Use of beta-lactoglobulin and pectin complexes in alternative dairy

Abstract

The present invention provides alternative dairy compositions comprising a plurality of particles comprising a complex of a non-animal Beta-lactoglobulin (BEG) protein and a non-animal pectin. The application further provides methods for preparation of such alternative dairy composition, and food products comprising said alternative dairy compositions.

Description

[0001] USE OF BETA-LACTOGLOBULIN AND PECTIN COMPLEXES IN ALTERNATIVE DAIRY

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to alternative, non-animal dairy products comprising stable complexes of beta-lactoglobulin and pectin.

[0004] BACKGROUND OF THE INVENTION

[0005] A primary challenge in developing alternative milk beverages is ensuring their shelflife stability, specifically the stability of the protein phase, and avoiding phase separation. These biomolecules, usually having globular fold and structure, are prone to aggregation and precipitation as the pH of their solution approaches their isoelectric point. This challenge is particularly evident when a neutral pH beverage, such as milk, is mixed with the acidic environment of coffee, causing the pH to drop near to the protein isoelectric point, ultimately leading to protein destabilization and aggregation.

[0006] One solution to this issue involves the incorporation of buffering agents. These buffers stabilize the beverage’s pH, preventing protein aggregation. However, the use of buffering agents presents inherent limitations. While they effectively prevent protein precipitation, they often impart undesirable off flavors, such as metallic or astringent notes, to the final edible product. Additionally, the presence of buffering agents on the ingredient list may conflict with consumer preferences for clean-label products. Regulatory restrictions further complicate their use, with limitations varying across different regions.

[0007] For instance, phosphates and citrates are two widely used buffering agents in the food industry. In the European Union, phosphates are prohibited in milk alternative beverages made from milk proteins produced through precision fermentation. Although citrates are allowed, stabilizing proteins often requires high concentrations of citrates, which can lead to strong metallic tastes in the final beverage.

[0008] In addition to flavor concerns, buffering agents come with another trade-off. While they prevent protein aggregation by maintaining a stable pH, they simultaneously preserve the small particle size of proteins, typically ranging from a few nanometers to several tens of nanometers. This small size limits the ability of protein particles to scatter light effectively, reducing the whitening effect of the beverage in coffee. Consequently, plant- based beverages, such as soy or oat milk, often appear darker than regular milk when added to coffee.

[0009] Given these challenges, the development of stable, appealing milk alternatives without heavy reliance on buffering agents remains a significant hurdle.

[0010] SUMMARY OF THE INVENTION

[0011] To overcome challenges mentioned in the background, a unique approach has been developed, building upon the concept of protein-polysaccharide complexation, specifically using recombinant beta-lactoglobulin (rBLG), a whey protein. Several studies have demonstrated that BLG can form stable complexes with polysaccharides, such as pectin, through electrostatic attraction.

[0012] Without being limited to any particular theory it is assumed that the two biopolymers (BLG and pectin) when mixed under specific conditions that promote electrostatic interactions and upon application of heat that cause to BLG unfolding and formation of electrostatic bonds with the polysaccharide eventually form a core of stable particles. The result is the formation of large particles, typically ranging from several hundred nanometers to a few microns, which exhibit excellent resistance to aggregation in both pH fluctuations and / or extreme temperatures.

[0013] The inventors exploited the unique characteristics of these protein-polysaccharide complexes to create alternative BLG-based milk-like beverages without the need for any pH regulators, phosphates, polyphosphates or divalent ion chelators. The formation of large particles not only stabilizes the protein but also enhances the final color of coffee when the beverage is added. This approach resolves three critical issues: resistance to aggregation in extreme pH changes, resistance to aggregation in high temperatures, and improved whitening capability in coffee due to enhanced light scattering by the larger protein particles. By employing the methods and particles provided herein, the inventors provide a comprehensive solution that enhances both the stability and sensory appeal of alternative milk beverages.

[0014] According to one aspect, the present invention provides an alternative dairy composition, comprising a plurality of particles, wherein the particles comprise a complex of a non-animal Beta-lactoglobulin (BLG) protein and a non-animal pectin. In some examples, the particles comprise a core consisting of the non-animal BLG protein and a nonanimal pectin. In other examples, each particle comprises a core consisting of the non-animal BLG protein and a non-animal pectin. In some examples, the composition comprises comprising a plurality of particles as described herein in water. According to some examples, the alternative dairy composition further comprises one or more of water, a non-animal lipid, a non-animal sweetener, NaCl, L-Cystine, calcium, or any combination thereof. According to other examples, alternative dairy composition further comprises water, non-animal lipid, non-animal sweetener, NaCl, L-Cystine, and calcium. According to some examples, the core consists of the non-animal BLG, a non-animal pectin, and non-animal lipid.

[0015] According to some examples, the alternative dairy composition comprises from 0.1% to 3.5% w / w of a non-animal BLG or from 0.5% to 2% w / w non-animal BLG.

[0016] According to some examples, the non-animal pectin is selected from a high methoxly pectin (HM), high methoxyl pectin amidated (HMA) low methoxyl pectin (LM), low methoxyl pectin amidated (LMA), and any combination thereof. According to some examples, the non-animal pectin is a mixture of LM and HM. According to some examples, the non-animal pectin is a mixture of LM(A) and HM(A). According to some examples, the non-animal pectin is a mixture of LM and HMA. According to some examples, the ratio between LM and HM(A) is from 1: 10 to 1:2 or from 1:2.5 to 1:7.5, respectively. According to some examples, the ratio between LM or LMA and HM is about 1:5.25. According to some examples, the ratio between LM(A) and HM(A) is from 1:2 to 1:20 or from 1:4 to 1: 18, or from 1:2.5 to 1:7.5, respectively. According to some examples, wherein the ratio between LM and HM is about 1:5.25. According to some examples, wherein the ratio between LM and HMA is about 1:5.25.

[0017] According to some examples, the alternative dairy composition comprises between 0.2% to 0.4% w / w total non-animal pectins. According to some examples, the alternative dairy composition comprises about 0.3% w / w total non-animal pectins.

[0018] According to some examples, the particles have an average particle size of 700 nm to 1400 nm. According to some examples, the particles have an average particle size of 900 nm to 2000 nm.

[0019] According to some examples, the alternative dairy composition comprises: i. from about 0.1 w / w% to about 3.5 % w / w BLG, ii. from about 0.25 % w / w to about 0.45 % w / w total pectins, iii. from about 0.1 % w / w to about 5 % w / w non-animal lipid, iv. from about 0.1 % w / w to about 6 % w / w sweetener, v. from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), vi. from about 0.005 % w / w to about 0.1 % w / w L-Cystine, and vii. from about 0.15 % w / w to about 0.25 % w / w calcium, and viii. up to 100 w / w water.

[0020] According to some examples, the alternative dairy composition comprises: i. from about 0.5 % w / w to about 1.5 % w / w BLG, ii. from about 0.2 % w / w to about 0.4 % w / w total pectins, iii. from about 0.5 % w / w to about 3.5 % w / w non-animal lipid, iv. from about 1 % w / w to about 1.75 % w / w sweetener, v. from about 0.05 % w / w to about 0.15 % w / w NaCl (edible salt), vi. from about 0.01 % w / w to about 0.05 % w / w L-Cystine, and vii. from about 0.4 % w / w to about 0.6 % w / w calcium, and viii. up to 100 % w / w water.

[0021] According to some examples, the alternative dairy composition has a pH of from 5 to 8. According to some examples, the alternative dairy composition is devoid of Gum Arabic, Carrageenan Lambda, Gellan, sodium hexametaphosphate (SHMP), mono-potassium phosphate, di-potassium phosphate, a phosphate buffer, polyphosphates, divalent ion chelators and / or a citrate buffer.

[0022] According to some examples, the alternative dairy composition described above is formulated as an alternative milk composition.

[0023] According to another aspect, the present invention provides a food product comprising the alternative dairy composition as described in any one of the herein provided aspects, examples, and embodiments. According to some examples the alternative dairy composition is an alternative milk composition. According to some examples, the food product is instant coffee.

[0024] According to some examples, the present invention provides an instant coffee comprising the alternative milk composition as described herein. According to some examples, the food product has: a. a pH of 5 to 6, and / or b. a Color (L, Hunter units) of 15 to 40 when the food product comprises from 3 to 20% v / v of the alternative milk composition.

[0025] According to yet another aspect, the present invention provides a method of preparation of an alternative dairy composition, the method comprising: i. Combining a composition comprising a hydrated pectin with a non-animal Betalactoglobulin; ii. Setting a pH of the composition obtained in step (i) to a value of from 3 to 6, thereby obtaining a complex of the pectin and the non-animal Betalactoglobulin; iii. Optionally adding a non-animal lipid to the composition obtained in previous step; iv. Optionally homogenizing the composition obtained in the previous step; and v. Pasteurizing the composition obtained in the previous step via ultra-high temperature pasteurization.

[0026] According to some examples, the method comprises adding (a) a non-animal lipid at step (iii), (b) homogenizing at step (iv), or (c) both (a) and (b).

[0027] According to some examples:

[0028] (a) Step (i) comprises a complete hydration of the non-animal BLG;

[0029] (b) The method comprises hydrating the pectin prior to step (i);

[0030] (c) Both (a) and (b).

[0031] According to some examples, the method comprises adjusting the pH of the composition to the value of from 6.5 to 7.5 in an additional step (vi) or after any one of steps (iii)-(v), optionally the pH adjustment is carried out by adding calcium carbonate.

[0032] According to some examples, the method comprising adding at least one of a sweetener, L-cystine, and NaCl at step (i). According to some examples: i. The BLG is a recombinant BLG, ii. The pectin is selected from high methoxly pectin (HM); high methoxyl pectin amidated (HMA)_low methoxyl pectin (LM), low methoxyl pectin amidated (LMA), and any combination thereof, or iii. Both (i) and (ii). According to some examples, the pectin is a mixture of LM(A) and HM(A), preferably a mixture of LM and HMA. According to some examples, the ratio between LM and HMA and is from 1: 10 to 1:2 or from 1:2.5 to 1:7.5 or about 1:5.25.

[0033] According to some examples, the method comprises adding from 0.1 to 1.5% BLG.

[0034] According to some examples, the method comprises: i. optionally dissolving in water from about 0.1 % w / w to about 6 % w / w sweetener, from about 0.01 % w / w to about 0.3 % w / w NaCl, from about 0.005 % w / w to about 0.1 % w / w L-Cystine, ii. hydrating from about 0.25 % w / w to about 0.45 % w / w total pectins in the composition obtained in previous step, if present, preferably wherein the pectins are a mixture of LM and HMA, iii. adding about 0.1 w / w% to about 3.5 % w / w BLG to the composition obtained in previous step and fully hydrating the BLG, iv. setting the pH of the composition obtained in previous step to a value of from 4.5 to 5, preferably to a value of 4.75 v. optionally adding and dispersing from about 0.1 % w / w to about 5 % w / w of non-animal lipid in the composition obtained in the previous step, optionally with preheating of the composition to from 60 to 70°C. vi. optionally homogenizing the composition obtained in the previous step, and vii. pasteurizing the composition the composition obtained in the previous step via an ultra-high temperature pasteurization.

[0035] According to some examples, the resulting alternative dairy composition comprises a plurality of particles comprising a core consisting of the non-animal dairy protein and pectin. According to some examples, the resulting particles have an average particle size of 900 nm to 2000 nm.

[0036] According to some examples, the method is devoid of adding one or more compounds selected from Gum Arabic, Carrageenan Lambda, Gellan, SHMP, phosphate buffer, polyphosphates, divalent ion chelators and citrate buffer.

[0037] According to some examples, the ultra-high temperature pasteurization comprises heating to from 110 °C to 125 °C for from 1 to 10 sec. According to some examples, the method further comprises heating the composition at step (ii) to from 70 °C to 90 °C.

[0038] According to another example, the present invention provides an alternative food composition prepared by a method as described herein.

[0039] BRIEF DESCRIPTION OF DRAWINGS

[0040] Fig. 1 shows the effect of particle sedimentation - on the left: final formulation of a stable alternative milk (1.5 % BLG + 0.3 % pectin), on the right: final formulation of an unstable alternative milk (3% BLG + 0.3% pectin).

[0041] DETAILED DESCRIPTION OF THE INVENTION

[0042] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. In case of conflict, the patent specification, including definitions, will control.

[0043] The present invention is based on an unexpected observation that alternative dairy product comprising particles comprising as a core complexes of BLG and pectin are stable at high temperature and low pH without forming a precipitate. According to one aspect, the present invention provides an alternative dairy composition, comprising a plurality of particles, wherein the particles comprise a complex of a non-animal milk protein and a nonanimal pectin. According to some embodiments, the alternative dairy composition is a liquid composition. According to some embodiments, the alternative dairy composition is a beverage. According to some embodiments, the alternative dairy composition is an alternative milk. According to some embodiments, the non-animal milk protein is a nonanimal whey protein. According to some embodiments, the non-animal milk protein is beta-lactoglobulin (BLG). According to some embodiments, BLG is a recombinant BLG. According to some embodiments, the BLG is a deaminated BLG. According to some embodiments, the BLG is a partially deaminated BLG. According to some embodiments, the BLG is an enzymatically deaminated BLG. According to some embodiments, the BLG is a partially enzymatically deaminated BLG. According to some embodiments, BLG is enzymatically deaminated by protein glutaminase. According to some embodiments, the present invention provides an alternative dairy composition, comprising a plurality of particles, wherein the particles comprise a complex of a non-animal beta-lactoglobulin (BLG) protein and a non-animal pectin. According to some embodiments, BLG is a recombinant BLG.

[0044] The terms "alternative dairy composition", "alternative dairy product", “dairy substitute" and "dairy alternative", are used herein interchangeably and refer to any consumable / edible product or foodstuff, which is not made from or derived from animals’ milk. Such products may replace animal-based products in one’s diet by having the nutritional and / or rheologic and / or organoleptic and / or physicochemical properties of the corresponding traditional animal-milk-based products. According to some embodiments, the alternative dairy composition is an alternative milk composition. In this context, the terms “milk alternative composition”, “alternative milk composition” and “alternative milk" relates to any non- animal derived composition, which can be used as a milk substitute, and / or as a dairy product substitute of milk traditionally provided from animals (e.g., cheese or ice cream). Thus, alternative milk compositions may be incorporated into dairy alternative products which are the final products provided to consumers.

[0045] The term “beta-lactoglobulin” (BLG) refers to a beta-lactoglobulin protein that is typically present in cow's milk. As used in the present invention, the term BLG further refers to isoform B of the BLG, i.e., beta-Lactoglobulin B (P-LG B), which is a small protein of 162 amino acids with a molecular mass of 18.2 kDa and optimum pH of 5.2 (UniProt D6QX31). Nevertheless, in some specific embodiments, the term BLG may refer to BLG-A isoform or to a combination of BLG-A and BLG-B. According to some embodiments, the BLG and / or the rBLG have the amino acid sequence SEQ ID NO: 1. The term "BLG protein" refers to intact protein. According to some embodiments, the BLG is a recombinant BLG. The term “BLG” encompasses known BLG variants, for example, known bovine BLG variants, and also analogs and chimera of the BLG. The term "analog”, “analog” and “sequence analog” are used herein interchangeably and refer to an analog of a peptide, polypeptide or protein having at least 70% sequence identity with the original peptide, wherein the analog retains the activity of the original peptide or protein. Thus, the terms “analog” and “active analog” may be used interchangeably. In some examples, the analog has at least 99%, 98%, 97%, 96% or 95% sequence identity with the original sequence. The term “analog” refers to a peptide, polypeptide or protein which contains substitutions, rearrangements, deletions, additions and / or chemical modifications in the amino acid sequence of the parent peptide. The substitutions of the amino acids may be conservative or non-conservative substitution. The non-conservative substitution encompasses substitution of one amino acid by any other amino acid. In one particular embodiment, the amino acid is substituted by a non-natural amino acid. The term “conservative substitution” as used herein denotes the replacement of an amino acid residue by another, without altering the overall conformation and biological activity of the peptide, including, but not limited to, replacement of an amino acid with one having similar properties (such as, for example, polarity, hydrogen bonding potential, acidic, basic, shape, hydrophobic, aromatic, and the like). Amino acids with similar properties are well known in the art. For example, according to one table known in the art, the following six groups each contain amino acids that are conservative substitutions for one another: (1) Alanine (A), Serine (S), Threonine (T); (2) Aspartic acid (D), Glutamic acid (E); (3) Asparagine (N), Glutamine (Q); (4) Arginine (R), Lysine (K); (5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and (6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

[0046] According to some embodiments, the BLG protein comprises an amino acid sequence selected from SEQ ID NOs: 1-10. According to some embodiments, the term BLG also encompasses analogs thereof. According to some embodiments, the BLG analog comprises an amino acid sequence selected from SEQ ID NOs: 11-20. Examples of recombinant BLG proteins are provided herein in SEQ ID NOs: 1-20.

[0047] According to some embodiments, the alternative dairy composition, e.g. alternative milk composition comprises from 0.1% to 3.5% w / w BLG. According to some embodiments, the alternative dairy composition, e.g. alternative milk composition comprises from 0.1% to 3.5% w / w, from 0.2% to 3.4% w / w, from 0.3% to 3.3% w / w, from 0.4% to 3.2% w / w, from 0.5% to 3.1% w / w, from 0.6% to 3.0% w / w, from 0.7% to 2.9% w / w, from 0.8% to 2.8% w / w, from 0.9% to 2.7% w / w, from 1.0% to 2.6% w / w, from 1.0% to 2.5%, from 1.1% to 2.5% w / w, from 1.2% to 2.4% w / w, from 1.3% to 2.3% w / w, or from 1.4% to 2.2% w / w BLG. According to some embodiments, the alternative dairy composition, e.g. alternative milk composition comprises from 0.25% to 3 % w / w BLG. According to some embodiments, the alternative dairy composition, comprises from 0.25% to 2.5 % w / w BLG. According to some embodiments, the alternative dairy composition, e.g. alternative milk composition comprises from 0.5% to 2 % w / w BLG. According to some embodiments, the alternative dairy composition, e.g. alternative milk composition comprises from 0.5% to 1.5 % w / w BLG.

[0048] The alternative dairy composition is a non-animal alternative dairy composition. The terms "non-animal" and "animal-free" refers to a product that is entirely free of animal- derived, and specifically free of milk-derived, components, such as BLG or other milk proteins. In this context, the term "milk" refers to milk from mammal animals such as cow's, goat's and sheep's milk. While all components of such products are non-animal, the present invention specifically relates to products comprising at least one recombinant component or ingredient. The term "recombinant dairy ingredient" refers to any ingredient, found in mammal dairy, that is recombinantly produced. According to one embodiment, the recombinant dairy ingredient is selected from a recombinant dairy protein, a recombinant dairy fat, and a recombinant dairy carbohydrate. According to some embodiments, the recombinant dairy ingredient is a recombinant dairy protein. According to some embodiments, the recombinant dairy protein is a recombinant whey protein. According to some embodiments, the recombinant dairy protein is P-Lactoglobulin (BLG). According to some embodiments, the recombinant dairy protein is isoform B of the BLG.

[0049] According to some embodiments, from 10 to 100% of particles comprise particles comprising a complex of a non-animal beta-lactoglobulin (BLG) protein and a non-animal pectin. According to some embodiments, from 20% to 100%, from 30% to 100%, from 40% to 100%, from 50% to 100%, from 60% to 100%, from 70% to 100%, from 80% to 100%, from 90% to 100%, from 20% to 95%, from 30% to 95%, from 40% to 95%, from 50% to 95%, from 60% to 95%, from 70% to 95%, from 80% to 95%, from 90% to 95%, from 20% to 90%, from 30% to 90%, from 40% to 90%, from 50% to 90%, from 60% to 90%, from 70% to 90%, or from 80% to 90% of particles comprise a complex of a non-animal beta- lactoglobulin (BLG) protein and a non-animal pectin.

[0050] According to some embodiments, the particles comprise a core comprising a complex of a non-animal Beta-lactoglobulin (BLG) protein and a non-animal pectin. According to some embodiments, from 20% to 100%, from 30% to 100%, from 40% to 100%, from 50% to 100%, from 60% to 100%, from 70% to 100%, from 80% to 100%, from 90% to 100%, from 20% to 95%, from 30% to 95%, from 40% to 95%, from 50% to 95%, from 60% to 95%, from 70% to 95%, from 80% to 95%, from 90% to 95%, from 20% to 90%, from 30% to 90%, from 40% to 90%, from 50% to 90%, from 60% to 90%, from 70% to 90%, or from 80% to 90% of particles comprise a core comprising a complex of a non-animal Betalactoglobulin (BLG) protein and a non-animal pectin. According to some embodiments, each particle comprises a core comprising a complex of a non-animal Beta-lactoglobulin (BLG) protein and a non-animal pectin. According to any one of the above embodiments, the core consisting of the non-animal BLG protein and non-animal pectin.

[0051] The term “pectin” refers to a polysaccharide that is present in fruits and vegetables and that has galacturonic acid segments with rhamnose side chains. Exemplary pectins include, but are not limited to, apple pectins, citrus pectins, grape pectins, carrot pectins, and combinations thereof. The term refers collectively to pectin in its normal sense, as well as fractions and derivatives thereof, as well as modified pectins (e.g., chemically modified pectins and enzymatically modified pectins). By way of example, pectin can be a derivatized pectin, a degraded (such as partially degraded) pectin or a modified pectin. An example of a chemically modified pectin and / or a pectin derivative is pectin that has been chemically treated, e.g., amidated.

[0052] The carboxyl group of galacturonic acid constituting the pectin or pectin derivative may be esterified. Therefore, the pectin may be low-methoxyl (LM) pectin, high-methoxyl (HM) pectin, amidated high-methoxyl pectin, amidated low-methoxyl pectin, or a combination thereof. The terms "LM(A)" refers to LM or amidated form thereof collectively, and the term "HM(A)" refers to HM or amidated form thereof collectively. The low- methoxyl pectin refers to pectin in which less than about 50% of the carboxyl groups are esterified. The high-methoxyl pectin means pectin in which about 50% or greater of the carboxyl groups are esterified. In some example embodiments, about 20% or greater, for example about 25% or greater, or about 30% or greater, and about 60% or greater, and about 55% or less, about 50% or less, or about 45% or less of the carboxyl groups are esterified. The phrase “degree of esterification” (DE) refers to the amount or percentage of esterified galacturonic acid units within a pectin structure.

[0053] According to some embodiments, the pectin has high methoxyl pectin content. According to some embodiments, the pectin has low methoxyl pectin content. According to some embodiments, the pectin is amidated. According to some embodiments, the pectin is not amidated. According to some embodiments, the pectin is selected from a high methoxly pectin (HM), high methoxyl pectin amidated (HMA) low methoxyl pectin (LM), low methoxyl pectin amidated (LMA), and any combination thereof. According to some embodiments, the pectin is a mixture of LM and HM. According to some embodiments, the pectin is a mixture of LMA and HM. According to some embodiments, the pectin is a mixture of LM and HMA. According to some embodiments, the pectin is a mixture of LMA and HMA.

[0054] According to some embodiments, the LMA has a degree of esterification of from 22 to 32%, preferably about 27+3%, a degree of amidation of from 15 to 28%, preferably about 22+3% and a content of galacturonic acid of more than 60%, preferably more than 65%. According to some embodiments, the LMA has a degree of esterification of about 27+3%, a degree of amidation of about 22+3% and a content of galacturonic acid of more than 65%. According to some embodiments, the content of galacturonic acid is from 60% to 100%, from 65% to 100% or from 65% to 95%.

[0055] According to some embodiments, the HM has a degree of esterification of more than

[0056] 62%. According to some embodiments, the HM has a degree of esterification of more than

[0057] 65%. According to some embodiments, the HM has a degree of esterification of more than

[0058] 68%. According to some embodiments, the HM has more than 60%, preferably more than

[0059] 65% galacturonic acid. According to some embodiments, the content of galacturonic acid is from 60% to 100%, from 65% to 100% or from 65% to 95%.

[0060] According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:2 to about 1:20. According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:4 to about 1: 18. According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:4 to about 1: 16. According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:2 to about 1: 10. According to some embodiments, the ratio between LM and HM is from about 1:2 to about 1:8. According to some embodiments, the ratio between LM and HM is from about 1:25 to about 1:75. According to some embodiments, the ratio between LM and HM is from about 27:72 to 16:84 or about 16:84. According to some embodiments, the ratio between LM and HM is about 1:5.25. According to some embodiments, the pectin is amidated. According to some embodiments, the pectin is a mixture of LMA and HM and the ratio is as stated in any one of the above embodiments. According to some embodiments, the pectin is a mixture of LM and HMA, and the ratio between the LM and the HMA is as stated in any one of the above embodiments. According to some embodiments, the DE value of HM (either HM or HMA) is 60% and more. According to some embodiments, the DE value of HM (either HM or HMA) is higher than 65%. According to some embodiments, the DE value of HM (either HM or HMA) is from 60 to 75% or from 62 to 70% or from 65 to 70%. According to some embodiments, the DE value of HM (either HM or HMA) is about 67%.

[0061] According to some embodiments, the DE value of the LM (either LM or LMA) is from 15 to 35%. According to some embodiments, the DE value of the LM (either LM or LMA) is from 20 to 30% or from 25% to 30%. According to some embodiments, the DE value of the LM (either LM or LMA) is about 27%.

[0062] According to any one of the above embodiments, the alternative dairy composition comprises from 0.1 to 0.45% w / w pectin. According to some embodiments, the alternative dairy composition comprises from 0.2% to 0.45% w / w of pectin. According to some embodiments, the alternative dairy composition comprises from 0.1% to 0.3% w / w of pectin. According to some embodiments, the alternative dairy composition comprises from 0.15% to 0.25% w / w of pectin. According to some embodiments, the alternative dairy composition comprises from 0.10% to 0.20% w / w of pectin. According to some embodiments, the alternative dairy composition comprises from 0.2% to 0.4% w / w of pectin. The amount of pectin refers collectively to all types of pectins in the composition. According to some embodiments, the alternative dairy composition comprises from about 0.22 to about 0.38% w / w of pectin. According to some embodiments, the alternative dairy composition comprises from about 0.25 to about 0.35% w / w of pectin. According to some embodiments, the alternative dairy composition comprises about 0.3% w / w of pectin.

[0063] According to some embodiments, the particles of the plurality of particles have an average particle size of 700 nm to 1400 nm. According to some embodiments, the particles have an average particle size of from 750 to 1200 nm.

[0064] As used herein, the term "particle size" refers to the longest dimension of the particles. For spherical particles, the term particle size refers to the diameter of the particle. The particle size may be determined by any known method, for example, by a laser scattering particle size distribution analyzer.

[0065] According to some embodiments, the alternative dairy composition such as alternative dairy milk of the present invention further comprises a lipid, a sweetener, NaCl, L-Cystine, calcium, or any combination thereof. According to some embodiments, the alternative dairy product of the present invention further comprises a lipid. According to some embodiments, the alternative dairy product of the present invention further comprises a sweetener. According to some embodiments, the alternative dairy product of the present invention further comprises a NaCl. According to some embodiments, the alternative dairy product of the present invention further comprises L-Cystine. According to some embodiments, the alternative dairy product of the present invention further comprises calcium. According to some embodiments, the alternative dairy product of the present invention further comprises a lipid, a sweetener, NaCl, L-Cystine, water and calcium.

[0066] According to some embodiments, the lipid is a non-animal lipid. The term "non-animal lipid" refers to any lipid, fat and oil that does not originate from an animal and / or milk. According to some embodiment, the lipid is plant-derived lipid. According to some embodiments, the lipids comprise an oil. According to some embodiments, the lipids comprise a non-animal fat. According to some embodiment, the oil is selected from shea oil, sunflower oil, coconut oil, rapeseed oil, nut oil, palm oil, kernel oil, olive oil, soya oil, cotton oil, and cocoa butter (Theobroma oil). According to some embodiments, the alternative dairy product of the present invention comprises from 0.5 to 6 wt% of the non-animal lipid. According to some embodiments, alternative dairy product of the present invention comprises from 1 to 5 wt% of the non-animal lipid. According to some embodiments, alternative dairy product of the present invention comprises from 2 to 4 wt% of about 3 wt% of the non-animal lipid fat such as plant oil.

[0067] According to some embodiments, formulating the alternative dairy composition, such as alternative dairy milk, comprises adding a sweetener. The term "sweetener" refers to any natural and artificial substances that provides a sweet taste in foods and beverages. According to some embodiments, the term sweetener excludes lactose. The term "sweetener" also comprises sugars and carbohydrates. The term “sweetener” as used herein refers to an organic compound that is generally sweet in taste. For example, sweeteners are generally used to impart a sweet taste in edible products. A sweetener can include artificial sweeteners and natural sweeteners such as plant-derived sweeteners. A sweetener can be generally safe for consumption. A sweetener suitable for use according to the present disclosure can have a sweetness intensity that is lower, similar to or greater than that of sucrose depending on the desired sweetness in the final product. In some instances, the sweeteners have a sweetness intensity that is greater than that of sucrose. Those sweeteners can be high intensity sweeteners. Sweeteners are often classified as either nutritive (caloric) or nonnutritive (non-caloric), natural or synthetic. Examples of sweeteners include but are not limited to sucrose, dextrose, lactose, glucose, advantame, sorbitol, mannitol, liquid glucose, honey molasses, saccharin, sucralose, rebaudioside A stevia, rebaudioside M stevia, stevioside, mogroside IV, mogroside V, alitame, saccharin, neohesperidin dihydrochalcone, cyclamate, neotame, N- [3_(3 -hydroxy- 4-methoxybenzyl yl) propyl] -L-a- aspartyl] -L- phenylalanine 1 -methyl ester, N- [3- (3- hydroxy-4-methoxyphenyl) -3-methylbutan yl] -L- a - aspartyl] -L- phenylalanine 1 -methyl ester, N- [3- (3- methoxy-4-hydroxyphenyl) propyl] -L- a - aspartyl] -L- phenylalanine 1 -methyl ester, curculin, cyclamate, aspartame, acesulfame potassium and others or mixtures thereof.

[0068] In some embodiments, the sweetener is a sugar. The term "sugar" refers to any edible sugar, carbohydrate, or sugar substitute. According to some embodiments, the sugar is a nonanimal sugar. According to some embodiment, the sugar is plant-derived sugar. According to some embodiments, the sugar is selected from a monosaccharide, disaccharide, and polysaccharide. According to some embodiments, the sugar is selected from glucose, fructose, mannose, xylose, arabinose, sucrose, dextrose, maltose, and galactose. According to some embodiments, the sugar is dextrose. According to some embodiments, alternative dairy product of the present invention comprises from 0.5 to 6 wt% of a sweetener such as a sugar. According to some embodiments, the alternative dairy product of the present invention comprises from 1 to 5 wt% of a sweetener such as a sugar. According to some embodiments, the alternative dairy product of the present invention comprises from 2 to 4 wt% of about 3 wt% of a sweetener such as a sugar.

[0069] According to some embodiments, the alternative dairy composition, such as the alternative dairy milk of the present invention comprises from 0.01 to 0.3 wt% of NaCl. According to some embodiments, the alternative dairy product of the present invention comprises from 0.02 to 0.25 wt% of NaCl. According to some embodiments, the alternative dairy product of the present invention comprises from 0.03 to 0.2 wt% of NaCl. According to some embodiments, the alternative dairy product of the present invention comprises from 0.05 to 0.15 wt% of NaCl. According to some embodiments, the alternative dairy product of the present invention comprises from 0.08 to 0.12 wt% of NaCl. According to some embodiments, the alternative dairy product of the present invention comprises about 0.1 wt% of NaCl.

[0070] According to some embodiments, the alternative dairy composition, such as the alternative dairy milk of the present invention comprises from 0.1 to 0.6 wt% of calcium as calcium carbonate. According to some embodiments, the alternative dairy product of the present invention comprises from 0.3 to 0.5 wt% of calcium as calcium carbonate. According to some embodiments, the alternative dairy product of the present invention comprises from 0.1 to 0.3 wt% of calcium as calcium carbonate. According to some embodiments, the alternative dairy product of the present invention comprises from 0.15 to 0.25 wt% of calcium as calcium carbonate.

[0071] According to some embodiments, the alternative dairy composition, such as the alternative dairy milk of the present invention comprises from 0.005 to 0.1 wt% of L- Cystine. According to some embodiments, the alternative dairy product of the present invention comprises from 0.007 to 0.08 wt% of L-Cystine. According to some embodiments, the alternative dairy product of the present invention comprises from 0.01 to 0.05 wt% of L- Cystine.

[0072] According to any one of the above embodiments, the alternative dairy composition comprises water up to 100% w / w.

[0073] According to any one of the above embodiments, the particles of the alternative dairy composition comprising a lipid, a sweetener, NaCl, L-Cystine, and calcium have an average particle size of from 900 nm to 2000 nm. According to any one of the above embodiments, the particles have the average particle size of from 1000 to 1500 nm. According to any one of the above embodiments, the particles have the average particle size of from 1100 to 1300 nm.

[0074] According to any one of the above embodiments, the particles of the alternative dairy composition, comprising a lipid, a sweetener, NaCl, L-Cystine and calcium, have a median size of from 500 to 900 nm. According to some embodiments, the particles have a median size of from 500 to 850 nm. According to some embodiments, the particles have a median size of from 500 to 800 nm. According to some embodiments, the particles have a median size of from 600 to 850 nm. According to some embodiments, the particles have a median size of from 550 to 850.. According to some embodiments, the particles have a median size of from 600 to 850 nm. According to some embodiments, the particles have a median size of from 600 to 800 nm. According to some embodiments, 99% of the particles have a size of less than 1200 nm. According to some embodiments, 99% of the particles have a size of less than 1100 nm. According to some embodiments, 99% of the particles have a size of less than 1050 nm. According to some embodiments, 99% of the particles have a size of less than 1000 nm. According to some embodiments, 99% of the particles have a size of less than 950 nm. According to some embodiments, 99% of the particles have a size of less than 900 nm. According to some embodiments, 99% of the particles have a size of less than 850 nm.

[0075] According to some embodiments, the alternative dairy composition comprises: i. from about 0.1 w / w% to about 3.0 % w / w BLG, ii. from about 0.1 % w / w to about 0.3 % w / w total pectins, iii. from about 0.1 % w / w to about 5 % w / w fat, iv. from about 0.1 % w / w to about 6 % w / w sweetener, v. from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), vi. from about 0.005 % w / w to about 0.1 % w / w L-Cystine, and vii. from about 0.15 % w / w to about 0.25 % w / w calcium, and viii. up to 100 w / w water.

[0076] According to some embodiments, the alternative dairy composition comprises i. from about 0.5 % w / w to about 2 % w / w BLG, ii. from about 0.1 % w / w to about 0.3 % w / w total pectins, iii. from about 0.5 % w / w to about 3.5 % w / w fat, iv. from about 1 % w / w to about 1.75 % w / w sweetener, v. from about 0.05 % w / w to about 0.15 % w / w NaCl (edible salt), vi. from about 0.01 % w / w to about 0.05 % w / w L-Cystine, and vii. from about 0.4 % w / w to about 0.6 % w / w calcium, and viii. up to 100 % w / w water.

[0077] According to any one of the above embodiments, the pH of the alternative dairy composition is from 5 to 8. According to some embodiments, the pH of the alternative dairy composition is from 6 to 8. According to some embodiments, the pH of the alternative dairy composition is from 6.5 to 7.5. According to some embodiments, the pH of the alternative dairy composition is about 7. According to some embodiments, the alternative dairy composition is devoid of a divalent ion chelator. The terms "chelating agent", “chelating salt”, and "chelator" are used herein interchangeably and refer to agents capable of chelating cations, such as divalent ions. In some embodiments, the chelating agent chelates divalent ions. In some examples, the chelating agent is selected from mono- and di-potassium phosphate and sodium hexametaphosphate (SHMP). Therefore, according to some embodiments, the alternative dairy composition is devoid of from mono- and di-potassium phosphate and sodium hexametaphosphate (SHMP).

[0078] According to some embodiments, the alternative dairy composition is devoid of Gum Arabic, Carrageenan Lambda, Gellan, sodium hexametaphosphate (SHMP), monopotassium phosphate, di-potassium phosphate, a phosphate buffer, polyphosphates, divalent ion chelators and / or a citrate buffer According to some embodiments, the alternative dairy composition is devoid of phosphates. According to some embodiments, the alternative dairy composition is devoid of polyphosphates. According to some embodiments, the alternative dairy composition is devoid of chelators of divalent ions. According to some embodiments, the alternative dairy composition is devoid of a buffer. The terms “devoid”, “does not include” and “does not comprise” may be used interchangeably and refer to a composition that does not include, contain or comprise a particular component, e.g., said composition comprises less than 0.1 wt%, less than 0.01 wt%, or less than 0.001 wt% of the component.

[0079] According to some embodiments, the alternative dairy composition is formulated as an alternative milk composition. Therefore, according to some embodiments, the present invention provides an alternative milk comprising a plurality of particles, wherein the particles comprise a complex of a non-animal beta-lactoglobulin (BLG) protein and a nonanimal pectin. According to some embodiments, BLG is a recombinant BLG. According to some embodiments, the alternative milk comprises (a) from about 0.1 w / w% to about 3.5 % w / w BLG, (b) from about 0.25 % w / w to about 0.45 % w / w total pectins, (c) from about 0.1 % w / w to about 5 % w / w fat, (d) from about 0.1 % w / w to about 6 % w / w sweetener, (e) from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), (f) from about 0.005 % w / w to about 0.1 % w / w L-Cy stine, and (g) from about 0.15 % w / w to about 0.25 % w / w calcium, and (h) up to 100 w / w water. According to some embodiments, the particles comprise a core comprising a complex of a non-animal BLG and a non-animal pectin. According to some embodiments, alternative milk composition has a pH of from 6 to 8. According to some embodiments, the alternative milk comprises from 0.25 to 3 % w / w BLG. According to some embodiments, the alternative milk comprises from 0.5 to 2 % w / w BLG. According to some embodiments, the alternative milk comprises from 0.1 to 0.3 % pectins. According to some embodiments, the alternative dairy composition is devoid of Gum Arabic, Carrageenan Lambda, Gellan, sodium hexametaphosphate (SHMP), mono-potassium phosphate, dipotassium phosphate, a phosphate buffer, polyphosphates, divalent ion chelators and / or a citrate buffer.

[0080] According to some embodiments, the alternative dairy composition, such as an alternative milk composition, is stable at high temperatures and / or low pH. According to some embodiments, the alternative dairy composition is stable at a temperature of from 60 to 100°C. According to some embodiments, the alternative dairy composition is stable at a temperature of from 70 to 95°C. According to some embodiments, the alternative dairy composition is stable at a temperature of from 75 to 90°C. According to some embodiments, the alternative dairy composition is stable at a temperature of from 80 to 90°C. According to some embodiments, the alternative dairy composition is stable at pH from 5 to 7. According to some embodiments, the alternative dairy composition is stable at pH from 5 to 6. The term "stable" as used here have a meaning that no precipitation occurs within 1 hour after exposure to the heat and / or pH. According to some embodiments, no precipitation occurs within 2 hours after exposure to the heat and / or pH. According to some embodiments, no precipitation occurs within 3 hours after exposure to the heat and / or pH. According to some embodiments, no precipitation occurs within 6 hours after exposure to the heat and / or pH.

[0081] According to another aspect, the present invention provides a food product comprising the alternative dairy composition according to any one of the above aspects and embodiments. All terms, embodiments and definitions disclosed in any one of the above aspects apply and are encompassed herein as well. According to some embodiments, the alternative dairy composition is an alternative milk composition. According to some embodiments, the alternative milk comprising a plurality of particles, wherein the particles comprise a complex of a non-animal beta-lactoglobulin (BLG) protein and a non-animal pectin. According to some embodiments, BLG is a recombinant BLG. According to some embodiments, the alternative milk comprises (a) from about 0.1 w / w% to about 3.5 % w / w BLG, (b) from about 0.25 % w / w to about 0.45 % w / w total pectins, (c) from about 0.1 % w / w to about 5 % w / w fat, (d) from about 0.1 % w / w to about 6 % w / w sweetener, (e) from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), (f) from about 0.005 % w / w to about 0.1 % w / w L-Cystine, and (g) from about 0.15 % w / w to about 0.25 % w / w calcium, and (h) up to 100 w / w water. According to some embodiments, the alternative milk comprises (a) from about 0.25 w / w% to about 3.0 % w / w BLG, (b) from about 0.1 % w / w to about 0.3 % w / w total pectins, (c) from about 0.1 % w / w to about 5 % w / w fat, (d) from about 0.1 % w / w to about 6 % w / w sweetener, (e) from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), (f) from about 0.005 % w / w to about 0.1 % w / w L-Cystine, and (g) from about 0.15 % w / w to about 0.25 % w / w calcium, and (h) up to 100 w / w water. According to some embodiments, the alternative milk comprises (a) from about 0.5 w / w% to about 2.0 % w / w BLG, (b) from about 0.1 % w / w to about 0.3 % w / w total pectins, (c) from about 0.1 % w / w to about 5 % w / w fat, (d) from about 0.1 % w / w to about 6 % w / w sweetener, (e) from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), (f) from about 0.005 % w / w to about 0.1 % w / w L-Cystine, and (g) from about 0.15 % w / w to about 0.25 % w / w calcium, and (h) up to 100 w / w water. According to some embodiments, the particles comprise a core comprising a complex of a non-animal Beta-lactoglobulin (BLG) protein and a non-animal pectin. According to some embodiments, alternative milk composition has a pH of from 6 to 8.

[0082] According to any one of the above embodiments, the BLG is deaminated. According to some embodiments, the BLG is enzymatically deaminated. According to some embodiments, the BLG is enzymatically partially deaminated. According to some embodiments, the BLG is enzymatically partially deaminated by protein glutaminase. According to some embodiments, from 30 to 60% of the BLG is deaminated. According to some embodiments, the BLG is enzymatically partially deaminated by protein glutaminase. According to some embodiments, from 35 to 55% of the BLG is deaminated. According to some embodiments, the BLG is enzymatically partially deaminated by protein glutaminase. According to some embodiments, from 40 to 50% of the BLG is deaminated.

[0083] According to any one of the above embodiments, the alternative food product has a foaming ranking of 5 or more. According to any one of the above embodiments, the alternative food product has a foaming ranking of 5.5 or more. According to any one of the above embodiments, the alternative food product has a foaming ranking of 6.5 or more. According to any one of the above embodiments, the alternative food product has a foaming ranking of 7 or more. According to any one of the above embodiments, the alternative food product has a foaming ranking of from 6 to 10. According to any one of the above embodiments, the alternative food product has a foaming ranking of from 7 to 10. According to any one of the above embodiments, the alternative milk product has a foaming ranking substantially identical to that of milk. Foaming ranking is measured as described in the Examples. The ranking is assessed by subjective analysis, wherein 0 refers to no foam and 10 represents a milk-like foam.

[0084] According to some embodiments, the food product is selected from coffee, tea, Masala Chai, chocolate, matcha, milkshake, and alcoholic beverages. According to some embodiments, the food product is coffee. According to some embodiments, the food product is an instant coffee.

[0085] According to some embodiments, the food product composition comprising the food product is an instant coffee and the alternative dairy composition is an alternative milk composition. According to some embodiments, the instant coffee has pH of from 5 to 6. According to some embodiments, the instant coffee has a Color (L, Hunter units) of 15 to 40 when the content of the alternative milk composition is from 3 to 20% v / v of the coffee.

[0086] According to yet another aspect, the present invention provides a method of preparation of an alternative dairy composition of the present invention. According to some embodiments, the resulting alternative dairy composition is as described in any one of the above aspects and embodiments. All terms, embodiments and definitions disclosed in any one of the above aspects apply and are encompassed herein as well.

[0087] According to some embodiments, the method of preparation of an alternative dairy composition of the present invention comprises: i. Combining a composition comprising hydrated pectin with a non-animal dairy protein; ii. Setting a pH from 3 to 6, thereby obtaining a complex of the pectin and the nonanimal dairy protein; iii. Optionally adding a non-animal lipid; iv. Optionally homogenizing the composition; and v. Pasteurizing the composition via ultra-high temperature pasteurization. According to some embodiments, step (i) comprises stirring the hydrated pectin with the non-animal dairy protein at from 200 to 2000 RPM for at least 15, e.g. for from 15 to 90 min or for from 15 to 60 min or for about 30 min. According to some embodiments, step (i) is performed at a temperature below 80°C. According to some embodiments, step (i) is performed at a temperature of from 30 to 80°C. According to some embodiments, step (i) is performed at a temperature of from 40 to 70°C. According to some embodiments, step (i) is performed at a temperature of from 40 to 60°C. According to some embodiments, step (i) is performed at a temperature of about 50°C. According to some embodiments, step (i) is performed by stirring at 300 to 800 RPM for from 15 to 45 min at a temperature of from 40 to 60°C. According to some embodiments, the step comprises a compete hydration of the non-animal dairy protein. According to some embodiments, the step comprises a compete hydration of the non-animal BLG.

[0088] According to some embodiments, the method comprises adding from 0.1% to 3.5% w / w BLG. According to some embodiments, the method comprises adding from 0.25% to 3% w / w BLG. According to some embodiments, the method comprises adding from 0.1% to 3.5% w / w, from 0.2% to 3.4% w / w, from 0.3% to 3.3% w / w, from 0.4% to 3.2% w / w, from 0.5% to 3.1% w / w, from 0.6% to 3.0% w / w, from 0.7% to 2.9% w / w, from 0.8% to 2.8% w / w, from 0.9% to 2.7% w / w, from 1.0% to 2.6% w / w, from 1.0% to 2.5%, from 1.1% to 2.5% w / w, from 1.2% to 2.4% w / w, from 1.3% to 2.3% w / w, from 1.4% to 2.2% w / w, from 1.5% to 2.1% w / w, from 1.6% to 2.0% w / w, from 1.7% to 1.9% w / w BLG. According to some embodiments, the method comprises adding from 0.25% to 3 % w / w BLG. According to some embodiments, the method comprises adding from 0.25% to 2.5 % w / w BLG. According to some embodiments, the method comprises adding from 0.5% to 2 % w / w BLG. According to some embodiments, the method comprises adding from 0.5% to 1.5 % w / w BLG.

[0089] All concentrations disclosed with respect to a method disclosed herein also refer to the concentration in the final products disclosed herein.

[0090] According to some embodiments, the BLG is added to a composition comprising a hydrated pectin in water. According to some embodiments, the composition of hydrated pectin comprises from 0.1% to 0.3% w / w of pectin. According to some embodiments, the composition of hydrated pectin comprises from 0.15% to 0.25% w / w of pectin. According to some embodiments, the composition of hydrated pectin comprises from 0.2% to 0.45% w / w of pectin. According to some embodiments, the composition of hydrated pectin comprises from 0.2% to 0.4% w / w of pectin. The amount of pectin refers collectively to all types of pectins in the composition. According to some embodiments, the composition of hydrated pectin comprises from about 0.22 to about 0.38% w / w of pectin According to some embodiments, the composition of hydrated pectin comprises from about 0.25 to about 0.35% w / w of pectin. According to some embodiments, the composition of hydrated pectin comprises about 0.3% w / w of pectin. According to some embodiments, the pectin is a mixture of LM and HM. According to some embodiments, According to some embodiments, the pectin is a mixture of LMA and HM. According to some embodiments, the pectin is a mixture of LM and HMA. According to some embodiments, the pectin is a mixture of LMA and HMA. According to some embodiments, the ratio between LM and HM is from about 1:2 to about 1: 10. According to some embodiments, the ratio between LM and HM is from about 1:2 to about 1:8. According to some embodiments, the ratio between LM and HM is from about 1:25 to about 1:75. According to some embodiments, the ratio between LM and HM is from about 27:72 to 16:84 or about 16:84. According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:2 to about 1:20. According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:4 to about 1: 18. According to some embodiments, the ratio between LM(A) and HM(A) is from about 1:4 to about 1: 16. According to some embodiments, the ratio between LM and HM is about 1:5.25. According to some embodiments, the pectin is amidated. According to some embodiments, the pectin is a mixture of LMA and HM and the ratio is as stated in any one of the above embodiments. According to some embodiments, the pectin is a mixture of LM and HMA and the ratio is as stated in any one of the above embodiments.

[0091] According to some embodiments, step (ii) comprises setting a pH to a value of from 3 to 6. According to some embodiments, step (ii) comprises setting a pH to a value of from 3.5 to 5.5. According to some embodiments, step (ii) comprises setting a pH to a value of from 5 to 5.5. According to some embodiments, step (ii) comprises setting a pH to a value of from 4.25 to 5.25. According to some embodiments, step (ii) comprises setting a pH to a value of from 4.5 to 5. According to some embodiments, step (ii) comprises setting a pH to a value of about 4.75. According to some embodiments, setting the pH to the desired value comprises acidifying the composition. According to some embodiments, setting the pH to the desired value may be performed by any known method. According to some embodiments, setting the pH to the desired value is performed by adding any edible acid. According to some embodiments, setting the pH to the desired value is performed by adding HC1.

[0092] According to any one of the above embodiments, the method further comprises deaminating BLG prior to addition of pectin. According to some embodiments, the deamination is a partial deamination. According to some embodiments, the deamination comprises deamination of from 30 to 60% or from 35 to 55% or from 40 to 55% of BLG. According to some embodiments, the deamination is enzymatic deamination. According to some embodiments, the deamination is performed by protein glutaminase. According to some embodiments, the deamination is performed by adding 0.01 to 0.05% w / w protein glutaminase. According to some embodiments, the deamination is performed by adding 0.01 to 0.03% w / w of about 0.02% protein glutaminase. According to some embodiments, the deamination is performed at a temperature between 45 to 65°C.

[0093] All compounds are as described in any one of above aspects and embodiments.

[0094] According to some embodiments, the method comprises adding a non-animal lipid at step (iii). According to some embodiments, the non-animal lipid is a fat. According to some embodiments, prior to adding the lipid the composition is heated to a temperature between 60°C and 80°C. According to some embodiments, prior to adding the lipid the composition is heated to a temperature between 60°C and 70°C. According to some embodiments, prior to adding the lipid the composition is heated to a temperature about 65°C. According to some embodiments, upon adding the lipid the composition is further stirred for from 5 to 30 min at from 5 to 20,000 RPM. According to some embodiments, the lipid is a non-animal lipid. According to some embodiment, the lipid is plant-derived lipid. According to some embodiments, the lipids comprise an oil. According to some embodiment, the oil is selected from shea oil, sunflower oil, coconut oil, rapeseed oil, nut oil, palm oil, kernel oil, olive oil, soya oil, cotton oil, and cocoa butter (Theobroma oil). According to some embodiments, the method comprises adding from 0.5 to 6 wt% of the non-animal fat. According to some embodiments, the method comprises adding from 1 to 5 wt% of the non-animal fat. According to some embodiments, the method comprises adding from 2 to 4 wt% of about 3 wt% of the non-animal fat such as plant oil.

[0095] According to some embodiment, the method comprises homogenizing the composition at step (iv). The terms "homogenized" and "homogenization" refer to the process or to the product that passed the process of homogenization. Homogenization may be performed by any known method and / or device. According to some embodiments, the homogenization is performed in 1, 2, 3 or 4 stages. According to some embodiments, the homogenization is performed at from about 50 to about 400 bar. According to some embodiments, homogenization is performed for from 2 to 120 minutes. According to the principles of the present invention, any homogenization stage and any homogenization pressure found to homogenize the compositions and products of the present invention are included. According to some embodiments, homogenization may be performed in two steps. Non-limiting examples are stage homogenizing at 50 or 60 bar and then at 200 bar. According to some embodiments, the composition is heated prior to homogenization, e.g. heated up to 50°C or up to 60°C or up to 70°C. According to some embodiments, the composition is homogenized at about 600 bar and 65°C using a two-stage homogenization process.

[0096] According to some embodiments, the method comprises adding a non-animal lipid at step (iii) and homogenizing at step (iv).

[0097] According to some embodiments, the method comprises hydrating the pectin prior to step (i). According to some embodiments, hydrating the pectin comprises dispersing pectin in water and stirring at from 200 to 2000 RPM for at least 15, e.g. for from 15 to 90 min or for from 15 to 60 min or about 30min. According to some embodiments, hydrating the pectin is performed at a temperature below 80°C. According to some embodiments, hydrating the pectin is performed at a temperature of from 30 to 80°C. According to some embodiments, hydrating the pectin is performed at a temperature of from 40 to 70°C. According to some embodiments, hydrating the pectin is performed at a temperature of from 40 to 60°C. According to some embodiments, hydrating the pectin is performed at a temperature of about 50°C. According to some embodiments, hydrating the pectin is performed by stirring at 300 to 800 RPM for from 15 to 45 min at a temperature of from 40 to 60°C. According to some embodiments, hydrating the pectin comprises its compete hydration. According to some embodiments, the pectin is as described above.

[0098] According to some embodiments, the method further comprises adding at least one of: a sweetener, L-cystine, and NaCl at step (i) or before step (i), e.g. before hydrating pectin. According to some embodiments, the method comprises adding from 0.5 to 6 wt% of a sweetener such as a sugar. According to some embodiments, the method comprises adding from 1 to 5 wt% of a sweetener such as a sugar According to some embodiments, the method comprises adding from 2 to 4 wt% of about 3 wt% of a sweetener such as a sugar. According to some embodiment, the sugar is plant-derived sugar. According to some embodiments, the sugar is selected from a monosaccharide, disaccharide, and polysaccharide. According to some embodiments, the sugar is selected from glucose, fructose, mannose, xylose, arabinose, sucrose, dextrose, maltose, and galactose. According to some embodiments, the sugar is dextrose.

[0099] According to some embodiments, the method comprises adding from 0.005 to 0.1 wt% of L-Cystine. According to some embodiments, the method comprises adding from 0.007 to 0.08 wt% of L-Cystine. According to some embodiments, the method comprises adding from 0.01 to 0.05 wt% of L-Cystine.

[0100] According to some embodiments, the method comprises adding from 0.01 to 0.3 wt% of NaCl. According to some embodiments, the method comprises adding from 0.02 to 0.25 wt% of NaCl. According to some embodiments, the method comprises adding from 0.03 to 0.2 wt% of NaCl. According to some embodiments, the method comprises adding from 0.05 to 0.15 wt% of NaCl. According to some embodiments, the method comprises adding from 0.08 to 0.12 wt% of NaCl. According to some embodiments, the method comprises adding about 0.1 wt% of NaCl.

[0101] According to some embodiments, the method further comprises adjusting the pH of the composition to a value of from 6.5 to 7.5 in an additional step (vi). According to some embodiments, the method further comprises adjusting the pH of the composition to the value of from 6.5 to 7.5 after any one of stages (iii)-(v). According to some embodiments, pH adjustment is carried out by adding calcium carbonate.

[0102] According to some embodiments, the method of preparation of an alternative dairy composition, the method comprises:

[0103] (i) optionally dissolving in water from about 0.1 % w / w to about 6 % w / w of a sweetener, from about 0.01 % w / w to about 0.3 % w / w NaCl, from about 0.005 % w / w to about 0.1 % w / w L-Cystine, (ii) hydrating from about 0.1 % w / w to about 0.3 % w / w total pectins, preferably a mixture of LM and HMA,

[0104] (iii) adding about 0.1 w / w% to about 3 % w / w BLG and fully hydrating the BLG,

[0105] (iv) setting pH to a value of from 4.5 to 5, preferably to a value of 4.75

[0106] (v) optionally adding and dispersing from about 0.1 % w / w to about 5 % w / w of a non-animal lipid, optionally with preheating of the composition to from 60 to 70°C.

[0107] (vi) optionally homogenizing the composition, and

[0108] (vii) pasteurizing the composition via ultra-high temperature pasteurization.

[0109] According to some embodiments, the present invention provides a method of preparation of an alternative dairy composition, the method comprising:

[0110] (i) dissolving in water from about 0.1% % w / w to about 6% w / w sweetener, from about 0.01 % w / w to about 0.3 % w / w NaCl, from about 0.005 % w / w to about 0.1 % w / w L-Cystine,

[0111] (ii) hydrating in the composition obtained in step (i) from about 0.1 % w / w to about 0.3 % w / w total pectins, preferably a mixture of LM(A) and HM(A),

[0112] (iii) adding about 0.2 w / w% to about 2 % w / w BLG to the composition of step (ii) and fully hydrating the BLG,

[0113] (iv) setting pH to a value of from 4.5 to 5, preferably to a value of 4.75

[0114] (v) adding and dispersing from about 0.1 % w / w to about 5 % w / w lipid in to composition obtained in step (iv), optionally with preheating of the composition to from 60 to 70°C.

[0115] (vi) homogenizing the composition obtained in step (v), and

[0116] (vii) pasteurizing the composition obtained in step (vi) via ultra-high temperature pasteurization.

[0117] According to some embodiments, the present invention provides a method of preparation of an alternative dairy composition, the method comprises:

[0118] (i) dissolving in water from about 1 % w / w to about 1.75 % w / w sweetener, from about 0.05 % w / w to about 0.153 % w / w NaCl, from about 0.01 % w / w to about 0.05 % w / w L-Cystine,

[0119] (ii) hydrating in the composition obtained in step (i) from about 0.1 % w / w to about 0.3 % w / w total pectins, preferably a mixture of LM(A) and HM(A), (iii) adding to the composition obtained in step (ii) about 0.5 w / w% to about 2 % w / w BLG and fully hydrating the BLG,

[0120] (iv) setting pH to a value of from 4.5 to 5, preferably to a value of 4.75

[0121] (v) adding and dispersing from about 1.0 % w / w to about 3.5 % w / w lipid in the composition obtained in step (iv), optionally with preheating of the composition to from 60 to 70°C.

[0122] (vi) homogenizing the composition obtained in step (v), and

[0123] (vii) pasteurizing the composition obtained in step (vi) via ultra-high temperature pasteurization.

[0124] According to some above embodiments, the method comprises partially deaminating BLG. According to any one of the above embodiments, hydration of pectin and / or nonanimal BLG is performed gradually. According to any one of the above embodiments, setting a pH at step (ii) is performed gradually.

[0125] According to some embodiments, the resulting alternative dairy composition comprises a plurality of particles, each particle comprises a core consisting of the non-animal dairy protein and pectin. According to some embodiments, the particles have an average particle size of 900 nm to 2000 nm. According to some embodiments, the particles have the average particle size of from 1000 to 1500 nm. According to some embodiments, the particles have the average particle size of from 1100 to 1300 nm. According to some embodiments, the median size of particles is from 500 to 900 nm. According to some embodiments, the median size of particles is from 500 to 850 nm, from 600 to 850 nm, or from 600 to 850 nm. According to some embodiments, 99% of the particles have a size of less than 1200 nm. According to some embodiments, the particles have a size as describe in any one of the above aspects and embodiments.

[0126] According to any one of the above embodiments, the alternative dairy composition has a foaming ranking of 5 or more. According to any one of the above embodiments, the alternative dairy composition has a foaming ranking of 5.5 or more. According to any one of the above embodiments, the alternative dairy composition has a foaming ranking of 6.5 or more. According to any one of the above embodiments, the alternative dairy composition has a foaming ranking of 7 or more. According to any one of the above embodiments, the alternative dairy composition has a foaming ranking of from 6 to 10. According to any one of the above embodiments, the alternative dairy composition has a foaming ranking of from 7 to 10. According to any one of the above embodiments, the alternative dairy composition has a foaming ranking substantially identical to that of milk.

[0127] According to some embodiments, the method is devoid of adding one or more of the compound selected from Gum Arabic, Carrageenan Lambda, Gellan, SHMP, phosphate buffer, polyphosphates, divalent ion chelators and citrate buffer. According to some embodiments, the method is devoid of adding a buffer. According to some embodiments, the method is devoid of adding a phosphate. According to some embodiments, the method is devoid of adding a divalent ion chelators. The term "divalent ion chelators" and "chelators of divalent ions" are used herein interchangeably.

[0128] According to some embodiments, the ultra-high temperature pasteurization comprises heating to from 110 to 125°C for from 1 to 10 sec. According to some embodiments, the ultra-high temperature pasteurization comprises heating to from 115 to 125 °C for from 1 to 5 sec. According to some embodiments, the ultra-high temperature pasteurization comprises heating to from 120 to 125°C for from 1 to 3 sec.

[0129] According to some embodiments, the method further comprising heating the composition at step (ii) to from 70 to 90°C.

[0130] According to some embodiments, the present invention provides an alternative dairy composition prepared by a method according to any one of the above aspects and embodiments. According to some embodiments, the alternative dairy composition is an alternative dairy composition. According to some embodiments, the alternative dairy composition is an alternative milk. Therefore, according to some embodiments, the present invention provides an alternative milk prepared by the methods according to any one of the above embodiments.

[0131] The terms “a,” “an,” and “the” ” are used herein interchangeably and mean one or more.

[0132] The term “and / or” is used to indicate one or both stated cases may occur, for example A and / or B includes, (A and B) and (A or B).

[0133] The term “or,” as used herein, denotes alternatives that may, where appropriate, be combined; that is, the term “or” includes each listed alternative separately as well as their combination if the combination is not mutually exclusive. The terms “comprising”, "comprise(s)", "include(s)", "having", "has" and "contain(s)," are used herein interchangeably and have the meaning of “consisting at least in part of’. When interpreting each statement in this specification that includes the term “comprising”, features other than that or those prefaced by the term may also be present. Related terms such as “comprise” and “comprises” are to be interpreted in the same manner. The terms “have”, “has”, having” and “comprising” may also encompass the meaning of “consisting of’ and “consisting essentially of’, and may be substituted by these terms. The term “consisting of’ excludes any component, step or procedure not specifically delineated or listed. The term “consisting essentially of’ means that the composition or component may include additional ingredients, but only if the additional ingredients do not materially alter the basic and novel characteristics of the claimed compositions or methods. Throughout this specification, unless the context requires otherwise, the word “comprise” or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0134] As used herein, the term “about”, when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of + / -10%, or + / -5%, + / -1%, or even + / -0.1% from the specified value.

[0135] Having now generally described the invention, the same will be more readily understood through reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the present invention.

[0136] EXAMPLES

[0137] Abbreviations

[0138] BLG - Beta-lactoglobulin; DA - degree of amidation; DE - Degree of esterification; DPP - Di-potassium phosphate; HC1 - Hydrochloric acid; HM - High methoxyl pectin; HMA - High methoxyl pectin amidated; LM - Low methoxyl pectin; LMA - Low methoxyl pectin amidated; MPP - Mono-potassium phosphate; NaOH - Sodium hydroxide; SHMP - Sodium hexametaphosphate; UHT - Ultra high temperature.

[0139] Materials & Methods

[0140] Remilk recombinant beta-lactoglobulin Bos taunts, Variant B). Pectins purchased from CP Kelco: SLENDID® Specialty Pectin Type 200, GENU® BETA Pectin, GENU® pectin type DF, GENU® Pectin type LM-106 AS -YA, GENU® Pectin type YM-115-H, GENU® Pectin YA-720, GENU® Pectin YA-700. In addition, Pectin Classic CM 201, Pectin APC147 and APC201 were obtained from Ingredion Co. Ltd. and DSM (respectively). Gum Arabic was purchased from Nexira (INSTANTGUM AA). Lambda carrageenan was obtained from CP-Kelco (GENU). Gellan gum was acquired from Danisco (GRINDSTED® Gellan DAI 200). Fat (Crokvitol™) was acquired from BUNGE. Table sugar and salt (Sugat) were purchased from a local supermarket (Rehovot, Israel). L-Cystine, di-potassium phosphate (DPP), mono-potassium phosphate (MPP) Sodium hydroxide (NaOH) and hydrochloric acid (HC1) were purchased from Sigma (Rehovot, Israel). Sodium hexa-metaphosphate was acquired from ICL (JOHA®, Israel). Calcium carbonate (Calcipur® 110 KP) was purchased from Omya. Instant coffee powder (Nescafe Tasters Choice) was purchased from a local supermarket (Rehovot, Israel).

[0141] Methods

[0142] Preparation of BLG-polysaccharide complexes

[0143] In the development of stable and useful BLG-polysaccharide complexed that may be used in an alternative milk and would be stable in high temperature several factors were tested.

[0144] The process of preparation of the complexes is sequential and includes several steps

[0145] Step a

[0146] To prepare BLG and polysaccharide complexes (specifically with pectin, Arabic gum, carrageenan, or gellan), the polysaccharide (0.1-1.0% w / w) was gradually dissolved in distilled water at 50°C and mixed thoroughly at 500 RPM for 45 minutes to ensure complete hydration. Following this, BLG was added to the solution (1.0, 1.5, or 3.0% w / w), and the mixture was further stirred for 45 minutes at 50 °C. As a control, a BLG solution was prepared under the same conditions without the addition of a polysaccharide.

[0147] Optionally, additional ingredients were incorporated, including L-cysteine (0.02% w / w) to prevent the release of volatile sulfur compounds after the heat treatment, sugar and salt (1.25% and 0.1% w / w, respectively) for taste adjustment. Step b

[0148] To facilitate the complexation of the two components, the pH was gradually lowered to 4.75 using 5M HC1 while maintaining the temperature at 50 °C and stirring at 400 RPM. The pH of the solution was monitored using a pH meter (Mettler Toledo). During this process, the solution turned from clear to turbid around pH~5.2 (the iso-electric point of BLG), signaling the formation of larger particles.

[0149] Step c

[0150] Next, the temperature was raised to 60 °C, and a vegetable fat (3.0% w / w) was added to the mixture, which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA- TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently (1) homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy), and (2) pasteurized with a tubular heat exchanger (121 °C, 2-second holding time; HTST / UHT Mini Pilot System, Armfield) or alternatively using a Vorwerk Thermomix TM 6 blender cooker (2L capacity; Vorwerk & Co Thermomix GmbH, Wuppertal, Germany) for 2-5 min at 85 °C. This treatment is referred to as “pasteurization heat treatment”.

[0151] Step d

[0152] In both cases, the pasteurized milks were bottled, cooled to room temperature and the final pH was raised to pH=6.8-7.0 using 5M NaOH or alternatively, calcium carbonate (0.5% w / w). Finally, the milk was stored at 4 °C for further evaluation.

[0153] Alternative step b

[0154] In addition to the method described above, an alternative process was tested with the aim of preventing pH adjustment after pasteurization (which is less desirable and less common in the food industry due to the risk of contamination after pasteurization). After reducing the pH to 4.75, the solution was heated to 85 °C for a duration of from 1 to 18 minutes using Vorwerk Thermomix TM 6 blender cooker. This treatment is referred to as “fixation heat step”. Then, the solution was cooled to room temperature, and the pH was adjusted to 6.8-7.0 via 5M NaOH. Then Step c as described above is performed - i.e. The remaining ingredients were added, and the solution was heated to 60 °C. Fat was introduced and homogenized as described above. The emulsion was then pasteurized and bottled as described above.

[0155] Evaluation of mouthfeel and time-dependent sedimentation of BLG-polysaccharide complex based-milk

[0156] A qualitative assessment of the BLG-polysaccharide complex stability during refrigeration, along with a sensory evaluation of the milk's taste, focusing on mouth-drying sensation, was conducted three days after preparation. Unstable and highly insoluble complexes tend to settle in the bottle after 24-48 hours, forming a distinct semi-solid phase. This was documented and rated on a scale of 0 to 3 or 0 to 5, where 0 represents no visible sedimentation, and 3 and 5 indicate a prominent sediment phase easily observed. These unstable complexes also contribute to a notable mouthfeel, often described as dry or astringent. This sensation was quantitatively evaluated by trained tasters (n > 3), who rated the mouth-drying effect on a scale from 0 to 3 or 0 to 5, with 0 representing no dryness and 3 or 5 indicating a significant dry mouth sensation. In addition, the final milk prototype was tasted by 11 tasters, comprising naive and professional milk tasters, which ranked the mouth drying sensation from a scale of 1 to 7.

[0157] Evaluation of the alternative milk comprising BLG-polysaccharide complexes performance in instant coffee

[0158] To evaluate the physical stability and color profile of the prepared milk samples in instant coffee, the following procedure was carried out: milk samples were added to instant coffee (2 g of Nescafe Taster's Choice mixed with 150 g of boiling tap water) in three different volumes: 5, 15, and 25 mL (3.22% v / v, 9% v / v, 14.3% v / v). After preparation, the coffee was left to stand at room temperature for at least 30 minutes, during which the stability of the BLG-polysaccharide complex (specifically, aggregation and sedimentation) was observed and recorded along with the final pH of the coffee.

[0159] Color measurements

[0160] To evaluate the color of different coffee samples, aliquots were transferred to a designated measurement cell and placed on a benchtop colorimeter (CR5; Konica Minolta Bench-top, Japan) utilizing the CIE L*a*b* color space. All measurements were repeated at least two individual repetitions. Particle size measurements

[0161] To elucidate the effect of different process or / and formulation adjustments on the average particle size of BLG-pectin complexes, samples were introduced to 4 mL plastic cuvettes and the average particle size was determined using photon cross-correlation spectroscopy technique (Nanophox, Sympatec GmbH, Germany). Laser intensity was adjusted to -84%, desired temperature was 25 °C, measuring mode was cross correlation. Size measurements were repeated three times for each sample in two individual repetitions (at least) and results are shown as Xso.3 andX99,3.

[0162] Viscosity measurements

[0163] The viscosity of BLG-pectin based milks were measured via a viscometer, using a VOL-SP-6.7 spindle at 100 RPM and 25°C. All measurements were repeated at least two individual repetitions and reported in Cp units.

[0164] Foaming evaluation

[0165] To evaluate the impact of the different ratios of BLG, pectin and internal ratios of LM:HMA on the foaming properties of the formed milk, cappuccino’s type beverages were prepared using a semi-professional coffee machine. Milk was frothed (150 gr, up to -67 °C) and poured to freshly prepared coffee made from ground Arabica coffee beans. The foaming of each sample was ranked between 1-10, where 10 represents foaming properties of bovine milk. The foaming ranking comprises three specific attributes - foaming, pouring, and formation of latte art.

[0166] Example 1. Initial screening of BLG complexation with different polysaccharides

[0167] To understand which polysaccharides can form stable complexes with BLG (3.0% w / w), five anionic polysaccharides were selected and screened. The formulation contained only BLG and a polysaccharide, as described in Table 1. The solution was pasteurized at pH=4.75 at 85 °C for 18 min via Vorwerk Thermomix TM 6 blender cooker. Once cooled to room temperature, the solutions were added to coffee and the complex stability was visually evaluated.

[0168] As shown in Table 1, both high and low ester amidated pectins exhibited complex stability. Gum Arabic, carrageenan and gellan were unstable and formed aggregates. Table 1.

[0169] * Pectin LMA used was - GENU® Pectin type LM-106 AS-YA (DE=23%, DA=24%), Pectin HMA used was - GENU® Pectin YA-700 (Unknown DE and DA).

[0170] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0171] Example 2. Initial screening of conditions of BLG complexation with pectin

[0172] Pectin HMA (GENU® Pectin YA-700) at four different concentrations (0.3. 0.5. 0.7 and 0.9% w / w), was selected for further examination. Immediately after pH was set to 4.75, a fixation heat step was applied via Thermomix TM 6 blender cooker by heating at 85 °C for 18 min. Subsequently, the solutions were cooled, and the pH was increased using 5M NaOH to pH=7.0. Next, fat (3% w / w) was added to the solution, dispersed and homogenized at 65° C. The solutions were pasteurized to 85° C for 2 sec using Vorwerk Thermomix TM 6 blender cooker to simulate a final moderate pasteurization process. Overall, the solutions comprised of SHMP (0.1% w / w, hydrated during the initial stage of preparation), pectin, BLG and fat (3.0% w / w).

[0173] Results detailed in Table 2 show that none of the samples created stable complexes. In addition, pectin concentrations above 0.5% lead to a highly viscous texture which is undesirable for milk-like products. Table 2.

[0174] *Pectin HMA used was - GENU® Pectin YA-700.

[0175] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0176] Example 3. Impact of buffering agents on BLG-pectin complexation

[0177] A similar protocol preparation protocol to Example 2 was repeated with two changes: the concentration of Pectin HMA (GENU® Pectin YA-700) was fixed to 0.3%, and after heat treatment (via Thermomix TM 6 blender cooker at 85 °C for 18 min), the pH was set to ~7.0 using DPP (0.3, 0.5, 0.7 and 0.9% w / w). Overall, the solutions comprised of SHMP (0.1% w / w, hydrated during the initial stage of preparation), pectin, BLG and fat (3.0% w / w).

[0178] Results are detailed in Table 3. In addition, complex stability in coffee was evaluated when adding reduced volumes of the solutions prepared (5 and 15 mL, not shown in Table 3). In such cases, all solutions exhibited instability in coffee, forming large aggregates.

[0179] Overall, the results indicate that while the addition of DPP may help maintain protein stability, it does not provide stability across different volumes. Furthermore, at higher concentrations of DPP, the solution became highly viscous, which is undesirable. Table 3.

[0180] *Pectin HMA used was - GENU® Pectin YA-700 (Unknown DE and DA).

[0181] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0182] Example 4. Impact of shortened heat fixation duration on BLG-pectin complexation

[0183] This experiment was aimed to assess the impact of reducing fixation heat step duration from 18 minutes to either 6 minutes, 3 minutes or 2-second holding on complex stability in coffee. A basic milk formulation was prepared with 0.3% HMA, 3.0% BLG, 3.0% fat, 0.7% DPP (added post-heat treatment to adjust the pH to -7.0, as described previously), and 0.1% SHMP (0.1% w / w, hydrated during the initial stage of preparation). After fat homogenization, the solution was pasteurized using UHT treatment (121 °C, 2- second holding time). As shown in Table 4, reducing the fixation heat step duration after pH adjustment does not appears to have a stabilizing effect on the complex (exhibited in coffee prepared with 15 mL milk. In 25 mL milk all samples showed complex stability).

[0184] Table 4.

[0185] *Pectin HMA used was - GENU® Pectin YA-700 (Unknown DE and DA).

[0186] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0187] Example 5. The effect of pasteurization type on BLG-pectin complexation

[0188] The objective of this trial was to assess the impact of two pasteurization protocols (UHT treatment (121 °C, 2-second holding time), moderate pasteurization (85°C, 2-second holding time)) on the stability and color of the solution. The formulation was identical to that used in the Example 4 above, with the fixation heat step following pH reduction set to 4 minutes at 85°C.

[0189] As shown in Table 5, the moderate pasteurization resulted in highly stable complexes that did not aggregate in any of the tested volumes, while the UHT pasteurization produced unstable complexes. In terms of mouth drying perception, there was a slight improvement in both samples compared to the previous samples (18-minute fixation heat step). However, an unpleasant astringency persisted in the sample (n=4 tasters). Table 5.

[0190] *Pectin HMA used was - GENU® Pectin YA-700 (Unknown DE and DA).

[0191] Overall, we concluded here that the fixation heat step followed by increasing pH to 7.0 and pasteurizing once again can have devastating effects on complex stability. Therefore, in the following examples, the solution was UHT pasteurized in pH=4.75 without any fixation steps.

[0192] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0193] Example 6. The effect of pH on BLG-pectin complex stability

[0194] Heat fixation (heating at 85°C for 4 minutes) was tested following pH reduction to one of five different pH values: 4.50, 4.75, 5.00, 5.20 and 5.40 Next, fat was introduced to the solution (at 65°C), dispersed and homogenized as described previously. The formulation contained pectin (GENU® Pectin YA-700), BLG and fat 3% w / w. Following the creation of a stable emulsion, each sample was pasteurized by Thermomix TM 6 blender cooker at 85° C for 4 min and immediately cooled on ice to room temperature. Finally, the pH was adjusted to pH=6.8 using 5M NaOH.

[0195] Results shown in Table 6 and demonstrate that all samples were unstable in instant coffee, exhibiting an undesirable protein aggregation and sedimentation. However, it was noticed that pH between 5.0 to 4.75 resulted the less protein aggregation in comparison with higher / lower pH values. Therefore, further experimentations were applied by complexing at pH=4.75. In addition, all samples showed instability when added to coffee. This suggests that pasteurization duration has a higher impact on complex stability compared to temperature. Thus, further tests were conducted via UHT pasteurization (120°C, 2 sec holding).

[0196] Table 6.

[0197] *Pectin HMA used was - GENU® Pectin YA-700 (Unknown DE and DA).

[0198] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0199] Overall, it can be seen that the system is very complex, and the its development is not linear or straightforward. From the analysis of previous examples, it was cumulatively concluded that heat fixation step is not required when a pasteurization step is present, and may even damage the composition. Therefore, in the future examples the heat fixation step was omitted.

[0200] Example 7. Fabricating stable BLG-pectin complexes via UHT at pH=4.75 without fat

[0201] A formulation comprising HMA (GENU® Pectin YA-700) 0.3% w / w, BLG 3% w / w and SHMP 0.1% w / w (without fat) was examined. The pH was set to pH=4.75 and pasteurized via UHT heat exchanger as described above (121 °C, 2-second holding time). Subsequently, the pH was increased by 5M NaOH (Table 7) or increasing concentrations of DPP (0.1-0.7%, %w / w, Table 8).

[0202] The preparation method is as follows: SHMP (0.1% w / w) was hydrated in water for 20 min at ambient temperature. Next, temperature was raised to 50°C and 0.3% w / w pectin HMA (GENU® Pectin YA-700) was introduced to the solution and stirred at 500 RPM until fully hydrated. Subsequently, BLG (3.0% w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. In this experiment fat was not included and no heat fixation was used. Next, the solution was pasteurized via UHT pasteurization (121 °C, 2-second holding time). Finally, the pH was set to pH=7.0 using 5M NaOH or alternatively the pH was increased using DPP (0.1-0.7%, %w / w) as detailed in Table 7.0.

[0203] Results show that UHT pasteurization at pH=4.75 created complexes stable in instant coffee regardless of the method in which the pH was increased (DPP or NaOH). Moreover, results suggest that the interaction between BLG and pectin in these conditions are highly heat- and pH- stable, thus eliminating the need for buffering agents such as DPP.

[0204] Table 7.

[0205] Table 8. For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown). Example 8. Fabricating stable BLG-pectin complexes via UHT at pH=4.75 with fat

[0206] As a continuation of the Example 7 above, the same formulation was tested, but with the addition of fat 3% w / w. Additionally, as a control, pectin was removed from the formulation to verify its role in stabilizing the complex.

[0207] The preparation method is as follows: SHMP (0.1% w / w) was hydrated in water for 20 min at ambient temperature. Next, temperature was raised to 50°C and 0.3% w / w pectin HMA (GENU® Pectin YA-700) was introduced to the solution and stirred at 500 RPM until fully hydrated. Subsequently, BLG (3.0% w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and 3.0% w / w fat was added to the solution which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA-TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy). Next, the solution was pasteurized via UHT pasteurization (121 °C, 2-second holding time). Finally, the pH was set to pH=7.0 using 5M NaOH.

[0208] The results, presented in Table 9 indicate that the incorporation of fat did not compromise the complex's stability, and was successful in increasing the overall lightness of the coffee (L value) by approximately 4.5 units. As can be seen from the control, removal of the pectin had a detrimental effect on formulation stability, and it is required for the stability of BLG composition. From the results of the control, it is observed that a composition of BLG cannot be pasteurized as the protein aggregated and clomped in the pasteurizer.

[0209] Table 9.

[0210] *Pectin HMA used was - GENU® Pectin YA-700 (Unknown DE and DA).

[0211] In addition, pH and average particle diameter were monitored during every step of the process, with and without fat addition, as shown in Table 10. Overall, results indicate that the addition of fat increased the particle size by approximately 500 nm. Table 10.

[0212] For different reasons, particles produced by the materials and methods described in this example were inadequate to be included in high quality alternative dairy products (data not shown).

[0213] Example 9. Screening of different types of pectins The aim of this experiment was to explore the performance of different types of pectins in the same process and formulation as described previously in Example 8).

[0214] The preparation method is as follows: SHMP (0.1% w / w) was hydrated in water for 20 min at ambient temperature. Next, temperature was raised to 50°C and 0.3% w / w pectin was introduced to the solution and stirred at 500 RPM until fully hydrated. Overall, 12 different pectins or pectin combinations were screened as detailed in Table 11. Further details on the brand name, company, chemical specification and source of every pectin are detailed in Table 12. The overall amount of pectin in all samples was equal to 0.3% (%w / w). When combining HM and LMA pectins (APC 147 + APC 201), the ratio was set to be 0.215% HM and 0.085% LMA. Subsequently, BLG (3.0% w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and 3.0% w / w fat was added to the solution which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA- TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy). Next, the solution was pasteurized via UHT pasteurization (121 °C, 2-second holding time). Finally, the pH was set to pH=7.0 using 5M NaOH.

[0215] Results show that applying HM or LMA alone produce unstable complexes, however their combination gave the desirable attributes, in terms of stability, color, particle sedimentation and dry mouth feeling. Table 11.

[0216] * Dry mouth sensation was ranked between 1 to 3 (*, **, ***), where * denotes low dry mouth-feel sensation and *** denotes very dry mouth-feel sensation (according to n>3 tasters). ** Particle sedimentation was ranked between 1 to 3 (*, **, ***), where * denotes low particle sedimentation in the bottle and *** denotes high particle sedimentation in the bottle.

[0217] Table 12.

[0218] For different reasons, particles produced by the materials and methods described in this example were adequate to be included in high quality alternative dairy products. Example 10. Screening of the optimal ratio between HM(A):LM(A) pectins

[0219] An experiment was conducted to find the optimal ratio between the two types of pectin (LM and HMA (APC 201+APC 147), total concentration of pectin was set to 0.3%). Thus, the following ratios were screened: 16:84, 28:72, 35:65, 40:60, 50:50 (LM:HMA, respectively). The formulation and process are similar to previous experiments (BLG, pectin, SHMP, fat).

[0220] Overall, results indicate that increasing the amount of LM and reducing the amount of HMA negatively affected the stability of the complex. Moreover, LMA increment caused protein aggregation during the UHT treatment, which in turn resulted in pasteurization instability and fouling in the heat exchanger. While both 16:84 and 28:72 ratios possessed desirable attributes, 16:84 showed less sedimentation in the bottle, which indicates that the complex formed was highly soluble and stabilized.

[0221] Table 13A.

[0222] For different reasons, particles produced by the materials and methods described in this example were adequate to be included in high quality alternative dairy products.

[0223] The effect of a ratio between LM and HMA was further examined as follows. Alternative milk was prepared as described above, where the total pectin was kept at 0.3% w / w, and the total amount of BLG was kept at 1.5% w / w. The ratio between LM and HMA was adjusted as following: 1:99, 5:95, 10:90 and 16:84, LM and HMA, respectively. The results are shown in Tables 13B - 13D. Table 13B. The effect of different LM:HMA ratios on the physical parameters of milk

[0224] Table 13C. The effect of different LM:HMA ratios on the physical parameters of coffee

[0225] Table 13D. The effect of different LM:HMA ratios on the physical parameters of coffee

[0226] As shown in Tables 13B-13D, increasing LM and decreasing HMA resulted in only minor differences in instant coffee color and stability. However, the performance of foaming may be influenced by higher HMA levels, as indicated by the ranking results. It is apparent that the LM:HM ratio of 5:95 to 20:80 provides enhanced foaming effect.

[0227] Example 11. Screening of the optimal concentration of LM:HMA: pectin combination

[0228] Following the identification of the optimal type and ratio of pectins, the total concentration of pectin was investigated. Thus, LM:HMA (16:84 ratio) in three different concentrations were evaluated, 0.2%, 0.3% and 0.4% (% w / w). The preparation method is as follows: SHMP (0.1% w / w) was hydrated in water for 20 min at ambient temperature. Next, temperature was raised to 50°C and pectin (0.2, 0.3 or 0.4% w / w) was introduced to the solution and stirred at 500 RPM until fully hydrated. Subsequently, BLG (3.0% w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and 3.0% w / w fat was added to the solution which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA-TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy). Next, the solution was pasteurized via UHT pasteurization (121 °C, 2-second holding time). Finally, the pH was set to pH~7.0 using 5M NaOH.

[0229] As shown in Table 14, increasing the total amount of pectin to 0.4% caused complex instability, which in turn created protein aggregation and clomping during the pasteurization. On the other hand, reducing the total pectin concentration to 0.2% increased particle sedimentation and complex instability, which is reflected in the low L values shown in coffee in comparison to 0.3% pectin. Therefore, 0.3% pectin was selected for further analyses.

[0230] Table 14A.

[0231] In yet additional experiment the overall concentration of pectin was further varied and tested as follows.

[0232] Milk was prepared as described above, where the total pectin was 0.1, 0.3 or 0.5% w / w. The total amount of BLG was kept at 1.5% w / w and the ratio between LM:HMA was kept at 16:84. Results are shown in Tables 14B, 14C, and 14D. Table 14B. The effect of different pectin percentages on the physical parameters of milk

[0233] As shown in Table 14B, increasing the pectin concentration leads to a corresponding rise in milk viscosity. This affects sensory perception, as higher pectin levels may negatively impact viscosity, texture, and mouthfeel. Conversely, lower pectin concentrations appear to reduce the dryness sensation, which is considered favorable from a sensory perspective.

[0234] Table 14C. The effect of different pectin percentages on the physical parameters of coffee Table 14D. The effect of different pectin percentages on the physical parameters of coffee

[0235] Table 14E. The effect of different pectin percentages on foaming ranking

[0236] As shown in Table 14C-14E, a pectin concentration of 0.3% w / w resulted in greater lightness in instant coffee compared to concentrations of 0.1% and 0.5% w / w. Concentration between 0.1% and 0.3% is the preferred. Regarding stability and foaming, the results indicate comparable performance across all tested pectin levels.

[0237] For different reasons, particles produced by the materials and methods described in this example were adequate to be included in high quality alternative dairy products.

[0238] Example 12. Adapting formulation to reduced BLG content

[0239] The experiment tested the pectin concentration in a formulation with a reduced BLG content of 1.0% and 1.5% (w / w). The preparation method is as follows: SHMP (0.1% w / w) was hydrated in water for 20 min at ambient temperature. Next, temperature was raised to 50°C and pectin (0.1- 0.4% w / w, LM:HMA ratio equal to 16:84) was introduced to the solution and stirred at 500 RPM until fully hydrated. Subsequently, BLG (1.0 or 1.5% w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and 3.0% w / w fat was added to the solution which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA-TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy). Next, the solution was pasteurized via UHT pasteurization (121°C, 2-second holding time). Finally, the pH was set to pH~7.0 using 5M NaOH.

[0240] As demonstrated in Table 15, to achieve a soluble and stable complex between the LM+HMA pectin combination and BLG, the ratio between the two components should be between 10-15:3 (BLG:pectin). Furthermore, lowering the BLG concentration to 1.0-1.5% effectively addressed particle sedimentation while preserving a high L value.

[0241] Table 15A.

[0242] In another experiment the effect of total amount of BLG (different BLG percentages) on the physical and sensory attributes of milk was assessed.

[0243] Milk was prepared as described above, where the total BLG was 0.25, 0.5, 1.0 or 1.5% w / w. The total amount of pectin was kept at 0.3% w / w and the ratio between LM:HMA was kept at 16:84. Results are shown in Tables 15B - 15E.

[0244] Table 15B. The effect of different BLG percentages on the physical parameters of milk

[0245] As shown in Table 15B, increasing the BLG protein concentration from 0.25% to 1.5% w / w had minimal impact on the milk's pH and viscosity. However, the sample containing 0.5% w / w BLG was notably different, exhibiting an average particle size 150- 200 nm lower than the others and producing a reduced dryness sensation compared to the other BLG concentrations tested.

[0246] Table 15C.The effect of different BLG percentages on the physical parameters of coffee Table 15D. The effect of different BLG percentages on the physical parameters of coffee

[0247] Table 15E. The effect of different BLG percentages Foaming ranking of coffee As shown in Table 15C-15D, a linear correlation was observed between BLG concentration and the lightness of instant coffee, based on colorimeter measurements. Despite this, all BLG levels demonstrated stability against protein aggregation under the coffee's acidic and high-temperature conditions. Regarding foaming properties (Table 15E), the lowest BLG concentration resulted in poor foam quality, characterized by large bubbles and low overrun. Increasing the BLG content improved foam volume; however, in the 0.5- 1.5% range, the foam became overly dense, preventing the formation of latte art. From the above data we conclude that the most suitable range of BLG is 0.5-2%.

[0248] Particles produced by the materials and methods described in this example were very adequate to be included in high quality alternative dairy products. Example 13. The effect of SHMP on BLG-pectin complex stability in a milk formulation

[0249] In earlier experiments, SHMP was added as a precautionary measure to chelate divalent ions, like calcium, which could interact with BLG and / or pectin and lead to complex instability during UHT treatment. In this experiment, SHMP was removed from the formulation to test this assumption and understand if the addition of a chelator is necessary in terms of stability.

[0250] The preparation method is as follows: Pectin (0.3% w / w, LM:HMA ratio equal to 16:84 (—1:5)) was introduced to water and stirred at 500 RPM until fully hydrated at 50°C. Subsequently, BLG (1.5 w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and 3.0% w / w fat was added to the solution which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA-TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy). Next, the solution was pasteurized via UHT pasteurization (121 °C, 2-second holding time). Finally, the pH was set to pH~7.0 using 5M NaOH.

[0251] Results in Table 16 show SHMP does not have an impact on complex stability.

[0252] Table 16.

[0253] Particles produced by the materials and methods described in this example were very adequate to be included in high quality alternative dairy products.

[0254] Example 14. BLG-pectin complex in a final milk prototype

[0255] To evaluate the physical and sensory properties of a milk prototype based on BLG- pectin complexes, the following method was applied: Sugar (1.25%), salt (0.1%) and L- Cystine (0.02%) were added to water at an ambient temperature and stirred at 500 RPM until fully hydrated. Next, pectin (0.3% w / w, LM:HMA ratio equal to 16:84) was introduced to water and stirred at 500 RPM until fully hydrated at 50°C. Subsequently, BLG (1.5 w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and 3.0% w / w fat was added to the solution which was then dispersed using an Ultra-Turrax device (T 25 digital ULTRA-TURRAX, IKA, Germany) for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process (GEA, Lab Homogenizer Panda Plus 2000, Italy). Next, the solution was pasteurized via UHT pasteurization (121°C, 2-second holding time). Finally, the pH was set to pH~7.0 using 5M NaOH.

[0256] Table 17 shows the final stability parameters of the milk prototype; Fig. 1 shows a long-term stability test (several days) of the final formulation of the alternative milk. In terms of complex stability in coffee, the addition of these components had a minor effect on particle sedimentation and color. In addition, a taste panel was conducted to evaluate the dry mouthfeel sensation of the milk prototype in comparison to an equivalent milk product stabilized by buffering agents. In both formulations, milk flavoring was added to the final sample. Eleven milk tasters ranked the dry mouth feeling in a scale of 1-7 (1 - not dry at all, 7- extremely dry). The average score given to the BLG -pectin complex milk was 3.2 while the control was ranked 2.2.

[0257] There is no difference in the appearance between instant coffee prepared with a local cow milk and alternative milk prepared in this example.

[0258] Furthermore, the incorporation of calcium was evaluated in this milk prototype. Calcium is a crucial nutrient in milk, and as a result, alternative beverage producers often add insoluble calcium compounds (such as calcium carbonate, calcium phosphate, or calcium citrate) to their formulations. However, these calcium-based compounds become increasingly soluble as the pH decreases, presenting a challenge in this case, as the milk solution enters the pasteurizer at a pH of 4.75. Therefore, calcium carbonate (0.5%) was added to the milk after pasteurization, instead of adding sodium hydroxide. This raised the pH to 7.0.

[0259] The addition of calcium did not affect the flavor profile, allowing us to address two issues: enriching our milk with calcium and avoiding the addition of sodium hydroxide, which is less preferred as a food ingredient. Table 17.

[0260] The final milk prototype has the following characteristics (compared to 3% fat Cow’s milk, purchased from a local shop):

[0261] 1. Color of milk prototype (HunterLab units, L units): L=88.45 (vs. Cow’s milk L=89.06);

[0262] 2. Color of coffee with milk prototype (HunterLab units): 25 mL: L=34.13 (vs. Cow’s milk L=38.13), 15 mL: L=27.56 (vs. Cow’s milk L=32.64), and 5 mL: L= 16.87 (vs. Cow’s milk L=21.22);

[0263] 3. Viscosity of milk prototype (cP, Spindle T-SP-1, 100 RPM): cP=5.01 (vs. Cow’s milk cP=5.6);

[0264] 4. Foam attributes of milk prototype: Overrun (%) =46.7 (vs. Cow’s milk overrun (%) =46+3.7), Stability (%) =75 (vs. Cow’s milk stability (%) =21+3.7).

[0265] Particles produced by the materials and methods described in this example were very adequate to be included in high quality alternative dairy products. Table 18 summarizes the BLG-pectin-complex milk prototypes produced by the inventors. Example 15

[0266] The preparation method is as follows: Pectin (0.05, 0.1, 0.3 and 0.5% w / w, LM:HMA ratio equal tol:99, 5:95, 10:90 and 16:84) was introduced to water and stirred at 500 RPM until fully hydrated at 50°C. Subsequently, BLG (0.25, 0.5, 1.0 and 1.5% w / w) was added to the solution and stirred for 30 min at 50°C. After full hydration, pH was set to pH=4.75 using 5M HC1. Following pH reduction, the temperature was raised to 65°C and the following materials were added to the solution: 3.0% w / w fat, 1.25% sucrose, 0.1% salt, 0.02% L-Cystine were added to the solution which was then dispersed using an Ultra-Turrax device for 10 minutes at 12,000 RPM. The emulsion was subsequently homogenized at 600 bar and 65 °C using a two-stage homogenization process. Next, the solution was pasteurized via UHT pasteurization (121 °C, 2-second holding time). Finally, the pH was set to pH~7.0 using 5M NaOH and refrigerated until further analyzed. Final pH and dry matter (%) were measured.

[0267] In the case of protein that has undergone a reaction (deamination) by protein glutaminase (PG), the preparation process is identical to that described above with the following changes: BLG was hydrated in water for 30 minutes at room temperature. Subsequently, the temperature was increased to 55°C and 0.02% of PG was added to the solution where the enzymatic reaction was set to 1 hour. Following this phase, pectin was added to the deamidated BLG solution and next steps were as described above.

[0268] Results

[0269] To improve the foaming ability of the BLG-pectin complexation milk, the protein underwent a reaction (deamidation treatment) using protein-glutaminase (PG), as described in the method section. The treatment was applied to a 1.5% w / w of BLG. Subsequently, pectin was added at a concentration of 0.3%, in a 16:84 ratio LM:HMA. Results are shown in Table 4.7 and 4.8.

[0270] Table 19. The effect of protein glutaminase on the physical parameters of milk As shown in Table 19, deamination had a minimal impact on the physical properties of the milk. However, a notable finding was that the tasting panel reported a stronger dryness sensation in the sample treated with PG compared to the control.

[0271] Table 20. The effect of protein glutaminase on the physical parameters of coffee

[0272] Table 21. The effect of protein glutaminase on the physical parameters of coffee

[0273] Tables 20-21 highlight the impact of protein deamination on both the lightness of instant coffee and, more notably, on foam characteristics. The resulting foam was less dense, making it easier to create latte art. It is concluded therefore that use of partially deaminated BLG is beneficial for preparation of coffee with alternative milk of the present invention.

[0274] Although the present invention has been described herein above by way of preferred embodiments thereof, it can be modified, without departing from the spirit and nature of the subject invention as defined in the appended claims.

Claims

CLAIMS1. An alternative dairy composition, comprising a plurality of particles, wherein the particles comprise a complex of a non-animal Beta-lactoglobulin (BLG) protein and a nonanimal pectin.

2. The alternative dairy composition of claim 1, wherein each particle comprises a core consisting of the non-animal BLG protein and a non-animal pectin.

3. The alternative dairy composition of claim 1 or claim 2, wherein the alternative dairy composition further comprises water, a non-animal lipid, a non-animal sweetener, NaCl, L- Cystine, calcium, or any combination thereof.

4. The alternative dairy composition of claim 3, wherein the alternative dairy composition further comprises water, non-animal lipid, non-animal sweetener, NaCl, L- Cystine, and calcium.

5. The alternative dairy composition of claim 3 or claim 4, wherein each particle comprises a core consisting of the non-animal BLG, a non-animal pectin, and non-animal lipid.

6. The alternative dairy composition of any one of claims 1 to 5, comprising 0.1% to 3.5% w / w non-animal BLG or from 0.5% to 2 % w / w non-animal BLG.

7. The alternative dairy composition of any one of claims 1 to 6, wherein the non-animal pectin is selected from a high methoxly pectin (HM), high methoxyl pectin amidated (HMA) low methoxyl pectin (LM), low methoxyl pectin amidated (LMA), and any combination thereof.

8. The alternative dairy composition of claim 7, wherein the non-animal pectin is a mixture of LM(A) and HM(A).

9. The alternative dairy composition of claim 8, wherein the non-animal pectin is a mixture of LMA and HM pectins or a mixture of LM and HMA pectins.

10. The alternative dairy composition of claim 8 or claim 9, wherein the ratio between LM(A) and HM(A) is from 1:2 to 1:20 or from 1:4 to 1:18, or from 1:2.5 to 1:7.5, respectively.

11. The alternative dairy composition of claim 10, wherein the ratio between LM and HM(A) is about 1:5.25.

12. The alternative dairy composition of any one of claims 1 to 11, comprising between 0.1% to 0.3% w / w total non-animal pectins.

13. The alternative dairy composition of claim 12, comprising about 0.3% w / w total nonanimal pectins.

14. The alternative dairy composition of any one of claims 1 to 3, wherein the plurality of particles has an average particle size of 700 nm to 1400 nm.

15. The alternative dairy composition of any one of claims 3 to 13, wherein the plurality of particles has an average particle size of 900 nm to 2000 nm and / or a median particle size of 500 to 900 nm.

16. The alternative dairy composition of any one of claims 1 to 15, comprising: i. from about 0.1 w / w% to about 3.5 % w / w BLG, ii. from about 0.1 % w / w to about 0.3 % w / w total pectins, iii. from about 0.1 % w / w to about 5 % w / w non-animal lipid, iv. from about 0.1 % w / w to about 6 % w / w sweetener, v. from about 0.01 % w / w to about 0.3 % w / w NaCl (edible salt), vi. from about 0.005 % w / w to about 0.1 % w / w L-Cystine, and vii. from about 0.15 % w / w to about 0.25 % w / w calcium, and viii. up to 100 w / w water.

17. The alternative dairy composition of claim 16, comprising: i. from about 0.5 % w / w to about 1.5 % w / w BLG, ii. from about 0.2 % w / w to about 0.4 % w / w total pectins, iii. from about 0.5 % w / w to about 3.5 % w / w non-animal lipid, iv. from about 1 % w / w to about 1.75 % w / w sweetener, v. from about 0.05 % w / w to about 0.15 % w / w NaCl (edible salt), vi. from about 0.01 % w / w to about 0.05 % w / w L-Cystine, and vii. from about 0.4 % w / w to about 0.6 % w / w calcium, and viii. up to 100 % w / w water.

18. The alternative dairy composition of any one of claims 1 to 17, having a pH of from 5 to 8.

19. The alternative dairy composition of any one of claims 1 to 18, devoid of Gum Arabic, Carrageenan Lambda, Gellan, sodium hexametaphosphate (SHMP), monopotassium phosphate, di-potassium phosphate, a phosphate buffer, polyphosphates, divalent ion chelators and / or a citrate buffer.

20. The alternative dairy composition of any one of claims 1 to 19, formulated as an alternative milk composition.

21. The alternative dairy composition of any one of claims 1 to 19, wherein the BLG is partially deaminated.

22. The alternative dairy composition of any one of claims 1 to 21, wherein the foaming ranking of the composition is 5.5 or more.

23. A food product comprising the alternative dairy composition of any one of claims 1 to 22.

24. The food product of claim 23, wherein the alternative dairy composition is an alternative milk composition.

25. The food product of claim 23 or claim 24, wherein the food product is instant coffee.

26. A food product composition comprising the alternative dairy composition of any one of claims 1 to 25, wherein the food product is an instant coffee, and wherein the alternative dairy composition is an alternative milk composition.

27. The food product of claim 25, wherein the food product has a. a pH of 5 to 6, and / or b. a Color (L, Hunter units) of 15 to 40 when the food product comprises from 3 to 20% v / v of the alternative milk composition.

28. A method of preparation of an alternative dairy composition, the method comprising: i. Combining a composition comprising a hydrated pectin with a non-animal Betalactoglobulin;ii. Setting a pH of the composition obtained in step (i) to a value of from 3 to 6, thereby obtaining a complex of the pectin and the non-animal Betalactoglobulin; iii. Optionally adding a non-animal lipid to the composition obtained in previous step; iv. Optionally homogenizing the composition obtained in the previous step; and v. Pasteurizing the composition obtained in the previous step via ultra-high temperature pasteurization.

29. The method of claim 28, comprising adding (a) a non-animal lipid at step (iii), (b) homogenizing at step (iv), or (c) both (a) and (b).

30. The method of claim 28 or claim 29, wherein:(a) Step (i) comprises a complete hydration of the non-animal BLG;(b) The method comprises hydrating the pectin prior to step (i);(c) Both (a) and (b).

31. The method of any one of claims 28 to 30, comprising adjusting the pH of the composition to the value of from 6.5 to 7.5 in an additional step (vi) or after any one of steps (iii)-(v), optionally the pH adjustment is carried out by adding calcium carbonate.

32. The method of any one of claims 28 to 31, further comprising adding at least one of a sweetener, L-cystine, and NaCl at step (i).

33. The method of any one of claims 28 to 32, wherein:(a) The BLG is a recombinant BLG,(b) The pectin is selected from high methoxly pectin (HM); high methoxyl pectin amidated (HMA)_low methoxyl pectin (LM), low methoxyl pectin amidated (LMA), and any combination thereof, or(c) Both (a) and (b).

34. The method of claim 33, wherein the pectin is a mixture of LM(A) and HM(A).

35. The method of claim 34, wherein the ratio between LM(A) and HM(A) and is from 1:2 to 1:20 1: 10 to 1:2 or from 1:4 to 1: 18, or from 1:2.5 to 1:7.5.

36. The method of any one of claims 28 to 35, comprising adding from 0.1 to 1.5% BLG.

37. The method of any one of claims 30 to 36, comprising: i. optionally dissolving in water from about 0.1 % w / w to about 6 % w / w sweetener, from about 0.01 % w / w to about 0.3 % w / w NaCl, from about 0.005 % w / w to about 0.1 % w / w L-Cystine, ii. hydrating from about 0.1 % w / w to about 0.3 % w / w total pectins in the composition obtained in previous step, if present, preferably wherein the pectins are a mixture of LM and HMA, iii. adding about 0.1 w / w% to about 3.5 % w / w BLG to the composition obtained in previous step and fully hydrating the BLG, iv. setting the pH of the composition obtained in previous step to a value of from 4.5 to 5, preferably to a value of 4.75 v. optionally adding and dispersing from about 0.1 % w / w to about 5 % w / w of non-animal lipid in the composition obtained in the previous step, optionally with preheating of the composition to from 60 to 70°C. vi. optionally homogenizing the composition obtained in the previous step, and vii. pasteurizing the composition the composition obtained in the previous step via an ultra-high temperature pasteurization.

38. The method according to any one of claims 28 to 37, wherein the resulting alternative dairy composition comprises a plurality of particles comprising a core consisting of the nonanimal dairy protein and pectin.

39. The method according to claim 38, wherein the particles have an average particle size of 900 nm to 2000 nm and / or median particle size of from 500 to 900 nm.

40. The method according to any one of claims 28 to 39, wherein the method is devoid of adding one or more compounds selected from Gum Arabic, Carrageenan Lambda, Gellan, SHMP, phosphate buffer, polyphosphates, divalent ion chelators and citrate buffer.

41. The method according to any one of claims 28 to 40, wherein the ultra-high temperature pasteurization comprises heating to from 110 °C to 125 °C for from 1 to 10 sec.

42. The method according to any one of claims 28 to 41, further comprising heating the composition at step (ii) to from 70 °C to 90 °C.

43. The method according to any one of claims 28 to 42, further comprising deaminating BLG prior to addition of pectin.

44. The method according to claim 43, wherein the deamination comprises addition of protein glutaminase.

45. The method according to claim 44, wherein protein glutaminase is added in the amount of from 0.01 to 0.05%.

46. An alternative food composition prepared by a method according to any one of claims 28 to 45.

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