Theobroma cacao cell biomass and compositions comprising same

Abstract

The present invention provides a cocoa powder including at least one compound selected from the group consisting of: an essential amino acid, a sugar, a neurotransmitter, and any combination thereof, wherein the at least one compound is enriched in a biomass of Theobroma cacao cells compared to a control cocoa powder, and is derived therefrom.

Description

THEOBROMA CACAO CELL BIOMASS AND COMPOSITIONS COMPRISING SAMECROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of IL Patent Application No. 316367, entitled “THEOBROMA CACAO CELL BIOMASS AND COMPOSITIONS COMPRISING SAME”, filed 14 October 2024, the contents of which are incorporated herein by reference in their entirety.FIELD OF INVENTION

[0002] The present invention is directed to, inter alia, edible products and / or ingredients thereof, such as including T. cacao cell biomass and methods of preparing the same.BACKGROUND

[0003] The cacao industry and cell culture community have long sought innovative methods to enhance the production and quality of cacao-derived products. Traditional agricultural practices often face limitations in terms of scalability, consistency, and environmental impact. These challenges necessitate the exploration of alternative approaches to cacao cultivation and processing. Current methods for producing cacao products involve complex processes that can lead to variability in flavor and nutritional content. Additionally, the reliance on conventional farming techniques poses challenges in meeting the growing demand for cacao while maintaining sustainability.

[0004] There is still a great need for a streamlined approach that allows for controlled production with consistent quality and enhanced nutritional profiles.SUMMARY

[0005] According to the first aspect, there us provided a cocoa powder comprising at least one compound selected from the group consisting of an essential amino acid, a sugar, a neurotransmitter, and any combination thereof, wherein the at least one compound is enriched in a biomass of Theobroma cacao cells compared to a control cocoa powder, and is derived therefrom.

[0006] According to another aspect, there is provided an edible composition comprising the cocoa powder of the invention.

[0007] In some embodiments, the essential amino acid is lysine, threonine, or both.

[0008] In some embodiments, the essential amino acid is present in the cocoa powder in an amount being greater by at least 1.5 compared to the amount of the essential amino acid in the control powder cocoa powder.

[0009] In some embodiments, the sugar is selected from the group consisting of: sucrose, fructose, glucose, galactose, D-glucopyranose, and any combination thereof.

[0010] In some embodiments, the sugar is present in the cocoa powder in an amount being greater by at least 1.4 compared to the amount of the sugar in the control powder cocoa powder.

[0011] In some embodiments, the cocoa powder is characterized by a total sugar content being greater by at least 2 compared to a total sugar content in the control powder cocoa powder.

[0012] In some embodiments, the neurotransmitter is dopamine, serotonin, or both.

[0013] In some embodiments, the cocoa powder is obtained or being derived from the biomass of T. cacao cells.

[0014] In some embodiments, the cocoa powder is obtained or prepared by a method comprising fermenting the biomass of T. cacao cells.

[0015] In some embodiments, the biomass of T. cacao cells comprises Alanine and Valine in a weight per weight ratio (w / w) ranging between 1 : 1 (w / w) and 200: 1 (w / w).

[0016] In some embodiments, the biomass of T. cacao cells comprises Valine and Serine in a w / w ranging between 6: 1 (w / w) and 1 :25 (w / w).

[0017] In some embodiments, the biomass of T. cacao cells comprises pyroglutamic acid and lactic acid in w / w ratio ranging between 1 : 1 (w / w) and 25: 1 (w / w).

[0018] In some embodiments, the biomass of T. cacao cells comprises glucose and erythritol in a w / w ratio of at least 5: 1 (w / w).

[0019] In some embodiments, the biomass of T. cocoa cells further comprises a total sugar content of at least 20 mg / gr of the biomass.

[0020] In some embodiments, the biomass of T. cocoa cells further comprises an antioxidant activity being at least 1.2-fold greater compared to a control biomass of T. cocoa cells.

[0021] In some embodiments, the biomass of T. cacao cells further comprises an amount of at least one catechin selected from the group consisting of: Epigall ocatechin 3 -gallate (EGCG), Gallocatechin 3 -gallate, Epigallocatechin, Catechin gallate, and any combination thereof, being at least 3 -fold greater by weight compared to the amount of the at least one catechin in a control biomass of T. cocoa cells.

[0022] In some embodiments, the biomass of T. cocoa cells is devoid of caffeine, theobromine, or both.

[0023] In some embodiments, cells of the biomass of T. cocoa cells are obtained or derived from an epicotyl, hypocotyl, cotyledon, or any combination thereof, of a T. cocoa seed.

[0024] In some embodiments, the edible composition is a foodstuff or an ingredient thereof.

[0025] In some embodiments, the foodstuff comprises any one of: chocolate, cake, brownie, and cookie, or any combination thereof.

[0026] In some embodiments, the foodstuff is a beverage.

[0027] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0028] 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

[0029] Fig. 1 includes a bar graph comparing the concentration of various amino acids and compounds in different samples, including cacao powder, NIBS, and several Theobroma cacao cell lines.

[0030] Figs. 2A-2D include bar graphs showing the concentration of essential amino acids, including Lysine (2A), Phenylalanine (2B), Threonine (2C), and Histidine (2D) across different T. cacao cell lines and cacao powder.

[0031] Fig. 3 includes a bar graph comparing the concentration of various acids in different samples, including cacao powder, NIBS, and several T. cacao cell lines.

[0032] Figs. 4A-4B include graphs showing total sugar content (4A) and the percentage composition of various sugars (4B) in different T. cacao cell lines of the invention.

[0033] Fig. 5 includes a vertical bar graph showing the antioxidant capacity, measured in mmol Trolox equivalent per mg cacao, across different T. cacao cell lines of the invention, and the controls: Nibs and cacao powder.

[0034] Figs. 6A-6D include vertical bar graph showing the concentrations of the catechins: Epigallocatechin 3-gallate (EGCG; 6A), Gallocatechin 3-gallate (6B), Epigallocatechin (6C), and Catechin gallate (6D) being enriched in different cultured T. cacao cell lines of the invention compared to controls: NIBS and cacao powder. Chemical structure of the different catechins are also included.

[0035] Fig. 7 includes a graph showing that glutamic acid is present in an amount of tens of micrograms per gr sample of a T. cacao cell line of the invention (1632). Further, this high level is maintained in a processed cocoa powder derived from the 1632 line (DS001 -160225) and surprisingly exceeds the amount of glutamic acid in control commercial cocoa powders (P001, P002, P008, and P009).

[0036] Fig. 8 includes a graph showing that the cocoa powder of the invention is enriched with essential amino acids, compared to control commercial cocoa powders. Specifically, lysine and threonine were observed in cocoa powder derived from a T. cacao cell line of the invention (1632) in at least 1.5-fold greater amounts compared to the control cocoa powders (P001, P002, P008, and P009 (see Table 1)).

[0037] Figs. 9A-9B include graphs showing that the cocoa powder of the invention is enriched with sugars compared to control commercial cocoa powders. (9 A) A graph showing the levels of fructose, galactose, D-glucopyranose, glucose, and sucrose in cocoa powder derived from a T. cacao cell line of the invention (1632) (as well as in the line itself before processing, e.g., fermentation, etc.) compared to the control cocoa powders (P001, P002, P008, and P009). (9B) A graph showing that cocoa powder of the invention includes a total sugar content in an amount at least 2-fold greater compared to the control cocoa powders (P001, P002, P008, and P009).

[0038] Figs. 10A-10B include vertical bar graphs showing that different T. cacao cell lines of the invention are enriched with neurotransmitters or precursors thereof, i.e., dopamine (10A) and the serotonin precursor 5 -Hydroxy try ptophan (5-HTP; 10B), compared to controls, e.g., NIBS and / or commercial cocoa powder (P002 as disclosed herein).DETAILED DESCRIPTIONCocoa powder

[0039] A cocoa powder comprising at least one compound selected from the group consisting of: an amino acid, a sugar, a neurotransmitter, and any combination thereof, wherein the at least one compound is enriched in a biomass of Theobroma cacao cells compared to a control cocoa powder, and is derived therefrom.

[0040] In some embodiments, enriched is endogenously, inherently, naturally, any equivalent thereof, or any combination thereof, in the biomass of T. cocoa cell from which the cocoa powder is derived or obtained. In some embodiments, enriched does not include supplemented with. In some embodiments, the at least one compound is not supplemented to the cocoa powder, the biomass of T. cocoa cells, or both. In some embodiments, the at least one compound is not exogenously supplemented and / or applied to the cocoa powder, the biomass of T. cocoa cells, or both.

[0041] As used herein, the term “endogenously” refers to compounds that are produced, synthesized, or naturally occurring within the Theobroma cacao cells themselves, rather than being added from external sources. Endogenous compounds may originate from the cellular metabolism, biosynthetic pathways, or natural physiological processes of the T. cacao cells during growth, development, or cultivation conditions. This is in contrast to “exogenously” referred to herein to compounds that are added, applied, or introduced from external sources outside of the Theobroma cacao cells. Exogenous compounds may include substances that are artificially supplemented, externally applied during processing, or added through external treatment methods rather than being naturally produced or synthesized within the cellular metabolism or biosynthetic pathways of the T. cacao cells themselves.

[0042] The terms “endogenously”, “inherently”, and “naturally” are used herein interchangeably.

[0043] In some embodiments, the cocoa powder is derived or obtained from the biomass of T. cocoa cells by means or methods including fermentation, roasting, drying, grinding, or any combination thereof. Means and methods of fermentation so as to obtain cocoa powders are common and would be apparent to one of ordinary skill in the art, such as described by De Vuyst et al., “Functional role of yeasts, lactic acid bacteria and acetic acid bacteria in cocoa fermentation processes”, FEMS Microbiology Reviews, Volume 44, Issue 4, (2020), Pages 432-453; and Racine et al., “Development and characterization of a pilot-scale model cocoa fermentation system suitable for studying the impact of fermentation on putative bioactive compounds and bioactivityof cocoa”. Foods, 8(3), 102 (2019), both of which are incorporated herein by reference in their entirety.

[0044] In some embodiments, the amino acid is an essential amino acid. In some embodiments, the amino acid is a non-essential amino acid. In some embodiments, the essential amino acid comprises a plurality of essential amino acids. In some embodiments, the non-essential amino acid comprises a plurality of non-essential amino acids. In some embodiments, the amino acid comprises an essential amino acid, a non-essential amino acid, plurality of thereof, or any combination thereof (such as one or more essential amino acids and one or more non-essential amino acids).

[0045] In some embodiments, a plurality refers to any integer being equal to or greater than 2.

[0046] In some embodiments, the essential amino acid comprises or is lysine. In some embodiments, the essential amino acid comprises or is threonine. In some embodiments, the essential amino acid comprises or is lysine and threonine.

[0047] In some embodiments, the non-essential amino acid comprises or is isoleucine. In some embodiments, the essential amino acid comprises or is glycine. In some embodiments, the essential amino acid comprises or is glutamic acid. In some embodiments, the essential amino acid comprises or is isoleucine, glycine, and glutamic acid.

[0048] In some embodiments, the amino acid is present in the cocoa powder in an amount being greater by at least 1.5, 1.7, 2.0, 2.5, 3.0, 3.5, or 4.0 compared to the amount of the amino acid in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the amino acid is present in the cocoa powder in an amount being greater by 1.5-10.0, 1.7-9.0, 2.0- 8.0, 2.5-10.0, 3.0-10.0, 3.5-9.5, or 4.0-10.0 compared to the amount of the amino acid in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0049] In some embodiments, the amino acid comprises or is lysine and is present in the cocoa powder in an amount being greater by at least 2.2, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 compared to the amount of the amino acid in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the amino acid comprises or is lysine and is present in the cocoa powder in an amount being greater by 1.5-10.0, 2.0-10.0, 2.2-9.5, 3.0-10.0, 3.5-9.5, or 4.0-10.0 compared to the amount of the amino acid in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0050] In some embodiments, the amino acid comprises or is threonine and is present in the cocoa powder in an amount being greater by at least 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 compared to the amount of the amino acid in a control powder cocoa powder, or any value and rangetherebetween. In some embodiments, the amino acid comprises or is threonine and is present in the cocoa powder in an amount being greater by 1.5-10.0, 2.0-10.0, 2.2-9.5, 3.0-10.0, 3.5-9.5, or 4.0-10.0 compared to the amount of the amino acid in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0051] In some embodiments, the amino acid comprises or is glycine and is present in the cocoa powder in an amount being greater by at least 1.5, 2.0, 5.0, 7.0, 10.0, 15.0, 20.0, or 30.0 compared to the amount of the amino acid in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the amino acid comprises or is glycine and is present in the cocoa powder in an amount being greater by 1.5-50.0, 2.0-45.0, 10.0-40.0, 10.0-30.0, or 15.0-45.0 compared to the amount of the amino acid in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0052] In some embodiments, the amino acid comprises or is glutamic acid and is present in the cocoa powder in an amount being greater by at least 5, 7, 10, 15, 20, 25, 30, or 50 compared to the amount of the amino acid in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the amino acid comprises or is glycine and is present in the cocoa powder in an amount being greater by 5-50, 10-50, 15-50, 20-45, 25-35, or 25-45 compared to the amount of the amino acid in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0053] In some embodiments, a sugar comprises a plurality of types of sugars. In some embodiments, a sugar comprises at least one sugar. In some embodiments, a sugar comprises one or more types of sugars. In some embodiments, the sugar is selected from: sucrose, fructose, glucose, galactose, D-glucopyranose, or any combination thereof. In some embodiments, the sugar is or comprises a combination of: sucrose, fructose, glucose, galactose, and D-glucopyranose.

[0054] In some embodiments, the sugar is present in the cocoa powder in an amount being greater by at least 1.4, 1.5, 2.0, 3.0, 5.0, 6.0, 8.0, 10.0, or 20.0 compared to the amount of the sugar in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the sugar is present in the cocoa powder in an amount being greater by at least 1.4-50.0, 1.5-40.0, 2.0- 30.0, 3.0-50.0, 5.0-35.0, 6.0-30.0, 8.0-40.0, or 10.0-40.0 compared to the amount of the sugar in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0055] In some embodiments, the sugar comprises or is sucrose and is present in the cocoa powder in an amount being greater by at least 2.5, 2.7, 3.0, 5.0, 10.0, 15.0, 20.0, or 30.0 compared to the amount of the sugar in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the sugar comprises or is sucrose and is present in the cocoa powder in an amount being greater by 2.5-50.0, 2.7-45.0, 3.0-40.0, 3.5-30.0, or 5.0-45.0 compared to the amountof the sugar in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0056] In some embodiments, the sugar comprises or is fructose and is present in the cocoa powder in an amount being greater by at least 2.0, 2.1, 2.5, 5.0, 10.0, 15.0, 20.0, or 30.0 compared to the amount of the sugar in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the sugar comprises or is fructose and is present in the cocoa powder in an amount being greater by 2.0-50.0, 2.2-45.0, 2.5-40.0, 2.5-10.0, or 2.5-5.0 compared to the amount of the sugar in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0057] In some embodiments, the sugar comprises or is glucose and is present in the cocoa powder in an amount being greater by at least 2.0, 2.1, 2.5, 5.0, 10.0, 15.0, 20.0, or 30.0 compared to the amount of the sugar in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the sugar comprises or is glucose and is present in the cocoa powder in an amount being greater by 2.0-50.0, 2.2-45.0, 2.5-40.0, 2.5-10.0, or 2.5-50.0 compared to the amount of the sugar in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0058] In some embodiments, the sugar comprises or is galactose and is present in the cocoa powder in an amount being greater by at least 1.3, 1.4, 1.5, 2.0, 3.0, 4.0, 5.0, or 7.0 compared to the amount of the sugar in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the sugar comprises or is galactose and is present in the cocoa powder in an amount being greater by 1.3-20.0, 1.5-20.0, 2.0-15.0, 2.5-20.0, or 3.0-25.0 compared to the amount of the sugar in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0059] In some embodiments, the sugar comprises or is D-glucopyranose and is present in the cocoa powder in an amount being greater by at least 3.4, 3.5, 4.0, 5.0, 6.0, 10.0, 20.0, or 40.0 compared to the amount of the sugar in a control powder cocoa powder, or any value and range therebetween. In some embodiments, the sugar comprises or is D-glucopyranose and is present in the cocoa powder in an amount being greater by 3.5-50.0, 4.0-45.0, 5.0-50.0, 6.5-40.0, or 8.0-50.0 compared to the amount of the sugar in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0060] In some embodiments, the cocoa powder comprises or is characterized by a total sugar content being greater by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 7- fold, or at least 10-fold compared to a total sugar content in a control powder cocoa powder, or any value and range therebetween. Each possibility represents a separate embodiment of theinvention. In some embodiments, the cocoa powder comprises or is characterized by a total sugar content being greater by 2-10-fold, 3-10-fold, 4-10-fold, 5-10-fold, or 7-10-fold compared to a total sugar content in a control powder cocoa powder. Each possibility represents a separate embodiment of the invention.

[0061] In some embodiments, the neurotransmitter comprises dopamine, serotonin, or both.

[0062] As used herein, the term “neurotransmitter” refers to a chemical messenger that transmits signals across synapses between neurons or from neurons to target cells. Neurotransmitters include, but are not limited to, monoamines such as dopamine, serotonin, norepinephrine, and epinephrine; amino acids such as gamma-aminobutyric acid (GABA), glutamate, and glycine; acetylcholine; and other signaling molecules that facilitate communication within nervous systems. In the context of Theobroma cacao cells, neurotransmitters refer to compounds that have similar chemical structures or biological activities to those found in animal nervous systems, even though they serve different physiological functions in plant cells.

[0063] According to another aspect, there is provided a composition comprising the cocoa powder of the invention. In some embodiments, the composition is an edible composition. In some embodiments, an edible composition is or comprises a foodstuff. In some embodiments, the composition is a nutraceutical. In some embodiments, the composition is a functional ingredient. In some embodiments, the composition is a beverage.

[0064] As used herein, the term “nutraceutical” refers to a product derived from food sources that provides health and medical benefits in addition to basic nutritional value. Nutraceuticals may include dietary supplements, functional foods, or medicinal foods that contain bioactive compounds with physiological benefits or protective effects against disease. In some aspects, nutraceuticals may encompass vitamins, minerals, herbs, amino acids, enzymes, probiotics, antioxidants, or other naturally occurring substances that may promote health, prevent disease, or support normal physiological functions. The term may apply to products that bridge the gap between food and pharmaceuticals, offering therapeutic or health-enhancing properties while being derived from natural food sources.

[0065] As used herein, the term “functional ingredient” refers to a component or substance that is incorporated e.g., into edible or consumable products, such as food products, to provide specific health benefits or physiological functions beyond basic nutrition. These ingredients may include bioactive compounds, nutrients, or other substances that contribute to improved health outcomes, disease prevention, or enhanced physiological performance when consumed as part of the diet. In some aspects, functional ingredients may include probiotics, prebiotics, antioxidants, vitamins, minerals, omega-3 fatty acids, plant sterols, proteins, or other bioactive molecules. The functionalingredient may be added during food processing or formulation to enhance the nutritional or health-promoting properties of the final food product.

[0066] As used herein, the term “cocoa powder" refers to a product and / or a powder that is made from cacao beans or cells thereof that have been fermented, roasted, and processed to at least partially remove the cocoa butter. The remaining solids are ground into a fine powder. It is commonly used in baking and beverages, providing a rich chocolate flavor and containing antioxidants and other nutrients.

[0067] As used herein, the term “food grade carrier” refers to a substance used to deliver or stabilize active ingredients, such as the biomass of the invention, in food products. It is safe for consumption and complies with food safety regulations. These carriers help in the formulation of food products by ensuring the even distribution of ingredients, enhancing texture, or improving shelf life.

[0068] In some embodiments, the foodstuff comprises: chocolate, a cake, a brownie, a cookie, or any combination thereof.

[0069] In some embodiments, the foodstuff comprises a beverage.Theobroma cacao cell biomass

[0070] According to another aspect, there is provided a biomass of Theobroma cacao cells (hereinafter may be referred to as “the biomass of the invention”).

[0071] As used herein, the term “biomass” refers to a mass of Theobroma cacao cells cultivated through cell culture techniques. This biomass is characterized by specific biochemical properties, such as amino acid ratios, sugar content, and antioxidant activity, which distinguish it from traditional cacao products. The biomass can be used as an ingredient in edible products, offering consistent quality and enhanced nutritional profiles.

[0072] In some embodiments, the biomass of the invention is characterized by an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 2:1 (w / w), 1 : 1 (w / w) and 20: 1 (w / w), 1 :1 (w / w) and 50: 1 (w / w), 1 : 1 (w / w) and 70: 1 (w / w), 1 : 1 (w / w) and 90: 1 (w / w), 1 : 1 (w / w) and 100:1 (w / w), 1 : 1 (w / w) and 150: 1 (w / w), or 1 : 1 (w / w) and 200:1 (w / w). Each possibility represents a separate embodiment of the invention.

[0073] In some embodiments, the biomass of the invention is characterized by a Valine to Serine w / w ranging between 6: 1 (w / w) and 1 :25 (w / w), 4: 1 (w / w) and 1 :25 (w / w), 2: 1 (w / w) and 1 :25 (w / w), 1 : 1 (w / w) and 1 :25 (w / w), 6: 1 (w / w) and 1 :20 (w / w), 6: 1 (w / w) and 1 : 15 (w / w), 6: 1 (w / w) and 1 : 10 (w / w), 6: 1 (w / w) and 1 :6 (w / w), 6:1 (w / w) and 1 : 1 (w / w), 4: 1 (w / w) and 1 :20 (w / w), 2: 1(w / w) and 1 :20 (w / w), 3 :1 (w / w) and 1 :5 (w / w), or 5: 1 (w / w) and 1 : 10 (w / w). Each possibility represents a separate embodiment of the invention.

[0074] In some embodiments, the biomass of the invention is characterized by a pyroglutamic acid to lactic acid w / w ratio between 1 : 1 (w / w) and 2: 1 (w / w), 1 : 1 (w / w) and 5 : 1 (w / w), 1 : 1 (w / w) and 7:1 (w / w), 1 :1 (w / w) and 10: 1 (w / w), 1 : 1 (w / w) and 12:1 (w / w), 1 : 1 (w / w) and 15: 1 (w / w), 1 : 1 (w / w) and 20: 1 (w / w), 1 : 1 (w / w) and 22: 1 (w / w), or 1 : 1 (w / w) and 25: 1 (w / w). Each possibility represents a separate embodiment of the invention.

[0075] In some embodiments, the biomass of the invention is characterized by a glucose to erythritol w / w ratio of at least 5 : 1 (w / w), at least 7 : 1 (w / w), at least 9 : 1 (w / w), at least 10:1 (w / w), at least 15: 1 (w / w), at least 20: 1 (w / w), at least 25: 1 (w / w), at least 50:1 (w / w), or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the biomass of the invention is characterized by a glucose to erythritol w / w ratio of 5:1 (w / w) to 500: 1 (w / w), 5: 1 (w / w) to 50: 1 (w / w), 5: 1 (w / w) to 25: 1 (w / w), 6: 1 (w / w) to 200: 1 (w / w), 8:1 (w / w) to 150: 1 (w / w), or 10: 1 (w / w) to 300: 1 (w / w). Each possibility represents a separate embodiment of the invention.

[0076] In some embodiments, the biomass of the invention is further characterized by a total sugar content of at least 20 mg / gr of the biomass, at least 25 mg / gr of the biomass, at least 30 mg / gr of the biomass, at least 50 mg / gr of the biomass, at least 70 mg / gr of the biomass, at least 90 mg / gr of the biomass, at least 100 mg / gr of the biomass, at least 120 mg / gr of the biomass, at least 150 mg / gr of the biomass, at least 175 mg / gr of the biomass, at least 200 mg / gr of the biomass, at least 220 mg / gr of the biomass, at least 250 mg / gr of the biomass, at least 300 mg / gr of the biomass, or at least 350 mg / gr of the biomass, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the biomass of the invention is further characterized by a total sugar content of 20 mg / gr to 350 mg / gr, 50 mg / gr to 350 mg / gr, 70 mg / gr to 350 mg / gr, 100 mg / gr to 350 mg / gr, 120 mg / gr to 350 mg / gr, 170 mg / gr to 350 mg / gr, 200 mg / gr to 350 mg / gr, 250 mg / gr to 350 mg / gr, 20 mg / gr to 200 mg / gr, 20 mg / gr to 150 mg / gr, or 50 mg / gr to 150 mg / gr, of the biomass. Each possibility represents a separate embodiment of the invention.

[0077] In some embodiments, the weight of the biomass or reference to the weight of the biomass as used herein refers to the dry weight of the biomass. In some embodiments, the weight of the biomass or reference to the weight of the biomass as used herein refers to the wet weight of the biomass.

[0078] In some embodiments, the biomass is further characterized by a total sugar content being at least 1.05-fold greater, at least 1.2-fold greater, at least 1.3-fold greater, at least 1.5-fold greater,at least 2-fold greater, at least 5-fold greater, at least 10-fold greater, at least 20-fold greater, or at least 50-fold greater, by weight compared to the total sugar content in a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the biomass is further characterized by a total sugar content being 1.05-fold to 50- fold greater, 1.2-fold to 50-fold greater, 1.5-fold to 50-fold greater, 2-fold to 50-fold greater, 5- fold to 50-fold greater, 10-fold to 50-fold greater, 15-fold to 50-fold greater, 20-fold to 50-fold greater, 2-fold to 30-fold greater, 5-fold to 30-fold greater, 10-fold to 50-fold greater, or 15-fold to 50-fold greater, by weight compared to the total sugar content in a control. Each possibility represents a separate embodiment of the invention.

[0079] In some embodiments, the biomass is further characterized by an amount of at least one essential amino acid being increased compared to the amount of the at least one essential amino acid in a control.

[0080] As used herein, the term “essential amino acid(s)” refers to amino acids that cannot be synthesized by the body and must be obtained through diet. They are crucial for various bodily functions, including protein synthesis, tissue repair, and nutrient absorption. Generally, amino acids regarded as “essential” include: Lysine, Phenylalanine, Threonine, Histidine, Leucine, Isoleucine, Valine, Methionine, and Tryptophan.

[0081] As used herein, essential amino acids such as lysine, phenylalanine, threonine, and histidine are highlighted for their increased presence in the Theobroma cacao cell biomass compared to traditional cacao products.

[0082] In some embodiments, increased is an increase of at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, 750%, 1,000%, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0083] In some embodiments, increased or an increase is compared to a control.

[0084] In some embodiments, the essential amino acid is selected from: lysine, phenylalanine, threonine, histidine, or any combination thereof.

[0085] In some embodiments, the biomass is further characterized by an amount of leucine being reduced by at least 30%, at least 35%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or 90% by weight compared to the amount of leucine in a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the biomass is further characterized by an amount of leucine being reduced by 30% to 99%, 40% to 99%, 50% to 99%, 60% to 99%, 70% to 99%, 80% to 99%, or 90% to 99% byweight compared to the amount of leucine in a control. Each possibility represents a separate embodiment of the invention.

[0086] In some embodiments, the biomass is further characterized by an amount of at least one fatty acid being stearic acid (also known as or termed "octadecanoic acid"), palmitic acid (also known as or termed "hexadecenoic acid"), or both, being greater by weight compared to the amount of the at least one fatty acid in a control. In some embodiments, the biomass is further characterized by an amount of at least one fatty acid being stearic acid, palmitic acid, and both, being at least 1.5-fold, 2-fold, 5-fold, 7-fold, 10-fold, 15-fold, or 20-fold, greater by weight compared to the amount of the at least one fatty acid in a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the biomass is further characterized by an amount of at least one fatty acid being stearic acid, palmitic acid, and both, being 1.5-fold to 20-fold, 2-fold to 20-fold, 5-fold to 20-fold, 7-fold to 20-fold, 10-fold to 20-fold, 12-fold to 20-fold, or 15-fold to 20-fold, greater by weight compared to the amount of the at least one fatty acid in a control. Each possibility represents a separate embodiment of the invention.

[0087] In some embodiments, the biomass is further characterized by an increased antioxidant activity compared to control.

[0088] As used herein, the term “antioxidant activity” refers to " the ability of a substance to neutralize free radicals or prevent oxidative damage in the body. As used herein, it measures the capacity of the T. cacao cell biomass to provide health benefits by reducing oxidative stress, which is linked to various diseases and aging processes. As exemplified herein, increased dopamine levels have been demonstrated over control(s), which indicates increased antioxidant activity.

[0089] In some embodiments, the antioxidant activity of the biomass of the invention is at least 1.05-fold greater, 1.2-fold greater, 1.5-fold greater, 2-fold greater, 3-fold greater, 5-fold greater, 10-fold greater, 12-fold greater, 15-fold greater, or 20-fold greater, compared to a control, or any value and range therebetween. Each possibility represents a separate embodiment of the invention. In some embodiments, the antioxidant activity of the biomass of the invention is 1.05-fold to 20- fold, 2-fold to 20-fold, 5-fold to 20-fold, 7-fold to 20-fold, 10-fold to 20-fold, 12-fold to 20-fold, or 15-fold to 20-fold, compared to a control. Each possibility represents a separate embodiment of the invention.

[0090] In some embodiments, the biomass is further characterized by an amount of at least one catechin being at least 1.5-fold greater, at least 2-fold greater, at least 3-fold greater, at least 5-fold greater, at least 10-fold greater, or at least 20-fold greater, by weight compared to the amount of the at least one catechin in a control, or any value and range therebetween. Each possibilityrepresents a separate embodiment of the invention. In some embodiments, the biomass is further characterized by an amount of at least one catechin being 1.5-fold to 100-fold greater, 2-fold to 100-fold greater, 5-fold to 100-fold greater, 10-fold to 100-fold greater, 15-fold to 100-fold greater, 25-fold to 100-fold greater, or 50-fold to 100-fold greater, by weight compared to the amount of the at least one catechin in a control. Each possibility represents a separate embodiment of the invention.

[0091] As used herein, the term “catechin” refers to a type of natural phenolic compound and antioxidant found in various plants, including cacao. Catechins are known for their health benefits, such as reducing oxidative stress and providing cardiovascular support. Generally, catechins include: Epicatechin, Epicatechin gallate (ECG), Epigallocatechin (EGC), Epigallocatechin gallate (EGCG), Gallocatechin, Gallocatechin gallate (GCG), Catechin, and Catechin gallate. As used herein, catechins like Epigallocatechin 3 -gallate (EGCG), Gallocatechin 3 -gallate, Epigallocatechin, and Catechin gallate are highlighted for their increased presence in the Theobroma cacao cell biomass compared to traditional cacao products.

[0092] In some embodiments, the at least one catechin is selected from: Epigallocatechin 3 -gallate (EGCG), Gallocatechin 3-gallate, Epigallocatechin, Catechin gallate, or any combination thereof.

[0093] Methods for determining the type and / or amount of any compound as recited herein, including, amino acids, acid, fatty acids, sugars, and antioxidants, are common and would be apparent to a skilled artisan. Non-limiting examples for such methods include but are not limited to, HPLC, GC-MS, antioxidant assay, as exemplified herein.

[0094] In some embodiments, the biomass is further characterized by being devoid of caffeine, theobromine, or both. In some embodiments, the biomass comprises no caffeine. In some embodiments, the biomass does not comprise caffeine. In some embodiments, the biomass comprises no theobromine. In some embodiments, the biomass does not comprise theobromine. In some embodiments, the biomass comprises no caffeine and no theobromine. In some embodiments, the biomass does not comprise caffeine and theobromine. In some embodiments, the biomass comprises negligible or undetected amount of caffeine, theobromine, or both.

[0095] In some embodiments, a control comprises or is cocoa nibs, cocoa powder, or both.

[0096] As exemplified herein, control, such as 'cocoa powder control' comprises Organic Raw Cacao Powder which was sourced from Peru, and purchased from nizat.com. Per 100 gr, the control cacao powder is characterized by: energy - 435 calories; total fats - 10 gr; saturated fatty acids - 6.5 gr; trans fatty acids 0.5 gr cholesterol 0 mg; sodium - 46 mg; total carbohydrates - 28gr; sugars - 2 gr; dietary fibers - 33 gr; proteins - 25 gr; potassium - 1532 mg; iron - 37 mg; calcium - 164 mg; and zinc - 5 mg.

[0097] As exemplified herein, a control, such as NIBS or cocoa nibs, comprises organic raw cacao beans having their dry peel removed, after which they were broken to pieces before grinding to fine powder for analysis. The organic raw cacao beans were sourced from Peru, and purchased from nizat.com. Per 100 gr, the control NIBS or cacao nibs is characterized by: energy - 640 calories; total fats - 51 gr; saturated fatty acids - 31 gr; trans fatty acids - less than 0.5 gr cholesterol 0 mg; total carbohydrates - 30 gr; dietary fibers - 27.5 gr; and proteins - 14 gr.

[0098] As used herein, the term “cocoa nibs (as well as NIBS or Nibs) refers to " small pieces of crushed cacao beans that have been fermented, dried, and roasted. They are the purest form of cacao for consumption used as a base ingredient in chocolate products. Cocoa nibs retain the natural flavors and nutrients of the cacao bean, including antioxidants, fiber, and healthy fats.

[0099] As used herein, the term “cocoa powder" refers to a product and or a powder that is made from cacao beans that have been fermented, roasted, and processed to at least partially remove the cocoa butter. The remaining solids are ground into a fine powder. It is commonly used in baking and beverages, providing a rich chocolate flavor and containing antioxidants and other nutrients.

[0100] In some embodiments, the cells of the biomass of the invention are obtained or derived from an epicotyl, hypocotyl, cotyledon, or any combination thereof, of a T. cocoa seed. In some embodiments, the cells of the biomass of the invention are of epicotyl, hypocotyl, cotyledon, or any combination thereof, origin of a T. cocoa seed. In some embodiments, the cells of the biomass are solitary cells or of cell clusters of not more than about 60 pm, also referred to herein as “fine culture”. In some embodiments, the cells of the biomass are clustered cells and / or cells in a cluster(s) and / or in an aggregate(s) of more than 60 pm, also referred to herein as “granular culture”. In some embodiments, the cells of the biomass of the invention are from callus- or calli- origin of a T. cocoa. In some embodiments, the cells of the biomass of the invention originate from callus or calli generated from a seed origin of a T. cocoa. In some embodiments, the cells of the biomass of the invention originate from callus or calli generated from of an epicotyl origin of a T. cocoa. In some embodiments, the cells of the biomass of the invention originate from callus or calli generated from a hypocotyl origin of a T. cocoa. In some embodiments, the cells of the biomass of the invention originate from callus or calli generated from a cotyledon origin of a T. cocoa.

[0101] Methods for generating callus, isolating cells therefrom, and propagating cells thereof, as provided herein, are common and would be apparent to a skilled artisan.

[0102] In some embodiments, the biomass is in a dried form. In some embodiments, the biomass is in a form of a powder.

[0103] Methods of drying and / or grinding are common and would be apparent to one of ordinary skill in the art. Non-limiting examples for such processing methods include, but are not limited to, Freeze-drying (lyophilization), air drying, spray drying, drum drying, vacuum drying, convection drying, ball milling, hammer milling, jet milling, and roller milling, to name a few.Theobroma cacao lines

[0104] According to another aspect, there is provided a biomass of cells of Theobroma cacao accession number RUQ 847 (‘Donor collection’ CIRAD (Centre de Cooperation Internationale en Researche Agronomique pour le Developpment), France). In some embodiments, the biomass of cells of Theobroma cacao accession number RUQ 847 is referred to herein as "the 847 biomass". In some embodiments, the 847 biomass comprises cells originating from the epicotyl of a germinated seedling of Theobroma cacao accession number RUQ 847, and is referred to herein as "the 847-E biomass".

[0105] In some embodiments, the 847-E biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 3 : 1 (w / w); (ii) a Valine to Serine w / w ranging between 2:1 (w / w) and 1 : 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 5: 1 (w / w) and 10:1 (w / w); or (iv) any combination of (i) to (iii).

[0106] In some embodiments, the 847-E biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 10: 1 (w / w) to 30: 1 (w / w); (ii) a total sugar content of 10 mg / gr to 40 mg / gr; (iii) a total sugar content being 1.05-fold to 3-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 15% or by 15% to 30% compared to control; (v) an amount of threonine being increased by at 15% or by 15% to 30% compared to control; (vi) an amount of histidine ranging between 0.35 mg / gr and 0.7 mg / gr; (vii) an amount of leucine being reduced by at least 70% or by 60% to 80% by weight compared to control; (viii) an amount of stearic acid being reduced by at least 1.2-fold or between 1.2-fold and 2-fold compared to control; (ix) an amount of palmitic acid being reduced by at least 1.2-fold or between 1.2-fold and 2-fold compared to control; (x) an antioxidant activity being at least 1.5-fold greater or between 1.5-fold to 4-fold compared to a control; or (xi) any combination of (i) to (x). In some embodiments, the 847-E biomass is further characterized by increased 5-HTP levels (e.g., at least 2, 5, 10-fold increase) compared to a control.

[0107] In some embodiments, the 847 biomass comprises cells originating from the cotyledon of a germinated seedling of Theobroma cacao accession number RUQ 847, and is referred to herein as "the 847-C biomass".

[0108] In some embodiments, the 847-C biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 4: 1 (w / w); (ii) a Valine to Serine w / w ranging between 2: 1 (w / w) and 1 : 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 1.5:1 (w / w) and 5: 1 (w / w); or (iv) any combination of (i) to (iii).

[0109] In some embodiments, the 847-C biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 15 : 1 (w / w) to 50: 1 (w / w); (ii) a total sugar content of 20 mg / gr to 80 mg / gr; (iii) a total sugar content being 1.5 -fold to 6-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 15% or by 15% to 30% compared to control; (v) an amount of threonine being increased by at 15% or by 15% to 30% compared to control; (vi) an amount of histidine ranging between 0.05 mg / gr and 0.3 mg / gr; (vii) an amount of leucine being reduced by at least 70% or by 60% to 90% by weight compared to control; (viii) an amount of stearic acid being reduced by at least 1.05-fold or between 1.05-fold and 2-fold compared to control; (ix) an amount of palmitic acid being reduced by at least 1.05- fold or between 1.05-fold and 2-fold compared to control; (x) an antioxidant activity being at least 1.5-fold greater or between 1.5-fold to 4-fold compared to a control; or (xi) any combination of (i) to (x). In some embodiments, the 847-C biomass is further characterized by increased 5-HTP levels (e.g., at least 2, 3, 4, 5-fold increase) compared to a control.

[0110] In some embodiments, the 847 biomass comprises cells originating from the hypocotyl of a germinated seedling of Theobroma cacao accession number RUQ 847, and is referred to herein as "the 847-H biomass".

[0111] In some embodiments, the 847-H biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 4: 1 (w / w); (ii) a Valine to Serine w / w ranging between 2: 1 (w / w) and 1 : 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 1.5:1 (w / w) and 5: 1 (w / w); or (iv) any combination of (i) to (iii).

[0112] In some embodiments, the 847-H biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 15 : 1 (w / w) to 50: 1 (w / w); (ii) a total sugar content of 20 mg / gr to 80 mg / gr; (iii) a total sugar content being 1.5 -fold to 6-fold greater by weight compared to the total sugar content in a control; (iv) an amount of histidine ranging between 0.01 mg / gr and 0.05 mg / gr; (v) an amount of leucine being reduced by at least 40% or by 40% to 60% by weight compared to control; (vi) an amount of stearic acid being reduced by at least 1.05-fold or between 1.05-fold and 2-fold compared to control; (vii) an amount of palmitic acid being reduced by at least 1.05-fold orbetween 1.05-fold and 2-fold compared to control; (viii) an antioxidant activity being at least 1.5- fold greater or between 1.5-fold to 4-fold compared to a control; or (ix) any combination of (i) to (viii). In some embodiments, the 847-H biomass is further characterized by increased 5-HTP levels (e.g., at least 2, 3, 4, 5-fold increase) compared to a control.

[0113] According to another aspect, there is provided a biomass of cells of Theobroma cacao accession number RUQ 235 (‘Donor collection’ International Cocoa Genebank, Trinidad (ICG,T), Trinidad and Tobago). In some embodiments, the biomass of cells of Theobroma cacao accession number RUQ 235 is referred to herein as "the 235 biomass".

[0114] In some embodiments, the 235 biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 4: 1 (w / w); (ii) a Valine to Serine w / w ranging between 2: 1 (w / w) and 1 : 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 1.5: 1 (w / w) and 5: 1 (w / w); or (iv) any combination of (i) to (iii).

[0115] In some embodiments, the 235 biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 15: 1 (w / w) to 50: 1 (w / w); (ii) a total sugar content of 90 mg / gr to 160 mg / gr; (iii) a total sugar content being 4-fold to 12-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 30% or by 25% to 60% compared to control; (v) an amount of phenylalanine being increased by at 25% or by 25% to 60% compared to control (vi) an amount of threonine being increased by at 35% or by 35% to 60% compared to control; (vii) an amount of leucine being reduced by at least 60% or by 60% to 80% by weight compared to control; (viii) an amount of stearic acid being increased by at least 1.5-fold or between 1.5-fold and 4-fold compared to control; (ix) an amount of palmitic acid being increased by at least 1.2-fold or between 1.2-fold and 2.5-fold compared to control; (x) an antioxidant activity being at least 2.5-fold greater or between 2.5-fold to 4.5-fold compared to a control; or (x) any combination of (i) to (x).

[0116] According to another aspect, there is provided a biomass of cells of Theobroma cacao accession number RUQ 137 (‘Donor collection’ International Cocoa Genebank, Trinidad (ICG,T), Trinidad and Tobago). In some embodiments, the biomass of cells of Theobroma cacao accession number RUQ 137 is referred to herein as "the 137 biomass". In some embodiments, the 137 biomass comprises cells originating from the cotyledon of a germinated seedling of Theobroma cacao accession number RUQ 137, and is referred to herein as "the 137-C biomass".

[0117] In some embodiments, the 137-C biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 4: 1 (w / w); (ii) a Valine to Serine w / w ranging between 2: 1 (w / w) and 1 : 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 3: 1 (w / w) and 10:1 (w / w); or (iv) any combination of (i) to (iii).

[0118] In some embodiments, the 137-C biomass is further characterized by: (i) a glucose to erythritol w / w ratio of at least 100:1; being devoid of erythritol, or both; (ii) a total sugar content of 90 mg / gr to 160 mg / gr; (iii) a total sugar content being 4-fold to 12-fold greater by weight compared to the total sugar content in a control; (iii) an amount of phenylalanine being increased by at 35% or by 35% to 60% compared to control; (iv) an amount of leucine being reduced by at least 25% or by 25% to 50% by weight compared to control; (v) an amount of stearic acid being increased by at least 1.5-fold or between 1.5-fold and 4-fold compared to control; (vi) an amount of palmitic acid being increased by at least 1.5-fold or between 1.5-fold and 4-fold compared to control; or (vii) any combination of (i) to (vi).

[0119] In some embodiments, the 137 biomass comprises cells originating from the hypocotyl of a germinated seedling of Theobroma cacao accession number RUQ 137, and is referred to herein as "the 137-H biomass".

[0120] In some embodiments, the 137-H biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 10:1 (w / w) and 30: 1 (w / w); (ii) a Valine to Serine w / w ranging between 2: 1 (w / w) and 9: 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 1.5:1 (w / w) and 5: 1 (w / w); or (iv) any combination of (i) to (iii).

[0121] In some embodiments, the 137-H biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 25:1 (w / w) to 55: 1 (w / w); (ii) a total sugar content of 170 mg / gr to 350 mg / gr; (iii) a total sugar content being 7-fold to 25-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 70% or by 70% to 95% compared to control; (v) an amount of phenylalanine being increased by at 35% or by 35% to 70% compared to control (vi) an amount of threonine being increased by at 50% or by 50% to 70% compared to control; (vii) an amount of leucine being reduced by at least 75% or by 75% to 95% by weight compared to control; (viii) an amount of stearic acid being increased by at least 2.5-fold or between 2.5-fold and 7-fold compared to control; (ix) an amount of palmitic acid being increased by at least 2-fold or between 2-fold and 6-fold compared to control; or (x) any combination of (i) to (ix).

[0122] According to another aspect, there is provided a biomass of cells of Theobroma cacao accession number RUQ 1637 (‘Donor collection’ Centro Agronomico Tropical de Investigacion y Ensenanza (CATIE), Costa Rica). In some embodiments, the biomass of cells of Theobroma cacao accession number RUQ 1637 is referred to herein as "the 1637 biomass".

[0123] In some embodiments, the 1637 biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 8: 1 (w / w) and 20: 1 (w / w); (ii) a Valine to Serinew / w ranging between 2: 1 (w / w) and 6: 1; (iii) a pyroglutamic acid to lactic acid w / w ratio between 1.5: 1 (w / w) and 5: 1 (w / w); or (iv) any combination of (i) to (iii).

[0124] In some embodiments, the 1637 biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 30: 1 (w / w) to 80: 1 (w / w); (ii) a total sugar content of 90 mg / gr to 160 mg / gr; (iii) a total sugar content being 4-fold to 12-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 35% or by 35% to 60% compared to control; (v) an amount of phenylalanine being increased by at 40% or by 40% to 60% compared to control (vi) an amount of threonine being increased by at 15% or by 15% to 30% compared to control; (vii) an amount of leucine being reduced by at least 85% or by 85% to 99% by weight compared to control; (viii) an amount of stearic acid being increased by at least 2.5-fold or between 2.5-fold and 6-fold compared to control; (ix) an amount of palmitic acid being increased by at least 2-fold or between 2-fold and 6-fold compared to control; (x) an antioxidant activity being at least 3-fold greater or between 3-fold to 9-fold compared to a control; or (xi) any combination of (i) to (x).

[0125] According to another aspect, there is provided a biomass of cells of Theobroma cacao accession number RUQ 1632 (‘Donor collection’ International Cocoa Genebank, Trinidad (ICG,T), Trinidad and Tobago). In some embodiments, the biomass of cells of Theobroma cacao accession number RUQ 1632 is referred to herein as "the 1632 biomass".

[0126] In some embodiments, the 1632 biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 4: 1 (w / w); (ii) a Valine to Serine w / w ranging between 2: 1 (w / w) and 1 :2; (iii) a pyroglutamic acid to lactic acid w / w ratio between 5:1 (w / w) and 15: 1 (w / w); or (iv) any combination of (i) to (iii).

[0127] In some embodiments, the 1632 biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 20: 1 (w / w) to 50: 1 (w / w); (ii) a total sugar content of 90 mg / gr to 160 mg / gr; (iii) a total sugar content being 4-fold to 12-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 70% or by 70% to 90% compared to control; (v) an amount of phenylalanine being increased by at 45% or by 45% to 70% compared to control (vi) an amount of threonine being increased by at 35% or by 35% to 55% compared to control; (vii) an amount of histidine ranging between 1.5 mg / gr and 3 mg / gr; (viii) an amount of leucine being reduced by at least 35% or by 35% to 55% by weight compared to control; (ix) an amount of stearic acid being increased by at least 2-fold or between 2-fold and 6-fold compared to control; (x) an amount of palmitic acid being increased by at least 2-fold or between 2-fold and 4.5-fold compared to control; (xi) an antioxidant activity being at least 1.2-fold greater or between 1.2-fold to 2.5-fold compared to a control; or (xii) any combination of (i) to (xi). Insome embodiments, the 1632 biomass is further characterized by increased 5-HTP levels (e.g., at least 2, 3, 4, 5 -fold increase) compared to a control.

[0128] According to another aspect, there is provided a biomass of cells of Theobroma cacao accession number RUQ 905 (‘Donor collection’ International Cocoa Genebank, Trinidad (ICG,T), Trinidad and Tobago). In some embodiments, the biomass of cells of Theobroma cacao accession number RUQ 905 is referred to herein as "the 905 biomass".

[0129] In some embodiments, the 905 biomass of the invention is characterized by: (i) an Alanine to Valine weight per weight ratio (w / w) between 50: 1 (w / w) and 300: 1 (w / w); (ii) a Valine to Serine w / w ranging between 1 : 1 (w / w) and 1 :30; (iii) a pyroglutamic acid to lactic acid w / w ratio between 1 : 1 (w / w) and 1.5: 1 (w / w); or (iv) any combination of (i) to (iii).

[0130] In some embodiments, the 905 biomass is further characterized by: (i) a glucose to erythritol w / w ratio of 10:1 (w / w) to 30: 1 (w / w); (ii) a total sugar content of 150 mg / gr to 400 mg / gr; (iii) a total sugar content being 12-fold to 30-fold greater by weight compared to the total sugar content in a control; (iv) an amount of lysine being increased by at 80% or by 80% to 99% compared to control; (v) an amount of phenylalanine being increased by at 55% or by 55% to 80% compared to control; (vi) an amount of leucine being reduced by at least 85% or by 85% to 99% by weight compared to control; (vii) an amount of stearic acid being increased by at least 3-fold or between 3-fold and 9-fold compared to control; (viii) an amount of palmitic acid being increased by at least 2.5-fold or between 2.5-fold and 7-fold compared to control; (ix) an antioxidant activity being at least 1.2-fold greater or between 1.2-fold to 2.5-fold compared to a control; or (x) any combination of (i) to (ix).

[0131] The above-mentioned accession number are available and identifiable in icgd . reading . ac . uk / i cqc / li st . php .

[0132] In some embodiments, a seed comprises or is a seedling germinating therefrom.

[0133] In some embodiments, the dried biomass of Theobroma cacao cells of the invention is characterized by an ash content of not more than 0.1 mg / gr, 0.5 mg / gr, 1 mg / gr, 1.5 mg / gr, 2 mg / gr, 3 mg / gr, or 5 mg / gr by weight of the dried biomass of Theobroma cacao cells, or any value and range therebetween. Each possibility represents a separate embodiment of the invention.

[0134] Methods for determining % ash are common and would be apparent to one of ordinary skill in the art. Non-limiting examples for such methods include but are not limited to dry ashing and wet ashing, which can be accomplished by conventional means or by the use of microwave systems. Conventional dry ashing is based upon sample incineration at high temperatures (500- 600°C) in a muffle furnace. Wet ashing (acid-facilitated oxidation) is often used as a preparationfor specific elemental analysis by atomic absorption, inductively coupled plasma, and / or mass spectrometry. Microwave ashing (dry or wet) is faster than conventional methods and requires little additional equipment or space, but sample throughput may be a limiting factor.General

[0135] 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 1,000 nanometers (nm) refers to a length of 1,000 nm ± 100 nm.

[0136] 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. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as an 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.

[0137] 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."

[0138] 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 subcombination. 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 sub-combinations of the various embodiments and elements thereof are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0139] 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, whichare 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.EXAMPLES

[0140] Various embodiments and aspects of the present invention as delineated herein above and as claimed in the claims section below find experimental support in the following examples. Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non-limiting fashion.

[0141] Generally, the nomenclature used herein, and the laboratory procedures utilized in the present invention include chemical, molecular, biochemical, and cell biology 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); "Cell Biology: A Laboratory Handbook", Volumes I-III Cellis, J. E., ed. (1994); The Organic Chemistry of Biological Pathways by John McMurry and Tadhg Begley (Roberts and Company, 2005); Organic Chemistry of Enzyme-Catalyzed Reactions by Richard Silverman (Academic Press, 2002); Organic Chemistry (6th Edition) by Leroy "Skip" G Wade; Organic Chemistry by T. W. Graham Solomons and, Craig Fryhle.Materials and MethodsCocoa strains and calli

[0142] Cacao beans were obtained from the International Cocoa Quarantine Centre, at the University of Reading (ICQC,R) (icgd.reading.ac.uk / icqc / list.php).

[0143] Seeds were wiped with clean tissue paper and placed in a sterile culture box, soaked in 70% ethanol for 1 min, followed by rinsing with DDW. Then, seeds are incubated in sterilization solution (2% NaOCl, 0.1% Tween and 0.2% PPM) for 30 min with gentle agitation. Following the incubation, seeds were washed with a washing solution (0.2% PPM) 3 times, 10 min each rinse. Finally, the seeds were incubated in sterile 0.2% PPM for 3 hours.

[0144] To obtain a culture from specific plant parts, seeds were placed in sterile culture boxes lined with cotton wool and covered with 50 ml of imbibing solution (0.2% PPM), and incubated in low light 16 h light photoperiod, 25 °C, until germinated and desired organ was developed. Otherwise, the seed coat is cut with Bayonet forceps and a sterile scalpel and peeled off with jewelers forceps. Exposed seeds are cut and covered with a sterilization solution for 30 min and washed 3 times with sterile DDW, 5 min each time.

[0145] Seeds or explants are chopped into small pieces (~ 0.5 cm), using a sterile scalpel knife, and place in a callus induction medium plate (CIM; 1XDKW, 3% sucrose, 0.8% plant agar, 0.2 mg / ml zeatin, 300 mg / ml antibiotics, 0.2% PPM, and 200 mg / ml vancomycin). In certain cases, wherein the formed calli was not friable, 2 mg / ml AgNCL was added to the media. Plates were sealed and incubated in low light until calli started to form. Plates were routinely refreshed.

[0146] Once callus has been generated for a given tissue type (~ 1-2 grams fresh weight), it was transferred into a sterile plastic 125 ml Erlenmeyer flask with vented cap containing a callus induction medium (CIM). Flasks were then shaken at 90 rpm under low light at 25 °C. Every 10- 14 days cultures were fed by removing 80% of the spent medium (without taking the cells), which was replaced with fresh media. When the culture reached sufficient confluence, the volume was gradually increased. Fine suspension (e.g., solitary cells and / or cell clusters of not more than about 60 pm in diameter) was separated from the clusters in several rounds, by sub-culturing a saturated culture, allowing the fines cell culture to take over the suspension and establish a defined culture. Cell lines were routinely sub cultured and fed, as required.

[0147] For biomass collection, cells were filtered and washed with running tap water, freeze-dried by lyophilization until dry, and then stored in -20 °C until used.Trolox-equivalent antioxidant capacity (TEAC) assay

[0148] Twenty (20) mg of dried tissue were ground in 400 pL of absolute ethanol and incubated in room temperature for 30 minutes. The solution was centrifuged for 1 minute at maximum speed, and the supernatant was separated.

[0149] For Trolox-equivalent antioxidant capacity (TEAC) assay, 5 pl of the ethanol extract was analyzed as described in Pellegrini et al. (“Total antioxidant capacity of plant foods, beverages and oils consumed in Italy assessed by three different in vitro assays.” The Journal of nutrition vol. 133,9 (2003): 2812-9).High performance liquid chromatography (HPLC)

[0150] For metabolic profiling through HPLC analysis, freeze-dried tissues were weighed, transferred into 1.5 ml Eppendorf vials, and ground and shaken in presence of 80% MeOH containing 0.5 ppm Biochanin A as internal standard. Samples were then centrifuged and supernatant was filtered through Acrodisc® Syringe Filters with GHP Membrane, 13 mm with 0.2 pm (PALL, USA) and transferred to vials for liquid chromatography / quadrupole time-of- flight / mass spectrometry (LC-qTOF-MS) analysis.

[0151] The LC-MS analysis was carried out on an Agilent 1290 Infinity series liquid chromatograph coupled with an Agilent 6530C Quadrupole Time of Flight (qTOF) massspectrometer (MS) (Agilent Technologies, Santa Clara, USA). Compounds were separated on a InfLab Poroshell 120 EC-C18; 2.1 x 100 mm 2.7 micron (Agilent). The gradient elution mobile phase consisted of 1% ACN:99% H2O with 0.1% (v / v) formic acid (eluent A) and 99% ACN containing 0.1% (v / v) formic acid (eluent B). The column was equilibrated with 96.5% A at a flow rate of 0.4 mL / min for 1 min. Eluent B was then increased to 10% B at 2-3.5 min and increase gradually to 80 % B from 3.5 min to 14-16.5 min, then raised further to 100 % B by 16.5 min to wash column and restored to 96.5 % solvent A by 18.5 for re-equilibration. Eluting compounds were subjected to Jet Stream electrospray ionization interface operated in positive mode with the following settings: gas temp of 25 °C with a flow of 9 L / min and nebulizer set to 40 psi, sheath gas temp of 250 °C at 10 L / min flow. VCap set to 3000 V, Nozzle Voltage of 0 V, fragmentor set to 110 V and the skimmer to 65 V. Data were acquired with a 90-1200 mass / charge (m / z) range.

[0152] The exact mass of products was compared and matched to published data and expected values. The [M+H]; [M+Na] ions of target compounds were detected using the 'find compound by formula' function and analyzed by Masshunter qualitative and quantitative analysis software versions B.07.00 and B.10.00 accordingly (Agilent technologies). When standards were available, retention time (RT) of purchased authentic standards were also compared (AS).Gas chromatography mass spectrometry (GC-MS)

[0153] For metabolic profiling through GC-MS analysis, about 60 mg of the freeze-dried ground powder were mixed with pre-cooled mix (1 mL) that contain methanol / chloroform / water (5:2:2 v / v) along with 200 pl of Ribitol (Img / ml), vortexed and followed by addition of 100 pl of precooled methanol. The samples were incubated for 10 minutes at 25 °C on an orbital shaker, Sonicated for 10 min in ultra-sonication bath at room temperature, and then centrifuged at full speed for 10 min. The supernatant was transferred into a new 2 ml safe-lock Eppendorf tube. Three hundred (300) pL of doubly distilled water (ddEEO) and 300 pL of chloroform were added to the supernatant, vortexed for 10 sec and centrifuged for 5 min at full speed. Six hundred and fifty (650) pl of the upper phase were transferred into a new 2 ml safe-lock tube and stored in -80 °C after bubbled with argon gas until used.

[0154] For derivatization, 200 pL of the sample was dried in vacuum. Residues were redissolved and derivatized for 120 min at 37 °C in 40 pL of 20 mg / mL methoxyamine hydrochloride in pyridine) followed by a 30-min treatment with 70 pL N-methyl-N- (trimethylsilyl)trifluoroacetamide at 37 °C. Seven (7) pL of a retention time standard mixture (0.029% v / v n-dodecane, n-pentadecane, n-nonadecane, n-docosane, n-octacosane, n- dotracontane, and n-hexatriacontane dissolved in pyridine) was added prior to trimethylsilylation for analysis with a splitless mode. A 1 pL aliquot of the MSTFA / Pyridine extract was injected intoan Agilent 7890B / HES-5977B GC-MSD system (Agilent Technologies, Inc., Santa Clara, CA, USA). The instrument was equipped with Rtx-5 SIL-ms column (30 m length x 0.25 mm i.d., 0.25 pm film thickness, stationary phase 95% dimethyl- 5% diphenyl polysiloxane). Gradient of injection temperature (MMI) was from 70 °C to 300 °C in 14.5 °C / sec, the transfer line was 350 °C, and the ion source adjusted to 250 °C. The carrier gas used was helium set at a constant flow rate of 1 ml / min. The temperature program was 1 min isothermal heating at 70 °C, followed by a 1 °C / min oven temperature ramp to 76 °C, followed by a 6 °C / min oven temperature ramp to 340 °C, and a final 5 min heating at 340 °C. Mass spectra were recorded at 1.6 scans per second with a mass-to-charge ratio 70 to 550 scanning range. Post-column back-flushing was used during the post-run time in every injection to keep detector clean, column flow was reversed for few minutes to remove high-boiling components to inlet split vent. JetClean procedure was used after 1 pL set (15 injections) to keep the ion source clean, based on reductive hydrogen cleaning principle.

[0155] Spectral searching in the Masshunter software (Quantitative and Unknown analyses) against RI libraries downloadable from the Max -Planck Institute for Plant Physiology in Golm, (gmd.mpimp-golm.mpg.de) and finally normalized by the internal standard ribitol and tissue mass.

[0156] During cell establishment, feed was performed every 10-14 days by removing 80% of the medium without taking cells. Then, the culture was fed with the same volume of fresh culture media. If culture is growing, the volume was gradually increased to 50 ml, from feeding to feeding.

[0157] During maintenance, fine cultures are sub-cultured (‘subbed’) every 7 days, once reached to packed cell volume (PCV) = ± 7, and diluted to PCV=2, in a 250 ml flask to 50 ml liquid CCSM media (viability >80%). Aggregated cultures are subbed every 4 feedings.Feeding

[0158] Remove 20 ml of media and add 25 ml of fresh media. Once reached PCV=±5, and diluted to PCV=2, in a 250ml flask to 50ml liquid CCSM media (viability >80%).PCV measurement

[0159] Remove 10 ml of suspension while swirling the flask and place the 10 ml into a sterile 15 ml conical tube. Allow to settle for exactly 30 min for consistency, and record the volume of the settled cells (=PCV).Table 1. Control cocoa powders used in the current studyResults

[0160] The cocoa powder of the invention was found to include increased amounts of amino acids, such as essential amino acids, sugars, including total sugar content, and neurotransmitters, compared to control commercial cocoa powders.

[0161] Specifically, the inventors show that the cocoa powder of the invention includes increased amounts of glutamic acid (Fig. 7). Moreover, the inventors show that the cocoa powder of the invention includes increased amounts of the essential amino acids, lysine and threonine (Fig. 8).,

[0162] Further, the inventors compared the amounts of sugars in the cocoa powder of the invention to control commercial powders. Surprisingly, the results show that the cocoa powder of the invention includes an increased amount of total sugar content compared to the control (Fig. 9B). Specifically, cocoa powder the invention was found to include increased amounts of fructose, galactose, D-glucopyranose, glucose, and sucrose compared to the control cocoa powders (Fig. 9A). The biomass of T. cacao cells from which the cocoa powder is derived or obtained included the above-mentioned sugars in levels equivalent to or greater than in the control powder.Surprisingly, the equivalent or increased levels of these particular sugars were not lost in processing of the biomass of T. cocoa cells so as to obtain the cocoa powder of the invention.

[0163] Further, it was found that the biomass of T. cocoa cells and cocoa powder of the invention include neurotransmitters in an amount greater than control cocoa powder (Fig. 10). Specifically, dopamine and serotonin were found to be enriched in the biomass of T. cacao cells of the invention. Of notion is the finding that serotonin levels exceeded the serotonin content of commercial cocoa powder by ~20-fold (Fig. 10B). Dopamine levels were essentially maintained in the cocoa powder (processed from the biomass of T. cacao cells of the invention; Fig. 10A).

[0164] The current analysis of T. cacao cell lines revealed significant variations in amino acid and compound concentrations compared to traditional cacao products. Figure 1 illustrates the concentration of various amino acids, as analyzed by GC-MS, including Alanine, Valine, and Lysine, across different samples. Notably, certain cell lines exhibited higher concentrations of these amino acids compared to cacao powder and NIBS. Specifically, the T. cacao cell biomass of the invention was found to have an Alanine to Valine weight per weight ratio (w / w) between 1 : 1 (w / w) and 200: 1 (w / w), wherein an inverse correlation of these amino acids characterizes the controls (NIBS, and commercial cacao powder). Further, the T. cacao cell biomass of the invention was also found to have a Valine to Serine w / w ranging between 6: 1 (w / w) and 1 :25 (w / w). In this regard, the controls are characterized by a Valine content that exceeds the Serine content by at least 7-fold.

[0165] Further, the T. cacao cell lines of the invention were found to have increased concentrations of essential amino acids compared to control. Specifically, the concentrations of Lysine, Phenylalanine, and Threonine, were either equivalent or higher in most T. cacao cell lines of the invention (Figs. 2A-2C). Of notion are T. cacao cell lines: 847-C, 847-H, and 847-E, which showed Histidine levels of 0.05 to 0.5 mg / gr (Fig. 2D). In sharp contrast, this essential amino acid was not detected in the controls.

[0166] The inventors have further examined acids’ profile of the T. cacao cell lines of the invention. The T. cacao cell lines demonstrated a diverse acid profile, as observed with a GC-MS analysis, with some lines showing elevated levels compared to cacao powder and NIBS (controls). Specifically, the T. cacao cell biomass of the invention was found to have a pyroglutamic acid to lactic acid w / w ratio between 1 :1 (w / w) and 25: 1 (w / w), wherein an inverse correlation of these acids characterizes the controls (NIBS, and commercial cacao powder), i.e., about 1 : 1.4 (w / w) (Fig. 3). In this regard, pyroglutamic acid is required for the production / recycling of glutathione, which is one of the most potent anti-oxidants in the body, and is especially important in getting rid of toxins.

[0167] The inventors further examined and compared the sugar content in the T. cacao cell lines of the invention to controls (NIBS, and commercial cocoa powder), through GC-MS analysis. The results showed that all T. cacao cell lines of the invention comprise higher total sugar concentrations than the controls (Fig. 4A). Specifically, the T. cacao cell lines of the invention were found to be characterized by a glucose to erythritol w / w ratio of at least 5: 1 (w / w) (Fig. 4B). In sharp contrast, the controls were found to be characterized by glucose to erythritol w / w ratio of not more that 4: 1, or even inversed, for NIBS and cocoa powder, respectively (Fig. 4B).

[0168] Further to the above, the inventors showed that most T. cacao cell lines of the invention are superior to the controls with respect to antioxidant activity. Of notion are lines 847-E, 847-C, 847F-r, 847-H, 235, and 1637, with an antioxidant activity at least 2-fold greater than the controls (Fig- 5).

[0169] In that regard, T. cacao cell lines of the invention were found to be enriched with catechins, a group of secondary metabolites, as found through HPLC analysis. Catechins are known to have antioxidative activities in plants and other living systems. In particular, either fine or granular cultures of the T. cacao cell lines of the invention were found to include greater levels of one or more types of catechins, inclusive of: EGCG, Gallocatechin 3 -gallate, Epigallocatechin, and Catechin gallate (Figs. 6A-6D, respectively).

[0170] 5-Hydroxytryptophan (5-HTP) is a natural amino-acid derivative and is a direct precursor of serotonin, known to support mood balance, sleep quality, and stress regulation. 5-HTP has not been detected in conventional cacao beans, whereas 5-HTP levels were significantly high in the T. cacao cell lines compared to commercial cacao (Fig. ). In particular, 847 epicotyl line (the 847-E biomass) showed 22-fold higher 5-HTP levels compared to commercial cacao; 847 cotyledon (the 847-C biomass) showed 8-fold higher 5-HTP levels compared to commercial cacao; 847 hypocotyl (the 847-H biomass) showed 7-fold higher 5-HTP levels compared to commercial cacao; and 1632 line showed 5-fold higher 5-HTP levels compared to commercial cacao.

[0171] To this end, the T. cacao cells biomass of the invention were found to have increased amounts of essential amino acids, as well as antioxidant activity, compared to NIBS and commercial cocoa powder, therefore, the implementation of which in food products and / or ingredients thereof, seems to be advantageous.

[0172] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

CLAIMSWhat is claimed is:

1. A cocoa powder comprising at least one compound selected from the group consisting of: an essential amino acid, a sugar, a neurotransmitter, and any combination thereof, wherein said at least one compound is enriched in a biomass of Theobroma cacao cells compared to a control cocoa powder, and is derived therefrom.

2. The cocoa powder of claim 1, wherein said essential amino acid is lysine, threonine, or both.

3. The cocoa powder of claim 1 or 2, wherein said essential amino acid is present in said cocoa powder in an amount being greater by at least 1.5 compared to the amount of said essential amino acid in said control powder cocoa powder.

4. The cocoa powder of any one of claims 1 to 3, wherein said sugar is selected from the group consisting of: sucrose, fructose, glucose, galactose, D-glucopyranose, and any combination thereof.

5. The cocoa powder of any one of claims 1 to 4, wherein said sugar is present in said cocoa powder in an amount being greater by at least 1.4 compared to the amount of said sugar in said control powder cocoa powder.

6. The cocoa powder of any one of claims 1 to 5, characterized by a total sugar content being greater by at least 2 compared to a total sugar content in said control powder cocoa powder.

7. The cocoa powder of any one of claims 1 to 6, wherein said neurotransmitter is dopamine, serotonin, or both.

8. The cocoa powder of any one of claims 1 to 7, obtained or being derived from said biomass of T. cacao cells.

9. The cocoa powder of any one of claims 1 to 8, obtained or prepared by a method comprising fermenting said biomass of T. cacao cells.

10. The cocoa powder of claim 9, wherein said biomass of T. cacao cells comprises Alanine and Valine in a weight per weight ratio (w / w) ranging between 1 : 1 (w / w) and 200: 1 (w / w).

11. The cocoa powder of claim 9 or 10, wherein said biomass of T. cacao cells comprises Valine and Serine in a w / w ranging between 6: 1 (w / w) and 1 :25 (w / w).

12. The cocoa powder of any one of claims 9 to 11, wherein said biomass of T. cacao cells comprises pyroglutamic acid and lactic acid in w / w ratio ranging between 1 : 1 (w / w) and 25: 1 (w / w).

13. The cocoa powder of any one of claims 9 to 12, wherein said biomass of T. cacao cells comprises glucose and erythritol in a w / w ratio of at least 5:1 (w / w).

14. The cocoa powder of any one of claims 9 to 13, wherein said biomass of T. cocoa cells further comprises a total sugar content of at least 20 mg / gr of said biomass.

15. The cocoa powder of any one of claims 9 to 14, wherein said biomass of T. cocoa cells further comprises an antioxidant activity being at least 1.2-fold greater compared to a control biomass of T. cocoa cells.

16. The cocoa powder of any one of claims 9 to 15, wherein said biomass of T. cacao cells further comprises an amount of at least one catechin selected from the group consisting of: Epigallocatechin 3 -gallate (EGCG), Gallocatechin 3 -gallate, Epigallocatechin, Catechin gallate, and any combination thereof, being at least 3 -fold greater by weight compared to the amount of said at least one catechin in a control biomass of T. cocoa cells.

17. The cocoa powder of any one of claims 9 to 16, wherein said biomass of T. cocoa cells is devoid of caffeine, theobromine, or both.

18. The cocoa powder of any one of claims 9 to 17, wherein cells of said biomass of T. cocoa cells are obtained or derived from an epicotyl, hypocotyl, cotyledon, or any combination thereof, of a T. cocoa seed.

19. An edible composition comprising the cocoa powder of any one of claims 1 to 18.

20. The edible composition of claim 19, being a foodstuff or an ingredient thereof.

21. The edible composition of claim 20, wherein said foodstuff comprises any one of: chocolate, cake, brownie, and cookie, or any combination thereof.

22. The edible composition of claim 21, wherein said foodstuff is a beverage.

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