Coloring material composition

Abstract

The present invention provides food colorant compositions, in particular the present invention provides compositions or colorant compositions comprising at least one phycobilin and at least one peptide, polypeptide and / or protein. In some embodiments, the phycobilin (such as phycocyanin) and the at least one peptide, polypeptide and / or protein form a complex.

Description

[Technical field]

[0001] TECHNICAL FIELD OF THEINVENTION The present invention relates to a food colorant composition, and in particular the present invention provides a composition or colorant composition comprising at least one phycobilin and at least one peptide, polypeptide and / or protein. In a preferred embodiment, the phycobilin (such as phycocyanin) and the at least one peptide, polypeptide and / or protein form a complex. The present invention also relates to the use of the food colorant compositions described herein in coloring food products, and to food products comprising the food colorant compositions described herein. [Background technology]

[0002] 2. Background of the Invention C-phycocyanin is a natural pigment-protein complex, extracted primarily from the biomass of Spirulina platensis. This natural blue pigment has a vivid blue color and is the closest natural alternative to the dye Brilliant Blue FCF, making it an attractive candidate to replace a wide range of synthetic blue pigments. Phycocyanins host linear tetrapyrrole chromophores, also called phycobilins, which are covalently bound to the protein subunits and are the source of the blue color. Thanks to its clean label, phycocyanin is used in a variety of food applications such as toppings and coatings for confectionery, ice cream and desserts.

[0003] However, phycocyanin from Spirulina is known to have very poor stability to acidity, causing aggregation and precipitation at pH levels below 4. Furthermore, phycocyanin is known to be easily denatured by heat treatments (e.g., pasteurization) during food processing, causing serious discoloration (denaturation T = 47°C, (Chaiklahan et al, Process biotechnology, 2012, 47: 659-664)). Therefore, many attempts have been made to improve the stability of phycocyanin used as a colorant.

[0004] WO15090697 reports a method for stabilizing phycocyanobilin with polyphenols to obtain a stabilized complex, but this stabilization requires first cleaving the native phycocyanin under heat or acid treatment and then reacting the chromophore with polyphenols. In Food Hydrocolloids, Selig et al., January 2018, (74:46-52) and more recently Li et al., October 2021 (119:106852) described the enhancement of color stability of Spirulina-based phycocyanin via polysaccharide complexes. Another approach was reported by Zhang et al. in Food Hydrocolloids August 2020, where they described that the colloidal stability of phycocyanin in acidic solutions was improved to some extent using whey protein, but the thermal stability was not improved.

[0005] Patent document CN111317142 discloses a stable functional phycocyanin multiple emulsion, its preparation method and application. The method includes the following steps: (1) mixing tannic acid, glycerinum, sodium alginate and phycocyanin to prepare an inner aqueous phase; (2) adding an emulsifier or biosurfactant to sunflower seed oil and emulsifying to obtain an oil phase; (3) mixing the inner aqueous phase and the oil phase, emulsifying to obtain a W / O emulsion, and solidifying to obtain a solidified W / O emulsion, where the W / O emulsion or the solidified W / O emulsion is regarded as the first phase; and (4) adding the first phase to the outer aqueous phase and emulsifying to obtain a phycocyanin W / O / W multiple emulsion. The phycocyanin W / O / W multiple emulsion disclosed in this invention has high thermochemical stability and light stability, is sensitive to acid, and has good emulsion stability.

[0006] WO2020 / 239913 describes a stabilized phycocyanin composition comprising a complexing agent and an encapsulating agent as a stabilizer. Despite these numerous attempts, stabilization of phycocyanin has not been successfully achieved and there is still a growing demand to identify efficient solutions to improve its stability. DISCLOSURE OF THEINVENTION

[0007] Summary of the Invention In this invention we introduce a new alternative method for phycocyanin stabilization using water-soluble plant-based proteins, yeast and animal proteins and peptides or mixtures thereof, characterized in that they are soluble under aqueous and acidic conditions and optionally have an isoelectric point higher than 4.5. This stabilization significantly improves the thermal and acidic stability of phycocyanin by preventing aggregation at acidic pH and protects the protein conformation during heat treatment. Applicants have surprisingly and unexpectedly found that phycocyanin is stabilized and does not lose color even after acidic conditions and heat treatment when complexed with water-soluble plant-based or animal proteins and peptides, characterized as being soluble under acidic conditions and optionally having an isoelectric point higher than 4.5. For example, but not limited to, the combination of phycocyanin with potato proteins improves the stability of phycocyanin. The stability of phycocyanin is further increased when soluble proteins (such as albumin-rich potato proteins) are combined with soluble peptides, for example from mung bean, soybean, rice and / or pea.

[0008] This improvement stabilizes phycocyanin in beverages and confectioneries, resulting in a vibrant and stable blue color. Using this blend, consumers can benefit from a clean, non-GMO label, excellent nutritional value, and neutral taste and off-notes in their end use. Thus, in a first aspect, the present invention provides a composition comprising at least one phycobilin (such as phycocyanin) and at least one protein, polypeptide and / or peptide, wherein the at least one protein, polypeptide and / or peptide is water-soluble, optionally has an isoelectric point (IEP) greater than 4, and / or is soluble under acidic conditions.

[0009] In a second aspect, the present invention provides a complex of at least one phycobilin (such as phycocyanin) and at least one protein, polypeptide and / or peptide, obtainable by mixing in an aqueous solution at least one protein, polypeptide and / or peptide with a composition comprising at least one phycobilin. The invention further provides in a third aspect a process for the formation of a composition or complex of any of the preceding aspects, the process comprising the step of mixing at least one peptide, polypeptide and / or protein, or a protein extract comprising at least one peptide, polypeptide and / or protein, with a composition comprising at least one phycobilin in an aqueous solution.

[0010] In a fourth aspect, the present invention further provides a method of stabilizing a phycobilin (such as phycocyanin), the method comprising the steps of: i) contacting phycobilins in an aqueous solution with at least one protein, polypeptide and / or peptide, or a protein extract comprising at least one protein, polypeptide and / or peptide; ii) Optionally, adding sugar. In a fifth aspect, the present invention further provides a stabilized phycobilin (such as phycocyanin) obtainable using the method of the third or fourth aspect.

[0011] In a sixth aspect, the invention further provides an encapsulated composition comprising a composition or complex according to either the first or second aspect, or a stabilized phycobilin according to the fifth aspect, said encapsulated composition being encapsulated using a cross-linked polymer derived from the group consisting of alginate, carrageenan and pectin, derivatives thereof, or combinations thereof. In a seventh aspect, the invention relates to a colourant comprising a composition or complex according to either the first or second aspect, a stabilized phycobilin according to the fifth aspect, or an encapsulated composition according to the sixth aspect, and optimally another pigment. In a further aspect, the invention relates to a food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product comprising a composition or complex according to any of aspects 1 or 2, a stabilized phycobilin according to the fifth aspect, an encapsulated composition according to the sixth aspect and / or a colorant according to the seventh aspect. [Brief description of the drawings]

[0012] [Figure 1]Figure 1. Evaluation of the stability of phycocyanin upon heating in a sugar syrup matrix, pH 3. Figure 1A shows color photographs of the samples, and Figure 1B shows black and white photographs. Sample 1A: 0.025% Spirulina extract (25% phycocyanin w:w) + 99.97% sugar syrup Brix (B) ° 60, pH 3. Sample 1B: 0.025% Spirulina extract + 0.15% yeast protein extract + 99.82% sugar syrup B ° 60, pH 3. Sample 1C: 0.025% Spirulina extract + 0.15% whey protein isolate + 99.82% sugar syrup B ° 60, pH 3. Sample 1D: 0.025% Spirulina extract + 0.15% potato protein extract + 99.82% sugar syrup B ° 60, pH 3. Sample 1E: 0.025% Spirulina extract + 0.075% Potato protein extract + 0.075% Rice peptide + 99.82% Sugar syrup B ° 60, pH 3. Sample 1F: 0.025% Spirulina extract + 0.15% Rice peptide + 99.82% Sugar syrup B ° 60, pH 3. Sample 1G: 0.025% Spirulina extract + 0.15% Mung bean peptide + 99.82% Sugar syrup B ° 60, pH 3. Sample 1H: 0.025% Spirulina extract + 0.075% Mung bean peptide + 0.075% Potato protein + 99.82% Sugar syrup B ° 60, pH 3. Sample 1I: 0.025% Spirulina extract + 0.15% soybean peptide + 99.82% sugar syrup B ° 60, pH 3. Sample 1j: + 0.025% Spirulina extract + 0.15% pea peptide + 99.82% sugar syrup B ° 60, pH 3. Sample R: Reference sample (untreated). Sample T: Treated at 80 ° C for 30 minutes.

[0013] [Diagram 2]Figure 2: Figure 2A. Stability of phycocyanin (A: control: 0.025% spirulina phycocyanin, B: 0.025% spirulina phycocyanin + 0.075% mung bean peptides + 0.075% potato protein) in pH 3 sugar syrup without heat treatment (T0) and after heat treatment at 80 °C for 5, 10, 20, and 30 min (60 °Brix, pH 3) (1) visually shows loss of color as a function of time; fucocyanin degradation rate is shown in a graph (Figure 2B): triangles: control, circles: spirulina extract + mung bean peptides and potato protein. [Diagram 3] Figure 3. Whey proteins. A: 0.1% Spirulina extract + 0.2% whey protein isolate. B: 0.1% Spirulina extract. C: 0.1% Spirulina extract + 0.2% potato protein. AT, BT, and CT: Samples A, B, and C heat-treated at 95°C for 5 min.

[0014] [Figure 4] Figure 4.4A: Thermal and colloidal stability (pH 3) results after heat treatment. Color (top) and black and white (bottom) photographs of the samples. A: 0.15% Spirulina extract + 0.05% potato protein extract. B: 0.15% Spirulina extract + 0.025% potato protein extract + 0.025% mung bean peptides. C: 0.15% Spirulina extract. (AT, BT, and CT: samples A, B, C after heat treatment). Figure 4B. Changes in the photostability of samples A, B, and C after 13 days of heat treatment. Figure 4C. Samples AT, BT, and CT (heat-treated samples) were exposed to light for 13 days. Color (top) and black and white (bottom) photographs of samples AT, BT, and CT before (day 0) and after (day 13) exposure.

[0015] [Diagram 5]Figure 5A. Samples: 5A: 0.2% bovine serum albumin + 0.025% Spirulina extract, 5B: 0.1% bovine serum albumin + 0.025% Spirulina extract. 5C: 0.05% bovine serum albumin + 0.025% Spirulina extract. 5D: 0,025% Spirulina extract / control. 5E: 0.05% chicken egg white + 0.025% Spirulina extract. 5F: 0.1% chicken egg white + 0.025% Spirulina extract. 5G: 0.2% chicken egg white + 0.025% Spirulina extract. Figure 5B: Variation of phycocyanin retention as a function of bovine serum concentration (left) and phycocyanin retention as a function of chicken egg albumin concentration (right), conditions: in sugar syrup at 60°brix, pH 3, heated at 80°C for 30 min. Figure 6. Peptides identified from pea. Figure 7. Peptides identified from soybean.

[0016] definition As used herein, the term "colorant composition" refers to any substance that imparts color by absorbing or scattering light of different wavelengths. The term "color" refers to color characteristics such as hue, chroma, purity, saturation, intensity, vividness, value, lightness, and darkness, as well as color model system parameters used to describe these characteristics, such as the L*a*b* values ​​of the Commission Internationale de l'Eclairage CIE 1976 CIELAB color space. The term "hue" refers to the property of a color that gives the color its name, such as red, blue, and brown.

[0017] Detailed Description In the following text, many different aspects of the present disclosure are broadly described. This description should be construed as illustrative only and does not describe all possible aspects, since describing all possible aspects is impractical, if not impossible. Any feature, characteristic, component, composition, ingredient, product, step or methodology described herein can be deleted in whole or in part, and can be combined or substituted with any other feature, characteristic, component, composition, ingredient, product, step or methodology described herein. Many alternative embodiments can be implemented using current technology or technology developed after the filing date of this patent, and would still fall within the scope of this patent claim.

[0018] Preferred and / or optional features of the present invention are described herein. Any aspect of the present invention may be combined with any other aspect of the present invention, unless the context requires otherwise. Any preferred or optional feature of any aspect may be combined alone or with any aspect of the present invention, as well as with any other preferred or optional feature, unless the context requires otherwise.

[0019] composition Surprisingly, it has been found that when a water-soluble peptide, polypeptide and / or protein is combined with at least one phycobilin (such as phycocyanin), the phycobilin (such as phycocyanin) is stable to acid, heat treatment and exposure to light. The present invention thus provides a composition or colorant composition comprising at least one phycobilin and at least one peptide, polypeptide and / or protein. In some embodiments, the phycobilin (such as phycocyanin) and the at least one peptide, polypeptide and / or protein form a complex.

[0020] Without being bound to any particular theory, complexes between phycobilins (such as phycocyanin) and soluble proteins, polypeptides and / or peptides improve the stability of phycobilins (such as phycocyanin) to heat and acidity by preventing aggregation at acidic pH and protecting protein conformation during heat treatment. Thus, the colorant compositions described herein provide colors, such as blue colors, that are stable to light, heat and / or acidic conditions. The present invention provides compositions (or "compositions of the invention" or "colorant compositions of the invention") comprising at least one phycobilin and at least one protein, polypeptide and / or peptide, where the at least one protein, polypeptide and / or peptide is water-soluble and, optionally, has an IEP greater than 4 and / or is soluble under acidic conditions.

[0021] The present invention also provides a complex comprising at least one phycobilin and at least one peptide, polypeptide and / or protein, obtainable by mixing in an aqueous solution at least one peptide, polypeptide and / or protein with a composition comprising at least one phycobilin. In certain embodiments, at least one protein, polypeptide and / or peptide is water soluble, has an IEP greater than 4, and / or is soluble under acidic conditions.

[0022] Phycobilin Phycobilins are light-harvesting pigments found in cyanobacteria but absent from higher plants. The basic structure of phycobilins consists of a tetrapyrrole unit, in which the four pyrrole rings form an open chain. Four major phycobilins exist in photosynthetic organisms: phycoerythrobilin, phycocyanobilin, phycoviolobilin, and phycourobilin. Differences in the degree of pi-electron conjugation are responsible for the unique absorption spectral properties and coloration of the chromophores. Phycoerythrobilins appear red, phycocyanobilins blue, phycoviolobilins purple, and phycourobilin yellow. Thus, the phycobilins used in the present invention may be selected from phycoerythrobilins, phycocyanobilins, phycoviolobilins, phycourobilins, and any mixtures thereof.

[0023] In certain embodiments, the phycobilin is a phycocyanobilin, such as phycocyanin, allophycocyanin, and phycoerythrin. In one embodiment, the phycobilin of the present invention may be a phycocyanobilin, which optionally has a blue color. Phycobiliproteins consist of two subunits (alpha and beta) that have a protein backbone with one or two linear tetrapyrrole chromophores covalently attached. In some embodiments, the phycobilin is phycocyanin. In certain embodiments, the phycocyanin is R-phycocyanin and / or C-phycocyanin.

[0024] The phycobilins (such as phycocyanins) used in the present invention may be of natural or synthetic origin. Phycocyanin in the present invention is obtained or can be obtained from any source containing proteins, such as certain cyanobacteria (also called blue-green algae). In certain embodiments, the phycocyanin is obtained from Arthrospira platensis (also called spirulina), Arthrospira fusiformis, Arthrospira maxima, Galdieria daedala, Galdieria sulphuraria, Galdieria maxima, Galdieria partita, Cyanidioschyzon merolae 10D, Cyanidioschyzon merolae DBV201, Cyanidium caldarium, Cyanidium rumpens, Cyanidium daedalum, Cyanidium maximum, Cyanidium partitum, and mixtures thereof.

[0025] In one embodiment, the phycocyanin is a cyanobacterial phycocyanin. In one embodiment, the phycobilin (such as phycocyanin) is phycocyanin from Spirulina (Arthrospira platensis). In one embodiment, the phycobilin (such as phycocyanin) is derived from extremophilic microalgae, such as phycocyanin from Galdieria sulphuraria or phycocyanin from Cyanidioschyzon merolae.

[0026] For example, but not limited to, phycocyanins from the Cyanidioschyzon merolae UniProt entry (alpha and beta chains) are Q85G43 (https: / / www.uniprot.org / uniprotkb / Q85G43 / entry), Q85G42 A0A5P9RV70, A0A5P9RVI1. For example, but not limited to, phycocyanins from the Galdieria sulphuraria UniProt entries are P00306 (https: / / www.uniprot.org / uniprotkb / P00306 / entry), P00311. The structures of phycocyanins from Cyanidioschyzon merolae and Galdieria sulphuraria are similar (e.g., using Uniprot BLAST, there is 84% ​​similarity between Uniprot entries Q85G43 and P00306). In one embodiment, phycobilin (0.1% in water) (such as phycocyanin from spirulina) has an L* value of 66.23±5%, an a* value of −33.1±5%, and a b* value of −47.52±5%. [ka]

[0027] The basic structure of phycocyanobilin In certain embodiments, the phycobilin (such as phycocyanin) can be an extract containing said phycobilin (such as phycocyanin), or a purified phycobilin (such as purified phycocyanin). By purified phycobilin (such as purified phycocyanin), in the present invention is meant phycobilin (such as phycocyanin) that is at least 80% w / w pure, such as at least 85%, such as at least 90%, such as at least 95%, or such as at least 99% w / w. Natural extracts can be obtained from the aforementioned organisms, such as Arthrospira platensis extract or Spirulina extract, Galdieria daedala extract, Galdieria sulphuraria extract, Galdieria maxima extract, and the like.

[0028] Extraction can be carried out by any method known in the art that allows for a degree of purity of the phycobilin (such as phycocyanin). Phycobilins (such as phycocyanin) can be extracted using water as a solvent. Various buffer systems can be used to improve the extraction yield. For example, a 1.5% CaCl2 aqueous solution can be used to obtain a higher extraction yield when compared to distilled water and sodium phosphate buffer (pH 7.0). Sodium phosphate buffer (pH 7.0), distilled water, NaCl solution (0.15M), and CaCl2 solution (10g / L) can also be used (Silveira, S.et al. 2007. Optimization of phycocyanin extraction from Spirulina platensis using factorial design. Bioresource Technology, 98(8), 1629-1634). In some embodiments, acetate buffer (pH 5.0) may also be used (Ilter, I., et al. 2018. Optimization of phycocyanin extraction from Spirulina platensis using different techniques. Journal of Food Composition and Analysis, 70, 78-88.).

[0029] Aqueous extraction can be assisted by further processing such as homogenization, bead milling, high pressure homogenization, microwave treatment, ultrasonic treatment, pulsed electric fields and moderate electric fields (DP. Jaeschke, et al. 2021. Phycocyanin from Spirulina: A review of extraction methods and stability. Journal of Food Research International, 143, 110314). A phycobilin extract (such as a phycocyanin extract) may contain at least 2% (w / w), such as at least 5% (w / w), such as at least 10% (w / w), such as at least 20% (w / w), such as at least 30% (w / w), such as 40% (w / w), or such as at least 50% (w / w) of phycocyanin. In certain embodiments of the invention, the amount of phycobilin (such as phycocyanin) present in the composition or complex is at least 0.5% (w / w), such as at least 0.7% (w / w), such as at least 0.9% (w / w), such as at least 1% (w / w), such as at least 1.5% (w / w), such as at least 2% (w / w), such as at least 2.5% (w / w), or such as at least 3% (w / w).

[0030] In a preferred embodiment of the invention, the amount of phycobilin (such as phycocyanin) present in the composition or complex of the invention is less than 50% (w / w), such as less than 40%, for example less than 25%, such as less than 20% (w / w), for example less than 15% (w / w), such as less than 13% (w / w), for example less than 10% (w / w), such as less than 9% (w / w), for example less than 8% (w / w), such as less than 7% (w / w), for example less than 6% (w / w), such as less than 5% (w / w), for example less than 4% (w / w), or such as less than 3% (w / w). In certain embodiments of the invention, the amount of phycocyanin present in the composition or complex is 0.5-30% (w / w), such as 0.5-20% (w / w), for example 0.5-12% (w / w), such as 0.5-5% (w / w), for example 0.8-3% (w / w), such as 1-20% (w / w), for example 2-12% (w / w), or for example 3-5% (w / w). In certain embodiments of the invention, the phycocyanin is not cleaved.

[0031] Phycobilin-stabilized peptides, polypeptides, and proteins The present invention shows how proteins, peptides, and combinations of proteins, peptides, and polypeptides form stabilizing complexes with at least one phycobilin (such as phycocyanin) that stabilize them. Without wishing to be bound by theory, it is believed that the at least one phycobilin (e.g., phycocyanin) and the at least one protein, polypeptide and / or peptide form a complex that protects the at least one phycobilin (e.g., phycocyanin) from precipitation under acidic conditions, during heat treatment and / or during exposure to light.

[0032] Such interactions may impart unexpected and unpredictable properties to the resulting compositions and complexes comprising at least one phycobilin (such as phycocyanin) and at least one protein, polypeptide and / or peptide. In some embodiments, additionally, certain amounts of free amino acids may be present. In one embodiment, the pH of the composition or complex according to the present invention is 2.0 to 5.0, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, and most preferably 2.4 to 3.3. Proteins are large biomolecules and macromolecules that contain one or more long chains of amino acid residues. A linear chain of amino acid residues (without a defined three-dimensional structure) is called a polypeptide. A protein contains at least one long polypeptide.

[0033] Protein is generally used to refer to an entire biological molecule that adopts a stable three-dimensional structure, whereas peptides and polypeptides generally refer to short amino acid oligomers that often lack stable 3D structure. Most proteins fold into unique 3D structures, and the shape into which a protein naturally folds is known as its native conformation.

[0034] The various structures of proteins are: - Primary structure: amino acid sequence. - Secondary structure: regularly repeated local structures stabilized by hydrogen bonds. The most common examples are α-helices, β-sheets and turns. Since secondary structure is local, many regions of different secondary structure can exist within the same protein molecule. - Tertiary structure: the overall shape of a single protein molecule; the spatial relationships of the secondary structures to each other. Tertiary structure is generally stabilized by non-local interactions, most commonly the formation of a hydrophobic core, but also by salt bridges, hydrogen bonds, disulfide bonds, and even post-translational modifications. The term "tertiary structure" is often used as a synonym for the term folding. Tertiary structure is what controls the basic function of a protein. - Quaternary structure: the structure formed by several protein molecules (polypeptide chains), in this context usually called protein subunits, functioning as a single protein complex.

[0035] In certain embodiments, a protein can have primary, secondary, tertiary, or quaternary structure. In certain embodiments, the protein is from animal, plant, yeast and / or bacterial origin. In some embodiments, the protein is completely or partially denatured (ie, has partially or completely lost secondary, tertiary or quaternary structure), thereby forming a polypeptide. In certain embodiments, proteins are hydrolyzed and the hydrolysis results in peptides and / or polypeptides of different sizes. In the present invention, the term "polypeptide" is understood as a linear chain of amino acid residues without a defined conformation. A polypeptide may be derived from one or more proteins that have been denatured and / or hydrolyzed.

[0036] In the present disclosure, the term "peptide" or "oligopeptide" is understood to denote a short chain of amino acids. A peptide in the present invention may be derived from one or more proteins, for example, by hydrolysis of one or more proteins. In one embodiment, a "peptide" or "peptide material" is understood to denote a protein hydrolysate, which may contain any type of peptide that may vary in length and a certain amount of free amino acids resulting from hydrolysis. The protein material may be hydrolyzed by any means known in the art, such as by using one or more hydrolases. In one example, an enzyme preparation with low exopeptidase activity is used to minimize the release of free amino acids and improve the taste profile of the protein hydrolysate. In one embodiment, the peptide material has a molecular weight of about 300 to about 10,000 daltons, and in another embodiment, about 300 to about 5,000 daltons, such as 300 to 2000 daltons, such as 300 to 1000 daltons, such as 500 to 1000 daltons, or 500 to 2000 daltons.

[0037] In some embodiments, the peptide contains fewer than 40 residues, such as fewer than 30 residues. Proteins, polypeptides and peptides of synthetic origin can also be used in the present invention. Protein hydrolysis can be carried out by chemical and enzymatic methods. Most of the enzymes used in protein hydrolysis are from animal sources (such as pancreatin and pepsin), vegetable sources (such as papain from papaya, ficin from fig, and bromelain from pineapple), and microbial sources (such as alcalase).

[0038] Proteolytic enzymes hydrolyze proteins at optimal temperatures and pH, usually targeting specific peptide cleavage bonds, resulting in digests consisting of amino acids and peptides of various sizes. Enzymes from animal sources are more specific for their site of action compared to plant enzymes, which are more broadly specific in their action. For example, the enzyme pepsin cleaves phenylalanine or leucine bonds. Papain has broad specificity and cleaves phenylalanine, arginine, and lysine bonds. Pancreatin cleaves tryptophan, arginine, tyrosine, leucine, phenylalanine, and lysine bonds. Hydrolysis of proteins can be carried out enzymatically, or by adding sufficient amounts of acid, or by a combination of enzymes (proteinase and peptidase activity) and acid. For example, when using hydrochloric acid, it is used at a final concentration of 5.5 M in the hydrolysis mixture. Other acids can be used in amounts that give a similar pH to 5.5 M hydrochloric acid.

[0039] For enzymatic hydrolysis, an enzyme or enzyme preparation containing one or more enzymes and having both proteinase and peptidase activity is used at a temperature appropriate for the enzyme or enzymes. The appropriate temperature is selected according to the temperature requirements of the enzyme. For example, UMAMIZYME™ can tolerate temperatures between about 40° C. and about 60° C., with an optimum temperature of about 55° C. A useful enzyme is a protease enzyme preparation, such as UMAMIZYME™ (Amano, Elgin, IL). Protease preparations contain two types of enzymes; proteinases, which hydrolyze proteins to form small peptides, and peptidases, which release amino acids from the termini of proteins and peptides. UMAMIZYME™ is derived from Aspergillus oryzae and is rich in Endo and Exo activities.

[0040] All enzymes used should be food grade. The amount or units of enzyme required is selected to ensure sufficient activity and avoid bitterness development. The amount of enzyme depends on the amount of protein and should have an enzyme:protein ratio of 0.5:20 to 3:20 (0.5 to 3 parts enzyme to 20 parts pea protein), e.g. 1:20 enzyme:protein (UMAMIZYME™ has 70 U / g at pH 7). Acid hydrolysis can use acids including, for example, one or more of hydrochloric acid, lactic acid, phosphoric acid, and citric acid. The hydrolysis is carried out at a suitable temperature, for example, from about 50° C. to about 70° C. In one preferred embodiment, the enzyme is pepsin.

[0041] Another alternative is a combination of acid hydrolysis and enzymatic hydrolysis. In this case, the acid, enzyme, pH and temperature must be selected so that they are compatible. Relevant data for many proteinase and peptidase enzymes are known and readily available. Acid hydrolysis is possible, but has the disadvantage of adding small amounts of salt when neutralizing the acid. The resulting salt enhancer may still be useful in reduced-salt products, but the potential for salt reduction may be slightly limited. More importantly, the contaminant 3-chloropropane-1,2-diol (3-MCPD) and other monochloropropanol and dichloropropanol contaminants may be produced, which is under discussion for its potential adverse health effects, leading consumers to avoid products that contain or may contain 3-MCPD.

[0042] In some embodiments, proteins, polypeptides and / or peptides can be obtained by a fermentation process. Various bacterial strains can be used to ferment proteins (e.g., plant proteins) to obtain products rich in proteins and peptides. In some embodiments, the majority of the carbohydrate molecules are then removed during fermentation to obtain high protein and / or peptide products. In some embodiments, fermented protein, polypeptide and / or peptide can be prepared by methods well known in the art. Protein source, for example, plant protein source described herein, such as tuber (such as potato), seed (such as pea, soybean), legume (such as mung bean), grain (such as rice), etc., can be cultured overnight, for example, 12 hours, at a temperature suitable for the microorganism used. For example, lactobacillus (lactic acid bacteria), such as L. plantarum, L. casei, L. brevis and L. helveticus, can be used. 37°C is the temperature suitable for L. plantarum. Any suitable medium can be selected, for example, MRS broth (Difco, United States of America).

[0043] Fermentation with a microorganism (such as Lactobacillus sp.) is initiated at a pH of at least 6 or higher, for example 6-7, using hydrolyzed protein as the fermentation broth and adding a sufficient amount of overnight protein source (potato, mung bean, etc.) culture. Fermentation is allowed to proceed until the pH drops to at least pH 5.5 or lower, for example from pH 5.5 to pH 4.5, which usually takes about 5-12 hours. The fermentation temperature is selected to accommodate the microorganism. Useful temperature ranges for Lactobacillus bacteria, particularly L. plantarum, include, for example, about 20°C to about 40°C, about 30°C to about 40°C, or about 35°C to about 40°C, with an optimum of approximately 36°C to 38°C. At lower temperatures the growth rate is reduced and at higher temperatures the microorganism is killed.

[0044] After fermentation (once the low pH is reached), the fermentation broth is pasteurized at 90° C. for 30 minutes to inactivate the microorganisms and enzymes. The pasteurized fermentation broth can then be filtered to remove large particles and concentrated, for example by evaporation, including boiling, for example up to 100°C. In certain embodiments, at least one protein, polypeptide and / or peptide is water soluble and / or has an IEP of greater than 4, such as 4.5, 5, 5.5, 6, 6.5, greater than 7, or such as greater than 7.5.

[0045] The term "water soluble" is understood to indicate that at least 50%, such as at least 60%, 70%, 80%, 90% or such as 99% of one or more proteins, polypeptides, peptides or mixtures thereof are water soluble, i.e. the proteins, polypeptides and / or peptides do not precipitate after a certain period of time, such as at least 1 day, at least 10 days, at least 30 days, at least 2 months or at least 9 months. In some embodiments, when a mixture of different proteins, polypeptides and / or peptides is used in the compositions or complexes of the invention, at least 50% of said proteins, polypeptides and / or peptides are water soluble, such as at least 60%, 70%, 80%, 90%, or for example at least 99%. In a preferred embodiment, at least 80% of the proteins, polypeptides and / or peptides used in the compositions or complexes of the invention are water soluble.

[0046] In certain embodiments, at least one protein, polypeptide and / or peptide is soluble under acidic conditions, e.g., soluble at a pH of less than 6, less than 5, less than 4, less than 3, or less than 2. In certain embodiments, when a mixture of different proteins, polypeptides and / or peptides is used in a composition or complex of the invention, at least 50% of said proteins, polypeptides and / or peptides are soluble under acidic conditions, e.g., at least 60%, 70%, 80%, 90%, or e.g., at least 99% are soluble. The isoelectric point (IEP) is the pH value at which the zeta potential of a protein or polypeptide is zero. The IEP can be measured using a Zetasizer, and when the change in zeta potential is displayed as a function of pH, the IEP is attributed to the zero value of zeta potential.

[0047] In some embodiments, the protein is selected from albumins, globulins (e.g., β-conglycisin, glycinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and any other soluble proteins from plant, animal or yeast origin, and any mixtures thereof. As mentioned above, preferably, the albumins, globulins (e.g., β-conglycisin, glycinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and any other soluble proteins from plant, animal or yeast origin, and any mixtures thereof, present in the composition of the complex of the present invention are water-soluble. In some embodiments, at least 50% of the albumins, globulins (e.g., β-conglycin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof are water soluble, e.g., at least 60%, 70%, 80%, 90%, or e.g., at least 99% of the albumins, globulins (e.g., β-conglycin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof are water soluble. In a preferred embodiment, at least 80% of the albumins, globulins (e.g., β-conglycinin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof used in the compositions or complexes of the present invention are water soluble.

[0048] In certain embodiments, the compositions or complexes of the present invention comprise essentially water-soluble albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and any other soluble proteins from plant, animal or yeast origin, and mixtures thereof, and optionally water-soluble polypeptides and / or water-soluble peptides derived therefrom. In one embodiment, the compositions or complexes of the present invention comprise water soluble albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof, any other soluble proteins from plant, animal or yeast origin, any mixtures thereof, and water soluble polypeptides and / or peptides derived therefrom (i.e., derived from albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof).

[0049] In certain embodiments, the compositions or complexes of the present invention comprise water soluble polypeptides and / or peptides derived from one or more of albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and any other proteins from plant, animal, or yeast origin. In some embodiments, the protein is not derived from or is essentially free of whey proteins, including β-lactoglobulin (abbreviated β-LG), α-lactalbumin (α-LA), immunoglobulins (IG), bovine serum albumin (BSA), bovine lactoferrin (BLF), and lactoperoxidase (LP).

[0050] In some embodiments, the albumin is not BSA. In some embodiments, the polypeptide or peptide is not derived from whey protein. Albumins are water-soluble globular proteins found in both animals and plants. Examples of albumin protein sequences are cited in the literature. For example, but not limited to, the potato albumin entries in UniProt are M0ZKG8_SOLTU and M0ZKI9_SOLTU. Currently, there are two major entries for mung bean albumin in the UniProt database; Q9FRT8 is a reviewed entry detailing a 10 kDa protein fragment, and Q43680 is an unreviewed entry detailing a 30 kDa protein. Also, several albumin-like proteins have been reported in the UniProt database: A0A1S3U7A2, A0A1S3W1Y8, A0A1S3W1L3, A0A1S3THU2, A0A1S3VX99, A0A1S3UZE0, A0A1S3VUC6, A0A1S3V3Y6, A0A1S3V3H6, A0A1S3V3D6, and A0A1S3UZ12.

[0051] Various pea (Pisum sativum) albumins have been reported in the UniProt database: P62931, P62929, P62928, P62927, P62926, D4AEP7, and P62930. In certain embodiments, albumins, globulins (e.g., β-conglycinin, glycinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadins, and any other proteins from plant, animal or yeast origin, and any mixtures thereof, are water-soluble, soluble under acidic conditions, and / or have an IEP greater than 4, e.g., greater than 4.5, 5, 5.5, 6, 6.5, 7, or e.g., greater than 7.5. By "at least one protein" in the present invention, it is understood that the composition of the present invention or the complex of the present invention may contain only one type of protein (e.g., only albumin or only albumin-like protein) or a complex mixture of proteins (e.g., a mixture of proteins such as albumin and glutelin).

[0052] It is understood that the compositions and complexes of the present invention can contain one or more proteins, polypeptides and / or peptides, and thus the mixture of proteins, polypeptides and / or peptides can contain one single type or different types of proteins, polypeptides and / or peptides. In some embodiments, the polypeptides and peptides can be derived from the same protein or different proteins (e.g., the protein is albumin and the peptides are derived from globulins and glutelins). According to one exemplary embodiment, the protein is one or more albumins, and optionally polypeptides and / or peptides derived from said albumins.

[0053] According to certain exemplary embodiments, the protein is one or more albumins, and optionally, the polypeptides and / or peptides are derived from different proteins; such as globulins (e.g., β-conglycinin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and mixtures thereof. As already mentioned above, the proteins, polypeptides and / or peptides may be of natural or synthetic origin and of plant or animal origin. In some embodiments, the proteins, polypeptides and / or peptides are plant-derived proteins, plant-derived polypeptides and / or plant-derived peptides. The proteins, polypeptides and / or peptides can be derived from any part of a plant that contains proteins (leaves, stems, grains, fruits, tubers, etc.), are water-soluble, optionally soluble under acidic conditions, and / or have an IEP greater than 4, for example greater than 4.5, 5, 5.5, 6, 6.5, 7, or 7.5.

[0054] According to certain exemplary embodiments, at least one plant-derived protein, polypeptide and / or peptide (e.g., albumin, albumin-derived polypeptides and / or peptides) may be derived from, without limitation, potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as, for example, russet potato (coarse brown skin), red potato, white potato, yellow potato (also called Yukon potato) and purple potato, tubers including sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, etc., and from rhizomes such as ginger, bamboo, etc. According to certain exemplary embodiments, at least one plant-derived protein, polypeptide and / or peptide (e.g., albumin, albumin-derived polypeptides and / or peptides) may be derived from legumes, including, without limitation, mung beans, black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, and mixtures thereof.

[0055] According to certain exemplary embodiments, at least one plant-derived protein, polypeptide and / or peptide (e.g., albumin, albumin-derived polypeptides and / or peptides) may be derived from nuts, including, without limitation, almonds, Brazil nuts, cashew nuts, peanuts, pecans, hazelnuts, pine nuts, walnuts, and mixtures thereof. According to certain exemplary embodiments, at least one plant-derived protein, polypeptide and / or peptide (e.g., albumin, albumin-derived polypeptides and / or peptides) may be derived from plant seeds, including, but not limited to, chia, flax, hemp, pumpkin, sesame, sunflower, and mixtures thereof. According to certain exemplary embodiments, at least one plant-derived protein, polypeptide and / or peptide (e.g., albumin, albumin-derived polypeptides and / or peptides) may be derived from a grain, including, but not limited to: oatmeal, wheat, barley, spelt, corn, rice and mixtures thereof.

[0056] According to certain exemplary embodiments, the at least one plant-derived protein, polypeptide and / or peptide (e.g., albumin, albumin-derived polypeptides and / or peptides) is water-soluble, soluble under acidic conditions, and / or has an IEP of greater than 4, e.g., 4.5, 5, 5.5, 6, 6.5, 7, or e.g., greater than 7.5, and may be derived from: a) tubers, including, without limitation, potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as russet (coarse brown skin), red, white, yellow (also called Yukon), and purple, sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, and the like; b) Legumes, including, without limitation: black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, mung beans, fava beans, and mixtures thereof;

[0057] c) Nuts, including, without limitation: almonds, Brazil nuts, cashews, peanuts, pecans, hazelnuts, pine nuts, walnuts, pistachios, and mixtures thereof; d) From the seeds of plants, including but not limited to: chia, flax, hemp, pumpkin, sesame, sunflower, quinoa, chickpea, green pea, pea, rapeseed / canola and mixtures thereof; e) From cereals, including but not limited to: oats, wheat, barley, spelt, corn, rice and mixtures thereof; and / or f) Any other part of a plant that is rich in protein.

[0058] According to certain exemplary embodiments, the at least one plant-derived protein is selected from albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins and / or gliadins (and polypeptides and / or peptides derived therefrom) and may be derived from or obtained from: a) tubers, including, without limitation, potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as russet (coarse brown skin), red, white, yellow (also called Yukon), and purple, sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, and the like;

[0059] b) Legumes, including, without limitation: black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, mung beans, fava beans, and mixtures thereof; c) Nuts, including, without limitation: almonds, Brazil nuts, cashews, peanuts, pecans, hazelnuts, pine nuts, walnuts, pistachios, and mixtures thereof; d) Seeds of plants, including but not limited to: chia, flax, hemp, pumpkin, sesame, sunflower, quinoa, chickpeas, green peas, peas, rapeseed / canola and mixtures thereof; e) Grains, including but not limited to: oats, wheat, barley, spelt, corn, rice and mixtures thereof.

[0060] In certain embodiments, the protein is a natural extract (e.g., of animal and / or plant origin) that comprises at least 30%, 40%, 50%, 60%, 70%, 80%, at least 90%, such as at least 99% of a single type of protein (such as albumin) or a mixture of proteins (such as albumin or albumin-like proteins and / or glutelins). In certain embodiments, the protein is one or more albumins from one or more of the aforementioned plant and / or animal sources (such as potato albumin). In certain embodiments of the compositions or complexes of the invention, the protein is obtained from potato and the polypeptides and / or peptides are obtained from mung bean, pea, soybean and / or rice.

[0061] In certain embodiments of the compositions or complexes of the invention, the at least one protein is selected from albumins, globulins (e.g., β-conglycinin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof. In certain embodiments of the compositions or complexes of the invention, the at least one protein is selected from albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilins, convicilins, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof, and optionally includes polypeptides and / or peptides derived therefrom.

[0062] In certain embodiments of the compositions or complexes of the invention, the at least one protein is an albumin, in particular a plant albumin, more particularly a potato albumin, including polypeptides and / or peptides derived therefrom. In certain embodiments of the compositions or complexes of the invention, the protein is an albumin, particularly a plant albumin, more particularly a potato albumin, and further includes polypeptides and / or peptides derived from albumins, globulins (e.g., β-conglycinin, glycychinin, vicilin, legumin A, legumin J), and / or glutelins (e.g., glutelins A1 and B1). In certain embodiments of the compositions or complexes of the invention, the albumin is obtained from potato, and optionally includes polypeptides and / or peptides derived from albumins, globulins (e.g., β-conglycinin, glycychinin, vicilin, legumin A, legumin J), and / or glutelins (e.g., glutelins A1 and B1) obtained from mung bean, pea, soybean, and / or rice.

[0063] In one embodiment, the composition of the invention or the complex of the invention comprises at least one animal and / or plant albumin, such as potato albumin, and optionally comprises polypeptides and / or peptides derived therefrom (i.e. polypeptides and / or peptides derived from albumin, such as potato albumin derived polypeptides and / or peptides). In one embodiment of the composition or complex of the present invention, the polypeptide and / or peptide has a molecular weight of 200 to about 6000 daltons, 300 to about 5,000 daltons, for example, 300 to 2000 daltons, for example, 300 to 1000 daltons, for example, 500 to 2000 daltons, for example, 500 to 1000 daltons.

[0064] In one embodiment, the composition of the invention or the complex of the invention comprises at least one animal and / or plant albumin, such as potato albumin, mung bean albumin, pea albumin, soybean albumin, egg albumin, etc., and optionally comprises polypeptides and / or peptides having a molecular weight of 300 to about 5,000 Daltons, such as 300 to 2000 Daltons, such as 300 to 1000 Daltons, such as 500 to 2000 Daltons, such as 500 to 1000 Daltons, derived from mung bean, pea, soybean and / or rice. In one embodiment, the composition of the invention or the complex of the invention comprises potato albumin and pea-derived polypeptides and / or peptides having a molecular weight in the range of 500 Da to 3000 Da, with the majority being between 800 and 2000 Da.

[0065] In one embodiment, the composition of the invention or the complex of the invention comprises potato albumin and soybean-derived polypeptides and / or peptides having a molecular weight in the range of 200 Da to 2000 Da, with the majority being between 300 and 1000 Da. In one embodiment, the composition of the invention or the complex of the invention comprises a protein-rich potato extract and a pea extract rich in polypeptides and / or peptides having a molecular weight in the range of 500 Da to 3000 Da, with the majority being between 800 and 2000 Da. In one embodiment, the composition of the invention or the complex of the invention comprises a protein-rich potato extract and a soy extract rich in polypeptides and / or peptides having a molecular weight in the range of 200 Da to 2000 Da, with the majority being between 300 and 1000 Da.

[0066] In certain embodiments of the compositions and complexes of the invention, the ratio of at least one protein to peptide is about 100:1 to 1:100, such as 90, 80, 70, 60, 50, 40, 30, 20, 10 to (:) 20, 30, 40, 50, 60, 70, 80, 90, such as 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, or 10:90. In certain embodiments of the compositions and complexes of the invention, the ratio of at least one phycobilin (such as phycocyanin) to at least one protein, polypeptide, and / or peptide is about 10:1 to about 1:10, such as about 5:1 to about 1:5, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:2. In preferred embodiments of the compositions and complexes of the invention, the ratio of at least one phycobilin (such as phycocyanin) to at least one protein, polypeptide and / or peptide is about 100:1 to 1:100, such as 90, 80, 70, 60, 50, 40, 30, 20, 10 to (:) 20, 30, 40, 50, 60, 70, 80, 90, such as 50:1 to about 1:50, such as about 40:1 to about 1:40, such as about 30:1 to about 1:30, such as about 20:1 to about 1:20, such as about 10:1 to about 1:10, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:2.

[0067] In some embodiments, the compositions or complexes of the present invention are provided as liquid or solid compositions. In certain embodiments, the compositions or complexes of the invention are provided as concentrated formulations comprising 1-95% w / w phycobilin (such as phycocyanin) and 1-95% w / w of at least one protein, polypeptide and / or peptide. In some embodiments, the compositions or complexes of the invention contain 1-90% w / w phycobilin (e.g., phycocyanin), for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 to 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 w / w phycobilin (e.g., phycocyanin).

[0068] In one embodiment, a composition or complex of the invention comprises 1-95% w / w protein, polypeptide and / or peptide, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 to 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1% w / w protein, polypeptide and / or peptide. In some embodiments, a composition or complex of the invention comprises, for example, 50% phycobilin (such as phycocyanin) and 50% protein, polypeptide and / or peptide. In one embodiment, the composition or complex of the present invention comprises about 5-15% w / w, such as about 9% w / w, of phycocyanin (such as Spirulina or Galdieria phycocyanin), about 5%-15% w / w, such as about 10% w / w, of potato protein, and about 5%-15% w / w, such as about 10% w / w, of mung bean peptides and water, and optionally, the mung bean peptides have a molecular weight of 300 to about 5,000 daltons, such as 300-2000 daltons, such as 300-1000 daltons, such as 500-2000 daltons, such as 500-1000 daltons, and the potato protein comprises albumin.

[0069] In some embodiments, the compositions or complexes of the present invention are provided as liquids that can be added to provide color to the final food product. In one embodiment, the composition or complex of the present invention comprises about 20-40% w / w, e.g., about 33% w / w, of phycocyanin (such as Spirulina or Galdieria phycocyanin), about 20-40% w / w, e.g., about 33% w / w, of potato protein, and about 20-40% w / w, e.g., about 33% w / w, of mung bean peptides, which then become a dry composition. In one embodiment, the composition or complex of the invention comprises about 5-15% w / w, e.g., about 9% w / w, of phycocyanin (such as Spirulina or Galdieria phycocyanin), about 5% to about 20% w / w, e.g., about 13% w / w, of potato protein, and about 20% to about 50% w / w, e.g., about 37% w / w, of rice peptides and water, optionally, the rice peptides having a molecular weight of 300 to about 5,000 daltons, e.g., 300-2000 daltons, e.g., 300-1000 daltons, e.g., 500-2000 daltons, e.g., 500-1000 daltons, and the potato protein comprises albumin.

[0070] In some embodiments, the compositions or composites of the present invention are provided as liquids that can be added to provide a desired color to the final food product. In one embodiment, the composition or complex of the invention comprises about 10% to about 20% w / w, e.g., about 14% w / w, of phycocyanin (such as Spirulina or Galdieria phycocyanin), about 10% to about 30%, e.g., about 20% w / w, of potato protein, and about 50% to about 70% w / w, e.g., about 66% w / w, of rice peptides, then in a dry composition, optionally wherein the rice peptides have a molecular weight of 300 to about 5,000 Daltons, e.g., 300 to 2000 Daltons, e.g., 300 to 1000 Daltons, e.g., 500 to 2000 Daltons, e.g., 500 to 1000 Daltons, and the potato protein comprises albumin. In one embodiment, the composition or complex of the present invention comprises about 5-15% w / w, e.g., about 9% w / w, of phycocyanin (such as Spirulina or Galdieria phycocyanin), about 5% to about 20% w / w, e.g., about 13% w / w, of potato protein, and about 20% to about 50% w / w, e.g., about 37% w / w, of soybean peptides and water. In one embodiment, the composition or complex of the present invention is provided as a liquid that can be added to provide a desired color to the final food product.

[0071] In one embodiment, the composition or complex of the invention comprises about 10% to about 20% w / w, e.g., about 14% w / w, of phycocyanin (such as Spirulina or Galdieria phycocyanin), about 10% to about 30%, e.g., about 20% w / w, of potato protein, and about 50% to about 70% w / w, e.g., about 66% w / w, of soybean peptides, which then form a dry composition. Those skilled in the art will understand that the stabilized compositions or complexes of the present invention may be formulated with additional carriers or other excipients, such as food approved carriers or excipients, and that the stabilized compositions or complexes may be used in an end use application to provide the characteristic color of the stabilized phycobilin (such as phycocyanin).

[0072] The present invention also relates to a composition comprising: i) at least one phycobilin (such as phycocyanobilin), and ii) at least one protein extract comprising one or more peptides, polypeptides and / or proteins, wherein at least one protein, polypeptide and / or peptide is water-soluble, has an IEP greater than 4 and / or is soluble under acidic conditions.

[0073] The present invention also relates to a complex comprising: i) at least one phycobilin (such as phycocyanobilin), and ii) at least one protein extract comprising one or more peptides, polypeptides and / or proteins, obtainable by mixing in an aqueous solution at least one protein extract with at least one composition comprising phycocyanobilin, wherein at least one protein, polypeptide and / or peptide is water-soluble, has an IEP greater than 4 and / or is soluble under acidic conditions. The protein extract may be of animal, plant, yeast and / or bacterial origin. In a preferred embodiment, the protein extract is of animal and / or plant origin.

[0074] By "plant protein extract" (e.g. "potato protein extract", "mung bean protein extract", "rice protein extract") is understood in the present invention an extract which is obtained or can be obtained from a plant or part of a plant (e.g. potato, mung bean, rice, pea and any other plant) and which comprises proteins, polypeptides and / or peptides. In certain embodiments, the protein extract (e.g., a plant and / or animal protein extract) comprises at least 30% w / w, 40% w / w, 50% w / w, 60% w / w, 70% w / w, 80% w / w, at least 90% w / w, such as at least 99% w / w of peptides, polypeptides and / or proteins. In preferred embodiments, the protein extract (e.g., a plant and / or animal protein extract such as potato protein extract, mung bean protein extract, rice protein extract, pea protein extract, etc.) is rich in protein. In some embodiments, the extract (e.g., a plant and / or animal protein extract) is rich in polypeptides and / or peptides.

[0075] In the present invention, "enriched in protein" or "rich in protein" means that the plant or animal protein extract (e.g. potato protein-enriched extract, mung bean protein-enriched extract, rice protein-enriched extract, pea protein-enriched extract) comprises at least 50%, such as at least 60%, 70%, 80%, or at least 90%, such as at least 99% w / w protein. In an embodiment, the plant or animal protein-enriched extract is water-soluble and / or soluble under acidic conditions, and optionally has an IEP of more than 4, such as more than 4.5, 5, 5.5, 6, 6.5, more than 7, or such as more than 7.5. In the present invention, "enriched in peptides" or "rich in peptides" means that the plant or animal protein extract (e.g. potato protein-enriched extract, mung bean protein-enriched extract, rice protein-enriched extract, pea protein-enriched extract) contains at least 50%, such as at least 60%, 70%, 80%, or at least 90%, such as at least 99% peptide and / or polypeptide protein.

[0076] In some embodiments, a protein extract (e.g., a protein-enriched protein extract or a peptide-enriched protein extract) contains at least 50%, 60%, 70%, or 80%, e.g., at least 90%, proteins, polypeptides and / or peptides that are water soluble and / or soluble under acidic conditions, and optionally have an IEP greater than 4, e.g., 4.5, 5, 5.5, 6, 6.5 or greater than 7, e.g., greater than 7.5. In some embodiments, a protein extract (e.g., a protein-enriched protein extract or a peptide-enriched protein extract) contains at least 50%, 60%, 70%, or 80%, such as at least 90%, of the proteins, polypeptides and / or peptides that have an IEP greater than 4, such as 4.5, 5, 5.5, 6, 6.5 or greater than 7, such as greater than 7.5.

[0077] In some embodiments, a protein extract (e.g., a protein-rich protein extract) contains at least 30%, 40%, 50%, 60%, 70%, 80%, or at least 90%, e.g., at least 99%, of one protein (e.g., albumin or albumin-like protein). In some embodiments, a protein extract (e.g., a protein-rich protein extract) comprises at least 30%, 40%, 50%, 60%, 70%, 80%, or at least 90%, e.g., at least 99%, of a mixture of proteins (e.g., albumin or albumin-like protein and glutelin). In certain embodiments, a protein extract comprises at least 30%, 40%, 50%, 60%, 70%, 80%, or at least 90%, such as at least 99%, of a protein (e.g., albumin or an albumin-like protein), and optionally polypeptides and / or peptides derived therefrom (e.g., albumin-derived peptides).

[0078] According to certain exemplary embodiments, the at least one protein extract comprises proteins, polypeptides and / or peptides, which may be derived from: a) tubers, including, without limitation, potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as russet (coarse brown skin), red, white, yellow (also called Yukon), and purple, sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, and the like; b) Legumes, including, without limitation: black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, mung beans, fava beans, and mixtures thereof;

[0079] c) Nuts, including, without limitation: almonds, Brazil nuts, cashews, peanuts, pecans, hazelnuts, pine nuts, walnuts, pistachios, and mixtures thereof; d) From the seeds of plants, including but not limited to: chia, flax, hemp, pumpkin, sesame, sunflower, quinoa, chickpea, green pea, pea, rapeseed / canola and mixtures thereof; e) From cereals, including but not limited to: oats, wheat, barley, spelt, corn, rice and mixtures thereof; f) eggs, milk, and other animal sources rich in protein; and mixtures thereof.

[0080] According to certain exemplary embodiments, at least one protein extract (e.g., a plant or animal protein extract) comprises proteins, polypeptides and / or peptides that are water soluble and / or soluble under acidic conditions, and optionally have an IEP greater than 4, e.g., greater than 4.5, 5, 5.5, 6, 6.5, or greater than 7, e.g., greater than 7.5; and may come from: a) tubers, including, without limitation, potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as russet (coarse brown skin), red, white, yellow (also called Yukon), and purple, sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, and the like;

[0081] b) Legumes, including, without limitation: black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, mung beans, fava beans, and mixtures thereof; c) Nuts, including, without limitation: almonds, Brazil nuts, cashews, peanuts, pecans, hazelnuts, pine nuts, walnuts, pistachios, and mixtures thereof; d) From the seeds of plants, including but not limited to: chia, flax, hemp, pumpkin, sesame, sunflower, quinoa, chickpea, green pea, pea, rapeseed / canola and mixtures thereof; e) From cereals, including but not limited to: oats, wheat, barley, spelt, corn, rice and mixtures thereof; f) eggs, milk, and other animal sources rich in protein; and mixtures thereof.

[0082] In certain embodiments, at least one protein extract (e.g., a plant or animal protein extract, such as a protein-rich protein extract) comprises proteins selected from albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and any mixtures thereof, and optionally polypeptides and / or peptides derived therefrom.

[0083] As mentioned above, preferably the one or more proteins selected from albumins, globulins, glutelins, prolamins, lectins, gliadins, tuberins, patatins, and any other proteins of plant, animal or yeast origin, and any mixtures thereof, are water-soluble. In certain embodiments, at least 50% of one or more proteins selected from albumin, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof are water soluble, such as at least 60%, 70%, 80%, 90%, or e.g. at least 99% of albumin, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadins, tuberins, patatins, and mixtures thereof are water soluble. In preferred embodiments, at least 80% of one or more proteins selected from albumins, globulins (e.g., β-conglycinin, glycinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and mixtures thereof are water soluble.

[0084] In certain embodiments, at least one protein extract (e.g., a plant or animal protein extract, such as a protein-rich protein extract) contains only water-soluble albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and any other soluble proteins of animal, plant, or yeast origin, and any mixtures thereof. In certain embodiments, at least one protein extract (e.g., a plant or animal protein extract, such as a protein-rich protein extract) comprises water-soluble proteins such as albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and mixtures thereof, as well as water-soluble polypeptides and / or water-soluble peptides derived therefrom (i.e., albumins, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, and mixtures thereof).

[0085] In certain embodiments, at least one protein extract (e.g., a plant or animal protein extract, such as a peptide-enriched extract) comprises water soluble polypeptides and / or peptides derived from, for example, albumins, globulins (e.g., β-conglycicin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelins (e.g., glutelins A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and mixtures thereof. In one embodiment, the peptide has a molecular weight of about 100 to about 10,000 daltons, for example, 100 to about 5,000 daltons, for example, 300 to 2000 daltons, for example, 200 to 500 daltons, for example, 300 to 1000 daltons, for example, 500 to 1000 daltons, for example, 500 to 2000 daltons, for example, 500 to 1000 daltons.

[0086] In certain embodiments, at least one protein extract (e.g., a plant or animal protein extract) contains at least 50%, 60%, 70%, 80%, 80%, or, for example, 99% of proteins, polypeptides and / or peptides that are soluble under acidic conditions, e.g., soluble at a pH of less than 6, less than 5, less than 4, less than 3, or less than 2. In certain embodiments, compositions and complexes of the invention comprise a potato protein extract (e.g., a protein-rich potato extract or a peptide-rich potato extract), and, optionally, a mung bean protein extract (e.g., a protein-rich and / or peptide-rich mung bean extract).

[0087] In one embodiment the composition or complex of the invention comprises a potato extract comprising at least 20% w / w, at least 30% w / w, 40% w / w, 50% w / w, 60% w / w, 70% w / w, 80% w / w, at least 90% w / w or such as at least 99% w / w potato albumin (e.g. a protein rich potato extract) and optionally a mung bean extract comprising at least 50% w / w, 60% w / w, 70% w / w, 80% w / w or such as at least 90% w / w polypeptides and / or peptides (peptide rich mung bean extract). In certain embodiments the natural extract comprises at least 20% w / w, at least 30% w / w, 40% w / w, 50% w / w, 60% w / w, 70% w / w, 80% w / w, at least 90% w / w, or such as at least 99% w / w potato albumin, and optionally polypeptides and / or peptides derived from mung bean.

[0088] Various plant protein extracts and methods for their production are described in the literature. Potato protein extracts and methods for their extraction are described, for example, in Peksa A. et al (Food 2009. 79-87 Global science books), which are incorporated herein by reference. Methods for producing plant protein extracts are known in the art. For example, when extracting soluble proteins, proteins, polypeptides and / or peptides, water or acidic water can be used as a solvent. Protein-rich plants or plant parts (e.g., potatoes, mung beans, rice, peas, etc.) can be processed prior to extraction, e.g., washed, dried, milled, or ground.

[0089] For example, the extraction process may include the following steps: (i) Extraction of protein-rich plants or plant parts (e.g. potato, mung bean, rice, pea, etc.) with a suitable solvent (e.g. water); (ii) evaporation of the solvent; and, if necessary, (iii) purification of the extract (e.g., by chromatography, membrane filtration, etc.); In certain embodiments, the extraction temperature ranges from about 20° C. to about 100° C. In certain embodiments, the extraction temperature ranges from about 50° C. to about 70° C. Typically, the ratio of plant material to solvent mixture used in the extraction process varies from about 1:1 to about 1:10, such as from about 1:3 to about 1:8, on a gram to milliliter basis. The incubation period (i.e., the period during which the plant material is in contact with the solvent) is typically from about 2 hours to about 24 hours.

[0090] Mechanical energy can be applied during the extraction process to promote homogenization of the mixture, alter the physical structure of the starting biological material, and increase the extraction yield. In some embodiments, the protein extract can be subjected to denaturation and / or hydrolysis (chemical, thermal or enzymatic) to obtain polypeptide and / or peptide extract. In some embodiments, the protein raw material can be hydrolyzed by one or more hydrolases. In one example, an enzyme preparation with low exopeptidase activity is used to minimize the release of free amino acids and improve the taste profile of the protein hydrolysate. The type of treatment (ie, the enzyme used), the time and temperature of the treatment can affect the length of the polypeptides and peptides.

[0091] Optionally, the enzyme after a specified incubation time can be destroyed by heat treatment. The peptide can further be separated by centrifugation or other techniques known in the art, and then purified, for example, using membrane filtration or other suitable techniques known in the art. An additional sterilization step can also be used. Optionally, the peptide preparation can be dried, for example, using spray drying. Animal proteins and peptides (polypeptides and / or peptide extracts) can be obtained using methods described previously. In one embodiment, the peptides from the peptide-enriched extract have a molecular weight of about 100 to about 10,000 daltons, such as 100 to about 5,000 daltons, for example, 300 to 2000 daltons, such as 200 to 500 daltons, for example, 300 to 1000 daltons, such as 500 to 1000 daltons, for example, 500 to 2000 daltons.

[0092] In some embodiments, compositions and complexes of the invention comprise a plant or animal protein-enriched extract (e.g., a potato protein extract, such as a potato protein extract containing albumin), and, optionally, a plant or animal peptide-enriched extract (e.g., a mung bean peptide-enriched extract, a rice peptide-enriched extract, and / or a pea peptide-enriched extract). In one embodiment the compositions and complexes of the invention comprise a potato protein extract enriched in proteins as described above, and optionally a mung bean peptide-enriched extract comprising peptides with a molecular weight of from about 100 to about 10,000 daltons, such as from 100 to about 5,000 daltons, for example from 300 to 2000 daltons, such as from 200 to 500 daltons, for example from 300 to 1000 daltons, such as from 500 to 1000 daltons, for example from 500 to 2000 daltons, for example from 500 to 1000 daltons.

[0093] In one embodiment the compositions and complexes of the invention comprise a potato protein-enriched extract rich in proteins as described above, and optionally a rice peptide-enriched extract comprising peptides with a molecular weight of from about 100 to about 10,000 daltons, such as from 100 to about 5,000 daltons, for example from 300 to 2000 daltons, for example from 200 to 500 daltons, for example from 300 to 1000 daltons, such as from 500 to 1000 daltons, for example from 500 to 2000 daltons, for example from 500 to 1000 daltons. In one embodiment the compositions and complexes of the invention comprise a potato protein-enriched extract rich in proteins as described above and, optionally, a pea peptide-enriched extract comprising peptides with a molecular weight of from about 100 to about 10,000 daltons, such as from 100 to about 5,000 daltons, for example from 300 to 2000 daltons, such as from 200 to 500 daltons, for example from 300 to 1000 daltons, such as from 500 to 1000 daltons, for example from 500 to 2000 daltons, such as from 500 to 1000 daltons, for example from 800 to 2000 daltons.

[0094] In one embodiment, the compositions and complexes of the present invention comprise a potato protein-enriched extract as described above, and optionally a mung bean peptide-enriched extract comprising peptides with a molecular weight of 300 to about 5,000 daltons, such as 300 to 2000 daltons, for example 300 to 1000 daltons, such as 500 to 2000 daltons, for example 500 to 1000 daltons, and a rice peptide-enriched extract comprising peptides with a molecular weight of 300 to about 5,000 daltons, such as 300 to 2000 daltons, for example 300 to 1000 daltons, such as 500 to 2000 daltons, for example 500 to 1000 daltons.

[0095] In a preferred embodiment, the potato protein extract is rich in proteins such as albumin, globulins (e.g. β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g. glutelin A1 and B1), and / or prolamins. In an embodiment, the potato protein extract comprises at least 50% w / w albumin, such as at least 60% w / w albumin, such as at least 70% w / w albumin, such as at least 80% w / w albumin, or such as at least 90% w / w albumin, and optionally one or more globulins (e.g. β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g. glutelin A1 and B1), and / or prolamins, and optionally polypeptides and / or peptides derived therefrom.

[0096] In a preferred embodiment, the mung bean peptides (e.g. a peptide-rich mung bean extract) comprise polypeptides and / or peptides having a molecular weight of 300 to about 5,000 Daltons, such as 300 to 2000 Daltons, such as 300 to 1000 Daltons, such as 500 to 2000 Daltons, such as 500 to 1000 Daltons, and optionally derived from one or more of the following proteins: albumin, globulins and glutelins. In a preferred embodiment, the peptides are predominantly derived from albumin (e.g. at least 60% w / w of the peptides are albumin peptides), such as more than 70% w / w, such as more than 80% w / w, such as more than 90% w / w).

[0097] In preferred embodiments, the rice peptides (e.g., peptide-rich rice extract) comprise polypeptides and / or peptides from glutelin, such as glutelin A1 and B1. In some embodiments, the rice peptides have a molecular weight of 300 to about 5,000 Daltons, such as 300 to 2000 Daltons, such as 300 to 1000 Daltons, such as 500 to 2000 Daltons, such as 500 to 1000 Daltons, and are optionally derived from one or more of albumin, globulin, and glutelin proteins. In preferred embodiments, the peptides are albumin peptides (e.g., about 10% w / w to 30% w / w or about 30% w / w to 70% w / w) and / or glutelin peptides, such as glutelin A1 and B1 (e.g., 30% to about 70% w / w). In some embodiments, the peptides are generated using pepsin enzymatic digestion.

[0098] In a preferred embodiment, the pea peptides (e.g. peptide-rich pea extract) comprise polypeptides and / or peptides from vicilin, legumin A, provicilin, convicilin, legumin J, and / or albumin 2. In an embodiment, the pea peptides have a molecular weight of 100 to about 10,000 daltons, such as 100 to about 5,000 daltons, for example 300 to 2000 daltons, such as 200 to 500 daltons, for example 300 to 1000 daltons, such as 500 to 1000 daltons, for example 500 to 2000 daltons, such as 500 to 1000 daltons, for example 800 to 2000 daltons. In an embodiment, the peptides have been generated using pepsin enzymatic digestion.

[0099] In a preferred embodiment, the soy peptides (e.g., peptide-rich soy extracts) comprise polypeptides and / or peptides from glycine G1 and / or β-conglycinin. In an embodiment, the soy peptides have a molecular weight of 100 to about 10,000 daltons, such as 100 to about 5,000 daltons, such as 300 to 2000 daltons, such as 200 to 500 daltons, such as 300 to 1000 daltons, such as 500 to 1000 daltons, such as 500 to 2000 daltons, such as 500 to 1000 daltons, such as 800 to 2000 daltons. In an embodiment, the peptides are produced using pepsin enzyme digestion.

[0100] In certain embodiments of the compositions and complexes of the invention, the ratio of protein-enriched extract (e.g., a potato protein-enriched extract) to peptide-enriched extract (e.g., a peptide-rich mung bean extract) is about 100:1 to 1:100, such as 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, or 10:90. In certain embodiments of the compositions and complexes of the invention, the ratio of the at least one phycobilin (such as phycocyanin) to the at least one plant protein extract (e.g., potato protein extract, mung protein extract, rice protein extract and / or pea protein extract) is about 50:1 to about 1:50, such as about 40:1 to about 1:40, such as about 30:1 to about 1:30, such as about 20:1 to about 1:20, such as about 10:1 to about 1:10, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:2. In one embodiment, the composition or complex of the invention comprises 0.025% Spirulina extract having 25% w / w total phycocyanin, 0.075% potato protein extract having at least 90% w / w protein, and 0.075% pea extract rich in peptides having at least 80% w / w peptides, optionally the pea peptides having a molecular weight in the range of 500 Da to 3000 Da, with the majority being between 800 and 2000 Da.

[0101] In one embodiment, the composition or complex of the invention comprises 0.025% Spirulina extract having 25% w / w total phycocyanin, 0.075% potato protein extract having at least 90% w / w protein, and 0.075% soy extract rich in peptides having at least 80% w / w peptides, optionally the soy peptides having a molecular weight in the range of 200 Da to 2000 Da, with the majority being between 300 and 1000 Da. In one embodiment, in a composition or complex of the invention, the phycobilin is phycocyanin, the protein is potato protein, and the peptides and / or polypeptides are from mung bean, soybean, rice and / or pea, and the ratio between them is about 0.008:1:1 to about 0.8:1:1.

[0102] In one embodiment, the composition or complex of the invention comprises a Spirulina extract containing at least 25% w / w phycocyanin, a potato protein extract having at least 90% w / w protein, and a peptide-rich pea extract having at least 80% w / w peptides, optionally the pea peptides having a molecular weight in the range of 500 Da to 3000 Da, with the majority being between 800 and 2000 Da; wherein the ratio of potato extract to pea extract is about 10:1 to about 1:10, such as about 5:1 to about 1:5, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:2, and wherein the ratio of phycocyanin:proteins and peptides is about 20:1 to about 1:20, such as about 10:1 to about 1:10, such as 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, or about 1:2.

[0103] In one embodiment, the composition or complex of the invention comprises 0.025% spirulina extract having 25% w / w total phycocyanin, 0.075% potato protein extract having at least 90% w / w protein, and a peptide-rich soy extract having at least 80% w / w peptides, optionally the soy peptides having a molecular weight in the range of 200 Da to 2000 Da, with the majority being between 300 and 1000 Da. and wherein the ratio of potato extract to pea extract is about 10:1 to about 1:10, such as about 5:1 to about 1:5, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:2, and wherein the ratio of phycocyanin:proteins and peptides is about 20:1 to about 1:20, such as about 10:1 to about 1:10, such as 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, or about 1:2. As demonstrated in the examples of the present application, the combination of phycocyanin with soluble peptides, polypeptides and proteins results in the stabilization of said phycocyanin. Thus, in some embodiments, the color provided by said composition or complex of the present invention is stable to heat, acidic conditions and / or exposure to light.

[0104] In certain embodiments of the compositions or complexes of the invention, the phycobilin (such as phycocyanin) present in said composition or complex and complexed with at least one soluble peptide, polypeptide and / or protein has a blue color that is stable to heat, acidic conditions, and / or exposure to light. It is known that dissolved phycocyanin (e.g., dissolved in water or other liquids such as syrup) will aggregate, precipitate, and lose color when exposed to acidic conditions, such as pH less than 5, e.g., pH less than 4.5. In the present invention, the term "stable to acidic conditions" means that the hue of at least one phycobilin (e.g., phycocyanin) present in the composition or complex of the present invention does not change or has a change of less than 30%, e.g., less than 20%, e.g., less than 10%, or e.g., less than 5%, relative to the hue measured before acidic treatment. "Acidic treatment or acidic conditions" in the present application means a pH less than 5, e.g., less than pH 4.5, e.g., less than pH 4, e.g., less than pH 3, or e.g., less than pH 2.

[0105] Thus, in certain embodiments, the color provided by the composition or complex of the present invention is stable at a pH of less than about 5, such as a pH of less than about 4.5, such as a pH of less than about 4, such as a pH of less than about 3, or such as a pH of less than about pH 2. Thus, in certain embodiments of the compositions or complexes of the invention, the phycobilin (such as phycocyanin) is stable at a pH below about 6, such as a pH below about 5, such as a pH below about 5, such as a pH below about 4, such as a pH below about 3, or such as a pH below about 2. In a preferred embodiment, the phycobilin (e.g., phycocyanin) present in the composition or complex of the present invention is stable at a pH of about 2.0 to about 5.0, about 3 to about 5, about 3 to about 4, preferably about 2.2 to about 4.0, more preferably about 2.3 to about 3.6, and most preferably about 2.4 to about 3.3.

[0106] Thus, in certain embodiments, the color provided by the composition or complex of the present invention is stable at a pH of about 2.0 to about 5.0, about 3 to about 5, about 3 to about 4, preferably about 2.2 to about 4.0, more preferably about 2.3 to about 3.6, and most preferably about 2.4 to about 3.3. In the present invention, the term "thermostable" means that the hue provided by at least one phycobilin (such as phycocyanin) does not change or has a hue change of less than 30%, such as less than 20%, such as less than 10%, or such as less than 5% relative to the hue measured before heat treatment.

[0107] Thus, in certain embodiments, the compositions or composites of the invention and the color provided by said compositions or composites of the invention are stable after heat treatment above 40° C., such as 50° C., 60° C., 70° C., 80° C., or even above 90° C., for at least 30 seconds, at least 1 minute, at least 5 minutes, at least 10 minutes, at least 30 minutes, at least 1 hour, at least 2 hours, or even for at least 5 hours. In certain embodiments, the color provided by said compositions or composites of the invention is stable after heat treatment at about 40° C. to 90° C., such as about 40° C. to 80° C., such as about 50° C. to 90° C., such as about 60° C. to 90° C., such as about 70° C. to 90° C. In certain embodiments, the heat treatment is for a period of 30 seconds to 5 hours, such as 1 minute to 1 hour, such as 10 minutes to 30 minutes.

[0108] Thus, in certain embodiments of compositions or complexes of the invention, the phycobilin (such as phycocyanin) is stable after heat treatment above 40°C, e.g., above 50°C, 60°C, 70°C, 80°C, or e.g., above 90°C, for at least 30 seconds, at least 1 minute, at least 5 minutes, at least 10 minutes, at least 30 minutes, at least 1 hour, at least 2 hours, or e.g., for at least 5 hours. In certain embodiments, the color provided by the composition or composite of the present invention is stable to exposure to light. In certain embodiments of the compositions or complexes of the invention, the color provided by the phycobilin (such as phycocyanin) is stable to exposure to light.

[0109] In the present invention, the term "stable to light" or "stable to light exposure" means that the hue provided by at least one phycobilin (such as phycocyanin) does not change or has a change of less than 30% relative to the hue measured in the dark. Thus, in certain embodiments of the compositions or complexes of the invention, the color provided by the phycobilin (such as phycocyanin) is stable after exposure to light for more than 5 minutes, such as more than 30 minutes, such as more than 1 hour, such as 2 hours, 3 hours, 4 hours, 5 hours, 10 hours, 12 hours, 24 hours, 2 days, 10 days, such as more than 30 days, or such as more than 60 days.

[0110] Thus, in certain embodiments of the compositions or complexes of the invention, the color provided by the phycobilin (e.g., phycocyanin) is stable after exposure to light for about 5 minutes to 60 days, e.g., about 30 minutes to 30 days, e.g., about 1 hour to 24 hours, e.g., about 2 days to 30 days, e.g., about 30 days to 60 days or more. In one embodiment, the colorant composition (such as a food colorant composition) of the present invention is in liquid form. Optionally, the colorant composition further comprises water. Optionally, the water in the colorant composition may be present in an amount of up to about 20% w / w. Optionally, the amount of water in the colorant composition is about 3.5%, 4.0%, 4.5% or 5.0% w / w. After the complex of at least one phycobilin (such as phycocyanin) and at least one protein, polypeptide and / or peptide (or the protein extract as described above) is formed, the composition of the present invention or the complex of the present invention can be dried by any method known in the art.

[0111] Thus, in one embodiment, the colorant composition (such as a food colorant composition) or complex of the present invention is in powder form. Optionally, the food colorant composition further comprises a carrier. Optionally, the carrier may be selected from the group consisting of maltodextrin, gum arabic, inulin, alginate, starch, modified starch, and combinations thereof. Optionally, the carrier may be maltodextrin. Optionally, the carrier may be present in the colorant composition in an amount of up to about 4.0% w / w. Optionally, the amount of carrier in the composition is about 1.0% to about 3.5% w / w. Optionally, the amount of carrier in the food composition is about 1.2%, 2.0%, 2.1% or 3.1% w / w. The present invention also relates to a process for the formation of a complex according to the present invention, which comprises the step of mixing at least one peptide, polypeptide and / or protein as defined herein, or a protein extract comprising at least one peptide, polypeptide and / or protein as defined herein, with a composition comprising at least one phycobilin in an aqueous solution.

[0112] In one embodiment of the process for complex formation of the present invention, the composition comprising at least one phycobilin in aqueous solution is mixed with at least one peptide, polypeptide and / or protein, followed by filtering to remove the insoluble fraction. The present invention also relates to a method for stabilizing phycobilins (or a method of the present invention) comprising the steps of: i) contacting phycobilins in an aqueous solution with at least one protein, polypeptide and / or peptide as defined above, or with a protein extract comprising at least one protein, polypeptide and / or peptide as defined above; ii) Optionally, adding sugar.

[0113] In certain embodiments, the method of the present invention further comprises filtering the mixture obtained in steps i) and / or ii) to remove the insoluble fraction. In certain embodiments, the method of the invention further comprises concentrating the result of step i) or ii). In certain embodiments, the methods of the present invention further comprise a drying step, such as spray drying. All the embodiments relating to the phycobilins, at least one protein, polypeptide and / or peptide, or protein extract described above for the inventive composition or complex also apply to the method of the invention.

[0114] In certain embodiments of the processes or methods of the invention, at least one protein, polypeptide and / or peptide is water soluble, has an IEP greater than 4, and / or is soluble under acidic conditions. In certain embodiments of the process or method of the present invention, the protein extract is a plant protein extract and / or an animal protein extract. In certain embodiments of the process or method of the present invention, the protein extract is a protein-rich extract. In certain embodiments of the process or method of the present invention, the protein extract is enriched in peptides.

[0115] In one embodiment of the process or method of the invention, the peptide has a molecular weight of about 100 to about 10,000 daltons, such as 100 to about 5,000 daltons, for example 300 to 2000 daltons, such as 200 to 500 daltons, for example 300 to 1000 daltons, such as 500 to 1000 daltons, for example 500 to 2000 daltons, such as 500 to 1000 daltons, or for example 800 to 2000 daltons. In certain embodiments of the process or method of the present invention, at least one protein, polypeptide and / or peptide, or protein extract is obtained from: a) tubers, such as potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as russet potato (coarse brown skin), red potato, white potato, yellow potato (also called Yukon potato), and purple potato, sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, etc.;

[0116] b) legumes, such as beans selected from black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, mung beans, broad beans, and mixtures thereof; c) nuts, such as nuts selected from almonds, Brazil nuts, cashews, peanuts, pecans, hazelnuts, pine nuts, walnuts, pistachios, and mixtures thereof; d) from plant seeds, for example seeds selected from chia, flax, hemp, pumpkin, sesame, sunflower, quinoa, chickpea, green pea, pea, rapeseed / canola and mixtures thereof; e) cereals, for example cereals selected from oats, wheat, barley, spelt, corn, rice and mixtures thereof, and / or f) Eggs, milk, and any other animal source rich in protein.

[0117] In certain embodiments of the processes or methods of the present invention, the phycobilin is selected from phycoerythrobilin, phycocyanobilin, phycoviolobilin, phycourobilin, and mixtures thereof. In certain embodiments of the process or method of the present invention, the protein extract comprises at least one protein selected from albumin, globulins (e.g., β-conglycisin, glycychinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamins, lectins, gliadin, tuberin, patatin, and mixtures thereof, and optionally peptides and / or polypeptides derived therefrom.

[0118] In certain embodiments of the process or method of the present invention, the protein extract contains at least one albumin selected from the following: potato albumin, yucca albumin, carrot albumin, radish albumin, beetroot albumin, mung bean albumin, black bean albumin, canelli bean albumin, kidney bean albumin, lentil albumin, lima bean albumin, pinto bean albumin, soybean albumin, white bean albumin, corn (Zea mays) albumin, corn albumin, corn sorghum ... mais) albumin, almond albumin, brazil nut albumin, cashew nut albumin, peanut albumin, pecan albumin, hazelnut albumin, pine nut albumin, walnut albumin, brazil nut albumin, cashew nut albumin, peanut albumin, pecan albumin, hazelnut albumin, pine nut albumin, walnut albumin, chia albumin, flax albumin, hemp albumin, pumpkin albumin, sesame albumin, sunflower albumin, green pea albumin, lentil albumin, egg albumin, bovine albumin, rapeseed / canola albumin, oat albumin, wheat albumin, barley albumin, spelt albumin, corn albumin, rice albumin and any mixtures thereof; and optionally peptides and / or polypeptides derived therefrom.

[0119] In one embodiment of the process or method of the invention, the protein is an extract from potato, and optionally includes peptide-enriched extracts from mung bean, pea and / or rice. In another aspect, the invention relates to a stabilized phycobilin obtainable using the method of the invention, or "the stabilized phycobilin of the invention". In one embodiment, the stable phycobilin (such as phycocyanin) is phycocyanin from Spirulina (Arthrospira platensis). In certain embodiments, the stable phycobilins (such as phycocyanins) of the present invention are stable under acidic conditions, heat stable, light stable, and / or light stable. The terms heat stable, light stable, and light stable have been previously explained.

[0120] In certain embodiments, the stabilized phycobilins (such as phycocyanins) of the present invention have a color with a maximum absorption band of 618 nm±5 nm. In certain embodiments, the stabilized phycobilins (such as phycocyanins) of the present invention have a color with a maximum absorption band of 565 nm±5 nm. In certain embodiments, the stabilized phycobilins (such as phycocyanins) of the present invention have a color with a maximum absorption band of 495 nm±5 nm. In certain embodiments, the stabilized phycobilins (such as phycocyanins) of the present invention have a color with a maximum absorption band of 550 nm±5 nm.

[0121] Color evaluation The colorant compositions and foods of the present disclosure can be analyzed using a spectrophotometer, and CIELAB L*a*b* values ​​can be calculated from the spectral data, as described in more detail below. The L*a*b* values ​​provide a means of characterizing the color and assessing the magnitude of difference between two colors. The L*a*b* values ​​also provide a means of characterizing the color and assessing the magnitude of difference between two colors of products as well as solutions. Measurement of colorant compositions and products in solid form is achieved using reflectance measurements from the surface of the product.

[0122] For example, the L*a*b* values ​​consist of a set of coordinate values ​​defined in a three-dimensional Cartesian coordinate system. L* is the lightness coordinate, providing a scale of lightness from black (0L* units) to white (100L* units) on the vertical axis. a* and b* are coordinates related to both hue and chroma. a* provides a scale from green (-a* units) to red (+a* units) on the horizontal axis, with the midpoint (0a* units) being neutral; b* provides a scale from blue (-b* units) to yellow (+b* units) on a second horizontal axis perpendicular to the first, with the midpoint (0b* units) being neutral. The three axes intersect at a value of 50 for L* and 0 for both a* and b*.

[0123] ΔE is a measure of the magnitude of the total color difference between two colors, expressed in the CIELAB L*a*b* color space. Experienced color observers have been reported to be unable to distinguish the difference between two colors when ΔE is approximately 2.3 or less. The ΔE of two different colors with L*a*b* values ​​L*1a*1b*1 and L*2a*2b*2 is calculated using Equation 1:

number

[0124] In one embodiment, the phycobilin (e.g., a Spirulina derived colour, such as a Spirulina extract) is present in the colourant compositions, complexes and colourant compositions of the invention in an amount ranging from about 0.5% to about 25.0% w / w. Optionally, the phycobilin-containing colour (e.g., a Spirulina derived colour, such as a Spirulina extract) is present in an amount ranging from about 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0%, 10.5%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, 21.0%, 22.0%, 23.0%, 24.0%, 25.0%, 26.0%, 27.0%, 28.0%, 29.0%, 30.0%, 31.0%, 32.0%, 33.0%, 34.0%, 35.0%, 36.0%, 37.0%, 38.0%, 39.0%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%, 49.0%, 50.0%, 51.0%, 52.0%, 53.0%, 54.0%, 55.0%, 56.0%, 57.0%, 58.0%, 59. 5%, 11.0%, 11.5%, 12.0%, 12.5%, 13.0%, 13.5%, 14.0%, 14.5%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%, 19.5%, 20.0%, 20.5% or 21.0% w / w. Optionally, the phycobilin-containing colorant (e.g., a Spirulina-derived pigment such as a Spirulina extract) is present in an amount of about 1.5%, 3.0%, 10%, 19.0% or 20.0% w / w. Optionally, the phycobilin-containing colorant (eg, a spirulina-derived pigment such as a spirulina extract) is present in an amount of about 1.7%, 2.7%, 10.2%, 18.7% or 20.0% w / w.

[0125] Mixing with other dyes The stabilized phycobilins (such as phycocyanins) present in the compositions, complexes and colorant compositions of the present invention can be combined with other pigments to produce a wide range of colors. For example, a composition of the present invention having a stable blue color will be mixed with a pigment having a yellow color to produce a stable green color. Thus, the compositions, composites and colorant compositions of the present invention may further comprise other pigments. The addition of further pigments may be carried out before the formation of a complex between the phycobilin (such as phycocyanin) and at least one protein, polypeptide and / or peptide (a complex of the invention) or after said complex is in place. Thus, in some embodiments, stabilized phycobilins (such as phycocyanins) can be mixed with other pigments. Thus, in some embodiments, the colorant compositions of the present invention can be mixed with other pigments. The pigments used may be of synthetic or natural origin. In a preferred embodiment, the pigments are natural pigments, such as colored food or natural dyes.

[0126] Pigments The main classes of plant pigments are chlorophylls, which give plants their green color; red, orange, or yellow carotenoids; anthocyanins, which appear red, purple, blue, or black depending on the pH; and red or yellow betalains. Thus, pigments suitable for use in the present invention are selected from the group consisting of anthocyanins, carotenoids, chlorophylls, betalains, anthraquinones, naphthoquinones and azaphilones. The pigments used in the present invention can be derived from synthetic sources or from natural sources such as plants, algae or fungi. Advantageously, pigments derived from natural sources will meet clean label requirements for food products.

[0127] Anthocyanin Anthocyanins are the sugar-free glycosides (aglycones) of anthocyanidins. The sugar molecules of anthocyanins are linked via O-glycosidic bonds to one or more hydroxyl groups that are typically present in the anthocyanidin molecule. Most naturally occurring anthocyanins are 3-O-glycosides. The most common types of anthocyanidins found in plants are cyanidin, delphinidin, pelargonidin, peonidin, petunidin, and malvidin, in which the hydroxy groups at positions 3, 5, 7, and at least one of the 3', 4', or 5' positions are sugar-substituted. Examples of natural anthocyanins that may be used in the present invention include, but are not limited to, pelargonidin, cyanidin, and peonidin-based anthocyanins.

[0128] Examples of sugar molecules found in the anthocyanin structure include arabinose, galactose, glucose, rhamnose, rutinose, sambubiose, sophorose, and xylose. Anthocyanins can be substituted with hydrogen, hydroxyl, and / or methoxyl groups at various positions. Anthocyanins can also be acylated, having one or more monosaccharides esterified to a sugar molecule at the 2-, 3-, 4-, and / or 6-positions of the monosaccharide. Many anthocyanins are acylated (usually at the C6-OH group of the glucose moiety) with either fatty acids (e.g., acetic, malic, malonic, oxalic, or succinic acids) or phenolic acids (e.g., p-hydroxybenzoic, caffeic, p-coumaric, ferulic, or sinapic acids). Thus, anthocyanins may be in the form of acylated glycoside anthocyanins, such as pelargonidin-based acylated anthocyanins, cyanidin-based acylated anthocyanins, and peonidin-based acylated anthocyanins, or structural analogues of pelargonidin-based acylated anthocyanins, cyanidin-based acylated anthocyanins, and peonidin-based acylated anthocyanins.

[0129] Red radish (Raphanus sativus L.) and red potato (Solanum tuberosum L.) provide color characteristics similar to FD&C Red #40. In one embodiment, the anthocyanin is a pigment derived from red radish. The main pigments of red radish and red potato are pelargonidin-3-sophoroside-5-glucoside acylated with either malonic acid, p-coumaric acid, and / or ferulic acid, and pelargonidin-3-rutinoside-5-glucoside acylated with p-coumaric acid, respectively (Rodriguez-Saona, LE et al., J. Food Sci. 1999, 64, 451-456, the disclosure of which is incorporated herein by reference). Thus, the anthocyanin used in the present invention may be pelargonidin-3-sophoroside-5-glucoside acylated with either malonic acid, p-coumaric acid, and / or ferulic acid, and / or pelargonidin-3-rutinoside-5-glucoside acylated with p-coumaric acid. In one embodiment, the anthocyanin is a pigment derived from black carrot.

[0130] Recently, cyanidin 3-xylosyl(glucosyl)galactoside acylated with sinapic acid, ferulic acid, and coumaric acid has been identified as the major anthocyanin in black carrot (Cuevas Montilla, E., et al, J. Agric. Food Chem. 2011, 59, 3385-3390, the disclosure of which is incorporated herein by reference). Therefore, the anthocyanin used in the present invention may be cyanidin 3-xylosyl(glucosyl)galactoside acylated with sinapic acid, ferulic acid, and coumaric acid. In one embodiment, anthocyanin can be present as the extract obtained or obtainable from Brassicaceae, Rosaceae, Solanaceae, Convolvulaceae, Umbelliferae or their mixture.The term mixture refers to the mixture obtained or obtainable when Brassicaceae, Rosaceae, Solanaceae, and / or Umbelliferae are extracted together using a single solvent, or when Brassicaceae, Rosaceae, Solanaceae, and Umbelliferae are extracted independently and the extracts obtained are combined.

[0131] The Brassicaceae plant can be Raphanus sativus L. (red radish). The Rosaceae plant can be Fragaria (strawberry). The Solanaceae plant can be Solanum tuberosum (red potato). The Convolvulaceae plant can be Ipomoea batatas (purple potato root). The Umbelliferae plant can be Daucus carota ssp. sativus var. atrorubens Alef. (black carrot). In one embodiment, the betalains may be present as an extract obtained or obtainable from a plant of the Amaranthaceae family. Optionally, the Amaranthaceae plant may be Beta vulgaris (beet). In one embodiment, an anthocyanin (0.1% in pH 3.0) derived pigment such as red radish (0.1% in water) has an L* value of 58.89±5%, an a* value of 69.81±5%, and a b* value of 51.43±5%. In one embodiment, a pigment derived from anthocyanins (0.1% in pH 3.0) such as black carrot has an L* value of 38.24±5%, an a* value of 62.95±5%, and a b* value of 25.24±5%.

[0132] Betalain Betalains are a class of red and yellow tyrosine-derived pigments found in plants of the Caryophyllaceae order, which replace the anthocyanin pigments. There are two categories of betalains: a) betacyanins, which appear reddish to purple in color. Examples of betacyanins present in plants include betanin, isobetanin, probetanin, and neobetanin; and b) Betaxanthins, which appear yellow to orange. Betaxanthins found in plants include vulgaxanthin, miraxanthin, portulaxanthin, and indicaxanthin. Thus, the betalains used in the present invention may be betacyanins, such as betanin, isobetanin, probetanin, and neobetanin; and / or betaxanthins, such as vulgaxanthin, miraxanthin, portulaxanthin, and indicaxanthin. Betalains are glycosides of betanidin aglycone, the core structure of which is betalaminic acid (i.e., 4-(2-oxoethylidene)-1,2,3,4-tetrahydropyridine-2,6-dicarboxylic acid).

[0133] Betanin is usually obtained from the juice extract of Beta vulgaris (red beet, beetroot). [ka] In one embodiment, the betalain is a beetroot derived pigment. Optionally, the betalain used in the present invention may be betanin. In one embodiment, betalains (0.1% in water) (e.g., a pigment derived from beetroot) have an L* value of 85.29±5%, an a* value of 26.66±5%, and a b* value of −5.76±5%.

[0134] In one embodiment, the betalain-containing colorant (e.g., beetroot-derived pigment, such as beetroot powder) is present in the food colorant composition in an amount ranging from about 25% to about 60% w / w. Optionally, the betalain-containing colorant (e.g., beetroot-derived pigment, such as beetroot powder) is present in an amount ranging from about 30% to about 50% w / w. Optionally, the betalain-containing colorant (e.g., beetroot-derived pigment, such as beetroot powder) is present in an amount of about 30.0%, 31.0%, 32.0%, 33.0%, 34.0%, 35.0%, 36.0%, 37.0%, 38.0%, 39.0%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%, 49.0% or 50.0 w / w. Optionally, the betalain-containing colorant (eg, a beetroot-derived pigment such as beetroot powder) is present in an amount of about 32.5%, 38.8%, 42.0% or 45.4% w / w.

[0135] In one embodiment, the anthocyanin-containing colorant (e.g., a black carrot-derived pigment, such as a black carrot concentrate) is present in the compositions, complexes, and colorant compositions of the present invention in an amount ranging from about 3.0% to about 8.5% w / w. Optionally, the anthocyanin-containing colorant (e.g., a black carrot-derived pigment, such as a black carrot concentrate) is present in an amount of about 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, or 8.5% w / w. Optionally, the anthocyanin-containing colorant (e.g., a black carrot-derived pigment, such as a black carrot concentrate) is present in an amount of about 3.5%, 4.0%, 4.5%, 6.0%, 7.0%, or 8.0% w / w. Optionally, the anthocyanin-containing colorant (e.g., a pigment derived from black carrot, such as a black carrot concentrate) is present in an amount of about 3.3%, 3.8%, 4.3%, 4.4%, 5.7%, 7.0%, 7.9% or 8.1% w / w.

[0136] In one embodiment, the anthocyanin-containing colorant (e.g., a red radish-derived pigment, such as red radish powder) is present in the food colorant composition in an amount ranging from about 0.1% to about 1.5% w / w. Optionally, the anthocyanin-containing colorant (e.g., a red radish-derived pigment, such as red radish powder) is present in an amount of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4% or 1.5% w / w. Optionally, the anthocyanin-containing colorant (e.g., a red radish-derived pigment, such as red radish powder) is present in an amount of about 0.5%, 0.6%, 0.9% or 1.3% w / w.

[0137] Yellow colorant Yellow colorants that can be used in the present invention include, but are not limited to, natural colorants such as carotenoids, safflower, gardenia yellow, riboflavin, and artificial colorants. Carotenoids are a class of natural pigments synthesized by plants, photosynthetic bacteria and algae and can be classified according to their chemical structure into: beta-carotene, a hydrocarbon that does not contain oxygen, and xanthophylls (containing oxygen), such as curcumin and lutein. The coloring properties of these pigments are provided by the conjugated double bond system that constitutes the light-absorbing chromophore. Absorption of light leads to a higher energy excited state of the molecule. In the case of carotenoids, the electronic transition that occurs is from π to π*, which does not require a high energy level to occur. The low required energy is associated with light in the visible range, 400-500 nm. Thus, yellow, orange and red colors are observed.

[0138] In one embodiment, carotenoids can be derived from fruits and vegetables such as carrots (including carrot juice), bacteria such as Blakeslea Tripora, and microalgae such as Duneliella Salina. In one embodiment, the carotenoid-containing colorant (such as carrot juice powder) is present in the food colorant composition in an amount ranging from about 25% to about 60% w / w. Optionally, the carotenoid-containing colorant (such as carrot juice powder) is present in an amount ranging from about 30% to about 50% w / w. Optionally, the carotenoid-containing colorant (such as carrot juice powder) is present in an amount of about 30.0%, 31.0%, 32.0%, 33.0%, 34.0%, 35.0%, 36.0%, 37.0%, 38.0%, 39.0%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%, 49.0% or 50.0% w / w.

[0139] In some embodiments, the stabilized phycobilins (such as phycocyanins), compositions, complexes, and colorant compositions of the present invention can be mixed with safflower, carrot juice, carotenoids, and other yellow pigments to produce a stable green color. The present invention also relates to kits comprising the various components of the compositions, composites, and colorant compositions of the present invention, and optionally instructions on how to mix, prepare, and / or use said components. In some embodiments, the kits contain components already mixed and ready to use (e.g., as a blend), and optionally instructions on how to use said components. In some embodiments, the kits contain other pigments, such as yellow or red.

[0140] Encapsulation The present invention also relates to an encapsulation composition comprising a composition or complex according to the present invention, or a stabilized phycobilin according to the present invention, said encapsulation composition being encapsulated using a cross-linked polymer from the group consisting of alginate, carrageenan and pectin, derivatives thereof, or combinations thereof. The present invention also relates to phycobilin (such as phycocyanin)-containing granules comprising a continuous medium of crosslinked polymers derived from the group consisting of alginate, carrageenan and pectin, derivatives thereof or combinations thereof, and additionally comprising at least one protein, polypeptide and / or peptide that is water soluble and optionally has an IEP greater than 4 and / or is soluble under acidic conditions, as described herein.

[0141] The present invention also relates to a method for preparing phycobilin (such as phycocyanin)-containing granules comprising: a) mixing in an aqueous solution a polymer from the group consisting of alginate, carrageenan and pectin, derivatives thereof or combinations thereof with at least one protein, polypeptide and / or peptide that is water soluble and optionally has an IEP greater than 4 and / or is soluble under acidic conditions; b) adding the mixture of a) in the form of droplets to a crosslinker; c) Collecting the formed granules.

[0142] Alginates, or polymers with the ability to bind relatively large amounts of water, or combinations of such polymers, can be used in the present invention. Alginates are high molecular weight carbohydrates obtained, for example, from seaweed and other algae. Alginates are linear copolymers of aL-guluronic acid (G) and bD-mannuronic acid (M). The alginate chain can be considered as a block copolymer, consisting of "G blocks" (homopolymeric regions of guluronic acid residues), "M blocks" (homopolymeric regions of mannuronic acid residues), and "MG blocks" (copolymeric regions of randomly alternating sequences of M and G) of various lengths. In addition to being heterogeneous in chemical composition, alginates have a very wide molecular weight distribution. Alginates are a collective term for a group of polymers. Their properties depend on the block structure and molecular weight. Above a certain critical molecular weight, the properties of alginates are mainly governed by the monomer composition and block structure. In general, increasing the content of guluronic acid results in mechanically stronger gels with improved stability in the presence of non-gelling / anti-gelling ions (Na+, Mg+, etc.) and calcium sequestrants. Gels with a high content of guluronic acid are more porous and have less shrinkage during gel formation. With a high content of mannuronic acid, the gels become softer and more elastic; they shrink more during gel formation and at the same time the porosity decreases.

[0143] Any kind of alginate can be used for encapsulation according to the invention. Higher molecular weight ones are generally preferred due to their greater mechanical stability in the tertiary emulsion process described below, although their higher viscosity is less critical. Lower molecular weight alginates are preferred for the spray drying process described below. Alginates form viscous solutions and retain large amounts of water. A preferred alginate is sodium alginate (CAS 9005-38-3), which is sold under the registered trademark PROTANAL by FMC Biopolymers, Philadelphia, USA. Useful alginates include iron, magnesium, potassium, ammonium, and calcium alginates.

[0144] Although alginates are particularly desirable for the preparation of granules according to the invention, other hydrophilic polymers having similar properties to alginates may be used in addition to or instead of alginates, and the term "alginates" as used herein includes such other hydrophilic polymers. Other examples of useful hydrophilic polymers are structurally related polysaccharides such as pectin and its derivatives. Pectin is a polymer consisting of polygalacturonic acid whose carboxylic acid groups are partially esterified with methanol. To obtain high thermal stability, it is preferred to use pectins with low degrees of esterification, particularly pectins with degrees of esterification less than 5%. Depending on the polymer used, alternative or additional crosslinking agents may be used, as is well known in the art.

[0145] Carrageenan may also be used. Granules (or particles) according to the invention are preferably prepared to give substantially round, roughly spherical matrix granules, as opposed to irregular agglomerates, or elongated thread-like matrix particles, which tend to form when the shear forces during preparation are too low or too high, respectively. The size of the granules may range from 5 to 2000 μm, preferably from 10 to 1000 μm, more preferably from 20 to 600 μm in diameter. The matrix granules are substantially insoluble in water. In the process according to the invention described herein, the size and size distribution range of the granules can be adjusted according to the desired application, depending on the selection of the process and the adjustment of the process parameters.

[0146] The particles of step b) are introduced into a solution of a crosslinker, for example a polyvalent cation (e.g. calcium, strontium, barium, iron, silver, aluminum, manganese, copper and zinc ions, preferably calcium ions), which may be 0.9-10%, preferably 0.9-2%, calcium chloride. The solution may be any suitable liquid, such as water or alcohol, preferably ethanol, or a mixture of water and alcohol. The ratio of water to alcohol depends on the application. The more water used, the more stable the resulting matrix particles are. Increasing the alcohol reduces the amount of oil on the surface, but may also extract oil from the interior of the particles. A water to alcohol ratio of 50:50 (wt / wt) is suitable for most applications.

[0147] The exposure time to the crosslinking agent (i.e., the multivalent cation solution) will vary depending on the ionic solution used, and the total amount and concentration of the solution. The exposure time and concentration of each ion can be easily determined by one of skill in the art. When using 0.9-10% calcium chloride at a ratio of 0.7:1-4:1, preferably 1.2:1-3:1, more preferably 1.5:1-2.5:1 saline:particles (wt / wt), about 1 hour is usually sufficient. The amount of ions required depends on the amount of alginate to be crosslinked and can be adjusted as known to those skilled in the art. Additional or alternative crosslinking agents may be used in place of saline, as is well known in the art.

[0148] The crosslinked particles obtained from step c) may be subjected to a drying step. This can be done by spray drying. In order not to destroy the crosslinked particles, low shear spray drying methods are preferred. Suitable methods will be immediately apparent to those skilled in the art. For example, a spray dryer consisting of a low shear positive displacement pump in combination with a rotating atomizer wheel can be used. Alternatively, many different pumps can be applied. A spray nozzle can also be used instead of the rotating wheel. Further examples of encapsulation methods are described in US2006292280A1 or US2010003521A1.

[0149] Uses and Products The present invention also relates to consumable or food, cosmetic or pharmaceutical or nutraceutical preparations comprising the stabilized phycobilins (such as phycocyanins) of the present invention, the compositions, complexes, and colorant compositions of the present invention. In one aspect, there is provided the use of the stabilized phycobilins (such as phycocyanins), compositions, complexes, and colorant compositions (food colorant compositions) of the present invention in coloring food. The stabilized phycobilins (such as phycocyanins), compositions, complexes, and food colorant compositions of the present invention can be added to food in an amount effective to increase, enhance, and / or modify the color properties of the food or a portion thereof.

[0150] Food includes the following general food categories as defined by the Food and Drug Administration (FDA): baked goods and baking mixes (including all ready-to-eat and ready-to-bake products), flours, and mixes that require preparation before serving; alcoholic beverages (including malt beverages, wines, spirits, and cocktail mixes); non-alcoholic beverages and beverage bases (including only special or spiced teas, soft drinks, coffee substitutes, and fruit- and vegetable-flavored gelatin drinks); breakfast cereals (including ready-to-eat instant and regular hot cereals); cheeses (including curd and whey cheeses, cream cheeses, natural cheeses, grated cheeses, processed cheeses, spreadable cheeses, dip cheeses, and cheeses with flavors other than 100% alcohol); cheeses, and other cheeses);chewing gum (in all forms);coffee and tea (including regular, decaffeinated and instant types);condiments and relishes (including plain seasoning sauces and spreads, olives, pickles and relishes but excluding spices and herbs);sugar confectioneries and frostings (including candy and flavoured frostings, marshmallows, cooking chocolate, as well as brown, lump, maple, powdered and raw sugars);dairy analogues (including non-dairy milk, frozen or liquid creamers, coffee whiteners, toppings and other non-dairy products);egg products (liquid, frozen or dried eggs and egg dishes made therefrom, i.e. egg rolls, Furong egg, egg salads, and frozen egg courses, but excluding raw eggs; fats and oils (including margarine, salad dressings, butter, salad oils, shortening, and edible oils); fish products (including prepared entrees, salads, appetizers, frozen courses, and spreads containing fish, shellfish, or other aquatic animals, but excluding raw fish); fresh eggs (including cooked eggs and egg dishes made only with fresh shell eggs); fresh fish (including only fresh and frozen fish, shellfish, and other aquatic animals); fresh fruit and fruit juices (only fresh fruit, citrus, melon, berries, and homemade "ades" and punches made therefrom) fresh meat (including only fresh or home-frozen beef or veal, pork, lamb or mutton and home-prepared fresh meat-containing dishes, salads, appetizers or sandwich spreads made therefrom);fresh poultry (including only fresh or home-frozen poultry and game and home-prepared fresh poultry-containing dishes, salads, appetizers or sandwich spreads made therefrom);fresh vegetables, tomatoes and potatoes (including only fresh home-prepared vegetables);frozen dairy desserts and mixes (including ice cream, ice milk, sorbets and other frozen dairy desserts and specialities);

[0151] Fruit and ice desserts (including all frozen fruit and ice desserts);Gelatins, puddings, and fillings (including flavored gelatin desserts, puddings, custards, parfaits, pie fillings, and gelatin-based salads);Grain products and pastas (including macaroni and noodle products, rice dishes, and frozen courses without meat or vegetables);Gravies and sauces (including all meat sauces and gravies, and tomato, milk, butter, and special sauces);Hard candies and cough drops (including all hard candies);Herbs, seeds, spices, seasonings, blends, extracts, and flavorings (including all natural and artificial spices, blends, and flavors);Homemade jams and jellies (including only homemade jams, jellies, fruit butters, preserves, and sweet spreads);Commercial jams and jellies (including only commercially processed jams, jellies, fruit butters, jams, and sweet spreads);Meat products (all meats and meat products prepared by commercial processing or prepared at home using commercially processed meats) including dishes, salads, appetizers, frozen meat courses, and sandwich ingredients that contain milk, whole milk, and skim milk (including whole milk, low fat milk, and skim milk only); dairy products (including flavored milk and dairy drinks, dry milk, toppings, snack dips, spreads, weight control dairy drinks, and other dairy-derived products); nuts and nut products (including whole or shelled tree nuts, peanuts, coconut, and nut and peanut spreads); vegetable protein products (including those defined as "reconstituted vegetable protein products" by the National Academy of Sciences / National Research Council) "food quality" category, and meat, poultry, and fish substitutes, look-alikes, and extenders made from vegetable proteins; poultry products (including all poultry and poultry-containing dishes, salads, appetizers, frozen poultry courses, and sandwich ingredients that are commercially processed or prepared at home using commercially processed poultry); processed fruits and fruit juices (including all commercially processed fruit, citrus, berry, and mixes); salads, juices and juice punches, concentrates, dilutions, "ades," and beverage substitutes made therefrom;Processed vegetables and vegetable juices (including all commercially processed vegetables, vegetable dishes, frozen multi-course vegetable dishes, and vegetable juices and blends); snack foods (including chips, pretzels, and other novelty snacks); soft candy (including candy bars, chocolate, fudge, mints, and other chewy or nougat candies); homemade soups (including meat, fish, poultry, vegetable, and combination homemade soups); soups and soup mixes (including commercially prepared meat, fish, poultry, vegetable, and combination soups and soup mixes); white sugar, granulated sugar (including white granulated sugar only); sugar substitutes (including granulated sugar, liquid sugar, and tablet sugar substitutes); and sweet sauces, toppings, and syrups (including chocolate, berry, fruit, corn syrup, and maple sweet sauces and toppings).

[0152] For example, the food colorant composition described above can be used to impart a blue color similar to that obtained when Brilliant Blue FCF (an artificial colorant) is used in foods such as dairy products, confectioneries, beverages, sauces / gravies, etc. Dairy products may refer to yogurt, custard, milk smoothies, milkshakes, and dairy ice cream. Confectionery products may refer to sweet or candy foods, such as chewing gum or hard and soft confectionery products, etc. Non-limiting examples of confectionery products include cakes, cookies, pies, chocolates, chewing gum, gelatin, ice cream, puddings, jams, jellies, gummies, hard candies, chewy candies, cereals and other breakfast foods, canned fruits and fruit sauces.

[0153] Beverage products may refer to beverages, beverage mixes and concentrates, including, but not limited to, alcoholic and non-alcoholic ready-to-drink beverages and dry powdered beverages. Non-limiting examples of beverages may include carbonated and non-carbonated beverages, such as sodas, fruit juices or vegetable juices. A sauce product may refer to a sweet or savory semi-solid composition used to add flavor, moisture, and / or visual appeal to a dish. Non-limiting examples of sauces include gravies and barbecue sauces. A meat analog product may refer to a food product made from vegetarian ingredients that resembles certain aesthetic qualities (e.g., texture, flavor, appearance) or chemical properties of a particular type of meat. A non-limiting example of a meat analog includes a vegetable (veggie) burger.

[0154] In yet another aspect, there is provided a food product comprising the food colorant composition described above. For example, the food product may be as described above. One of ordinary skill in the art will recognize that the optimal amount of food coloring composition present in a given food product will be determined by factors such as overall desired color, solubility, regulatory approval, etc. One of ordinary skill in the art can readily determine the optimal amount of color for a given product based on these factors. In one aspect, there is provided a use of the stabilized phycobilins (such as stabilized phycocyanins) of the present invention, the compositions, complexes, and colorant compositions (colorant compositions) of the present invention in coloring pharmaceuticals, cosmeceuticals, nutraceuticals, or cosmetics. The stabilized phycobilins (such as phycocyanins), the compositions, complexes, and colorant compositions of the present invention can be added to a product (pharmaceutical, nutraceutical, or cosmetic) in an amount effective to increase, enhance, and / or modify the color characteristics of the product or a portion thereof.

[0155] In another aspect, the present invention relates to a pharmaceutical, cosmeceutical, nutraceutical, or cosmetic composition comprising the stabilized phycobilins (such as stabilized phycocyanins) of the present invention, the compositions, complexes, and colorant compositions of the present invention as defined herein, and optionally a vehicle suitable for formulating the stabilized phycobilins (such as stabilized phycocyanins), the compositions, complexes, and colorant compositions of the present invention into the pharmaceutical, cosmeceutical, or cosmetic composition. The stabilized phycobilins (such as stabilized phycocyanins), the compositions, complexes, and colorant compositions of the present invention will provide the pharmaceutical, cosmeceutical, cosmetic, or nutraceutical formulation with a desired color (such as a stable green or blue color). As used herein, a "pharmaceutical composition" refers to a physiologically tolerable composition and molecular entity. Preferably, the term "pharmaceutical acceptable" means that it is approved by a state or federal regulatory agency for use in animals, more particularly in humans, or is included in the United States Pharmacopeia or other generally recognized pharmacopoeias.

[0156] As used herein, "cosmetic composition" refers to a composition that includes one or more products that provide such benefits and are suitable for use in the personal hygiene of humans or animals, or for use in enhancing natural beauty or altering the appearance of the human or animal body without affecting its structure or function. If desired, the cosmetic composition provided by the present invention can contain, in addition to the composition of the present invention, one or more cosmetic products, i.e., substances or mixtures intended to be in contact with the external parts of the human or animal body (epidermis, hair system, nails, lips, etc.) or the teeth and buccal mucosa, for the exclusive or primary purpose of cleaning, perfume, altering the appearance, protecting, keeping in good condition, or improving body odor. Examples of cosmetically acceptable vehicles include products included in the INCI (International Nomenclature of Cosmetic Ingredients) list. The compositions of the present invention can be added to a wide variety of products for cosmetic use, including make-up, creams for cleansing, protecting, treating, or caring for the skin, especially the face, hands, and feet (e.g., day and night creams, make-up remover creams, foundation creams, and sunscreens), liquid foundations, make-up remover lotions, body lotions for protection or skin care, sunscreen lotions, skin care lotions, gels, or foams, such as cleansing, sunscreen, artificial tanning lotions, bath additives, deodorant compositions, aftershave gels or lotions, hair removal creams, and compositions used for insect bites and sores.The compositions of the present invention can take any of a wide variety of forms, including, for example, dressings, lotions, solutions, sprays, creams, gels, ointments, and the like.

[0157] As used herein, the term "cosmeceutical" refers to a product suitable for use on the human or animal body that contains one or more cosmeceuticals (functional cosmetics, dermopharmaceuticals, or active cosmetics), i.e., topical hybrid products with cosmetic and pharmaceutical properties that contain higher and more effective concentrations of active ingredients that have a benefit on the user's skin, hair, and / or nails, and thus are intermediate between cosmetics and pharmaceuticals. Examples of cosmeceuticals include essential oils, ceramides, enzymes, minerals, peptides, vitamins, etc.

[0158] The term "nutraceutical" as used herein refers to a product suitable for human or animal use that contains one or more natural products that provide a health benefit or have a therapeutic action related to the prevention or alleviation of disease, including supplements, i.e., provided in a non-food matrix (e.g., capsules, powders, etc.) of concentrated natural bioactive substances normally present (or not) in food, which, when taken in higher doses than those present in food, have a greater positive effect on health than normal food may have. Thus, the term "nutraceutical" also includes isolated or purified foods, as well as additives or supplements provided in dosage forms normally used orally, e.g., capsules, tablets, sachets, drinking bottles, etc.; such products provide a physiological benefit or provide protection against disease, generally chronic disease. Optionally, the nutraceuticals provided by the present invention may contain, in addition to the compositions of the present invention, one or more nutraceuticals (products or substances related to the prevention or alleviation of disease), such as flavonoids, omega-3 fatty acids, etc., and / or one or more prebiotics (non-digestible food ingredients that stimulate probiotic activity and / or growth), such as oligofructose, pectin, inulin, galactooligosaccharides, lactulose, human milk oligosaccharides, dietary fiber, etc.

[0159] In some embodiments, the pH of the product (consumable or food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic) is less than 6, such as less than 5, such as less than 4, such as less than 3, or such as less than 2. In some embodiments, the product (consumable or food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic) has been heat treated and / or exposed to light. In some embodiments, the pH of the product (consumable or food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic) is less than 6, such as less than 5, such as less than 4, such as less than 3, or such as less than 2, has been heat treated, and / or has been exposed to light.

[0160] In some embodiments, the product (consumable or food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic) has been heat treated and / or exposed to light. The present invention relates to acidic food, cosmetic or pharmaceutical or nutraceutical preparations comprising the stabilized phycobilins (such as phycocyanins) of the present invention, the compositions, complexes, and colorant compositions of the present invention. In one embodiment of an acidic product (such as an acidic food product), the amount of protein, polypeptide, peptide or mixture thereof is 0.005-1 wt%.

[0161] In some embodiments of acidic products (such as acidic foods), the weight ratio of protein, polypeptide, peptide or mixtures thereof to phycocyanin ranges from 10:1 to 1:10. In one embodiment of an acidic product (such as an acidic food product), the pH of the product (such as a food product) is 2.0 to 4.6, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, and most preferably 2.4 to 3.3. In certain embodiments of an acidic product (such as an acidic food product), at least one protein, polypeptide and / or peptide is water soluble and / or soluble at a pH of 2-5, such as a pH of 2-4, such as a pH of 2-3.

[0162] In certain embodiments, the at least one protein, polypeptide and / or peptide is selected from one or more of potato, mung bean, rice and soy proteins, polypeptides and / or peptides. In some embodiments, the solvent consists of water and optionally ethanol if the food is liquid, or the food matrix in the case of a solid or semi-solid food. In some embodiments, the amount of solvent or food matrix ranges from 60-99 wt%. In certain embodiments, the wt % is based on the total weight of the liquid, solid, or semi-solid foodstuff. The acidic food composition of the present invention may be ready to use or may be in the form of a food additive, which is added in solid, pasty or liquid form to the preparation of ingestible food products.

[0163] In the case of food compositions, the acids are preferably selected from the list approved for acidification in feed, in particular carbonic acid, phosphoric acid, citric acid, malic acid, tartaric acid and lactic acid, more particularly citric acid. As regards the non-food acidic compositions according to the invention, they may be inter alia pharmaceutical, veterinary or cosmetic and may further comprise any additive and / or active agent known and used in compositions of this type. The acidic composition according to the present invention in the form of a solid, liquid or paste can incorporate the stabilized phycobilin (such as stabilized phycocyanin), the composition, complex and colorant composition (colorant composition) of the present invention, for example in powder form. The acidic composition, in particular the acidic food composition, can then be in any form of food composition commonly known, such as cream, gel, foam, paste, etc., particularly solids such as biscuits or cakes, dry foods to be cooked, powders to be diluted, compositions or "jelly" solid gelatins, foams, etc.

[0164] According to the present invention, the composition may be a liquid acidic aqueous composition in which the stabilized phycobilin (such as stabilized phycocyanin), the composition, the complex, and the colorant composition (colorant composition) of the present invention are dissolved. It may be in the form of a ready-to-use composition or in the form of a liquid concentrate to be diluted for ingestion or for addition to or preparation of solid foods, such as concentrated liquid composition coatings or "toppings" applied to color cakes. Among these concentrated compositions are syrups, with or without alcohol. The liquid acidic compositions according to the invention may be of variable viscosity and may optionally contain viscosity modifiers, gelling agents and other additives such as those known to those skilled in the art of structuring and conventionally used in the art for the preparation of food liquids.

[0165] According to a particular embodiment of the present invention, the liquid food composition may be a carbonated or non-carbonated acidic beverage, in particular water, juice, sports drinks, athletic drinks, recovery drinks, etc. The composition of these beverages is well known to those skilled in the art and may contain, in particular, sugars, inorganic salts, food additives, dissolved gases, etc. The beverage according to the present invention is a normal acidic beverage in which all or part of the dyes normally used are replaced by the stabilized phycobilins (such as stabilized phycocyanins) of the present invention, compositions, complexes and colorant compositions of the present invention (colorant compositions) which are resistant to acidic pH, heat treatment and light according to the present invention as described above. In the liquid acidic composition used in the present invention, the content of phycobilin (such as phycocyanin) may be 2.5 mg / L to 2500 mg / liter, preferably 25 mg / L to 300 mg / succica.

[0166] In ready-to-use liquid composition type beverages, the content of phycobilins (such as phycocyanin) can generally be 25 mg / L to 300 mg / L, preferably 50 mg / L to 100 mg / L. In concentrated liquid compositions that are diluted for use as syrups, the content of phycobilins (such as phycocyanin) may generally be from 250 mg / L to 2500 mg / L, preferably from 500 mg / L to 1000 mg / L. In the solid composition, the content of phycobilin (such as phycocyanin) can generally be 0.01 mg / g to 10 mg / g, preferably 0.1 mg / g to 5.0 mg / g, and most preferably 0.25 mg / g to 2.5 mg / g. The invention is further illustrated by the following non-limiting examples. EXAMPLES

[0167] material and method. Phycocyanin Extract Phycocyanin is extracted from Spirulina (Arthrospira platensis) with water, the extract is filtered, purified, and concentrated, and powdered phycocyanin is obtained by spray drying using maltodextrin as a carrier. The final extract contains 25% wt of total phycocyanin, including C-phycocyanin and allophycocyanin, calculated as described in Yoshikawa and Belay (2008) J AOAC Int. May-Jun 2008;91(3):524-9. Potato protein: Potato protein was purchased from AVEBE (The Netherlands). Protein was isolated from potatoes by aqueous extraction, followed by purification, concentration and finally drying. The protein content of the final extract is greater than 90% w / w (dry weight).

[0168] Mung bean peptides: The samples were purchased from Nutraonly (China), and the proteins were first extracted with water and then hydrolyzed with enzymes to obtain peptides. The peptide content is >90% (W:W). Rice peptides: Samples were purchased from Organicway (China). Rice proteins were first extracted from the raw material; then peptides were obtained by enzymatic hydrolysis, followed by sterilization, filtration, and concentration, and finally spray drying. The peptide content was >90% (by dry weight), with molecular weights ranging from 200 Da to 4000 Da, with the majority being 300-2000 / 1000 Da. Rice peptides were analyzed using LC-MS. The main proteins identified as the source of peptides are glutelin A1 and B1. These peptides were probably generated using pepsin enzymatic digestion.

[0169] Pea peptides: Pea peptides were purchased from Aromiens (USA). Pea proteins were first extracted with water, and macropeptides were obtained by enzymatic degradation of the protein extract, after which the enzymes were destroyed by heat treatment. Peptides were separated by centrifugation and then purified using membrane filtration. After sterilization, powdered extracts were obtained by spray drying. The peptide content was >80% (dry weight) with molecular weights ranging from 500 Da to 3000 Da, with the majority being between 800 and 2000 Da. Pea peptides were analyzed using LC-MS. The main proteins identified as the source of peptides are vicilin, legumin A, provicilin, convicilin, legumin J, and albumin 2. The peptides were probably generated using pepsin enzymatic digestion.

[0170] Soybean peptides: Soybean peptides were purchased from Aromiens (USA). Soybean proteins were first extracted with water and peptides were obtained by enzymatic hydrolysis of the protein extract. The enzymes were then heat denatured. Peptides were separated by centrifugation and subsequently purified using membrane filtration. After concentration, the extract was sterilized and finally formulated into powder form by spray drying. The peptide content was >80% and the molecular weight ranged from 200 Da to 2000 Da, with the majority being between 300 and 1000 Da. Soybean peptides were analyzed using LC-MS. The main proteins identified as the origin of peptides were: Glycine G1 and β-conglycinate (soy protein) These peptides were probably generated using pepsin enzyme digestion.

[0171] Analysis of peptide samples: Pea peptides sample preparation 20 mg of pea peptide sample (powder) was weighed and diluted with 100 mL of ACN / HO+0.1% formic acid (70:30). The solution was 0.2μ filtered before analysis by LC-HRMS. LC-MS conditions Column: AdvanceBio Peptide Map (250 x 2.1 mm; 2.7 μ) Temperature: 50°C Flow rate: 0.25ml / min Injection volume: 2μl Detection: Full scan with ESI+ (range m / z: 200-3000) dd-MS2: Data-dependent scanning with ESI+ Mobile phase: A: ACN, 0.1% formic acid B: Water, 0.1% formic acid

[0172] Pea peptide samples were analyzed by LC-MS (liquid chromatography mass spectrometry) and reprocessed with BioPharma Finder software (Thermo) to identify the primary sequences of the pea peptides and to identify which enzymes were used to hydrolyze the samples during sample preparation. Trypsin, chymotrypsin, and pepsin were searched with BioPharma Finder software. The sequences of major pea proteins (globulins, legumins, vicilins, and albumins) were downloaded from Uniprot and imported into BioPharma Finder according to several publications (Burger T., Zhang Y., 2019. “Recent Progress in the Utilization of Pea Protein as an Emulsifier for Food Applications”. Trends in Food Science & Technology. https: / / doi.org / 10.1016 / j.tifs.2019.02.007).

[0173] The best results were obtained with the enzyme pepsin, the least specific of the three enzymes investigated: it was with pepsin that the highest percentage of identified ions in the protein sequence and the highest sequence coverage were found. The six major proteins identified in pepsin in this pea sample were vicilin, legumin A, provicilin, convicilin, legumin J, and albumin 2, which accounted for 22.6% of the total ions detected in this sample with sequence coverage of 71.9%, 65.8%, 82.9%, 65.8%, 59.4%, and 81.8%, respectively. For vicilin, the most abundant protein identified in this sample, 300 ions were identified as peptide sequences for this protein, with a total sequence coverage of 71.9% (Figure 6). Several ions of glycinin (the major soybean protein), glutelin and prolamin (the major rice protein) were also identified with sequence coverage ranging from 26.5% to 67%, but at very low abundance. These proteins are probably also present in pea, but at very low concentrations.

[0174] Analysis of soybean and rice peptides. Sample preparation: 75 mg of soy and rice peptide samples (powder) were weighed and diluted with 50 mL of ACN / HO+0.1% formic acid (70:30). The solution was 0.2μ filtered prior to analysis by LC-HRMS. LC-MS results (Q Exactive Orbitrap): Soybean and rice peptide samples were analyzed by LC-MS and reprocessed with BioPharma Finder software (Thermo) to identify the predominant peptide sequence and to identify which enzyme was used to hydrolyze the samples during sample preparation. Trypsin, chymotrypsin, and pepsin were investigated with BioPharma Finder software. According to several publications, the sequences of major soybean proteins (glycinin and conglycinin), major rice proteins (glutelin and prolamin), and major pea proteins (globulins, legumin, vicilin, and albumin) were downloaded from Uniprot and imported into BioPharma Finder (Chatterjee et al., 2018; Rani et al.,2018 and Burger et al., 2019).

[0175] Soy Peptides: The best results were obtained with the enzyme pepsin, the least specific of the three enzymes investigated: it was with pepsin that the highest percentage of identified ions in the protein sequence and the highest sequence coverage were found. The five major proteins identified in pepsin in this soybean sample were glycinin G1, β-conglycinin alpha subunits 1 and 2, glycinin G5, and β-conglycinin beta subunit, which accounted for 21.4% of the total ions detected in this sample with sequence coverage of 73.9%, 73%, 85.2%, 62.6%, and 79.6%, respectively (Figure 6 ). For glycinin G1, the most abundant protein identified in this sample, 285 ions were identified as peptide sequences for this protein, with a total sequence coverage of 73.9% (Figure 7).

[0176] Rice Peptides: The best results were obtained with the pepsin enzyme, the least specific of the three enzymes investigated: it was with pepsin that the highest percentage of identified ions in the protein sequences and the highest sequence coverage were found, although these recoveries were lower than those observed in the soybean samples. The two major proteins identified with pepsin in this rice sample were glutelin A1 and glutelin B1, which accounted for 3.6% and 2.1% of the total ions detected in this sample, with sequence coverage of 65.5% and 65.5%, respectively. For glutelin A1, the most abundant protein identified in this sample, 200 ions were identified as peptide sequences for this protein, with a total sequence coverage of 65.5%.

[0177] Example 1. Evaluation of the stability of phycocyanin against heat in a sugar syrup matrix, pH 3 (for confectionery applications): 0.025g of Spirulina blue extract powder (25% total phycocyanin w / w) was dissolved in 1mL of distilled water. 0.150g of protein, peptide, or mixture thereof (1:1 W / W) was slowly poured into the blue Spirulina extract solution with continuous stirring for 30 minutes. The blend was then introduced into 99g of sugar syrup Brix°60, pH3 and mixed for 20 minutes. Sugar syrup was measured by spectrophotometer (Konica Minolta) for evaluation of initial L, a, and b parameters and set as the reference. The samples were then heat treated at 80° C. for 30 min using a water bath. After heating, the mixtures were measured again against the original standard and the differences in L, a, b and DE2000 parameters were assessed.

[0178] Sample 1A: 0.025% Spirulina extract (25% phycocyanin w:w) + 99.97% sugar syrup Brix (B) ° 60, pH 3. Sample 1B: 0.025% Spirulina extract + 0.15% Yeast protein extract + 99.82% Sugar syrup B ° 60, pH 3. Sample 1C: 0.025% Spirulina extract + 0.15% whey protein isolate + 99.82% sugar syrup B°60, pH3. Sample 1D: 0.025% spirulina extract + 0.15% potato protein extract + 99.82% sugar syrup B ° 60, pH 3. Sample 1E: 0.025% spirulina extract + 0.075% potato protein extract + 0.075% rice peptides + 99.82% sugar syrup B ° 60, pH 3. Sample 1F: 0.025% spirulina extract + 0.15% rice peptide + 99.82% sugar syrup B°60, pH3. Sample 1G: 0.025% spirulina extract + 0.15% mung bean peptide + 99.82% sugar syrup B°60, pH3. Sample 1H: 0.025% spirulina extract + 0.075% mung bean peptides + 0.075% potato protein + 99.82% sugar syrup B°60, pH3. 1I: 0.025% spirulina extract + 0.15% soybean peptide + 99.82% sugar syrup B°60, pH3. Sample 1J: + 0.025% Spirulina extract + 0.15% pea peptides + 99.82% sugar syrup B ° 60, pH 3. Sample 1K: 0.025% spirulina extract + 0.075% pea peptides + 0.075% potato protein + 99.82% sugar syrup B ° 60, pH 3. Sample 1L: 0.025% spirulina extract + 0.075% soy peptides + 0.075% potato protein + 99.82% sugar syrup B ° 60, pH 3. Sample 1M: 0.025% Spirulina extract + 0.075% Mung bean protein + 0.075% Potato protein + 99.82% Sugar syrup B°60, pH3.

[0179] [Table 1]

[0180] Compared to the control (1A: control containing only Spirulina extract), samples containing different protein candidates showed better color retention when the Spirulina-containing samples were exposed to heat treatment. Specifically, potato proteins and mung bean peptides, pea peptides and potato proteins, and soybean peptides and potato proteins improved the stability of phycocyanin. Interestingly, these mixtures significantly improved the color stability (1H, 1K, 1L, M), suggesting a synergistic effect between potato proteins and mung bean peptides, potato proteins and pea peptides, or potato proteins and soybean peptides.

[0181] Example 2: Kinetics of decomposition absorption measurements Figures 2A and 2B show the change in phycocyanin retention upon heat treatment. The PC retention of phycocyanin was measured by UV-Vis absorption and can be identified as the ratio of absorbance after heat treatment to that before heating. The results showed that the phycocyanin retention was significantly improved when combined with potato and mung bean peptides (Sample B: 0.025% Spirulina extract + 0.075% Mung bean peptide + 0.075% Potato protein + 99.82% Sugar syrup B ° 60, pH 3) compared to the control sample A (retention: 0.91 vs. 0.34) (Sample A: Sample 1A: 0.025% Spirulina extract (25% phycocyanin w:w) + 99.97% Sugar syrup B ° 60, pH 3).

[0182] Example 3: Precipitation of whey protein isolate 0.1 g of Spirulina blue liquid extract (25% total phycocyanin w / w) was dissolved in 1 mL of distilled water. 0.2 g of protein, peptide, or mixture thereof (1:1 W / W) was incorporated into the blue Spirulina extract solution with continuous stirring for 30 min. The blend was then added to 99 g of beverage, Brix°10, pH 3.5, and mixed for 10 min using a magnetic stirrer (300 rpm). Beverage composition: 89.96% water + 10% sucrose + 0.025% potassium sorbate + 0.015% sodium benzoate The final pH was fixed at 3.5 with citric acid. The beverages were measured with a spectrophotometer (Konica Minolta) for evaluation of initial L, a, b parameters and set as reference. The samples were then heat treated at 95°C for 5 minutes using a water bath. After heating, the samples were measured again against the original standard to assess the differences in the L, a, b and DE2000 parameters.

[0183] [Table 2]

[0184] result: [Table 3] As can be seen in Figure 3, compared to the control sample (B), whey proteins (A) did not prevent precipitation after heat treatment (AT), whereas potato proteins (CT) increased the colloidal stability of phycocyanin, even after prolonged heat treatment at high temperature (95 °C for 5 min).

[0185] Example 4. Improved colloidal, thermal and light stability in beverage applications 0.15 g of Spirulina blue liquid extract (7% total phycocyanin w / w) was dissolved in 1 mL of distilled water. 0.05 g of protein, peptide, or mixture thereof (1:1 W / W) was incorporated into the blue Spirulina extract solution with continuous stirring for 30 min. The blend was first filtered using a 0.125 filter and then added to 99 g of beverage, Brix°10, pH 3.5, and mixed for 10 min using a magnetic stirrer (300 rpm). Beverage composition: 89.96% water + 10% sucrose + 0.025% potassium sorbate + 0.015% sodium benzoate The final pH was fixed at 3.5 with citric acid.

[0186] The beverages were measured with a spectrophotometer (Konica Minolta) for evaluation of initial L, a, b parameters and set as reference. The samples were then heat treated at 90°C for 1 min using a water bath (AT, BT, CT). After heating, the samples were measured again against the initial standard and the differences in L, a, b and DE2000 parameters were assessed. To evaluate the photostability, the samples were continuously exposed to light. The color loss was measured by a spectrophotometer (Konica Minolta) for evaluation of the initial L, a, b parameters and set as the baseline.

[0187] Sample 4A: 0.1% Spirulina extract (7% total phycocyanin w / w) + 0.05% potato protein + 99.85% beverage, B°10, pH 3.5 Sample 4B: 0.1% Spirulina extract (7% total phycocyanin w / w) + 0.025% potato protein + 0.025% mung bean peptides + 99.85% beverage, B°10, pH 3.5 Sample 4C: 0.1% Spirulina extract (7% total phycocyanin w / w) + 99.90% beverage, B°10, pH 3.5 Samples AT, BT, and CT were obtained by subjecting samples A, B, and C to heat treatment (90°C, 1 minute). [Table 4]

[0188] Figure 4A and Table 4 showed that potato proteins (4AT and 4BT) and the mixture of potato proteins and mung bean peptides showed improved colloidal stability, preventing precipitation of phycocyanin at pH 3.5 (very close to the IEP of PC). Color retention was also significantly improved after heat treatment at 90°C, pH 3.5 for 1 min. Photostability was initiated (shelf life): Figures 4B and 4C.

[0189] Samples of beverages AT, BT and CT (after heat treatment) were measured with a spectrophotometer (Konica Minolta) to evaluate the initial L, a and b parameters, which were set as the baseline before light exposure. The samples were then exposed to light and different measurements were performed at different times. The results from this example (Figure 4B) showed that, compared to the control (sample 4CT), both 4AT and 4BT again showed a significant enhancement of colloidal stability, which was maintained in time after 13 days of light exposure. Moreover, sample 4BT, which contains mung bean peptides and potato proteins, showed the best color retention, even under successive heat treatments and light exposure (see Figure 4C, day 13). This result suggests that these ingredients, in addition to thermal stability, protected phycoxianin from photo-oxidation.

[0190] Example 5. Albumin protocol The effect of albumin protein on the stabilization of phycocyanin at high temperature and low pH was investigated. A sugar syrup matrix of pH 3 containing 0.025% Spirulina extract (25% phycocyanin W / W) was prepared as already done in Example 1. Albumin from chicken egg white and albumin from bovine serum (BSA) were added at different levels ranging from 0.05% to 0.2% (W:W). The initial absorbance of the mixtures at 618 nm was measured. The samples were then heat treated at 80 °C for 30 min using a water bath. After heating, the absorbance was measured again and the color retention was determined by the absorbance ratio:

number

[0191] sample: Sample 5A: 0.025% spirulina extract + 0.05% BSA + 99.92% sugar syrup B°60, pH3 Sample 5B: 0.025% spirulina extract + 0.1% BSA + 99.87% syrup B°60, pH3 Sample 5C: 0.025% spirulina extract + 0.2% BSA + 99.77% syrup B°60, pH3 Sample 5D: 0.025% Spirulina extract + 99.97% sugar syrup B°60, pH3 Sample 5E: 0.025% spirulina extract + 0.2% egg white + 99.77% syrup B°60, pH3 Sample 5F: 0.025% spirulina extract + 0.1% egg white + 99.87% syrup B°60, pH3 Sample 5G: 0.025% spirulina extract + 0.05% egg white + 99.92% syrup B°60, pH3

[0192] result: [Table 5]

[0193] As can be seen from Figure 5A and Table 5, albumin from chicken egg white improved the retention of phycocyanin. However, bovine serum albumin only slightly improved the retention of phycocyanin. This difference may be explained by changes in the albumin structure from these animal sources. As can be seen from Figure 5(A), the samples containing chicken egg albumin showed better color retention compared to the control sample. Increasing the albumin concentration improved the retention of phycocyanin.

[0194] These results suggest that the structure of albumin may effectively contribute to the stabilization of phycocyanin, and the high phycocyanin retention obtained in the potato protein and mungbean mixtures can be explained by the presence of other protein and peptide fractions, such as globulins, prolamins, etc., that can provide further improvements. Figure 5B shows the change in phycocyanin retention in function of bovine serum albumin concentration (left) and the change in phycocyanin retention in function of bovine serum albumin concentration (right).

[0195] 6. Encapsulation of the Complexes of the Invention (Phycobilins and Protein-Peptides) material and method. The phycocyanin extract used was Vegebrite™ Ultimate ex Naturex, France, a high-strength purified extract of Spirulina platensis with a phycocyanin content of 20-25% by weight. Potato protein as described in Example 1 was used. Mung bean peptides were obtained from Nutraonly (ShaanXi, PRC). The peptide content was >90% (W:W). A 60°B sugar syrup was prepared by adding enough citric acid to 40 parts by weight of distilled water to give a pH of 3. 60 parts by weight of sucrose was then added and dissolved by heating to 70°C with mixing to give a clear syrup.

[0196] Example 6.1 Preparation of phycocyanin granules containing potato proteins and mung bean peptides 60g of sodium alginate was weighed into 1900g of distilled water and allowed to hydrate with gentle stirring for 30 minutes. 40g of phycocyanin extract, 20g of potato protein, and 20g of mung bean peptides were then added to the solution. The mixture was allowed to hydrate for another 30 minutes. For the cross-linking solution, 60 g of calcium lactate pentahydrate was dissolved in 2940 g of distilled water. This solution was placed in a vessel equipped with a rotating disk with a diameter of 10 cm. The alginate / phycocyanin / potato protein and mung bean peptide isolate solutions were poured into the center of the disc and droplets were spun from the edge of the rotating disc into the lactate solution. The resulting granules were collected from the crosslinking solution at 5 and 30 minutes after formation, washed with water, and dried in a fluid bed dryer to a moisture content of 1-8 wt%. They were then freeze-milled to reduce particle size to less than 100 μm.

[0197] Thermal Stability Test Examples 6A-6F were prepared as described below: Example 6A was a comparative example. Comparative Example 6A: 0.15 g of phycocyanin extract powder was dissolved in 1 ml of distilled water. This solution was added to 99.85 g of 60°B sugar syrup and mixed for 20 minutes. Example 6B: 0.15g of phycocyanin extract powder was dissolved in 1ml of distilled water. 0.075g of potato protein and 0.075g of mung bean peptide were slowly added and dissolved, and stirred continuously for 30 minutes. This solution was added to 99.70g of 60°B sugar syrup and mixed for 20 minutes. Example 6C: 0.25 g of phycocyanin granules containing potato proteins and mung bean peptides (collected after 5 min cross-linking) prepared as described in Example 2 was dispersed in 1 ml of distilled water. The dispersion was stirred continuously for 30 min. The dispersion was then added to 99.75 g of 60°B sugar syrup and mixed for 20 min.

[0198] Example 6D: 0.25 g of phycocyanin granules containing potato proteins and mung bean peptides (collected after 30 min of cross-linking) prepared as described in Example 2 was dispersed in 1 ml of distilled water. The dispersion was stirred continuously for 30 min. The dispersion was then added to 99.75 g of 60°B sugar syrup and mixed for 20 min. The CIELAB color space parameters L*, a*, and b* were measured with a spectrophotometer (Konica Minolta). These values ​​serve as reference values. The samples were then heat treated in a water bath at 80°C for 30 minutes. After heating, L*, a*, and b* were measured again and the ΔE*ab values ​​were calculated. The ΔE*ab value is a measure of the color difference of the sample in the CIELAB color space before and after heating. The higher the ΔE*ab, the greater the color difference and the lower the color stability when the sample was subjected to heat treatment. The results are reported in the table below.

[0199] [Table 6]

[0200] Compared to Comparative Example 6A, which contained only phycocyanin extract, all samples containing proteins showed better color stability (i.e. lower ΔE*ab values). Encapsulation of samples containing a mixture of potato and mung bean proteins in alginate (Examples 6C and 6D) improved heat stability.

[0201] Example 7. Stabilization of phycocyanin from extremophilic microalgae (Cyanidioschyzon merolae) The aim of this example is to investigate the stabilization efficiency of the studied proteins and peptides against phycocyanins from extremophilic red microalgae such as Galdieria Sulphuraria and Cyanidioschyzon merolae. Crude phycocyanin extract (5% phycocyanin) from Cyanidioschyzon merolae was used in the beverage (the formulation of the same has already been described in detail). sample: Sample 7A: 0.3% Cyanidioschyzon merolae extract + 99.7% beverage, B°10, pH3.5 Sample 7B: 0.3% Cyanidioschyzon merolae extract + 0.15% soybean peptide + 99.55% beverage, B°10, pH3.5 Light stability: Samples were exposed to 350W / m light using Suntest (ATLAS). 2 The plants were then subjected to light stress for 20 hours at 40°C. Thermal stability: Samples were stored at 40°C for 2 days.

[0202] Color Change: The color loss was measured by a spectrophotometer (Konica Minolta), and the initial L, a, and b parameters were evaluated and set as the standard. (dE2000, (dE00)) [Table 7] The results showed that the addition of soybean peptides improved both the thermal and photostability of Cyanidioschyzon merolae phycocyanin, supporting the expanded use of peptides and proteins as stabilizers of phycocyanin from extremophilic microalgae.

Claims

1. A composition comprising at least one phycobilin and at least one protein, polypeptide and / or peptide, wherein the at least one protein, polypeptide and / or peptide is water-soluble and, optionally, has an IEP greater than 4 and / or is soluble under acidic conditions.

2. A complex of at least one phycobilin and at least one protein, polypeptide and / or peptide obtainable by mixing at least one protein, polypeptide and / or peptide with a composition comprising at least one phycobilin in an aqueous solution.

3. 3. The complex of claim 2, wherein at least one protein, polypeptide and / or peptide is water-soluble, has an IEP greater than 4, and / or is soluble under acidic conditions.

4. 2. The composition or complex of claim 1, wherein the pH of the composition or complex is 2.0 to 5.0, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, and most preferably 2.4 to 3.

3.

5. 2. The composition or complex of claim 1, wherein at least one protein, polypeptide and / or peptide is from a plant and / or from an animal origin.

6. 2. The composition or complex of claim 1, wherein the protein, polypeptide and / or peptide is obtained from: a) tubers, such as potato (Solanum tuberosum) and any of the 4,000 described potato varieties, such as russet potato (coarse brown skin), red potato, white potato, yellow potato (also called Yukon potato), and purple potato, sweet potato (Ipomoea butatas), manioc or yucca (Manihot esculenta), dahlia, carrot (Daucus carota subsp. sativus), radish, beetroot, etc.; b) beans selected from black beans, canelli beans, kidney beans, lentils, lima beans, pinto beans, soybeans, white beans, mung beans, broad beans, and mixtures thereof; c) nuts, such as nuts selected from almonds, Brazil nuts, cashews, peanuts, pecans, hazelnuts, pine nuts, walnuts, pistachios, and mixtures thereof; d) plant seeds, such as seeds selected from chia, flax, hemp, pumpkin, sesame, sunflower, quinoa, chickpea, green pea, pea, rapeseed / canola and mixtures thereof; e) derived from cereals, for example cereals selected from oats, wheat, barley, spelt, corn, rice and mixtures thereof, and / or f) Eggs.

7. 7. A composition or complex according to claim 6, wherein the protein is obtained from potato and the polypeptides and / or peptides are obtained from mung bean, pea, soybean and / or rice.

8. 2. The composition or complex of claim 1, wherein the at least one protein is selected from albumin, globulin (e.g., β-conglycinin, glycinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamin, lectin, gliadin, tuberin, patatin, and mixtures thereof.

9. 2. The composition or complex of claim 1, wherein the at least one protein is selected from albumin, globulin (e.g., β-conglycinin, glycinin, vicilin, legumin A, legumin J), provicilin, convicilin, glutelin (e.g., glutelin A1 and B1), prolamin, lectin, gliadin, tuberin, patatin, and mixtures thereof, and optionally comprises polypeptides and / or peptides derived therefrom.

10. 9. The composition or complex of claim 8, wherein at least one protein is an albumin, in particular a plant albumin, more in particular a potato albumin, and further comprises polypeptides and / or peptides derived therefrom.

11. The composition or complex of claim 8, wherein the protein is albumin, particularly plant albumin, more particularly potato albumin, and further comprises polypeptides and / or peptides derived from albumin, globulins (e.g., β-conglycinin, glycinin, vicilin, legumin A, legumin J), and / or glutelins (e.g., glutelins A1 and B1).

12. 12. The composition or complex of claim 11, wherein the albumin is obtained from potato, and optionally comprises polypeptides and / or peptides derived from albumin, globulins (e.g., β-conglycinin, glycinin, vicilin, legumin A, legumin J), and / or glutelins (e.g., glutelins A1 and B1) obtained from mung bean, pea, soybean, and / or rice.

13. 2. The composition or complex of claim 1, wherein the polypeptide and / or peptide has a molecular weight of 200 to about 6000 daltons, 300 to about 5,000 daltons, such as 300 to 2000 daltons, for example 300 to 1000 daltons, such as 500 to 2000 daltons, for example 500 to 1000 daltons.

14. 2. The composition or complex of claim 1, wherein the phycobilin is selected from phycoerythrobilin, phycocyanobilin, phycoviolobilin, phycourobilin, and mixtures thereof.

15. 2. The composition or complex of claim 1, wherein the phycocyanobilin is phycocyanin obtained from Arthrospira platensis (also known as Spirulina), Arthrospira fusiformis, Arthrospira maxima, Galdieria daedala, Galdieria sulphuraria, Galdieria maxima, Galdieria partita, Cyanidioschyzon merolae 10D, Cyanidioschyzon merolae DBV201, Cyanidium caldarium, Cyanidium rumpens, Cyanidium daedalum, Cyanidium maximum, Cyanidium partitum, and any mixture thereof.

16. 2. The composition or complex of claim 1, wherein the ratio of the at least one protein to the at least one polypeptide and / or peptide is about 100:1 to 1:100, such as 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, or 10:

90.

17. 2. The composition or complex of claim 1, wherein the ratio of phycobilin to the at least one peptide, polypeptide and / or protein is in the range of about 10:90 to about 90:10, or about 20:80 to about 80:20, or about 30:70 to about 70:30, or about 40:60 to about 60:40, or about 45:55 to about 55:45, about 50:1 to about 1:50, such as about 40:1 to about 1:40, for example about 30:1 to about 1:30, such as about 20:1 to about 1:20, for example about 10:1 to about 1:10, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:

2.

18. 2. The composition or complex of claim 1, wherein the ratio of the at least one phycobilin to the at least one protein, polypeptide and / or peptide is from about 20:1 to about 1:20, e.g., from about 10:1 to about 1:10, e.g., 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:

2.

19. 19. The composition or complex of claim 18, wherein the phycocyanin is phycocyanin obtained from Arthrospira platensis (also called Spirulina) or from Galdieria sulphuraria, the protein is potato protein, and the peptide is obtained from mung bean, and the ratio is 0.8:1:

1.

20. 2. The composition or complex of claim 1, characterized in that it has a stable color at a pH of less than about 6, such as a pH of less than about 5, such as a pH of less than about 5, such as a pH of less than about 4, such as a pH of less than about 3, or such as a pH of less than about 2.

21. 10. The composition or composite of claim 1, characterized in that it has a heat-stable and / or light-stable color.

22. 10. A process for the formation of a composition or complex as defined in claim 1, comprising the step of mixing at least one peptide, polypeptide and / or protein as defined in claim 1, or a protein extract comprising at least one peptide, polypeptide and / or protein, with a composition comprising at least one phycobilin as defined in claim 1 in an aqueous solution.

23. 23. The process of claim 22, further comprising filtering the mixture of the composition comprising at least one phycobilin in aqueous solution and at least one peptide, polypeptide and / or protein or protein extract to remove insoluble fractions.

24. 1. A method for stabilizing phycobilins, comprising the steps of: i) contacting phycobilins with at least one protein, polypeptide and / or peptide according to claim 1 in an aqueous solution; and ii) optionally adding sugars; Optionally, further comprising filtering the mixture obtained in step i) and / or ii) to remove the insoluble fraction; Optionally, further comprising concentrating the results of step i) or ii); Optionally, the method further comprises a drying step, such as spray drying.

25. 25. The process or method of any one of claims 22 to 24, wherein the phycobilin is selected from phycoerythrobilin, phycocyanobilin, phycoviolobilin, phycourobilin and mixtures thereof.

26. 26. The process or method of claim 25, wherein the phycocyanobilin is phycocyanin obtained from Arthrospira platensis (also known as Spirulina), Arthrospira fusiformis, Arthrospira maxima, Galdieria daedala, Galdieria sulphuraria, Galdieria maxima, Galdieria partita, Cyanidioschyzon merolae 10D, Cyanidioschyzon merolae DBV201, Cyanidium caldarium, Cyanidium rumpens, Cyanidium daedalum, Cyanidium maximum, Cyanidium partitum, and any mixture thereof.

27. 25. The process or method of claim 22 or 24, wherein the ratio of the at least one protein to the at least one polypeptide and / or peptide is about 100:1 to 1:100, such as 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, or 10:

90.

28. 25. The process or method of claim 22 or 24, wherein the ratio of phycobilin to the at least one peptide, polypeptide and / or protein is in the range of about 10:90 to about 90:10, or about 20:80 to about 80:20, or about 30:70 to about 70:30, or about 40:60 to about 60:40, or about 45:55 to about 55:45, about 50:1 to about 1:50, such as about 40:1 to about 1:40, for example about 30:1 to about 1:30, such as about 20:1 to about 1:20, for example in the range of about 10:1 to about 1:10, such as 1:1, 1:2, 1:3, 3:1, 2:1, 0.5:1, or about 0.8:

2.

29. 25. A stabilized phycobilin obtained using the process or method of claim 24.

30. 30. The stabilized phycobilin of claim 29, characterized in that it has a color that is stable under acidic conditions, stable to heat, and / or stable to light.

31. An encapsulated composition comprising the composition or complex of claim 1 or the stabilized phycobilin of claim 29 or 30, encapsulated using a cross-linked polymer derived from the group consisting of alginate, carrageenan and pectin, derivatives thereof, or combinations thereof.

32. A colorant comprising the composition or complex of claim 1 or the stabilized phycobilin of claim 29 or 30, and optimally other pigments.

33. A colorant comprising the encapsulated composition of claim 31, and optionally other pigments.

34. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product comprising the composition or complex of claim 1 or the stabilized phycobilin of claim 29 or 30.

35. 35. The food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product of claim 34, having a pH of less than 5, such as less than 4, such as less than 3, or such as less than 2.

36. 35. The food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product of claim 34, having a pH of 2.0 to 5.0, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, most preferably 2.4 to 3.

3.

37. 35. The food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product of claim 34, which has been subjected to a heat treatment.

38. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic comprising the encapsulated composition of claim 31.

39. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product as described in claim 38, having a pH of less than 5, such as less than 4, such as less than 3, or such as less than 2.

40. The food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product of claim 38, having a pH of 2.0 to 5.0, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, and most preferably 2.4 to 3.

3.

41. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product as described in claim 38, which has been subjected to heat treatment.

42. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic comprising the colorant of claim 32.

43. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product as described in claim 42, having a pH of less than 5, such as less than 4, such as less than 3, or such as less than 2.

44. The food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product of claim 42, having a pH of 2.0 to 5.0, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, and most preferably 2.4 to 3.

3.

45. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product as described in claim 42, which has been subjected to heat treatment.

46. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic comprising the colorant of claim 33.

47. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product as described in claim 46, having a pH of less than 5, such as less than 4, such as less than 3, or such as less than 2.

48. The food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product of claim 46, having a pH of 2.0 to 5.0, preferably 2.2 to 4.0, more preferably 2.3 to 3.6, and most preferably 2.4 to 3.

3.

49. A food, pharmaceutical, cosmeceutical, nutraceutical or cosmetic product as described in claim 46, which has been subjected to heat treatment.

50. A method for improving the shelf life of the blue color of a food, drug, cosmeceutical, nutraceutical, or cosmetic, comprising adding the composition or complex of claim 1 and / or the stabilized phycobilin of claim 29 or 30 to the base of the food, drug, cosmeceutical, nutraceutical, or cosmetic.

51. A method for improving the shelf life of the blue color of a food, drug, cosmeceutical, nutraceutical or cosmetic, comprising adding the encapsulating agent of claim 31 to the base of the food, drug, cosmeceutical, nutraceutical or cosmetic.

52. A method for improving the shelf life of the blue color of a food, drug, cosmeceutical, nutraceutical or cosmetic, comprising adding the colorant of claim 32 to the base of the food, drug, cosmeceutical, nutraceutical or cosmetic.

53. A method for improving the shelf life of the blue color of a food, drug, cosmeceutical, nutraceutical or cosmetic, comprising adding the colorant of claim 33 to the base of the food, drug, cosmeceutical, nutraceutical or cosmetic.

54. 10. A kit comprising at least one phycobilin as defined in claim 1 and at least one peptide, polypeptide and / or protein, or a protein extract containing at least one peptide, polypeptide and / or protein, and optionally instructions for mixing, preparing and / or using said components.

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