about 1 pm to about 1 mm.

[0085] Each of the one or more filaments may be at least about 100 nm in diameter.

[0086] The one or more filaments may agglomerate to form a particle which comprises two or more filaments associated with one another. For example the two or more filaments may be associated with one another via non-covalent interactions selected from one or more of: hydrogen bonding, hydrophobic interactions, ion-dipole interactions, and ionic bonds.

[0087] The particles may have a particle size of at least about 300 pm in diameter. The particles may have a particle size of at least about 400 pm in diameter. The particles may have a particle size of at least about 500 pm in diameter. The particles may have a particle size of at least about 1 mm in diameter.

[0088] The particle size may be no more than 100 mm in diameter. The particle size may be no more than 50 mm in diameter. The particle size may be no more than 25 mm in diameter. The particle size may be no more than 10 mm in diameter. The particle size may be no more than 1 mm in diameter.

[0089] The particle size may be in the range of from about 300 pm to about 100 mm in diameter. The particle size may be in the range of from about 300 pm to about 10 mm in diameter. The particle size may be in the range of from about 300 pm to about 1 mm in diameter.

[0090] In a preferred embodiment, the protein is potato protein and the biopolymer is xanthan gum. In a further preferred embodiment, the protein is potato protein, the biopolymer is xanthan gum and the mineral is calcium.Method of product no the formulation

[0091] The solution comprising the protein and the mineral may comprise water. Thus, the solution may be prepared by adding the protein and mineral to water.

[0092] The step of providing the solution comprising the protein and the mineral may involve adding the protein and mineral to the solution (e.g. water) in quantities to achieve a weight percentage of protein in the formulation and a concentration of mineral in the formulation as defined in any of the embodiments relating to the first aspect and the second aspect.

[0093] The step of providing the solution comprising the protein and the mineral may further comprise stirring the solution of the protein and mineral until complete solubilisation occurs.The stirring may be performed for at least about 5 minutes, at least about 10 minutes, at least about 20 minutes, at least about 30 minutes, at least about 40 minutes, at least about 50 minutes, at least about 1 hour, at least about 1 .5 hours, at least about 2 hours, or at least about 2.5 hours.

[0094] Preferably, the stirring is performed for about 2 hours.

[0095] The step of providing the solution comprising the protein and the mineral may further comprise adjusting the pH to provide the solution having a pH of less than about 7.0. For example, the adjusting the pH may comprise adding an acid to the solution comprising the protein and the mineral. The acid may be selected from: hydrochloric acid, sulfuric acid, citric acid, tartaric acid, acetic, malic acid, fumaric acid, vinegar, phosphoric acid, or their respective deprotonated species, i.e. sulphate, citrate, tartarate, acetate, malate, fumarate, chloride, and phosphate. Preferably, the acid is hydrochloric acid (e.g. 6M hydrochloric acid).

[0096] The solution comprising the protein and the mineral may have a pH of less than about 6.5. The solution comprising the protein and the mineral may have a pH of less than about 6.0. The solution comprising the protein and the mineral may have a pH of less than about 5.5. The solution comprising the protein and the mineral may have a pH of less than about 5.0. The solution comprising the protein and the mineral may have a pH of less than about 4.5.

[0097] The solution comprising the protein and the mineral may have a pH in the range of from about 2.0 to about 4.5. The solution comprising the protein and the mineral may have a pH in the range of from about 2.0 to about 4.0. The solution comprising the protein and the mineral may have a pH in the range of from about 2.0 to about 3.5. The solution comprising the protein and the mineral may have a pH in the range of from about 2.0 to about 3.0. The solution comprising the protein and the mineral may have a pH in the range of from about 3.0 to about 4.0.

[0098] Preferably, the solution comprising the protein and the mineral has a pH of about 2.0.

[0099] The protein may be selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein, duckweed protein, lactoferrin, lysozyme, gelatin, milk protein, bovine serum albumin, whey protein, casein, caseinate, egg protein, albumin, gluten, gelatin Type B, rice protein, soy protein, wheat protein, millet protein, legume, legumin, gluten, corn protein, zein, almond protein, peanut protein, pistachio protein, cashew nut protein, walnut protein, hazelnut protein, pine nut protein, coconut protein, insect protein, algal protein, microbial protein, spirulina, fungalprotein (mycoprotein), single cell protein, leaf protein, mung bean protein, red kidney bean protein, tomato seed protein, pumpkin seed protein, oat protein, flax seed protein, chickpea protein, hemp protein, rapeseed protein, quinoa protein, amaranth protein, barley protein, rye protein, sorghum protein and sesame protein in the form of isolates, concentrates or hydrolysates.

[0100] The protein may be selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein, lactoferrin, lysozyme, gelatin, milk protein, bovine serum albumin, whey protein, casein, caseinate, egg protein, albumin, gluten, gelatin Type B, rice protein, soy protein, wheat protein, millet protein, legume, legumin, gluten, corn protein, zein, almond protein, peanut protein, pistachio protein, cashew nut protein, walnut protein, hazelnut protein, pine nut protein, coconut protein, insect protein, algal protein, microbial protein, spirulina, fungal protein (mycoprotein), single cell protein, leaf protein, mung bean protein, red kidney bean protein, tomato seed protein, pumpkin seed protein, oat protein, flax seed protein, chickpea protein, hemp protein, rapeseed protein, quinoa protein, amaranth protein, barley protein, rye protein, sorghum protein and sesame protein in the form of isolates, concentrates or hydrolysates.

[0101] The protein may be selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein and duckweed protein.

[0102] The protein may be selected from the group comprising: potato protein, lupin protein, oat protein, sunflower protein, fava bean protein, and almond protein. The protein may be selected from potato protein and lupin protein. Preferably, the protein is potato protein.

[0103] The mineral may comprise one or more essential minerals for human biochemical processes. The mineral may comprise: calcium, magnesium, iron, zinc, potassium, sodium, iodine, selenium, or any combination thereof. The mineral may comprise: calcium, magnesium, iron, zinc, potassium, sodium, or any combination thereof. The mineral may comprise: calcium, magnesium, zinc or iron. The mineral may comprise: calcium, magnesium, or zinc. The mineral may comprise calcium or iron. The mineral may comprise calcium.

[0104] The mineral may be added to the solution in the form of a mineral salt. For example, the mineral may comprise: a calcium salt, a magnesium salt, an iron salt, a zinc salt, a potassium salt, a sodium salt, an iodine salt, a selenium salt, or any combination thereof, the mineral may comprise: a calcium salt, a magnesium salt, an iron salt, a zinc salt, a potassium salt, a sodium salt, or any combination thereof. The mineral may comprise: acalcium salt, a magnesium salt, a zinc salt or an iron salt. The mineral may comprise: a calcium salt, a magnesium salt, or a zinc salt. The mineral may comprise: a calcium salt or an iron salt. The mineral may comprise a calcium salt.

[0105] The salt may be selected from any pharmaceutically acceptable salt. Suitable salts include, but are not limited to, halide salts, salts of acceptable inorganic acids such as hydrochloric, sulfuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids, or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, malic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulfonic, toluenesulfonic, benzenesulfonic, salicylic, sulfanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric acids.

[0106] The mineral salt may be selected from: calcium chloride, magnesium citrate, zinc citrate and iron sulfate. The mineral salt may be selected from: calcium chloride, magnesium citrate and zinc citrate. The mineral salt may be selected from: calcium chloride and iron sulfate. The mineral salt may be calcium chloride.

[0107] The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 50 °C (e.g. at least about 55 °C, at least about 60 °C, at least about 65 °C, at least about 70 °C, at least about 75 °C, or at least about 80 °C). The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 60 °C (e.g. at least about 65 °C, at least about 70 °C, at least about 75 °C, or at least about 80 °C, at least about 85 °C, at least about 90 °C). The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 70 °C (e.g. at least about 75 °C, at least about at least about 80 °C, at least about 85 °C, at least about 90 °C, at least about 95 °C or at least about 100 °C). The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 80 °C (e.g. at least about 85 °C, at least about 90 °C, at least about 95 °C, or at least about 100 °C).

[0108] The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 50 °C and no more than about 150 °C. The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 60 °C and no more than about 125 °C. The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 65 °C and no more than about 100 °C. The step of heating the solution to form the protein-mineral intermediate may be performed at a temperature of at least about 70 °C and no more than about 90 °C. The step of heating the solution to form the protein-mineralintermediate may be performed at a temperature of at least about 75 °C and no more than about 85 °C.

[0109] Preferably, the step of heating the solution to form the protein-mineral intermediate is performed at about 80 °C.

[0110] The step of heating the solution to form the protein-mineral intermediate may comprise heating the mixture for at least about 5 minutes, at least about 10 minutes, at least about 20 minutes, at least about 30 minutes, at least about 40 minutes, at least about 50 minutes, at least about 1 hour, at least about 1.5 hours, at least about 2 hours, or at least about 2.5 hours.

[0111] The step of heating the solution to form the protein-mineral intermediate may comprise heating the mixture for no more than 3 hours. The step of heating the solution to form the protein-mineral intermediate may comprise heating the mixture for no more than 2 hours.

[0112] It may be that the step of heating the solution to form the protein-mineral intermediate comprises heating the mixture for about 2 hours.

[0113] Without wishing to be bound by theory, it is thought that the heating time may play a role in the degree of aggregation of the filaments of the formulation. For example, it is thought that limiting the heating time of the solution to form the protein-mineral intermediate results in partial unfolding of the protein (i.e. the protein is not fully denatured), which may lead to the exposure of hydrophobic or aggregation-prone regions of the protein-mineral intermediate. After introduction of the biopolymer to form the filament, the presence of the hydrophobic or aggregation-prone regions allows for aggregation with neighbouring filaments. As a result of this aggregation, the formulations of the invention can form an interconnected network. The resulting particles demonstrate desirable viscosity, boundary film and lubrication properties.

[0114] The step of combining the protein-mineral intermediate with the biopolymer may involve adding the biopolymer to the protein-mineral intermediate in a quantity to weight percentage of biopolymer in the formulation as defined in any of the embodiments relating to the first aspect and the second aspect.

[0115] The step of combining the protein-mineral intermediate with the biopolymer may comprise adding the biopolymer to the protein-mineral intermediate while shearing the mixture to form the formulation of the invention.

[0116] The step of combining the protein-mineral intermediate with the biopolymer may be performed under stirring.

[0117] The step of combining the protein-mineral intermediate with the biopolymer may be performed at a temperature of at least about 10 °C, at least about 15 °C, at least about 20 °C, or at least about 25 °C. Preferably, the step of combining the protein-mineral intermediate with the biopolymer is performed at around ambient room temperature (i.e. 20- 30 °C).

[0118] The step of combining the protein-mineral intermediate with the biopolymer may be performed for at least 2 hours. The step of combining the protein-mineral intermediate with the biopolymer may be performed for at least 4 hours. The step of combining the proteinmineral intermediate with the biopolymer may be performed for at least 8 hours. The step of combining the protein-mineral intermediate with the biopolymer may be performed for at least 12 hours. The step of combining the protein-mineral intermediate with the biopolymer may be performed for at least 24 hours.

[0119] The biopolymer may be a polysaccharide. The biopolymer may be selected from the group comprising: xanthan gum, K-carrageenan, i-carrageenan, A-carrageenan, agar, agarose, alginate, pectin, dextran sulphate, cellulose and modified cellulose (ethyl cellulose, microcrystalline cellulose, carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose), locust bean gum, gum Arabic, gum ghatti, konjac gum (glucomannan), Karaya gum, Cassia gum, gellan gum, guar gum, modified starches, chitosan, inulin, beta-glucans, gum tragacanth, and any negatively-charged polysaccharide. Preferably, the biopolymer is xanthan gum.

[0120] The formulation and / or components of the formulation may be as described in any of the embodiments relating to the first aspect and the second aspect.Uses

[0121] In an aspect of the invention, there is provided the formulation of the invention for use as a medicament.

[0122] In another aspect of the invention, there is provided the formulation of the invention for use in treating a disease or condition caused by or associated with a mineral deficiency in a patient.

[0123] The disease or condition caused by or associated with the mineral deficiency is selected from: iron deficiency anaemia, osteoporosis, osteopenia, osteomalacia, hypocalcaemia, rickets, impaired immune function, muscle weakness, hypokalaemia, hypothyroidism, goitre, and cognitive impairments.

[0124] Thus, the mineral deficiency may be one or more of: iron deficiency, calcium deficiency, zinc deficiency, magnesium deficiency, potassium deficiency, sodium deficiency, iodine deficiency, and selenium deficiency.

[0125] In another aspect of the invention, there is provided a use of the formulation of the invention as a mineral supplement, wherein the use is not a method of treatment of the human or animal body.

[0126] In another aspect of the invention, there is provided a method for the treatment of a disease or condition caused by or associated with a mineral deficiency in a patient, the method comprising administering a formulation of the invention to a patient in need thereof.

[0127] The disease or condition caused by or associated with the mineral deficiency is selected from: iron deficiency anaemia, osteoporosis, osteopenia, osteomalacia, hypocalcaemia, rickets, impaired immune function, muscle weakness, hypokalaemia, hypothyroidism, goitre, and cognitive impairments.

[0128] Thus, the mineral deficiency may be one or more of: iron deficiency, calcium deficiency, zinc deficiency, magnesium deficiency, potassium deficiency, sodium deficiency, iodine deficiency, and selenium deficiency.EXAMPLESMaterials and methods

[0129] Potato protein isolate (> 90% protein content) was purchased from Sosa Ingredients (Barcelona, Spain). Lupin protein isolate (> 90% protein content) was purchased from Prolupin GmbH (Grimmen, Germany). Fava bean protein isolate Tendra® (> 85% protein content) was purchased from Consun Protein (Dinteloord, The Netherlands). Sunflower protein Heliaflor® (> 55% protein content) and Almond protein (> 50% protein content) were both obtained from All Organic Trading GmbH (Kempten, Germany). Oat protein PrOatein® (> 55% protein content) was obtained from Lantmannen Oats (Kimstand, Sweden). Xanthan gum was obtained from Sigma Aldrich (UK). Calcium chloride was obtained from Sigma Aldrich (UK). Magnesium citrate and zinc citrate were obtained from Peak Supps (Bridgend, UK). Solutions were made with ultrapure (Type 1) water having a resistivity of 18.2 MO. cm at 25 °C (Avidity Duo™ II. I, Thame, UK) and the pH adjusted as per requirement using HCI or NaOH.CharacterisationRheology

[0130] A modular compact rheometer MCR 302 stress-controlled (Anton Paar, Austria) equipped with a parallel plate geometry (PP50, 50 mm diameter plate) was used to measure the apparent viscosity of samples. The gap was fixed at 1.0 mm, and the temperature was controlled at 37 °C for the entire duration of the measurement. Apparent viscosity of (variouscontrols and the protein-polysaccharide-mineral filaments), milks, yoghurts was measured at shear rates ranging between 0.01 and 1000 s-1.Tribology

[0131] A Mini-Traction Machine (MTM, PCS Instruments, UK) was used to measure the lubrication properties of samples. A ball-on-disk configuration was used with polydimethylsiloxane (PDMS) specimens (PCS Instruments, UK). The diameter of the ball and disc was 19 and 46 mm, respectively. The temperature was set to 37 °C and the normal load was fixed at 2.0 N for all measurements. Friction coefficients were measured for the samples (various controls and the protein-polysaccharide-mineral filaments), milks, yoghurts at speeds varying from 1 - 1000 mm / s.Confocal scanning laser microscopy (CLSM)

[0132] A Zeiss LSM 700 CLSM (Carl Zeiss MicroImaging GmbH, Jena, Germany) confocal microscope was used to measure the microstructure of the samples. A stock solution of Fast Green (1 mg mL-1in Milli-Q water) was used to stain the protein to a final concentration of 0.1 mg mL'1, which was excited at a wavelength of 633 nm. The emission filter was set at 660 — 710 nm. Samples were placed on a concave confocal microscope slide, secured with a glass coverslip and imaged using an oil immersion 63* lens and the pinhole diameter maintained at 1 Airy Unit to filter out the majority of the light scattering.Cryogenic-scanning electron microscopy (cryo-SEM) and energy dispersive x-ray spectroscopy (EDX)

[0133] The cryo-SEM images were acquired by mounting the sample into freezing rivets using carbon paint and optimum temperature cutting medium. The samples were then plunge into slushed nitrogen at approximately -210 °C and then transferred to the cryo prep chamber. The samples were freeze fractured using a cooled knife and etched at -90 °C for 3 min. Samples were sputter coated using Iridium (Ir) at 5 uA for 30 s. The samples were transferred to the SEM for imaging at -140 °C. Samples were imaged in an FEI Helios G4 CX Dualbeam with a Quorum Technologies PP3010 cryo-SEM preparation system.

[0134] EDX was performed on the samples using an oxford instrument 150 mm2 X-max SDD detector, using Aztec software and 15 kV accelerating voltage.Light microscopy

[0135] Samples were placed onto microscope glass slides, overlaid with a glass cover slip, and imaged using Olympus-BH-2 microscope (Olympus, Japan) equipped with a 4* objective. Images were acquired using a colour digital camera (Euromex sCMEX-3 digital 3.0 Mpix USB-2 sCMOS) and processed with ImageFocus Plus V2 software.

[0136] With this invention, the inventors demonstrate formulations comprising one or more filaments comprising protein, mineral and biopolymer. These formulations achieve improved mineral delivery and better lubrication performance than commercial mineral sources, and provide lowering of friction coefficients without the need for high viscosity.Example 1 : Manufacture of the protein-mineral-biopolymer filaments at pH 2.0

[0137] Protein and mineral solutions were prepared by adding the selected powdered protein and the selected mineral together in ultrapure (Type 1) water and stirred for 2 hours to ensure complete solubilisation. Then the pH of the solution was adjusted to pH 2.0 by adding hydrochloric acid (6 M), and finally the solution containing the protein and mineral is heated at 80 °C for 2 hours at pH 2.0 to denature and result in acid-induced hydrolysis of protein forming cross-linking with mineral ions. This microstructure containing protein and minerals was cooled down to room temperature (25 °C).

[0138] Biopolymer was added to the selected protein + mineral formulation at varying biopolymer : protein weight ratios (0.01 :1 - 1 :1 w / w) at room temperature whilst being sheared for 24 hours under constant stirring for a complete hydration. If necessary, said mixture may be heated to improve microbial stability or to initiate a Maillard reaction. This results in the alignment and formation of a protein-biopolymer-mineral filament.

[0139] Exemplary formulations 1-23 are provided in Table 1 below, in which the amount of each species employed in the method for synthesising each formulation (as described above) is disclosed.Table 1Example 2: Analysis of lubrication performance and stability of potato protein-xanthan qum- calcium filaments

[0140] The lubrication performance (represented by the friction coefficient as a function of speed) of exemplary formulation 4 of the invention was assessed relative to the mineral component on its own (Control 1), the mineral component in combination with the protein component in the absence of biopolymer (Control 2), and the mineral component in combination with the biopolymer in the absence of the protein component (Control 3) (Figure 2).

[0141] The exemplary formulation 4 achieved improved friction coefficients across all speeds tested relative to Controls 1-3. Notably, at lower entrainment speeds, the friction coefficients demonstrated by exemplary formulation 4 are almost an order of magnitude lower than those of Controls 1-3.

[0142] The stability of exemplary formulation 4 was assessed in a stability study following a month of storage. In particular, the lubrication performance of a fresh sample of the formulation was tested alongside that of a sample of the same formulation that has been stored for 1 month (Figure 3). As depicted in Figure 3, the stored sample achieved comparable lubrication properties to the fresh sample, thereby demonstrating the shelf-life and stability of the formulations of the invention.Example 3: Varying the amount of mineral

[0143] The lubrication performance of exemplary formulations 1 to 5 was assessed relative to one another, where each formulation comprised a different concentration of calcium in the range of from 5 mM to 150 mM. As depicted in Figure 4, the formulations comprising a concentration of calcium of between 5 mM and 100 mM achieved comparably low friction coefficients, demonstrating good lubrication properties. However, formulation 5 comprising a concentration of calcium of 150 mM achieved a notably higher friction coefficient. Accordingly, the inventors found that a concentration of less than 150 mM is required to achieve the desired bio-lubrication performance.Example 4: Varying the type and amount of protein

[0144] Figure 5 shows the lubrication performance achieved when varying the type of protein employed in the formulations of the invention. In particular, the friction coefficient as a function of entrainment speed was assessed for each of exemplary formulations 4 and 6- 12. As depicted in Figure 5, each of these formulations achieved comparably good friction coefficients across all entrainment speeds, thereby demonstrating the versality of the formulations of the invention using different protein components.

[0145] Figure 7 shows the lubrication performance achieved when varying the amount of protein present in the formulations (exemplary formulations 4 and 18 to 20). As depicted in Figure 7A, varying the amount of protein from 2 wt% to 4 wt% of the formulation did not have a significant effect on the bio-lubrication properties, with each of formulations 18 to 20 achieving good lubrication performance, while optimal lubrication performance was achieved with 1 wt% of protein (formulation 4). This trend is also reflected in the viscosity data presented in Figure 7B. This data demonstrates that the desired lubrication properties can be achieved using different amounts of protein.Example 5: Varying the amount of biopolymer

[0146] Figure 6 shows the lubrication performance achieved when varying the amount of biopolymer present in the formulations (exemplary formulations 4 and 13 to 17). As depicted in Figure 6A, formulations employing 0.01 wt% (formulation 13) of biopolymer achieve substantially lower viscosity than formulations employing between 0.1 wt% and 1 wt% (formulations 4 and 14-17) of biopolymer. This trend is mirrored by the tribology data provided in Figure 6B, in which the formulation employing 0.01 wt% (formulation 13) of biopolymer demonstrates a friction coefficient almost an order of magnitude higher than the formulations comprising higher levels of biopolymer. In contrast, employing between 0.1 wt% and 1 wt% (formulations 4 and 14-17) of biopolymer leads to favourable lubrication performance. Thus, it is thought that a minimum level of biopolymer of more than 0.01 wt% is required to achieve the desired bio-lubrication properties.Example 6: Effect of pH

[0147] The effect of varying the pH on the lubrication performance of formulations of the invention was assessed.

[0148] Figure 8A depicts the lubrication data for exemplary formulation 4 at pH 2, pH 4, pH 7 and pH 9, compared to the lubrication data previously generated for Control 1. As shown, at pH 7 and 9, the friction coefficients of exemplary formulation 4 were comparable to those obtained for Control 1 across all entrainment speeds. In contrast, as the pH of the formulation was reduced to pH 4 and pH 2, the value of the friction coefficients decreased, indicating improved lubrication properties at lower pH.

[0149] Figure 8B shows SEM images of the formulation at pH 2, pH 7 and pH 9. At low pH (i.e. pH 2), the filaments are clearly visible and intact. However, at higher pH, the filament structure appears to break down. Thus, without wishing to be bound by theory, it is thought that the formulations are not structurally stable at pH 7 or higher and, as a result, exhibit a loss in lubrication properties.

[0150] A similar trend is observed in Figure 9A, in which the friction coefficients for exemplary formulation 10 at pH 7 and pH 9 were comparable to those of Control 1 across all entrainment speeds, while the friction coefficients at pH 2 and 4 were significantly lower. Figure 9B also shows intact filament structures at pH 2 but a break down in the filament structure at pH 7 and pH 9.

[0151] Accordingly, this data demonstrates that the desired lubrication properties are achieved at a pH of less than 7.0.Example 7: Varying the mineral

[0152] The effect of varying the mineral on the lubrication performance was also assessed using exemplary formulations 4 and 21-22. As depicted in Figure 10, although formulationscontaining calcium were shown to have the best lubrication performance, replacing calcium with zinc or magnesium did not significantly impact the values of the friction coefficients across all entrainment speeds, with all exemplary formulations 4 and 21-22 achieving improved lubrication performance relative to the control.

[0153] The effect of varying the mineral on the lubrication performance was also assessed using exemplary formulation 23 alongside formulations 4 and 21-22. As depicted in Figure 10, formulations containing iron were shown to have the best lubrication performance. However, calcium, zinc and magnesium perform similarly in terms of the values of the friction coefficients across all entrainment speeds and therefore all minerals tested achieve improved lubrication performance relative to the control.

[0154] Accordingly, this data demonstrates the breadth of application of this technology to different mineral supplements.

Claims

CLAIMS1. A formulation comprising one or more filaments, the one or more filaments comprising: a protein in an amount of from about 0.1% to less than about 5% by weight; a mineral in a concentration of from about 1 mM to less than about 150 mM; and a biopolymer in an amount of more than about 0.01 % to about 5% by weight; wherein the weight ratio of biopolymer to protein in the formulation is in the range of from about x: 1 to about y: 1 , wherein x > 0.01 and y < 1 , and wherein the formulation has a pH of less than about 7.0; wherein the protein is ionically cross-linked by the mineral and / or the protein is cross-linked by the mineral via ion-dipole interactions.

2. The formulation of claim 1 wherein the protein is associated with the biopolymer via non-covalent interactions.

3. The formulation of claim 2, wherein the protein is associated with the biopolymer via non-covalent interactions selected from one or more of: hydrogen bonding, hydrophobic interactions, dipole-dipole interactions, ion-dipole interactions, and ionic bonds.

4. The formulation of any preceding claim, wherein the weight ratio of biopolymer to protein in the formulation is in the range of from about 0.1 :1 to about 1 :1.

5. The formulation of any preceding claim, wherein the protein is selected from the group comprising: potato protein, lupin protein, pea protein, oat protein, sunflower protein, fava bean protein, almond protein, duckweed protein, lactoferrin, lysozyme, gelatin, milk protein, bovine serum albumin, whey protein, casein, caseinate, egg protein, albumin, gluten, gelatin Type B, rice protein, soy protein, wheat protein, millet protein, legume, legumin, gluten, corn protein, zein, almond protein, peanut protein, pistachio protein, cashew nut protein, walnut protein, hazelnut protein, pine nut protein, coconut protein, insect protein, algal protein, microbial protein, spirulina, fungal protein (mycoprotein), single cell protein, leaf protein, mung bean protein, red kidney bean protein, tomato seed protein, pumpkin seed protein, oat protein, flax seed protein, chickpea protein, hemp protein, rapeseed protein, quinoa protein, amaranth protein, barley protein, rye protein, sorghum protein and sesame protein in the form of isolates, concentrates or hydrolysates.

6. The formulation of any preceding claim, wherein the protein is potato protein.

7. The formulation of any preceding claim, wherein the protein is present in an amount in the range of from about 1 % to about 4% by weight.

8. The formulation of any preceding claim, wherein the mineral comprises one or more essential minerals for human biochemical processes.

9. The formulation of any preceding claim, wherein the mineral is selected from the group comprising: calcium, magnesium, iron, zinc, potassium, sodium, iodine, selenium, or any combination thereof, optionally wherein the mineral is calcium.

10. The formulation of any preceding claim, wherein the mineral is present in a concentration in the range of from about 50 mM to less than about 150 mM.

11. The formulation of any preceding claim, wherein the biopolymer is a polysaccharide.

12. The formulation of claim 11 , wherein the biopolymer is selected from the group comprising: xanthan gum, K-carrageenan, i-carrageenan, A-carrageenan, agar, starch, maltodextrin, agarose, alginate, pectin, dextran sulphate, cellulose and modified cellulose (ethyl cellulose, microcrystalline cellulose, carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose), locust bean gum, gum Arabic, gum ghatti, konjac gum (glucomannan), Karaya gum, Cassia gum, gellan gum, guar gum, modified starches, chitosan, inulin, beta-glucans, gum tragacanth, and any negatively-charged polysaccharide.

13. The formulation of claim 12, wherein the biopolymer is xanthan gum.

14. The formulation of any preceding claim, wherein the biopolymer is present in an amount in the range of from about 0.1 % to about 1 % by weight.

15. The formulation of any preceding claim, wherein the formulation has a pH of less than about 5.0, optionally wherein the formulation has a pH in the range of from about 2.0 to about 4.0.

16. The formulation of any preceding claim, wherein each of the one or more filaments are at least about 1 pm in length, optionally wherein the one or more filaments are from at least about 1 pm to about 100 mm in length.

17. A method of producing the formulation of any of claims 1 to 16, the method comprising: providing a solution comprising the protein and the mineral, the solution having a pH of less than about 7.0; heating the solution to form a protein-mineral intermediate; and combining the protein-mineral intermediate with the biopolymer to form the formulation of any of claims 1 to 16.

18. The method of claim 17, wherein the step of heating the solution is performed for no more than about 2 hours.

19. A formulation obtainable or obtained by the method of claim 17 or claim 18.

20. The formulation of any of claims 1 to 16 for use as a medicament.

21. The formulation of any of claims 1 to 16 for use in treating a disease or condition caused by or associated with a mineral deficiency in a patient.

22. The formulation for use of claim 21 , wherein the disease or condition caused by or associated with the mineral deficiency is selected from: iron deficiency anaemia, osteoporosis, osteopenia, osteomalacia, hypocalcaemia, rickets, impaired immune function, muscle weakness, hypokalaemia, hypothyroidism, goitre, and cognitive impairments.

23. Use of a formulation of any of claims 1 to 16 as a mineral supplement, wherein the use is not a method of treatment of the human or animal body.

24. A method for the treatment of a disease or condition caused by or associated with a mineral deficiency in a patient, the method comprising administering a formulation of any of claims 1 to 16 to a patient in need thereof.