Betaxanthin pigment compositions

Abstract

Pigment compositions comprising betaxanthin pigment molecules are provided. Compositions wherein less than 5% of pigment molecules are betalamic acid are also provided as are compositions wherein at least 40% of betaxanthin molecules are indicaxanthin or iso-indicaxanthin and compositions comprising less than 5% sugars by weight. Methods of producing betaxanthins and pigment compositions are also provided.

Description

BETAXANTHIN PIGMENT COMPOSITIONSCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 635,728, filed April 18, 2024, the contents of which are all incorporated herein by reference in their entirety.REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (PHYT-P-003-PCT.xml; Size: 51,440 bytes; and Date of Creation: April 3, 2025) is herein incorporated by reference in its entirety.FIELD OF INVENTION

[0003] The present invention is in the field of pigment compositions.BACKGROUND OF THE INVENTION

[0004] Betalains are tyrosine-derived, red-violet and yellow plant pigments found in only one group of angiosperms, the Caryophyllales order, in which they occur in a mutually exclusive fashion with the chemically distinct and widespread anthocyanin pigments. The betalain class contains a wide array of compounds, which are generally classified into two groups; the red-violet betacyanins, and the yellow betaxanthins.

[0005] Due to their high water solubility, pH stability and antioxidative properties, betalains may be used as natural food colorants and dietary supplements. Betaxanthins can be found in several edible plants including red beets (beetroot - Beta vulgaris), yellow beet, yellow and red prickly pear (also known as cactus fruit, Barbary fig, Tuna, etc, fruits of plants from the Genus Opuntia sp..), Swiss chard, dragon fruit (Pitaya) and additional fruits, vegetables, flowers, and leaves of plants from the order Caryophyllales. Despite its betaxanthin content, fruit extracts have several drawbacks as a source of food colorants. First, extracts generally include a mix of betalains, often combining betacyanins and betaxanthins, but also can have a mix of different betaxanthins. This limits control over color and hue specificity. Moreover, field-to-field and season-to-season variation in concentration and composition of betalainsderived from plant extracts hinders the production of a food colorant with reproducible qualities. The seasonal nature of agricultural crops also results in a non-stable supply chain with occasional shortages leading to price hikes. In addition, the pigment concentration in the plant extract can be relatively low, negatively impacting its ability to color foods in high efficacy.

[0006] New methods of producing betalains in cultured microbial cells are provided in International Patent Publications WO2017 / 122189 and WO2022 / 253815 and Tinggaard Thomsen, “Beet red food colourant can be produced more sustainably with engineered Yarrowia lipolytica”, Nat Microbiol., 2023 Dec;8(12):2290-2303, Grewal et al., “Bioproduction of a betalain color palette in Saccharomyces cerevisiae”, Metab Eng. 2018 Jan:45:180-188, Hou et al., “Metabolic engineering of Escherichia coli for de novo production of betaxanthins”, Babaei et al., “Combinatorial engineering of betalain biosynthesis pathway in yeast Saccharomyces cerevisiae”, Biotechnol Biofuels Bioprod. 2023 Aug 17; 16( 1): 128. A new superior formulation of betaxanthin pigment is greatly needed.SUMMARY OF THE INVENTION

[0007] The present invention provides pigment compositions comprising betaxanthin pigment molecules. Compositions wherein less than 5% of pigment molecules are betalamic acid molecules or betacyanin molecules are also provided as are compositions wherein at least 40% of betaxanthin molecules are proline-betaxanthin (indicaxanthin), histidinebetaxanthin (muscaaurin VII), glutamic acid-betaxanthin (vulgaxanthin II), phenylalanine- betaxanthis, methionine -betaxanthin, leucine-betaxanthin (vulgaxanthin IV), serinebetaxanthin or glutamine -betaxanthin (vulgaxanthin I) and compositions comprising less than 5% sugars by weight. Methods of producing betaxanthins and pigment compositions are also provided.

[0008] According to a first aspect, there is provided a pigment composition comprising betaxanthin pigment molecules and wherein less than 5% of pigment molecules in the composition are betalamic acid molecules or betacyanin molecules.

[0009] According to another aspect, there is provide a pigment composition comprising betaxanthins, wherein at least 50% of the betaxanthins are proline-betaxanthin, at least 50% of the betaxanthins are histidine -betaxanthins, at least 50% of the betaxanthins are glutamicacid-betaxanthins, at least 50% of the betaxanthins are phenylalanine-betaxanthins, at least 50% of the betaxanthins are methionine-betaxanthins, at least 50% of the betaxanthins are leucine-betaxanthins, or at least 50% of the betaxanthins are serine-betaxanthins.

[0010] According to another aspect, there is provided a pigment composition comprising betaxanthin pigment molecules, wherein the composition comprises less than 5% mono-and di-saccharides by weight.[Oi l] According to some embodiments, at least 90% of the betaxanthins are prolinebetaxanthins or 90% of the betaxanthins are histidine -betaxanthins.

[0012] According to some embodiments, less than 5% of pigment molecules in the composition are betalamic acid molecules or betacyanin molecules.

[0013] According to some embodiments, the pigment composition comprises less than 1 mg / L or 1 mg / kg betalamic acid.

[0014] According to some embodiments, at least 50% of the betaxanthins are prolinebetaxanthin, at least 50% of the betaxanthins are histidine-betaxanthins, at least 50% of the betaxanthins are glutamic acid-betaxanthins, at least 50% of the betaxanthins are phenylalanine-betaxanthins, at least 50% of the betaxanthins are methionine-betaxanthins, at least 50% of the betaxanthins are leucine-betaxanthins, or at least 50% of the betaxanthins are serine-betaxanthins.

[0015] According to some embodiments, at least 90% of the betaxanthins are prolinebetaxanthins or 90% of the betaxanthins are histidine-betaxanthins.

[0016] According to some embodiments, at least 90% of all pigment molecules in the composition are indicaxanthin or iso-indicaxanthin.

[0017] According to some embodiments, the composition comprises less than 5% mono-and di-saccharides by weight.

[0018] According to some embodiments, the composition comprises less than 0.1 pg / kg geosmin, less than 0.01 g / L nitrates or both.

[0019] According to some embodiments, a ratio of indicaxanthin to iso-indicaxanthin is at least 3:1.

[0020] According to some embodiments, the composition is a liquid composition or a powder.

[0021] According to some embodiments, the pigment composition comprises less than 5% dietary carbohydrates by weight.

[0022] According to some embodiments, less than 10% of all pigment molecules in the composition are betacyanins.

[0023] According to some embodiments, less than 1% of all pigment molecules in the composition are betacyanins.

[0024] According to some embodiments, the pigment composition comprises less than 1 mg / L or 1 mg / kg betacyanin.

[0025] According to some embodiments, the pigment composition comprises at least 500 mg magnesium per liter of composition or at least 1000 mg magnesium per kilogram of composition.

[0026] According to some embodiments, the pigment composition is a liquid and comprising greater than 1.5 grams betaxanthin pigment molecules per liter of composition or is a powder and comprising greater than 1.5 grams betalain pigment molecules per kilogram of composition.

[0027] According to some embodiments, the pigment composition is a liquid and comprising at least 4 grams betaxanthin pigment molecules per liter of composition or is a powder and comprising at least 4 grams betalain pigment molecules per kilogram of composition.

[0028] According to some embodiments, the betacyanin molecules comprise betanin, betanidin, and stereoisomers thereof.

[0029] According to some embodiments, less than 5% of pigment molecules in the composition are betalamic acid molecules.

[0030] According to another aspect, there is provided a method of producing betaxanthin comprising an amino acid of interest, the method comprising culturing a cell comprising integrated into its genome at least one tyrosine hydroxylase gene that is not an L-DOPA oxidase gene, and at least one DOPA 4,5-dioxygenase (DOD) gene in a culture medium supplemented with the amino acid of interest and collecting the culture medium, thereby producing a betaxanthin comprising an amino acid of interest.

[0031] According to some embodiments, the amino acid is selected from, proline, histidine, methionine, glutamic acid, serine, leucine, phenylalanine and glutamine.

[0032] According to some embodiments, at least 40% of the betaxanthin produced comprises the amino acid.

[0033] According to some embodiments, the cell further comprises and at least one NADPH cytochrome P450 reductase gene integrated into its genome.

[0034] According to some embodiments, the method is a method of producing a pigment composition according to embodiments of the present invention.

[0035] According to some embodiments, the tyrosine hydroxylase gene encodes a protein comprising an amino acid sequence of SEQ ID NO: 2, 12, 14, 17, 19, 21 or 23 or a sequence with at least 85% identity thereto that converts tyrosine to L-DOPA and does not convert L- DOPA to cyclo-DOPA, and the DOD gene encodes a protein comprising an amino acid sequence of SEQ ID NO: 4 or 6 or a sequence with at least 85% identity thereto that converts L-DOPA to betalamic acid.

[0036] According to some embodiments, the NADPH cytochrome P450 reductase gene encodes a protein comprising an amino acid sequence of SEQ ID NO: 8 or 10 or a sequence with at least 85% identity thereto that comprises reductase activity.

[0037] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 Table of characteristics of betalain pigment composition according to one embodiment of the invention and previously known compositions.

[0039] Figure 2: Table of all detected pigment molecules in a composition according to one embodiment of the invention.

[0040] Figure 3: Images of the color of betalain pigment composition of the invention and previously known compositions.

[0041] Figure 4: Color Point measurement values as compared to Yellow Prickly Pear Juice extract.

[0042] Figure 5: Reaction of betalamic acid with proline to produce indicaxanthin, glutamic acid to produce glutamic acid-betaxanthin, phenylalanine to produce phenylalaninebetaxanthin, methionine to produce methionine-betaxanthin, leucine to produce leucinebetaxanthin, serine to produce serine -betaxanthin, histidine to produce histidine-betaxanthin and glutamine to produce glutamine -betaxanthin.

[0043] Figures 6A-6B: (6A) Bar graph of relative production of various betaxanthins produced by media supplementation with various amino acids. (6B) Table of percent purity of various amino acid-betaxanthins out of all betaxanthins produced by addition of the amino acid into the media.

[0044] Figures 7A-7B: (7A) Photograph of liquid pigment compositions produced by addition to media of various amino acids. (7B) Color Point measurement values as compared to media with no amino acids (set to zero) of the pigment compositions of 7A.DETAILED DESCRIPTION OF THE INVENTION

[0045] The present invention, in some embodiments, provides pigment compositions comprising betaxanthin pigment molecules. Compositions wherein less than 5% of pigment molecules are betalamic acid molecules or betacyanin molecules are also provided as are compositions wherein at least 40% of betaxanthin molecules are proline-betaxanthin, histidine-betaxanthin, glutamic acid— betaxanthin, phenylalanine-betaxanthin, methioninebetaxanthin, leucine -betaxanthin, serine-betaxanthin or glutamine -betaxanthin and compositions comprising less than 5% sugars by weight. Methods of producing betaxanthins and pigment compositions are also provided.

[0046] The invention is based, at least in part, on the surprising finding that by integrating the genes of the betaxanthin synthesis pathway into the genome of a non-betalain producing cell (e.g., yeast) very high yields, with very high purity, of betaxanthin can be produced. Compositions in which less than 5%, or even less than 1%, of pigment molecules were betalamic acid or betacyanins could be produced. Further, it was found that supplementing the cell culture media with an amino acid could produce betaxanthin comprising that amino acid (e.g., proline-betaxanthin). This could be done with very high purity, such that over 40%, (or even 50%, or even 60% or even 90%) of the betaxanthin in the composition was betaxanthin with that amino acid. In particular proline-betaxanthin and histidine-betaxanthin could be produced at greater than 90% purity.

[0047] By a first aspect, there is provided a composition comprising betalain pigment molecules.

[0048] In some embodiments, the composition is a pigment composition. In some embodiments, the pigment is a betalain pigment. In some embodiments, the pigment is a yellow pigment. In some embodiments, the pigment is within the yellow spectrum. In some embodiments, the pigment is within the yellow-orange spectrum. In some embodiments, the yellow pigment reflects light with a wavelength between 570 and 580 nanometers. In some embodiments, the yellow pigment reflects light with a wavelength between 560 and 590 nanometers. In some embodiments, the yellow pigment reflects light with a wavelength between 570 and 585 nanometers. In some embodiments, the yellow pigment reflects light with a wavelength between 560 and 580 nanometers. In some embodiments, the yellow pigment comprises lambda max (Xmax) of about 435-480 nanometers. In some embodiments, a lambda max is an absorption lambda max. In some embodiments, the lambda max is the lambda max in aqueous solution. It will be understood by a skilled artisan that measuring lambda max in an organic solvent will shift the lambda max, but the lambda max of yellow pigment in an organic solvent is well known to the skilled artisan. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 435-485 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 400-440 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 410-440 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 410-435 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 370-485 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 370-480 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 380-480 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 380-485 nanometers. In some embodiments, the yellow pigment comprises a lambda max (Xmax) of about 470-485 nanometers. It will be understood by a skilled artisan that the exact color of the composition is determined by the abundance of each betalain pigment and that the final color can be varied by varying the relative amounts of the various betalains in the composition.

[0049] In some embodiments, the composition is an edible composition. In some embodiments, the composition is organic. In some embodiments, the composition is fit for human consumption. In some embodiments, the composition is a dye. In some embodiments,the composition is a food coloring. In some embodiments, the composition is an additive. In some embodiments, the composition is a food supplement. In some embodiments, the composition is a food supplement with health claims. In some embodiments, the composition is a nutraceutical. In some embodiments, the composition is an antioxidant. In some embodiments, the composition is a reducing agent. In some embodiments, the composition is a food preservative. In some embodiments, the composition is a coloring composition. In some embodiments, the composition is a cookable composition. In some embodiments, the composition is thermostable. In some embodiments, the composition is a liquid composition. In some embodiments, the composition is aqueous. In some embodiments, the composition is a solid. In some embodiments, the solid is a powder. In some embodiments, the composition is a crystal. In some embodiments, the composition is a non-natural composition. In some embodiments, the composition comprises betalain pigment molecule combinations and ratios not found in nature. In some embodiments, the composition comprises a color not found in nature. Because the color of the pigment composition of the invention is not found in nature the composition of the invention cannot be considered a product of nature.

[0050] Betalains are water-soluble pigments derived from tyrosine present in vacuoles of plants of the order Caryophyllales and in mushrooms of the genera Amanita, Hygrocybe and Hygrophorus. Within the plant kingdom, betalains are found in only one group of angiosperms, the Caryophyllales. In this order, betalains and anthocyanins occur in a mutually exclusive fashion, i.e., no plant species produces both types of pigments. One of the most prominent features of the Caryophyllales order is its dominance in arid and semi- arid regions, and habitation of saline and alkaline soils. While some families within the Caryophyllales are distributed worldwide in a variety of habitats, members of several families are particularly adapted to arid or saline regions, including the Aizoaceae (ice-plant family), Portulacaceae (purslane family) and most notably, the Cactaceae (cactus family).

[0051] Betalains are divided into two classes of pigment molecules: red-purple colored betacyanins and yellow-orange colored betaxanthins. In some embodiments, the betalain pigment molecule is a betaxanthin. In some embodiments, the betalain pigment molecule is a betaxanthin pigment molecule. In some embodiments, the betalain pigment molecules comprise a betaxanthin. In some embodiments, the betalain pigment molecules comprise betacyanins and betaxanthins.

[0052] In some embodiments, the betalain pigment molecules comprise indicaxanthin. Indicaxanthin is a betaxanthin produced by the conjugation of proline to betalamic acid. Itis also known as proline-betaxanthin. It is a known antioxidant and has a bright yellow color. The condensation of betalamic acid with proline to produce indicaxanthin is provided in Figure 5. In some embodiments, indicaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises indicaxanthin. In some embodiments, the stereoisomer of indicaxanthin is iso-indicaxanthin. In some embodiments, indicaxanthin comprises indicaxanthin and iso-indicaxanthin. In some embodiments, the betalain pigment molecules comprise indicaxanthin and iso-indicaxanthin. In some embodiments, the composition comprises indicaxanthin and iso-indicaxanthin.

[0053] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betaxanthin molecules in the composition are indicaxanthin. In some embodiments, at least 85% of the betaxanthin molecules in the composition are indicaxanthin. In some embodiments, at least 90% of the betaxanthin molecules in the composition are indicaxanthin. In some embodiments, at least 95% of the betaxanthin molecules in the composition are indicaxanthin. In some embodiments, at least 98% of the betaxanthin molecules in the composition are indicaxanthin. In some embodiments, at least 99% of the betaxanthin molecules in the composition are indicaxanthin.

[0054] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are indicaxanthin or iso- indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betaxanthin molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, at least 85% of the betaxanthin molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, at least 90% of the betaxanthin molecules in the composition are indicaxanthin or iso- indicaxanthin. In some embodiments, at least 95% of the betaxanthin molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, at least 98% of the betaxanthin molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, at least 99% of the betaxanthin molecules in the composition are indicaxanthin or iso-indicaxanthin.

[0055] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at leastat least 85% of the betalain molecules in the composition are indieaxanthin. In some embodiments, at least 90% of the betalain molecules in the composition are indieaxanthin. In some embodiments, at least 95% of the betalain molecules in the composition are indieaxanthin. In some embodiments, at least 98% of the betalain molecules in the composition are indieaxanthin. In some embodiments, at least 99% of the betalain molecules in the composition are indieaxanthin.

[0056] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are indieaxanthin or iso- indieaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betalain molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 85% of the betalain molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 90% of the betalain molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 95% of the betalain molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 98% of the betalain molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 99% of the betalain molecules in the composition are indieaxanthin or iso-indieaxanthin.

[0057] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are indieaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the pigment molecules in the composition are indieaxanthin. In some embodiments, at least 85% of the pigment molecules in the composition are indieaxanthin. In some embodiments, at least 90% of the pigment molecules in the composition are indieaxanthin. In some embodiments, at least 95% of the pigment molecules in the composition are indieaxanthin. In some embodiments, at least 98% of the pigment molecules in the composition are indieaxanthin. In some embodiments, at least 99% of the pigment molecules in the composition are indieaxanthin.

[0058] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are indieaxanthin or iso- indieaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the pigment molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 85% of the pigment molecules in the composition are indieaxanthin or iso-indieaxanthin. In some embodiments, at least 90% of the pigment molecules in the composition are indieaxanthin or iso-indieaxanthin. In someembodiments, at least 95% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, at least 98% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, at least 99% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin.

[0059] In some embodiments, the betalain pigment molecules comprise histidinebetaxanthin. Histidine-betaxanthin is a betaxanthin produced by the conjugation of histidine to betalamic acid and is also known as muscaaurin VII. The condensation of betalamic acid with histidine to produce histidine-betaxanthin is provided in Figure 5. In some embodiments, histidine-betaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises histidine-betaxanthin. In some embodiments, the stereoisomer of histidine-betaxanthin is histidine-isobetaxanthin. In some embodiments, histidine-betaxanthin comprises histidine-betaxanthin and histidine-isobetaxanthin. In some embodiments, the betalain pigment molecules comprise histidine-betaxanthin and histidine- isobetaxanthin. In some embodiments, the composition comprises histidine-betaxanthin and histidine-isobetaxanthin.

[0060] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are histidine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betaxanthin molecules in the composition are histidine-betaxanthin. In some embodiments, at least 85% of the betaxanthin molecules in the composition are histidine-betaxanthin. In some embodiments, at least 90% of the betaxanthin molecules in the composition are histidine-betaxanthin. In some embodiments, at least 95% of the betaxanthin molecules in the composition are histidine-betaxanthin. In some embodiments, at least 98% of the betaxanthin molecules in the composition are histidine-betaxanthin. In some embodiments, at least 99% of the betaxanthin molecules in the composition are histidine-betaxanthin.

[0061] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betaxanthin molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 85% of the betaxanthin molecules in the composition are histidine-betaxanthin or histidine- isobetaxanthin. In some embodiments, at least 90% of the betaxanthin molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, atleast 95% of the betaxanthin molecules in the composition are histidine -betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 98% of the betaxanthin molecules in the composition are histidine -betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 99% of the betaxanthin molecules in the composition are histidinebetaxanthin or histidine-isobetaxanthin.

[0062] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are histidine -betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betalain molecules in the composition are histidine -betaxanthin. In some embodiments, at least 85% of the betalain molecules in the composition are histidinebetaxanthin. In some embodiments, at least 90% of the betalain molecules in the composition are histidine-betaxanthin. In some embodiments, at least 95% of the betalain molecules in the composition are histidine-betaxanthin. In some embodiments, at least 98% of the betalain molecules in the composition are histidine-betaxanthin. In some embodiments, at least 99% of the betalain molecules in the composition are histidine-betaxanthin.

[0063] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 85% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 90% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 95% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 98% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 99% of the betalain molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin.

[0064] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are histidine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, at least 85% of the pigment molecules in the composition are histidine- betaxanthin. In some embodiments, at least 90% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, at least 95% of the pigmentmolecules in the composition are histidine-betaxanthin. In some embodiments, at least 98% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, at least 99% of the pigment molecules in the composition are histidine- betaxanthin.

[0065] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 80% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 85% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 90% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 95% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 98% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, at least 99% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin.

[0066] In some embodiments, the betalain pigment molecules comprise glutamic acid- betaxanthin. Glutamic acid-betaxanthin is a betaxanthin produced by the conjugation of glutamic acid to betalamic acid and is also known as vulgaxanthin II. The condensation of betalamic acid with glutamic acid to produce glutamic acid-betaxanthin is provided in Figure 5. In some embodiments, glutamic acid-betaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises glutamic acid-betaxanthin. In some embodiments, the stereoisomer of glutamic acid-betaxanthin is glutamic acid- isobetaxanthin. In some embodiments, glutamic acid-betaxanthin comprises glutamic acid- betaxanthin and glutamic acid-isobetaxanthin. In some embodiments, the betalain pigment molecules comprise glutamic acid-betaxanthin and glutamic acid-isobetaxanthin. In some embodiments, the composition comprises glutamic acid-betaxanthin and glutamic acid- isobetaxanthin.

[0067] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are glutamic acid- betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 50% of thebetaxanthin molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin.

[0068] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are glutamic acid- betaxanthin or glutamic acid-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin.

[0069] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are glutamic acid-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are glutamic acid-betaxanthin.

[0070] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin.

[0071] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are glutamic acid-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are glutamic acid-betaxanthin.

[0072] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are glutamic acid- betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin.

[0073] In some embodiments, the betalain pigment molecules comprise phenylalaninebetaxanthin. Phenylalanine -betaxanthin is a betaxanthin produced by the conjugation of phenylalanine to betalamic acid. The condensation of betalamic acid with phenylalanine to produce phenylalanine-betaxanthin is provided in Figure 5. In some embodiments, phenylalanine-betaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises phenylalanine-betaxanthin. In some embodiments, the stereoisomer of phenylalanine-betaxanthin is phenylalanine-isobetaxanthin. In some embodiments, phenylalanine-betaxanthin comprises phenylalanine-betaxanthin and phenylalanine- isobetaxanthin. In some embodiments, the betalain pigment molecules comprisephenylalanine-betaxanthin and phenylalanine-isobetaxanthin. In some embodiments, the composition comprises phenylalanine-betaxanthin and phenylalanine-isobetaxanthin.

[0074] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are phenylalanine- betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin.

[0075] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are phenylalanine- betaxanthin or phenylalanine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin.

[0076] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are phenylalanine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are phenylalanine-betaxanthin.

[0077] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are phenylalanine -betaxanthin or phenylalanine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are phenylalanine- betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin.

[0078] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are phenylalanine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are phenylalanine- betaxanthin.

[0079] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are phenylalanine- betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin.

[0080] In some embodiments, the betalain pigment molecules comprise methioninebetaxanthin. Methionine -betaxanthin is a betaxanthin produced by the conjugation of methionine to betalamic acid. The condensation of betalamic acid with methionine to produce methionine-betaxanthin is provided in Figure 5. In some embodiments, methioninebetaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises methionine-betaxanthin. In some embodiments, the stereoisomer of methioninebetaxanthin is methionine-isobetaxanthin. In some embodiments, methionine-betaxanthin comprises methionine-betaxanthin and methionine-isobetaxanthin. In some embodiments, the betalain pigment molecules comprise methionine-betaxanthin and methionine- isobetaxanthin. In some embodiments, the composition comprises methionine-betaxanthin and methionine-isobetaxanthin.

[0081] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are methionine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are methionine-betaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are methionine-betaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are methionine-betaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are methionine-betaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are methionine- betaxanthin.

[0082] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are methionine- betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are methionine- betaxanthin or methionine-isobetaxanthin.

[0083] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are methionine -betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are methionine -betaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are methionine-betaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are methionine-betaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are methionine-betaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are methionine-betaxanthin.

[0084] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are methionine- betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin.

[0085] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are methionine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are methionine-betaxanthin.

[0086] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the compositionare methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are methionine- betaxanthin or methionine-isobetaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin.

[0087] In some embodiments, the betalain pigment molecules comprise leucine -betaxanthin. Leucine-betaxanthin is a betaxanthin produced by the conjugation of leucine to betalamic acid and is also known as vulgaxanthin IV. The condensation of betalamic acid with leucine to produce leucine-betaxanthin is provided in Figure 5. In some embodiments, leucinebetaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises leucine-betaxanthin. In some embodiments, the stereoisomer of leucine- betaxanthin is leucine-isobetaxanthin. In some embodiments, leucine-betaxanthin comprises leucine-betaxanthin and leucine-isobetaxanthin. In some embodiments, the betalain pigment molecules comprise leucine-betaxanthin and leucine-isobetaxanthin. In some embodiments, the composition comprises leucine-betaxanthin and leucine-isobetaxanthin.

[0088] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are leucine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are leucine-betaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are leucine-betaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are leucine-betaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are leucine-betaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are leucine-betaxanthin.

[0089] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition areleucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin.

[0090] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are leucine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are leucine-betaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are leucine- betaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are leucine-betaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are leucine-betaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are leucine-betaxanthin.

[0091] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are leucine- betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are leucine- betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are leucine- betaxanthin or leucine-isobetaxanthin.

[0092] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are leucine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are leucine- betaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are leucine-betaxanthin.

[0093] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are leucinebetaxanthin or leucine-isobetaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are leucine- betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are leucine- betaxanthin or leucine-isobetaxanthin.

[0094] In some embodiments, the betalain pigment molecules comprise serine-betaxanthin. Serine-betaxanthin is a betaxanthin produced by the conjugation of serine to betalamic acid. The condensation of betalamic acid with serine to produce serine-betaxanthin is provided in Figure 5. In some embodiments, serine-betaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises serine-betaxanthin. In some embodiments, the stereoisomer of serine-betaxanthin is serine-isobetaxanthin. In some embodiments, serine-betaxanthin comprises serine-betaxanthin and serine-isobetaxanthin. In some embodiments, the betalain pigment molecules comprise serine-betaxanthin and serine-isobetaxanthin. In some embodiments, the composition comprises serine-betaxanthin and serine-isobetaxanthin.

[0095] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are serine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are serine-betaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are serine- betaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are serine-betaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are serine-betaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are serine-betaxanthin.

[0096] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition areserine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are serine- betaxanthin or serine-isobetaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are serine- betaxanthin or serine-isobetaxanthin.

[0097] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are serine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are serine-betaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are serine- betaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are serine-betaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are serine-betaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are serine-betaxanthin.

[0098] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are serine-betaxanthin or serine- isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are serine- betaxanthin or serine-isobetaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are serine- betaxanthin or serine-isobetaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are serine- betaxanthin or serine-isobetaxanthin.

[0099] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are serine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are serine- betaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, at least 55% of the pigmentmolecules in the composition are serine-betaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are serine-betaxanthin.

[0100] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are serine-betaxanthin or serineisobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are serine- betaxanthin or serine-isobetaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are serine- betaxanthin or serine-isobetaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are serine- betaxanthin or serine-isobetaxanthin.

[0101] In some embodiments, the betalain pigment molecules comprise glutaminebetaxanthin. Glutamine-betaxanthin is a betaxanthin produced by the conjugation of glutamine to betalamic acid and is also known as vulgaxanthin I. The condensation of betalamic acid with glutamine to produce glutamine-betaxanthin is provided in Figure 5. In some embodiments, glutamine-betaxanthin is the molecule presented in Figure 5. In some embodiments, the composition comprises glutamine-betaxanthin. In some embodiments, the stereoisomer of glutamine-betaxanthin is glutamine-isobetaxanthin. In some embodiments, glutamine-betaxanthin comprises glutamine-betaxanthin and glutamine-isobetaxanthin. In some embodiments, the betalain pigment molecules comprise glutamine-betaxanthin and glutamine-isobetaxanthin. In some embodiments, the composition comprises glutamine- betaxanthin and glutamine-isobetaxanthin.

[0102] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are glutamine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are glutamine-betaxanthin.

[0103] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betaxanthin molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betaxanthin molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 45% of the betaxanthin molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 50% of the betaxanthin molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 55% of the betaxanthin molecules in the composition are glutamine- betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 60% of the betaxanthin molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin.

[0104] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are glutamine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are glutamine- betaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are glutamine-betaxanthin.

[0105] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the betalain molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the betalain molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 45% of the betalain molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, at least 50% of the betalain molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 55% of the betalain molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 60% of the betalain molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin.

[0106] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are glutamine-betaxanthin.Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are glutaminebetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are glutamine-betaxanthin.

[0107] In some embodiments, at least 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99 or 100% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, at least 40% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 45% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, at least 50% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 55% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, at least 60% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin.

[0108] In some embodiments, the composition is devoid of betacyanin molecules. In some embodiments, the composition is essentially devoid of betacyanin molecules. In some embodiments, essential devoid is a level that is below the threshold of detection. In some embodiments, detection is by mass spectrometry. In some embodiments, detection is by LC- MS. In some embodiments, less than 50, 40, 35, 30, 35, 30, 35, 10, 7, 5, 4, 3, 2 or 1% of all betalain molecule in the composition are betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, less than 25% of all betalain molecules in the composition are betacyanins. In some embodiments, less than 15% of all betalain molecules in the composition are betacyanins. In some embodiments, less than 10% of all betalain molecules in the composition are betacyanins. In some embodiments, less than 5% of all betalain molecules in the composition are betacyanins. In some embodiments, less than 1% of all betalain molecules in the composition are betacyanins.

[0109] In some embodiments, less than 50, 40, 35, 30, 35, 30, 35, 10, 7, 5, 4, 3, 2 or 1% of all pigment molecules in the composition are betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, less than 25% of all pigment molecules in the composition are betacyanins. In some embodiments, less than 15% of allpigment molecules in the composition are betacyanins. In some embodiments, less than 10% of all pigment molecules in the composition are betacyanins. In some embodiments, less than 5% of all pigment molecules in the composition are betacyanins. In some embodiments, less than 1% of all pigment molecules in the composition are betacyanins.

[0110] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2.5, 2, 1, 0.75, 0.5, 0.25 or 0.1 milligram per liter (mg / L) betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 2.5 mg / L betacyanins. In some embodiments, the composition comprises less than 2 mg / L betacyanins. In some embodiments, the composition comprises less than 1 mg / L betacyanins. In some embodiments, the composition comprises less than 0.5 mg / L betacyanins. In some embodiments, the composition comprises less than 0.1 mg / L betacyanins.

[0111] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2.5, 2, 1, 0.75, 0.5, 0.25 or 0.1 milligram per kilogram (mg / kg) betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 2.5 mg / kg betacyanins. In some embodiments, the composition comprises less than 2 mg / kg betacyanins. In some embodiments, the composition comprises less than 1 mg / kg betacyanins. In some embodiments, the composition comprises less than 0.5 mg / kg betacyanins. In some embodiments, the composition comprises less than 0.1 mg / kg betacyanins.

[0112] In some embodiments, less than 50, 40, 35, 30, 35, 30, 35, 10, 7, 5, 4, 3, 2 or 1% of all pigment molecules in the composition are betalamic acid molecules. Each possibility represents a separate embodiment of the invention. In some embodiments, less than 25% of all pigment molecules in the composition are betalamic acid molecules. In some embodiments, less than 15% of all pigment molecules in the composition are betalamic acid molecules. In some embodiments, less than 10% of all pigment molecules in the composition are betalamic acid molecules. In some embodiments, less than 5% of all pigment molecules in the composition are betalamic acid molecules. In some embodiments, less than 1% of all pigment molecules in the composition are betalamic acid molecules. In some embodiments, the composition is devoid of betalamic acid. In some embodiments, devoid is essentially devoid.

[0113] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, 0.25 or 0.1 milligram per liter (mg / L) betalamic acid. Each possibility representsa separate embodiment of the invention. In some embodiments, the composition comprises less than 3 mg / L betalamic acid. In some embodiments, the composition comprises less than 2 mg / L betalamic acid. In some embodiments, the composition comprises less than 1 mg / L betalamic acid. In some embodiments, the composition comprises less than 0.5 mg / L betalamic acid. In some embodiments, the composition comprises less than 0.1 mg / L betalamic acid.

[0114] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, 0.25 or 0.1 milligram per kilogram (mg / kg) betalamic acid. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 3 mg / kg betalamic acid. In some embodiments, the composition comprises less than 2 mg / kg betalamic acid. In some embodiments, the composition comprises less than 1 mg / kg betalamic acid. In some embodiments, the composition comprises less than 0.5 mg / kg betalamic acid. In some embodiments, the composition comprises less than 0.1 mg / kg betalamic acid.

[0115] In some embodiments, less than 50, 40, 35, 30, 35, 30, 35, 10, 7, 5, 4, 3, 2 or 1% of all pigment molecules in the composition are betacyanin molecules. Each possibility represents a separate embodiment of the invention. In some embodiments, a betacyanin comprises betanin. In some embodiments, a betacyanin comprises betanin or a stereoisomer thereof. In some embodiments, a betacyanin comprises betanin and a stereoisomer thereof. In some embodiments, a betacyanin comprises betanidin. In some embodiments, a betacyanin comprises betanidin or a stereoisomer thereof. In some embodiments, a betacyanin comprises betanidin and a stereoisomer thereof. In some embodiments, a betacyanin comprises betanin and / or betanidin. In some embodiments, a betacyanin comprises betanin, betanidin and / or a stereoisomer thereof. In some embodiments, the betacyanin is betanin and betanidin. In some embodiments, the betacyanin is betanin, betanidin and a stereoisomer thereof. In some embodiments, less than 25% of all pigment molecules in the composition are betacyanin molecules. In some embodiments, less than 15% of all pigment molecules in the composition are betacyanin molecules. In some embodiments, less than 10% of all pigment molecules in the composition are betacyanin molecules. In some embodiments, less than 5% of all pigment molecules in the composition are betacyanin molecules. In some embodiments, less than 1% of all pigment molecules in the composition are betacyanin molecules. In some embodiments, the composition is devoid of betacyanin. In some embodiments, devoid is essentially devoid.

[0116] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, 0.25 or 0.1 milligram per liter (mg / L) betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 3 mg / L betacyanins. In some embodiments, the composition comprises less than 2 mg / L betacyanins. In some embodiments, the composition comprises less than 1 mg / L betacyanins. In some embodiments, the composition comprises less than 0.5 mg / L betacyanins. In some embodiments, the composition comprises less than 0.1 mg / L betacyanins.

[0117] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, 0.25 or 0.1 milligram per kilogram (mg / kg) betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 3 mg / kg betacyanins. In some embodiments, the composition comprises less than 2 mg / kg betacyanins. In some embodiments, the composition comprises less than 1 mg / kg betacyanins. In some embodiments, the composition comprises less than 0.5 mg / kg betacyanins. In some embodiments, the composition comprises less than 0.1 mg / kg betacyanins.

[0118] In some embodiments, less than 50, 40, 35, 30, 35, 30, 35, 10, 7, 5, 4, 3, 2 or 1% of all pigment molecules in the composition are betalamic acid or betacyanin molecules. Each possibility represents a separate embodiment of the invention. In some embodiments, less than 50, 40, 35, 30, 35, 30, 35, 10, 7, 5, 4, 3, 2 or 1% of all pigment molecules in the composition are a pigment molecule selected from betalamic acid and betacyanin molecules. Each possibility represents a separate embodiment of the invention. In some embodiments, a betalamic acid or betacyanin comprises betanin. In some embodiments, a betalamic acid or betacyanin comprises betanidin. In some embodiments, a betalamic acid or betacyanin comprises betanin and / or betanidin. In some embodiments, the betalamic acid or betacyanin is betanin and betanidin. In some embodiments, less than 25% of all pigment molecules in the composition are betalamic acid or betacyanin molecules. In some embodiments, less than 15% of all pigment molecules in the composition are betalamic acid or betacyanin molecules. In some embodiments, less than 10% of all pigment molecules in the composition are betalamic acid or betacyanin molecules. In some embodiments, less than 5% of all pigment molecules in the composition are betalamic acid or betacyanin molecules. In some embodiments, less than 1% of all pigment molecules in the composition are betalamic acid or betacyanin molecules. In some embodiments, the composition is devoid of betalamic acid and betacyanin. In some embodiments, devoid is essentially devoid.

[0119] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, 0.25 and 0.1 milligram per liter (mg / L) betalamic acid and betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 3 mg / L betalamic acid and betacyanins. In some embodiments, the composition comprises less than 2 mg / L betalamic acid and betacyanins. In some embodiments, the composition comprises less than 1 mg / L betalamic acid and betacyanins. In some embodiments, the composition comprises less than 0.5 mg / L betalamic acid and betacyanins. In some embodiments, the composition comprises less than 0.1 mg / L betalamic acid and betacyanins.

[0120] In some embodiments, the composition comprises less than 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5, 0.25 or O.l milligram per kilogram (mg / kg) betalamic acid or betacyanins. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 3 mg / kg betalamic acid or betacyanins. In some embodiments, the composition comprises less than 2 mg / kg betalamic acid or betacyanins. In some embodiments, the composition comprises less than 1 mg / kg betalamic acid or betacyanins. In some embodiments, the composition comprises less than 0.5 mg / kg betalamic acid or betacyanins. In some embodiments, the composition comprises less than 0.1 mg / kg betalamic acid or betacyanins.

[0121] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85-97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92,87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the betaxanthins in the composition are indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments. 80-99% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 80- 98% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 85- 99% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 85- 98% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 90- 99% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 90- 98% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 95- 99% of the betaxanthins in the composition are indicaxanthin. In some embodiments, 95- 98% of the betaxanthins in the composition are indicaxanthin.

[0122] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80-99% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 80-98% of the betaxanthins in the composition are indicaxanthin or iso- indicaxanthin. In some embodiments, 85-99% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 85-98% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 90-99% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 90-98% of the betaxanthins in the composition are indicaxanthin or iso- indicaxanthin. In some embodiments, 95-99% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 95-98% of the betaxanthins in the composition are indicaxanthin or iso-indicaxanthin.

[0123] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the pigment molecules in the composition are indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80-99% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 80-98% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 85-99% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 85-98% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 90- 99% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 90-98% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 95-99% of the pigment molecules in the composition are indicaxanthin. In some embodiments, 95-98% of the pigment molecules in the composition are indicaxanthin.

[0124] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80- 99% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 80-98% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 85-99% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 85-98% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 90-99% of the pigment molecules in the composition are indicaxanthin or iso- indicaxanthin. In some embodiments, 90-98% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 95-99% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin. In some embodiments, 95-98% of the pigment molecules in the composition are indicaxanthin or iso-indicaxanthin.

[0125] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the betaxanthins in the composition are histidine -betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80-99% of the betaxanthins in the composition are histidine -betaxanthin. In some embodiments, 80-98% of the betaxanthins in the composition are histidine -betaxanthin. In some embodiments, 85-99% of the betaxanthins in the composition are histidine -betaxanthin. In some embodiments, 85- 98% of the betaxanthins in the composition are histidine -betaxanthin. In some embodiments, 90-99% of the betaxanthins in the composition are histidine-betaxanthin. In some embodiments, 90-98% of the betaxanthins in the composition are histidine-betaxanthin. In some embodiments, 95-99% of the betaxanthins in the composition are histidine-betaxanthin. In some embodiments, 95-98% of the betaxanthins in the composition are histidine-betaxanthin.

[0126] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the betaxanthins in the composition are histidine -betaxanthin or histidine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80- 99% of the betaxanthins in the composition are histidine-betaxanthin or histidine- isobetaxanthin. In some embodiments, 80-98% of the betaxanthins in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 85-99% of the betaxanthins in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 85-98% of the betaxanthins in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 90-99% of the betaxanthins in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 90-98% of the betaxanthins in the composition are histidine-betaxanthin or histidine- isobetaxanthin. In some embodiments, 95-99% of the betaxanthins in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 95-98% of the betaxanthins in the composition are histidine-betaxanthin or histidine-isobetaxanthin.

[0127] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the pigment molecules in the composition are histidine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80-99% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, 80-98% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, 85-99% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, 85-98% of the pigment molecules in the composition are histidine- betaxanthin. In some embodiments, 90-99% of the pigment molecules in the compositionare histidine-betaxanthin. In some embodiments, 90-98% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, 95-99% of the pigment molecules in the composition are histidine-betaxanthin. In some embodiments, 95-98% of the pigment molecules in the composition are histidine-betaxanthin.

[0128] In some embodiments, 80-100, 80-99, 80-98, 80-97, 80-95, 80-93, 80-92, 80-90, SO- 87, 80-85, 81-100, 81-99, 81-98, 81-97, 81-95, 81-93, 81-92, 81-90, 81-87, 81-85, 82-100, 82-99, 82-98, 82-97, 82-95, 82-93, 82-92, 82-90, 82-87, 82-85, 85-100, 85-99, 85-98, 85- 97, 85-95, 85-93, 85-92, 85-90, 85-87, 87-100, 87-99, 87-98, 87-97, 87-95, 87-93, 87-92, 87-90, 90-100, 90-99, 90-98, 90-97, 90-95, 90-93, 90-92, 92-100, 92-99, 92-98, 92-97, 92- 95, 92-93, 93-100, 93-99, 93-98, 93-97, 93-95, 95-100, 95-99, 95-98, or 95-97% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 80-99% of the pigment molecules in the composition are histidine-betaxanthin or histidine- isobetaxanthin. In some embodiments, 80-98% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 85-99% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 85-98% of the pigment molecules in the composition are histidine- betaxanthin or histidine-isobetaxanthin. In some embodiments, 90-99% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 90-98% of the pigment molecules in the composition are histidine- betaxanthin or histidine-isobetaxanthin. In some embodiments, 95-99% of the pigment molecules in the composition are histidine-betaxanthin or histidine-isobetaxanthin. In some embodiments, 95-98% of the pigment molecules in the composition are histidine- betaxanthin or histidine-isobetaxanthin.

[0129] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are glutamic acid-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-65% of the betaxanthins in the composition are glutamic acid-betaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are glutamic acid-betaxanthin. In some embodiments, 45-65% of the betaxanthins in the composition are glutamic acid-betaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are glutamic acid-betaxanthin. In some embodiments, 50-65% of the betaxanthins in the composition are glutamic acid- betaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are glutamic acid-betaxanthin. In some embodiments, 55-60% of the betaxanthins in the composition are glutamic acid-betaxanthin. In some embodiments, 55-65% of the betaxanthins in the composition are glutamic acid-betaxanthin.

[0130] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-65% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 45-65% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 50-65% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 55-65% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin. In some embodiments, 55-60% of the betaxanthins in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin.

[0131] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are glutamic acid-betaxanthin. Each possibility represents a separateembodiment of the invention. In some embodiments, 40-65% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, 45-65% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are glutamic acid- betaxanthin. In some embodiments, 50-65% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, 55-65% of the pigment molecules in the composition are glutamic acid-betaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are glutamic acid-betaxanthin.

[0132] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40- 65% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 45-65% of the pigment molecules in the composition are glutamic acid- betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 50-65% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid- isobetaxanthin. In some embodiments, 55-65% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin. In some embodiments, 55- 60% of the pigment molecules in the composition are glutamic acid-betaxanthin or glutamic acid-isobetaxanthin.

[0133] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40- 65% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 45-65% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 50-65% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 55-65% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin. In some embodiments, 55-60% of the betaxanthins in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin.

[0134] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are phenylalanine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-65% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, 45-65% of the pigment molecules in the composition are phenylalanine- betaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, 50-65% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, 55-65% of the pigment molecules in the composition are phenylalanine-betaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are phenylalanine-betaxanthin.

[0135] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40- 65% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 45-65% of the pigment molecules in the composition are phenylalanine- betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine- isobetaxanthin. In some embodiments, 50-65% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 50- 60% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 55-65% of the pigment molecules in the composition are phenylalanine-betaxanthin or phenylalanine-isobetaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are phenylalanine- betaxanthin or phenylalanine-isobetaxanthin.

[0136] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are methionine -betaxanthin or methionine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the betaxanthins in the composition are methionine -betaxanthin or methionine-isobetaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are methioninebetaxanthin or methionine-isobetaxanthin. In some embodiments, 45-55% of thebetaxanthins in the composition are methionine -betaxanthin or methionine-isobetaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are methioninebetaxanthin or methionine-isobetaxanthin. In some embodiments, 50-55% of the betaxanthins in the composition are methionine -betaxanthin or methionine-isobetaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are methioninebetaxanthin or methionine-isobetaxanthin. In some embodiments, 55-60% of the betaxanthins in the composition are methionine-betaxanthin or methionine-isobetaxanthin.

[0137] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are methionine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are methionine- betaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are methionine-betaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are methionine-betaxanthin.

[0138] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40- 55% of the pigment molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, 45-55%of the pigment molecules in the composition are methionine-betaxanthin or methionineisobetaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are methionine-betaxanthin or methionine-isobetaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are methionine-betaxanthin or methionine- isobetaxanthin.

[0139] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are leucine-betaxanthin or leucine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the betaxanthins in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are leucine-betaxanthin or leucine- isobetaxanthin. In some embodiments, 45-55% of the betaxanthins in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 50-55% of the betaxanthins in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 55- 60% of the betaxanthins in the composition are leucine-betaxanthin or leucine- isobetaxanthin.

[0140] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are leucine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules inthe composition are leucine-betaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, 45- 60% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are leucine-betaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are leucine- betaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are leucine-betaxanthin.

[0141] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are leucine- betaxanthin or leucine-isobetaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin. In some embodiments, 50- 60% of the pigment molecules in the composition are leucine-betaxanthin or leucine- isobetaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are leucine-betaxanthin or leucine-isobetaxanthin.

[0142] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are serine -betaxanthin or serine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the betaxanthinsin the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are serine-betaxanthin or serine- isobetaxanthin. In some embodiments, 45-55% of the betaxanthins in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 50-55% of the betaxanthins in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 55-60% of the betaxanthins in the composition are serine-betaxanthin or serine-isobetaxanthin.

[0143] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are serine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, 50- 55% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are serine-betaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are serine- betaxanthin.

[0144] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42- 65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45- 55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50- 70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53- 70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. Insome embodiments, 40-60% of the pigment molecules in the composition are serinebetaxanthin or serine-isobetaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are serine-betaxanthin or serine-isobetaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are serine- betaxanthin or serine-isobetaxanthin.

[0145] In some embodiments, 35-70, 35-65, 35-60, 35-57, 35-55, 35-53, 35-52, 35-50, 35- 47, 35-45, 35-40, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40-47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 35-45% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 35-50% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, 35-55% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 35-60% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 40-50% of the betaxanthins in the composition are glutamine- betaxanthin or glutamine-isobetaxanthin. In some embodiments, 40-55% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 40-60% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 45-50% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 45-55% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, 45-60% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 50-55% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 50-60% of the betaxanthins in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 55-60% of the betaxanthins in the composition are glutamine -betaxanthin or glutamine-isobetaxanthin.

[0146] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are glutamine-betaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are glutamine- betaxanthin. In some embodiments, 50-60% of the pigment molecules in the composition are glutamine-betaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are glutamine-betaxanthin.

[0147] In some embodiments, 40-70, 40-65, 40-60, 40-57, 40-55, 40-53, 40-52, 40-50, 40- 47, 40-45, 41-70, 41-65, 41-60, 41-57, 41-55, 41-53, 41-52, 41-50, 41-47, 41-45, 42-70, 42-65, 42-60, 42-57, 42-55, 42-53, 42-52, 42-50, 42-47, 42-45, 45-70, 45-65, 45-60, 45-57, 45-55, 45-53, 45-52, 45-50, 45-47, 47-70, 47-65, 47-60, 47-57, 47-55, 47-53, 47-52, 47-50, 50-70, 50-65, 50-60, 50-57, 50-55, 50-53, 50-52, 52-70, 52-65, 52-60, 52-57, 52-55, 52-53, 53-70, 53-65, 53-60, 53-57, 53-55, 55-70, 55-65, 55-60, or 55-57% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. Each possibility represents a separate embodiment of the invention. In some embodiments, 40-55% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, 40-60% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 45-55% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, 45-60% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine-isobetaxanthin. In some embodiments, 50-55% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin. In some embodiments, 50-60% of the pigment molecules in the compositionare glutamine -betaxanthin or glutamine-isobetaxanthin. In some embodiments, 55-60% of the pigment molecules in the composition are glutamine-betaxanthin or glutamine- isobetaxanthin.

[0148] As demonstrated hereinbelow, natural prickly pear extracts and juice contain measurable levels of betalamic acid and betacyanins and have a mix of different betaxanthin molecules (greater than 20% non indicaxanthin) (Fig. 1). Further, previous examples of betaxanthins produced by fermentation contained measurable levels of betalamic acid and a combination of different betaxanthins (International Patent Publication WO2017 / 122189, Table 2). The composition of the invention is thus the first to contain betaxanthins without measurable (or visible) contamination of betalamic acid or betacyanins and with highly pure levels of a specific betaxanthin (e.g., indicaxanthin, histidine-betaxanthin, etc.). Purity is used herein to refer to purity with respect to other betalain molecules.

[0149] In some embodiments, the composition comprises iso-indicaxanthin. In some embodiments, the composition comprises detectable levels of iso-indicaxanthin. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at least 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5: 1, 5:1, 5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1, 12:1, 15:1, 17:1, 20:1, or 23:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at least 1.5:1. In some embodiments, the ratio of indicaxanthin to iso- indicaxanthin in the composition is at least 2:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at least 3:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at least 4:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at least 4.5:1.

[0150] In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at most 5:1, 5.5:1, 6:1, 6.5:1, 7:1, 7.5:1, 8:1, 8.5:1, 9:1, 9.5:1, 10:1, 12:1, 15:1, 17:1, 20:1, 25:1, 50:1, 75:1, 100:1, 125:1 or 150:1. Each possibility represents a separate embodiment of the invention. In some embodiments, the ratio of indicaxanthin to iso- indicaxanthin in the composition is at most 5:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at most 10:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at most 50:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at most 100:1. In some embodiments, the ratio of indicaxanthin to iso-indicaxanthin in the composition is at most 150:1.

[0151] In some embodiments, the composition comprises a betalain pigment composition such as is provided in Figure 1 for PhytoYellow. In some embodiments, the composition comprises a betaxanthin pigment composition such as is provided in Figure 2 for PhytoYellow. In some embodiments, the composition comprises a yellow color such as is shown in Figure 3. In some embodiments, the composition is PhytoYellow.

[0152] In some embodiments, the composition is low in carbohydrates. In some embodiments, the composition is low in sugar. In some embodiments, the carbohydrate is sugar. In some embodiments, sugar is simple sugars. In some embodiments, simple sugars comprise monosaccharides and disaccharides. In some embodiments, simple sugars are monosaccharides. In some embodiments, simple sugars are disaccharides. In some embodiments, a composition low in carbohydrates comprises less than 25, 23, 20, 18, 15, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1, 0.05 or 0.01% carbohydrates. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 10% carbohydrates. In some embodiments, the composition comprises less than 5% carbohydrates. In some embodiments, the composition comprises less than 1% carbohydrates. In some embodiments, the composition comprises less than 10% sugar. In some embodiments, the composition comprises less than 5% sugar. In some embodiments, the composition comprises less than 1% sugar. In some embodiments, the percentage is by weight. In some embodiments, the composition is devoid of sugar. In some embodiments, the composition is essentially devoid of sugar. In some embodiments, the composition comprises less than 10 grams carbohydrates per 100 grams of composition. In some embodiments, the composition comprises less than 5 grams carbohydrates per 100 grams of composition. In some embodiments, the composition comprises less than 1 gram carbohydrates per 100 grams of composition. In some embodiments, the composition comprises less than 10 grams sugars per 100 grams of composition. In some embodiments, the composition comprises less than 5 grams sugars per 100 grams of composition. In some embodiments, the composition comprises less than 1 gram sugar per 100 grams of composition.

[0153] In some embodiments, the composition is low in carbohydrates. In some embodiments, carbohydrates are dietary carbohydrates. In some embodiments, carbohydrates do not comprise betalains. In some embodiments, carbohydrates are nonbetalain carbohydrates. In some embodiments, carbohydrates are carbohydrates that can be processed by humans to produce energy. It will be understood by a skilled artisan that while many molecules are technically carbohydrates, dietary carbohydrates are those that a humandigestive tract can digest and convert into energy (calories). In some embodiments, a composition low in carbohydrates comprises less than 40, 35, 30, 27, 25, 23, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1, or 0.05% carbohydrates. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 1% carbohydrates. In some embodiments, the composition comprises less than 5% carbohydrates. In some embodiments, the composition comprises less than 10% carbohydrates. In some embodiments, the percentage is by weight.

[0154] In some embodiments, the composition comprises at least 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1200, 1300, 1400 or 1500 mg magnesium per liter. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises at least 500 mg magnesium per liter. In some embodiments, the composition is a liquid and comprises at least 500 mg magnesium per liter. In some embodiments, the composition comprises at least 1000 mg magnesium per liter. In some embodiments, the composition is a liquid and comprises at least 1000 mg magnesium per liter. In some embodiments, the composition comprises at least 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 or 2000 mg magnesium per kilogram. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises at least 500 mg magnesium per kilogram. In some embodiments, the composition is a solid and comprises at least 500 mg magnesium per kilogram. In some embodiments, the composition comprises at least 1000 mg magnesium per kilogram. In some embodiments, the composition is a solid and comprises at least 1000 mg magnesium per kilogram.

[0155] In some embodiments, the composition comprises less than 5, 4, 3, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01 pg / kg geosmin. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 0.01 pg / kg geosmin. hr some embodiments, the composition comprises less than 0.1 pg / kg geosmin. In some embodiments, the composition comprises less than 1 pg / kg geosmin. In some embodiments, the composition is essentially devoid of geosmin. In some embodiments, the composition is a geosmin-free composition.

[0156] In some embodiments, the composition comprises less than 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0005 or 0.0001 g / L nitrates. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises less than 0.01 g / L nitrates. In some embodiments, the composition comprises lessthan 0.005 g / L nitrates. In some embodiments, the composition comprises less than 0.001 g / L nitrates. In some embodiments, the composition is essentially devoid of nitrates. In some embodiments, the composition is a nitrate-free composition.

[0157] In some embodiments, the composition is a high yield composition. In some embodiments, the composition is a high titer composition. In some embodiments, yield is concentration. In some embodiments, the composition comprises a higher titer of betalains than are found in nature. In some embodiments, in nature is in a prickly pear. In some embodiments, the prickly pear is a yellow prickly pear. In some embodiments, in a prickly pear is in a prickly pear extract. In some embodiments, in a prickly pear is in prickly pear juice. In some embodiments, a prickly pear extract is a betalain prickly pear extract. In some embodiments, the composition comprises a betalain concentration of greater than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 grams betalain pigment molecules per liter of composition. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises a betalain concentration of greater than 0.05 grams betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 0.5 grams betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 0.75 grams betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 1 gram betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 1.5 grams betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 3 grams betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 3.5 grams betalain pigment molecules per liter of composition. In some embodiments, the composition comprises a betalain concentration of greater than 4 grams betalain pigment molecules per liter of composition. It will be understood by a skilled artisan that for commercially relevant production of pigment a concentration of at least about 4 gr / 1 is needed. In some embodiments, betalain pigment molecules is betaxanthin pigment molecules. In some embodiments, betalain pigment molecules is indicaxanthin molecules. In some embodiments, betalain pigment molecules is indicaxanthin and iso-indicaxanthin molecules.

[0158] In some embodiments, the composition comprises a betalain concentration of greater than 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1 grams betalain pigment molecules per kilogram of composition. Each possibility represents a separate embodiment of the invention. In some embodiments, the composition comprises a betalain concentration of greater than 0.05 grams betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 0.5 grams betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 0.75 grams betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 1 gram betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 1.5 grams betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 3 grams betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 3.5 grams betalain pigment molecules per kilogram of composition. In some embodiments, the composition comprises a betalain concentration of greater than 4 grams betalain pigment molecules per kilogram of composition. In some embodiments, betalain pigment molecules is betaxanthin pigment molecules. In some embodiments, betalain pigment molecules is indicaxanthin molecules. In some embodiments, betalain pigment molecules is indicaxanthin and iso-indicaxanthin molecules.

[0159] In some embodiments, the composition is devoid of intact cells. In some embodiments, devoid of is essentially devoid of. In some embodiments, cells are plant cells. In some embodiments, cells are yeast cells. In some embodiments, the cells are bacterial cells. In some embodiments, the composition is devoid of plant proteins not of the betalain synthesis pathway.

[0160] In some embodiments, the composition comprises at least one yeast protein. In some embodiments, the protein is a secreted protein. It will be understood by a skilled artisan that even after filtering some yeast protein will end up in the final product. In nature, no composition exists that would have both a betalain and a yeast protein.

[0161] In some embodiments, the composition comprises an additive. In some embodiments, the additive is a preservative. In some embodiments, the additive is a color enhancer. In some embodiments, the preservative is an antimicrobial. In some embodiments, the preservative extends the shelf life of the composition. Examples of preservatives include,but are not limited to nitrite, sulfate, benzoic acid, sodium benzoate, and sodium propionate to name but a few.

[0162] By another aspect, there is provide a method of producing betaxanthin comprising an amine of interest, the method comprising culturing a cell comprising at least one tyrosine hydroxylase gene that is not an L-DOPA oxidase gene, and at least one DOPA 4,5- dioxygenase (DOD) gene, in a culture medium supplemented with the amine of interest and collecting the culture medium, thereby producing a betaxanthin comprising an amine of interest.

[0163] By another aspect, there is provided a method of producing betaxanthin comprising an amino acid of interest, the method comprising culturing a cell comprising at least one tyrosine hydroxylase gene that is not an L-DOPA oxidase gene, and at least one DOPA 4,5- dioxygenase (DOD) gene, in a culture medium supplemented with the amino acid of interest and collecting the culture medium, thereby producing a betaxanthin comprising an amino acid of interest.

[0164] By another aspect, there is provided a method of producing a pigment composition of the invention, the method comprising culturing a cell comprising integrated into its genome at least one tyrosine hydroxylase gene that is not an L-DOPA oxidase gene, and at least one DOPA 4,5-dioxygenase (DOD) gene in a culture medium and collecting the culture medium, thereby producing a pigment composition of the invention.

[0165] In some embodiments, the genes are transiently expressed in the cell. In some embodiments, the at least one tyrosine hydroxylase gene that is not an L-DOPA oxidase gene is transiently expressed in the cell. In some embodiments, the at least one DOPA 4,5- dioxygenase (DOD) gene is transiently expressed in the genome of the cell. In some embodiments, at least one NADPH cytochrome P450 reductase gene is expressed in the cell. In some embodiments, the cell comprises at least one NADPH cytochrome P450 reductase gene. In some embodiments, the at least one NADPH cytochrome P450 reductase gene is transiently expressed in the cell. In some embodiments, transient expression is transfection. In some embodiments, transient expression is transduction. In some embodiments, transient expression is viral infection.

[0166] In some embodiments, the genes are integrated into the genome of the cell. In some embodiments, the at least one tyrosine hydroxylase gene that is not an L-DOPA oxidase gene is integrated into a genome of the cell. In some embodiments, the at least one DOPA 4,5- dioxygenase (DOD) gene is integrated into the genome of the cell. In some embodiments, atleast one NADPH cytochrome P450 reductase gene is expressed in the cell. In some embodiments, the cell comprises at least one NADPH cytochrome P450 reductase gene. In some embodiments, the at least one NADPH cytochrome P450 reductase gene is integrated into a genome of the cell.

[0167] Methods of gene integration into a cell’s genome are well known in the art and any such method may be used. Examples include use of homologous recombination, CRISPR- Cas9, TALE nucleases, and zinc-finger nucleases as well as lentiviral and adenoviral integration. In some embodiments, the integration is viral integration. In some embodiments, the integration is adenoviral integration. In some embodiments, the integration is by homologous recombination. In some embodiments, the integration is by a guided nuclease. In some embodiments, the guided nuclease is CRISPR-Cas. In some embodiments, the guided nuclease is CRISPR-Cas9. In some embodiments, the integration is by CRISPR- Cas9.

[0168] In some embodiments, the genes are exogenous genes. In some embodiments, the genes are not endogenous to the cell. In some embodiments, the genes lack introns. In some embodiments, the genes are cDNAs integrated into the cell’s genome. In some embodiments, the genes are plant genes. In some embodiments, the cell is not a plant cell. In some embodiments, the cell is a plant cell. In some embodiments, the cell is a plant cell that does not express betalains. In some embodiments, the plant cell does not endogenously express betalains. In some embodiments, the cell is a bacterial cell. In some embodiments, the bacterial cell is E. coli. In some embodiments, the cell is a eukaryotic cell. In some embodiments, the cell is a fungal cell. In some embodiments, the fungus is a unicellular fungus. In some embodiments, the fungus is a filamentous fungus. In some embodiments, the fungus is not a filamentous fungus. In some embodiments, the fungus is yeast. In some embodiments, the cell is a yeast cell. In some embodiments, the yeast is Saccharomyces cerevisiae. Additional examples of fungal cells that can be used as the cell of the invention include, but are not limited to Yarrowia lipolytica cells, Pichia pastoris cells, and Fusarium veneatum cells.

[0169] In some embodiments, a gene is integrated at least once. In some embodiments, at least one copy of the gene is integrated. In some embodiments, at least one copy of a tyrosine hydroxylase gene that is not a L-DOPA oxidase is integrated. In some embodiments, at least one copy of a DOPA 4, 5-dioxygenase (DOD) gene is integrated. In some embodiments, at least one copy of a NADPH cytochrome P450 reductase gene is integrated.

[0170] In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase gene is a CYP76AD1 beta-clade gene. In another embodiment, the CYP76AD1-P clade gene is a CYP76AD1-P clade gene as set forth in Brockington et al. 2015 (New Phytol. 2015 Sep;207(4): 1170-80), which is incorporated herein by reference in its entirety). In one embodiment, the CYP76AD1-P clade gene is a CYP76AD1-P clade gene as set forth in Supplemental Figure 2 of Brockington et al. 2015. The beta-clade of CYP76AD1 genes is well known in the art and a large number of representative examples of this clade are provided in Supplemental Figure 2 of Brockington et al. 2015. While both the alpha and beta-clade are tyrosine hydroxylases, the alpha clade are also L-DOPA oxidases while the beta clade are not. 10 amino acids that define the beta-clade are disclosed in Brockington et al. (see Figure 5). These 10 amino acids are always present in beta-clade proteins and never present in alpha-clade proteins. These amino acids produce a unique function of the beta-clade which is not oxidizing L-DOPA. Those 10 amino acids are Dil l, SI 14, S 131, G186, M207, E213, Y241, T275, N450 and V486 (positions are given with respect to CYP76AD1, which is an alpha-clade protein). In light of the numerous examples of beta-clade proteins and the structural function correlation between these 10 amino acids and beta-clade function, this genus of genes is fully described. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase gene is selected from CYP76AD5, CYP76AD6, CYP76AD9, CYP76AD10, CYP76AD15, CYP76AD17, and CYP76AD19. In some embodiments, the tyrosine hydroxylase gene that is not an L-DOPA oxidase is CYP76AD5. In some embodiments, CYP76AD5 is Beta vulgaris CYP76AD5. In some embodiments, the nucleic acid sequence encoding CYP76AD5 comprises SEQ ID NO: 1. In some embodiments, the nucleic acid sequence encoding CYP76AD5 consists of SEQ ID NO: 1. In some embodiments, nucleic acid sequence is codon optimized. In some embodiments, codon optimized is optimized for expression in yeast. In some embodiments, the codon optimized nucleic acid sequence encoding CYP76AD5 comprises SEQ ID NO: 15. In some embodiments, the codon optimized nucleic acid sequence encoding CYP76AD5 consists of SEQ ID NO: 15. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the gene is a coding sequence of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 1 or SEQ ID NO: 15. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 1 or SEQ ID NO: 15. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 15 and hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, thetyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 15 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0171] In some embodiments, L-DOPA is 1-3,4-dihydroxyphenylalanine. In some embodiments, hydroxylating tyrosine comprises converting tyrosine to L-DOPA. In some embodiments, the tyrosine hydroxylase converts tyrosine to L-DOPA. In some embodiments, oxidizing L-DOPA comprises converting L-DOPA to cyclo-DOPA. In some embodiments, the tyrosine hydroxylase does not convert L-DOPA to cyclo-DOPA. In some embodiments, the tyrosine hydroxylase that is not a L-DOPA oxidase does not convert L-DOPA to cyclo-DOPA. In some embodiments, does not convert is cannot convert.

[0172] In some embodiments, the amino acid sequence of CYP76AD5 comprises SEQ ID NO: 2. In some embodiments, the amino acid sequence of CYP76AD5 consists of SEQ ID NO: 2. In some embodiments, the CYP76AD5 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 2. In some embodiments, the CYP76AD5 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 2. In some embodiments, the CYP76AD5 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 2 that hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0173] In some embodiments, the tyrosine hydroxylase gene that is not an L-DOPA oxidase is CYP76AD6. In some embodiments, CYP76AD6 is Beta vulgaris CYP76AD6. In some embodiments, the nucleic acid sequence encoding CYP76AD6 comprises SEQ ID NO: 11. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 11. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 11. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 11 and hydroxylizes tyrosine and does not oxidize L- DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 11 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0174] In some embodiments, the amino acid sequence of CYP76AD6 comprises SEQ ID NO: 12. In some embodiments, the amino acid sequence of CYP76AD6 consists of SEQ ID NO: 12. In some embodiments, the CYP76AD6 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 12. In some embodiments, the CYP76AD6 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 12. In some embodiments, the CYP76AD6 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 12 thathydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0175] In some embodiments, the tyrosine hydroxylase gene that is not an L-DOPA oxidase is CYP76AD15. In some embodiments, CYP76AD15 is Mirabilis jalapa CYP76AD15. In some embodiments, the nucleic acid sequence encoding CYP76AD15 comprises SEQ ID NO: 13. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 13. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 13. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 13 and hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 13 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0176] In some embodiments, the amino acid sequence of CYP76AD15 comprises SEQ ID NO: 14. In some embodiments, the amino acid sequence of CYP76AD15 consists of SEQ ID NO: 14. In some embodiments, the CYP76AD15 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 14. In some embodiments, the CYP76AD15 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 14. In some embodiments, the CYP76AD15 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 14 thathydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0177] In some embodiments, the tyrosine hydroxylase gene that is not an L-DOPA oxidase is CYP76AD9. In some embodiments, CYP76AD9 is Opuntia ficus CYP76AD9. In some embodiments, the nucleic acid sequence encoding CYP76AD9 comprises SEQ ID NO: 16. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 16. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 16. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 16 and hydroxylizes tyrosine and does not oxidize L- DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 16 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0178] In some embodiments, the amino acid sequence of CYP76AD9 comprises SEQ ID NO: 17. In some embodiments, the amino acid sequence of CYP76AD9 consists of SEQ ID NO: 17. In some embodiments, the CYP76AD9 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 17. In some embodiments, the CYP76AD9 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 17. In some embodiments, the CYP76AD9 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 17 that hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0179] In some embodiments, the tyrosine hydroxylase gene that is not an L-DOPA oxidase is CYP76AD10. In some embodiments, CYP76AD10 is Phytolacca americana CYP76AD10. In some embodiments, the nucleic acid sequence encoding CYP76AD10 comprises SEQ ID NO: 18. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 18. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 18. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 18 and hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises asequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 18 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0180] In some embodiments, the amino acid sequence of CYP76AD10 comprises SEQ ID NO: 19. In some embodiments, the amino acid sequence of CYP76AD10 consists of SEQ ID NO: 19. In some embodiments, the CYP76AD10 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 19. In some embodiments, the CYP76AD10 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 19. In some embodiments, the CYP76AD10 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 19 thathydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0181] In some embodiments, the tyrosine hydroxylase gene that is not an L-DOPA oxidase is CYP76AD17. In some embodiments, CYP76AD17 is Mollugo verticillate CYP76AD17. In some embodiments, the nucleic acid sequence encoding CYP76AD17 comprises SEQ ID NO: 20. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 20. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 20. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 20 and hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 20 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0182] In some embodiments, the amino acid sequence of CYP76AD17 comprises SEQ IDNO: 21. In some embodiments, the amino acid sequence of CYP76AD17 consists of SEQ ID NO: 21. In some embodiments, the CYP76AD17 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 21. In some embodiments, the CYP76AD17 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 21. In some embodiments, the CYP76AD17 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 21 thathydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In someembodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0183] In some embodiments, the tyrosine hydroxylase gene that is not an E-DOPA oxidase is CYP76AD19. In some embodiments, CYP76AD19 is Dianthus caryophyllus CYP76AD19. In some embodiments, the nucleic acid sequence encoding CYP76AD19 comprises SEQ ID NO: 22. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises SEQ ID NO: 22. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase consists of SEQ ID NO: 22. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 22 and hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the tyrosine hydroxylase gene that is not a L-DOPA oxidase comprises a sequence with at least 85, 90, 92, 95, 97, 99 or 100% sequence identity to SEQ ID NO: 22 and is a CYP76AD1 beta-clade gene. Each possibility represents a separate embodiment of the invention.

[0184] In some embodiments, the amino acid sequence of CYP76AD19 comprises SEQ ID NO: 23. In some embodiments, the amino acid sequence of CYP76AD19 consists of SEQ ID NO: 23. In some embodiments, the CYP76AD19 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 23. In some embodiments, the CYP76AD19 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 23. In some embodiments, the CYP76AD19 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 23 that hydroxylizes tyrosine and does not oxidize L-DOPA. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains hydroxylase activity. In some embodiments, the protein does not have oxidase activity.

[0185] In some embodiments, the cell does not comprise a CYP76AD1 alpha clade gene. In some embodiments, the cell is devoid of a CYP767AD1 alpha clade gene. In some embodiments, the cell does not express nor have integrated into its genome a CYP76AD1 alpha clade gene. In some embodiments, the cell does not comprise a CYP76AD1 gene. In some embodiments, the cell is devoid of a CYP76AD1 gene. In some embodiments, the cell does not express nor have integrated into its genome a CYP76AD1 gene.

[0186] Just as the beta-clade is well-known and defined, so too the alpha-clade is well- known and defined. Examples of alpha-clade proteins can be found in Supplemental Figure 2 of Brockington. Further, the 10 amino acids differentiating the alpha-clade from the beta-cladeare provided in Figure 5 of Brockington. They are Hl 11, T114, T131, L186, 1207, D213, N241, V275, T450 and T486.

[0187] In some embodiments, the DOD gene is Mirabilis jalapa DOD (MjDOD). In some embodiments, the DOD gene is Beta vulgaris DODA1 (BvDODAl). In some embodiments, the nucleic acid sequence encoding MjDOD comprises SEQ ID NO: 3. In some embodiments, the nucleic acid sequence encoding BvDODAl comprises SEQ ID NO: 5. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the DOPA 4,5-dioxygenase gene comprises SEQ ID NO:3. In some embodiments, the DOPA 4,5-dioxygenase gene consists of SEQ ID NO: 3. In some embodiments, the DOPA 4,5-dioxygenase gene comprises SEQ ID NO: 5. In some embodiments, the DOPA 4,5-dioxygenase gene consists of SEQ ID NO: 5. In some embodiments, the DOPA 4,5-dioxygenase gene comprises a sequence with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 3 while retaining dioxygenase activity. Each possibility represents a separate embodiment of the invention. In some embodiments, the DOPA 4,5-dioxygenase gene comprises a sequence with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 5 while retaining dioxygenase activity. Each possibility represents a separate embodiment of the invention. In some embodiments, retaining activity is encoding a protein that retains activity. Other examples of DOD genes include but are not limited to PgDOD from Portulaca grandiflora (Accession No. AJ580598), BgDOD from Bougainvillea glabra (Accession No. AB435373), and AmDOD from Amanita muscaria (Accession No. P87064). In some embodiments, dioxygenase activity comprises converting L-DOPA to betalamic acid. In some embodiments, the DOD enzyme converts L-DOPA to betalamic acid.

[0188] In some embodiments, the amino acid sequence of MjDOD comprises SEQ ID NO:4. In some embodiments, the amino acid sequence of MjDOD consists of SEQ ID NO: 4. In some embodiments, the MjDOD gene comprises a nucleic acid sequence that encodes SEQ ID NO: 4. In some embodiments, the MjDOD gene consists of a nucleic acid sequence that encodes SEQ ID NO: 4. In some embodiments, the MjDOD gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 4. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains dioxygenase activity.

[0189] In some embodiments, the amino acid sequence of BvDODAl comprises SEQ ID NO: 6. In some embodiments, the amino acid sequence of BvDODAl consists of SEQ ID NO: 6. In some embodiments, the BvDODAl gene comprises a nucleic acid sequence thatencodes SEQ ID NO: 6. In some embodiments, the BvDODAl gene consists of a nucleic acid sequence that encodes SEQ ID NO: 6. In some embodiments, the BvDODAl gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 6. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains dioxygenase activity. In some embodiments, the protein converts L-DOPA to betalamic acid.

[0190] In some embodiments, the NADPH cytochrome P450 reductase gene is NADPH — cytochrome P450 reductase 2 (ATR2) from Arabidopsis thaliana. In some embodiments, the NADPH cytochrome P450 reductase gene is NADPH — cytochrome P450 reductase 1 (ATR1) from Arabidopsis thaliana. In some embodiments, the cell comprises at least one copy of the ATR1 gene integrated into its genome. In some embodiments, the cell comprises at least one copy of the ATR2 gene integrated into its genome. In some embodiments, the nucleic acid sequence encoding ATR1 comprises SEQ ID NO: 7. In some embodiments, an optimized nucleic acid sequence encoding ATR1 comprises SEQ ID NO: 24. In some embodiments, optimized is codon optimized. In some embodiments, optimized is optimized to remove restriction sites. In some embodiments, optimized is optimized for expression in a cell. In some embodiments, the nucleic acid sequence encoding ATR2 comprises SEQ ID NO: 9. In some embodiments, an optimized nucleic acid sequence encoding ATR2 comprises SEQ ID NO: 25. In some embodiments, optimized is codon optimized. In some embodiments, optimized is optimized for expression in a cell. In some embodiments, the gene is without introns. In some embodiments, the gene is a cDNA of the gene. In some embodiments, the NADPH cytochrome P450 reductase gene comprises SEQ ID NO: 7. In some embodiments, the NADPH cytochrome P450 reductase gene consists of SEQ ID NO: 7. In some embodiments, the NADPH cytochrome P450 reductase gene comprises SEQ ID NO: 9. In some embodiments, the NADPH cytochrome P450 reductase gene consists of SEQ ID NO: 9. In some embodiments, the DOPA 4,5-dioxygenase gene comprises a sequence with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 7 while retaining reductase activity. Each possibility represents a separate embodiment of the invention. In some embodiments, the DOPA 4,5-dioxygenase gene comprises a sequence with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 24 while retaining reductase activity. Each possibility represents a separate embodiment of the invention. In some embodiments, the NADPH cytochrome P450 reductase gene comprises a sequence with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 9 while retaining reductase activity. Each possibility represents a separate embodiment of the invention. In some embodiments, theNADPH cytochrome P450 reductase gene comprises a sequence with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 25 while retaining reductase activity. Each possibility represents a separate embodiment of the invention. In some embodiments, retaining activity is encoding a protein that retains activity. Other examples of NADPH cytochrome P450 reductase genes include but are not limited to NADPH cytochrome P450 reductase 1 or 2 from Medicago truncatula (MtCPRl / MtCPR2) and NADPH cytochrome P450 reductase 1 or 2 from Lotus japonicus (LjCPRl / LjCPR2).

[0191] In some embodiments, the amino acid sequence of ATR1 comprises SEQ ID NO: 8. In some embodiments, the amino acid sequence of ATR1 consists of SEQ ID NO: 8. In some embodiments, the ATR1 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 8. In some embodiments, the ATR1 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 8. In some embodiments, the MjDOD gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 8. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains reductase activity.

[0192] In some embodiments, the amino acid sequence of ATR2 comprises SEQ ID NO: 10. In some embodiments, the amino acid sequence of ATR2 consists of SEQ ID NO: 10. In some embodiments, the ATR2 gene comprises a nucleic acid sequence that encodes SEQ ID NO: 10. In some embodiments, the ATR2 gene consists of a nucleic acid sequence that encodes SEQ ID NO: 10. In some embodiments, the ATR2 gene comprises or consists of a nucleic acid sequence that encodes a protein with at least 70, 75, 80, 85, 90, 95, 97 or 99% identity to SEQ ID NO: 10. Each possibility represents a separate embodiment of the invention. In some embodiments, the protein retains reductase activity.

[0193] In some embodiments, the cell does not comprise a cyclo-DOPA-5-O- glucosyltransferase gene. In some embodiments, the cell is devoid of a cyclo-DOPA-5-O- glucosyltransferase gene. In some embodiments, a cyclo-DOPA-5-O-glucosyltransferase gene is a 5-O-cyclo-DOPA-glucosyltransferase gene. In some embodiments, the cell does not comprise a 5-O-betanidin-glycosyltransferase gene. In some embodiments, the cell is devoid of a 5-O-betanidin-glycosyltransferase gene.

[0194] In some embodiments, the inserted genes are under the control of a strong promoter. In some embodiments, the genes are integrated into the genome with a promoter. In some embodiments, the promoter is operatively linked to the genes. The term “operably linked” is intended to mean that the nucleotide sequence of interest (i.e., the integrated genes) is linkedto the regulatory element or elements in a manner that allows for expression of the nucleotide sequence. In some embodiments, the strong promoter is the GAL1 / 10 promoter. In some embodiments, all integrated genes are operatively linked to a GAL1 / 10 promoter. In some embodiments, the GAL1 / 10 promoter comprises or consists of the nucleotide sequence provided in SEQ ID NO: 26. In some embodiments, the GAL1 / 10 promoter is an optimized sequence.

[0195] In some embodiments, the inserted genes are linked to a terminator sequence. In some embodiments, the terminator is a strong terminator. It will be understood that being linked to a terminator sequence is intended to mean that the nucleotide sequence of interest (i.e., the integrated genes) proceeds the termination sequence or terminator in a manner that allows for termination of transcription at the end or just after the sequence of interest. Examples of terminators that may be used are the terminators of CYC1, ADH2, ADH1, and TEF2.

[0196] In some embodiments, the media comprises the amine. In some embodiments, the media is supplemented with the amine. In some embodiments, supplementation is in addition to essential amine present in the media. In some embodiments, the media comprises the amino acid. In some embodiments, the media is supplemented with the amino acid. In some embodiments, supplementation is in addition to essential amino acids present in the media.

[0197] In some embodiments, the amine is an amino acid. In some embodiments, the amine is not an amino acid. Examples of amines that can be conjugated to betalamic acid include, but are not limited to Tyramine, 3-Methoxy Tyramine, Histamine, T- Aminobutyric Acid, Methylated Arginine, Ethanolamine, Putrescine, Phenethylamine, Tryptamine, Ethylamine, Propylamine, 2-Phenylethylamine, N-Methyl-Ethanamine, N-Methyl-N-Propylamine, Pyrrolidine, Aniline, N-Methyl- Aniline, N-Ethyl- Aniline, Indoline, (S)-Phenylalanine, and (S)-Indoline-2-Carboxylic Acid.

[0198] In some embodiments, the amino acid is any amino acid. In some embodiments, the amino acid is selected from proline, leucine, phenylalanine, serine, histidine, glutamic acid, methionine and glutamine. In some embodiments, the amino acid is selected from proline, leucine, phenylalanine, serine, histidine, glutamic acid, and methionine. In some embodiments, the amino acid is selected from proline, histidine, glutamic acid, serine and leucine and methionine. In some embodiments, the amino acid is selected from proline, histidine, glutamic acid, and methionine. In some embodiments, the amino acid is selected from proline, histidine, and methionine. In some embodiments, the amino acid is selectedfrom proline and histidine. In some embodiments, the amino acid is proline. In some embodiments, the amino acid is histidine. In some embodiments, the amino acid is methionine. In some embodiments, the amino acid is glutamic acid. In some embodiments, the amino acid is serine. In some embodiments, the amino acid is leucine. In some embodiments, the amino acid is glutamine. In some embodiments, the amino acid is phenylalanine.

[0199] In some embodiments, the culturing is for a time sufficient to produce betaxanthin. In some embodiments, the culture medium is collected after betaxanthin is produced. In some embodiments, the sufficient time is at least 1, 2, 3, 4, 5, 6 or 7 days. Each possibility represents a separate embodiment of the invention.

[0200] In some embodiments, the method is a method of producing a pigment composition. In some embodiments, the pigment composition comprises at least 40% of betathanin in the composition being the amino acid-betaxanthin. In some embodiments, the pigment composition comprises at least 40% of betathanin in the composition comprising the amino acid. In some embodiments, the pigment composition comprises at least 50% of betaxanthin in the composition being the amino acid-betaxanthin. In some embodiments, the pigment composition comprises at least 50% of betaxanthin in the composition comprising the amino acid. In some embodiments, the pigment composition comprises at least 60% of betaxanthin in the composition being the amino acid-betaxanthin. In some embodiments, the pigment composition comprises at least 60% of betaxanthin in the composition comprising the amino acid. In some embodiments, the pigment composition comprises at least 90% of betaxanthin in the composition being the amino acid-betaxanthin. In some embodiments, the pigment composition comprises at least 90% of betaxanthin in the composition comprising the amino acid. In some embodiments, the pigment composition comprises at least 95% of betaxanthin in the composition being the amino acid-betaxanthin. In some embodiments, the pigment composition comprises at least 95% of betaxanthin in the composition comprising the amino acid. In some embodiments, the pigment composition comprises at least 98% of betaxanthin in the composition being the amino acid-betaxanthin. In some embodiments, the pigment composition comprises at least 98% of betaxanthin in the composition comprising the amino acid. In some embodiments, the pigment composition is a pigment composition of the invention.

[0201] By another aspect, there is provided a pigment composition produced by the cells.

[0202] In some embodiments, the pigment composition is secreted by the cells. In some embodiments, the pigment composition is from lysing the cells. In some embodiments, the pigment composition is produced without lysing the cells. In some embodiments, the pigment composition is produced by processing the media in which the cells are grown.

[0203] As used herein, the term "about" when combined with a value refers to plus and minus 10% of the reference value. For example, a length of about 1000 nanometers (nm) refers to a length of 1000 nm+-100 nm.

[0204] It is noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a polynucleotide" includes a plurality of such polynucleotides and reference to "the polypeptide" includes reference to one or more polypeptides and equivalents thereof known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely," "only" and the like in connection with the recitation of claim elements, or use of a "negative" limitation.

[0205] In those instances where a convention analogous to "at least one of A, B, and C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, and C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and / or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase "A or B" will be understood to include the possibilities of "A" or "B" or "A and B."

[0206] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the invention are specifically embraced by the present invention and are disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all subcombinations of the various embodiments and elements thereof are also specificallyembraced by the present invention and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0207] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents, unless the context clearly dictates otherwise. The terms “a” (or “an”) as well as the terms “one or more” and “at least one” can be used interchangeably.

[0208] Furthermore, “and / or” is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” as used in a phrase such as “A and / or B” is intended to include A and B, A or B, A (alone), and B (alone). Likewise, the term “and / or” as used in a phrase such as “A, B, and / or C” is intended to include A, B, and C; A, B, or C; A or B; A or C; B or C; A and B; A and C; B and C; A (alone); B (alone); and C (alone).

[0209] Wherever embodiments are described with the language “comprising,” otherwise analogous embodiments described in terms of “consisting of’ and / or “consisting essentially of’ are included.

[0210] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

[0211] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.EXAMPLES

[0212] Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include molecular, biochemical, microbiological and recombinant DNA techniques. Such techniques are thoroughly explained in the literature. See, for example, "Molecular Cloning: A laboratory Manual" Sambrook et al., (1989); "Current Protocols in Molecular Biology" Volumes I-III Ausubel, R. M., ed. (1994); Ausubel et al., "Current Protocols in Molecular Biology", John Wiley and Sons, Baltimore, Maryland (1989); Perbal, "A Practical Guide to Molecular Cloning", John Wiley & Sons, New York (1988); Watsonet al., "Recombinant DNA", Scientific American Books, New York; Birren et al. (eds) "Genome Analysis: A Laboratory Manual Series", Vols. 1-4, Cold Spring Harbor Laboratory Press, New York (1998); methodologies as set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659 and 5,272,057; "Cell Biology: A Laboratory Handbook", Volumes I- III Cellis, J. E., ed. (1994); "Culture of Animal Cells -A Manual of Basic Technique" by Freshney, Wiley-Liss, N. Y. (1994), Third Edition; "Current Protocols in Immunology" Volumes LIII Coligan J. E., ed. (1994); Stites et al. (eds), "Basic and Clinical Immunology" (8th Edition), Appleton & Lange, Norwalk, CT (1994); Mishell and Shiigi (eds), "Strategies for Protein Purification and Characterization -A Laboratory Course Manual" CSHL Press (1996); all of which are incorporated by reference. Other general references are provided throughout this document.Example 1: Production of new Betaxanthin pigment composition

[0213] The Inventors disclosed in International Patent Publication WO2017 / 122189 methods of producing pigment compositions comprising betalains by transient expression of exogenous genes of the betalain production pathway in yeast. Herein, this work has been built upon to produce superior betalain containing pigment compositions. In specific, these compositions comprise very high concentrations of betaxanthin pigment molecules, contain no betacyanin contamination and have unique color expression.

[0214] Saccharomyces cerevisiae strain BY4742 (S288C derivative) was used for betalain production. Rather than transient expression of the genes of the betalain pathway as had been done previously, the exogenous genes were now added to the yeast cells by genomic integration. Genomic integration allowed for a more precise control of the expression of the various genes. The strain exogenously expressed a tyrosine hydroxylase that was not also a L-DOPA oxidase (CYP76AD5, a CYP76AD1 beta-clade protein) and a DOPA 4,5- dioxygenase (MjDOD). Additionally, the strains contained a genomic integration of Arabidopsis thaliana NADPH-cytochrome P450 reductase 2 (ATR2). The cells were also fed with high levels of proline in the fermentation media. Proline drives conversion of betalamic acid to indicaxanthin (also called proline-betaxanthin). Since the betalain products are secreted, no extraction is required, and the recovery process is initiated by biomass clearance, followed by concentration and stabilization of the secreted betalain pigments.

[0215] When the secreted pigments were analyzed by HPLC, it was determined that while the strain secreted large amounts of indicaxanthin (nearly the only betaxanthin produced), no betalamic acid or betacyanins were detected (Fig. 1). This is in contrast to InternationalPatent Publication WO2017 / 122189 in which transient expression of CYP76AD6 and D0DA1 in yeast produced a mix of betalamic acid, valine-betaxanthin and vulgaxanthin (glutamine-betaxanthin) (see Table 2, sample “h” of WO2017 / 122189). This is also in contrast to natural prickly pear extracts which contain a mixture of various betaxanthins, betacyanins and betalamic acid (Fig. 1).

[0216] The liquid preparation product contained 100% betaxanthin pigment molecules without any contamination of other pigments (Fig. 2) Betaxanthin chromatographic purity was assessed at -480 nm (the / .max for most betaxanthins), betacyanins at a range of 534- 540 nm and betalamic acid at -409 nm. Not only was the composition purely betaxanthins but it was over 98% indicaxanthin (including both of its isomers). Further, the composition comprised heretofore unheard of concentrations of the pigment. Measured titers for total betaxanthins were greater than 4.2 grams per liter. These yields are greatly increased as compared to the compositions produced in WO2017 / 122189. In that work only liquid formulations were produced and the highest concentration achieved was only 20 mg / 1, which is roughly the same concentration that can be achieved from fruit extracts. This formulation is thus more than 100 times more concentrated than what has been known in the art.

[0217] In the fermentation process, the production organism is grown under submerged fed- batch, batch or continuous fermentation for the production of the pigments. Working cell banks are prepared from the master cell bank after being tested for microbial purity, specific growth rate, final biomass (dry cell weight) and production yields, prior to the fermentation process. The microbial purity is also periodically validated by microscopy and microbial culture of the fermentation broth. The process parameters including pH, temperature, agitation, and dissolved oxygen are routinely controlled and monitored throughout the process. The broth containing the product is harvested by removing the production organism from the fermentation broth at the end of the fermentation process using filtration, microfiltration, centrifugation, filter press or any other method of cell removal. A fully chemically defined medium was used for this process. In previous works a commercial synthetic medium was used (standard SD media).

[0218] In the downstream processes (DSP), a series of ultrafiltration and / or nanofiltration and / or reverse osmosis (with or without diafiltrations) steps or other suitable processes are used to purify and concentrate the pigment composition. The production can also include an additional step of water evaporation to concentrate the material for a liquid preparation. The liquid preparation can be dried to a powder form using a dryer (spray / freeze) or a fluidizer(fluidized bed reactor). The molecules can also be purified using crystallization methods. These DSP were fully absent from the Inventors’ previous work.

[0219] The reported yield of greater than 4 gr / L was recorded after this processing. This is highly significant, as 4 gr / L is considered the minimum concentration needed for a commercial composition. Extracts that are insufficiently concentrated cannot be easily added to foods without altering taste / appearance and generating unwanted bulk. In contrast, Tinggaard Thomsen et al., which describes an optimized yeast strain for betalain production produced at best 1.2- 1.3 gr / L betacyanins.Example 2: Comparison of new formulation to known Prickly pear compositions

[0220] Prickly pear is the major commercially available source of betaxanthin pigments, produced by physical and / or chemical extraction of the pigments from the plant. The new formulation produced herein was termed Phy to Yellow and was compared to two known commercially available prickly pear juice compositions. The compositions were analyzed by liquid chromatography-mass spectrometry (LC-MS) and various aspects of the compositions were quantified. This includes their elemental composition, nutritional values, and pigment composition (Fig. 1). Interestingly, PhytoYellow contained a much lower carbohydrate content than pear extracts and was essentially devoid of simple sugars unlike pear products.

[0221] The pigments produced were also very different than what is found in pear. The differences in pigment composition were quantified not only by liquid chromatography (Fig. 1), but also by spectrophotometry (Fig. 3). Commercial pear juices contained a mix of betaxanthins, betacyanins and betalamic acid. Further, the commercial juices had almost no iso-indicaxanthin. In contrast, the Phyto Yellow composition contained no detectable betacyanins or betalamic acid. Further, the betaxanthins were over 98% indicaxanthin (both isomers).

[0222] Measurements of L*, a*, b* values of the CIELAB color space by chromameter and comparison of these values to other compositions also found the color of the new compositions to be distinct (score of >2.5) (Fig. 4). This demonstrates that the pigment composition produced by genomic integration of the betaxanthin synthesis pathway into yeast is distinct from the pigment composition produced by pear. Further, the color difference caused by the production of nearly pure indicaxanthin is sufficiently pronounced that it can be observed by eye. Even small changes in the amounts of different pigmentmolecules can affect the final color and the instant system provides a means for producing highly pure betaxanthin compositions.Example 3: Production of additional Betaxanthin pigment compositions

[0223] The same genomically integrated yeast strain was again used for production of betaxanthins, however, instead of adding high levels of proline to the media seven other amino acids were tested. Nine cultures were setup: one without amino acid supplementation, and the rest with supplementation of proline, leucine, phenylalanine, serine, histidine, glutamic acid, methionine or glutamine. The yield of betaxanthins produced varied considerably depending on the amino acid supplemented, with proline, histidine and methionine producing the highest yields, and leucine, serine, glutamic acid and glutamine producing significantly lower yields (Fig. 6A). Phenylalanine produced the lowest yields, which were nearly as low as when no amino acid was supplemented.

[0224] Proline-betaxanthin in the initial experiment had produced very high purity and once again in this repeat greater than 90% of all betaxanthins in the composition were prolinebetaxanthin (Fig. 6B). Histidine -betaxanthin was also found to be produced with very high purity (-95%), but all other amino acids showed lower purity (44-61%). It is notable that in all cases except for glutamine-betaxanthins at least 50% of all betaxanthin produced included the supplemented amino acid.

[0225] The addition of various amino acids also impacted the yellow color produced (Fig. 7A). CIELAB color space was again used to quantify these differences, and it was found that many of the colors produced were distinct (difference of greater than 2.5) (Fig. 7B). Thus, the addition of different amino acids produces unique composition that can even be differentiated by the human eye.

[0226] Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

Claims

CLAIMS:

1. A pigment composition comprising betaxanthin pigment molecules and wherein less than 5% of pigment molecules in said composition are betalamic acid molecules or betacyanin molecules.

2. A pigment composition comprising betaxanthins, wherein at least 50% of said betaxanthins are proline-betaxanthin, at least 50% of said betaxanthins are histidinebetaxanthins, at least 50% of said betaxanthins are glutamic acid-betaxanthins, at least 50% of said betaxanthins are phenylalanine-betaxanthins, at least 50% of said betaxanthins are methionine -betaxanthins, at least 50% of said betaxanthins are leucine-betaxanthins, or at least 50% of said betaxanthins are serine-betaxanthins.

3. A pigment composition comprising betaxanthin pigment molecules, wherein said composition comprises less than 5% mono-and di-saccharides by weight.

4. The pigment composition of claim 2, wherein at least 90% of said betaxanthins are proline-betaxanthins or 90% of said betaxanthins are histidine -betaxanthins.

5. The pigment composition of any one of claims 2 to 4, wherein less than 5% of pigment molecules in said composition are betalamic acid molecules or betacyanin molecules.

6. The pigment composition of any one of claims 1 to 5, comprising less than 1 mg / L or 1 mg / kg betalamic acid.

7. The pigment composition of any one of claims 1, 3 and 5 to 6, wherein at least 50% of said betaxanthins are proline-betaxanthin, at least 50% of said betaxanthins are histidine-betaxanthins, at least 50% of said betaxanthins are glutamic acid- betaxanthins, at least 50% of said betaxanthins are phenylalanine-betaxanthins, at least 50% of said betaxanthins are methionine-betaxanthins, at least 50% of said betaxanthins are leucine-betaxanthins, or at least 50% of said betaxanthins are serinebetaxanthins.

8. The pigment composition of claim 7, wherein at least 90% of said betaxanthins are proline-betaxanthins or 90% of said betaxanthins are histidine-betaxanthins.

9. The pigment composition of any one of claims 1 to 8, wherein at least 90% of all pigment molecules in said composition are indicaxanthin or iso-indicaxanthin.

10. The pigment composition of any one of claims 1 to 2 and 4 to 9, wherein said composition comprises less than 5% mono-and di-saccharides by weight.

11. The pigment composition of any one of claims 1 to 10, wherein said composition comprises less than 0.1 pg / kg geosmin, less than 0.01 g / L nitrates or both.

12. The pigment composition of claim 9, wherein a ratio of indieaxanthin to iso- indieaxanthin is at least 3:1.

13. The pigment composition of any one of claims 1 to 12, wherein said composition is a liquid composition or a powder.

14. The pigment composition of any one of claims 1 to 13, comprising less than 5% dietary carbohydrates by weight.

15. The pigment composition of any one of claims 1 to 14, wherein less than 10% of all pigment molecules in the composition are betacyanins.

16. The pigment composition of claim 15, wherein less than 1% of all pigment molecules in the composition are betacyanins.

17. The pigment composition of any one of claims 1 to 16, comprising less than 1 mg / L or 1 mg / kg betacyanin.

18. The pigment composition of any one of claims 1 to 17, comprising at least 500 mg magnesium per liter of composition or at least 1000 mg magnesium per kilogram of composition.

19. The pigment composition of any one of claims 1 to 18, being a liquid and comprising greater than 1.5 grams betaxanthin pigment molecules per liter of composition or being a powder and comprising greater than 1.5 grams betalain pigment molecules per kilogram of composition.

20. The pigment composition of claim 19, being a liquid and comprising at least 4 grams betaxanthin pigment molecules per liter of composition or being a powder and comprising at least 4 grams betalain pigment molecules per kilogram of composition.

21. The pigment composition of any one of claims 1 and 5 to 20, wherein said betacyanin molecules comprise betanin, betanidin, and stereoisomers thereof.

22. The pigment composition of any one of claims 1 and 5 to 21, wherein less than 5% of pigment molecules in said composition are betalamic acid molecules.

23. A method of producing betaxanthin comprising an amino acid of interest, the method comprising culturing a cell comprising integrated into its genome at least one tyrosinehydroxylase gene that is not an L-DOPA oxidase gene, and at least one DOPA 4,5- dioxygenase (DOD) gene in a culture medium supplemented with said amino acid of interest and collecting said culture medium, thereby producing a betaxanthin comprising an amino acid of interest.

24. The method of claim 23, wherein said amino acid is selected from, proline, histidine, methionine, glutamic acid, serine, leucine, phenylalanine and glutamine.

25. The method of claim 24, wherein at least 40% of the betaxanthin produced comprises said amino acid.

26. The method of claims 23 to 25, wherein said cell further comprises and at least one NADPH cytochrome P450 reductase gene integrated into its genome.

27. The method of any one of claims 24 to 26, wherein said method is a method of producing a pigment composition of any one of claims 2 and 4 to 22.

28. The method of any one of claims 23 to 27, wherein said tyrosine hydroxylase gene encodes a protein comprising an amino acid sequence of SEQ ID NO: 2, 12, 14, 17, 19, 21 or 23 or a sequence with at least 85% identity thereto that converts tyrosine to L-DOPA and does not convert L-DOPA to cyclo-DOPA, and said DOD gene encodes a protein comprising an amino acid sequence of SEQ ID NO: 4 or 6 or a sequence with at least 85% identity thereto that converts L-DOPA to betalamic acid.

29. The method of any one of claims 26 to 28, wherein said NADPH cytochrome P450 reductase gene encodes a protein comprising an amino acid sequence of SEQ ID NO: 8 or 10 or a sequence with at least 85% identity thereto that comprises reductase activity.

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