Methods of valorizing polysaccharides obtained from fruit-based starting material

Abstract

A method for increasing the value of a polysaccharide containing material obtained from a fruit that is not a pomegranate by generating a mixture of polysaccharide cleavage products. The method comprises (i) reacting a fruit-based starting material containing polysaccharide with a Fenton's reagent containing a peroxide agent and a metal, and (ii) cleaving the reacted polysaccharide with a cleavage agent to generate the mixture of polysaccharide cleavage products. The polysaccharide cleavage products generally contain higher amounts of oligosaccharides, lower average molecular weight, increased solubility, and decreased viscosity when dissolved in aqueous solution compared to the fruit-based starting material. Also provided are compositions and soluble fruit fiber comprising mixtures of polysaccharide cleavage products.

Description

METHODS OF VALORIZING POLYSACCHARIDES OBTAINED FROM FRUIT¬BASED STARTING MATERIALCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 561,002, filed March 4, 2024, which is hereby incorporated by reference in its entirety.FIELD

[0002] This invention relates generally to methods for valorizing polysaccharides obtained from fruit-based starting material by depolymerizing polysaccharides to generate mixtures of polysaccharide cleavage products.BACKGROUND

[0003] Many fruit types contain dietary fiber in their primary cell walls, and especially in the middle lamella which binds the primary cell walls of adjacent cells. The dietary fibers in the middle lamella usually include pectin-type fibers. Pectin-type fibers are rich in galacturonic acid but may take different forms. In one form, they are primarily homogalacturonans which are linear chains of a-(l-4)-linked D-galacturonic acid. In another form, the D-galacturonic acid residues in the a-(l-4)-linked chains have branches, such as for example D-xylose branches from a backbone of D-galacturonic acid residues.Rhamnogal acturonan I pectins (RG-I) are a further form. These have a backbone of the repeating disaccharide: 4)-a-D-galacturonic acid-(l,2)-a-L-rhamnose-(l). Usually, sidechains of various neutral sugars branch off from many of the rhamnose residues. The neutral sugars are mainly D-galactose, L-arabinose and D-xylose, with the types and proportions of neutral sugars varying with the origin of fruit. Another form of pectin is the rhamnogal acturonan II (RG-II) form. This is a complex, highly branched polysaccharide in which the backbone is made primarily of D-galacturonic acid units. Many pectin-type fibers obtained from fruits contain more than one of these forms and can contain them all. In all forms, many of the carboxyl groups of galacturonic acid are esterified with methanol. Fruit fibers also contain smaller amounts of hemicellulose fibers mainly comprised of xyloglucans, glucomannans and beta-glucans.

[0004] Significant amounts of fruit pomace are produced as a byproduct during the processing of fruits for jams, juices, fermented products, and the like. Often these pomacesare then used as animal feeds or fertilizers, even though they are a substantial source of functional components such as dietary fibers, polyphenols and the like. However, in certain instances, the pectins are extracted for uses in pharmaceutical, food, cosmetic, and other industries. These uses include acting as biopolymers, gelling agents, preservatives, antioxidants, etc. Key sources of pectin for these applications are apples and citrus fruits.

[0005] In certain fruits, pectin is found in the form of protopectin which is relatively insoluble with no gel-forming properties. Therefore, protopectin must be partially depolymerized to solubilize it sufficiently for extraction. A common method for solubilizing and extracting pectin from the pomace is by adding hot, dilute acid at pH values from 1.5 to 3.5. Various acids are used such as mineral acids (sulfuric, hydrochloric, nitric) and organic acids (citric, malic, oxalic). After several hours of treatment, the protopectin loses some of its branching and chain length and goes into solution. After filtering, the extract is concentrated, and the pectin is then precipitated by adding ethanol or isopropanol. Several other extraction techniques such as microwave-assisted extraction, ultrasound-assisted extraction, pulsed electric field extraction, subcritical water or CO2 extraction, and enzyme- assisted extraction, have also been proposed.

[0006] The pectins obtained from these extraction processes are in the form of large polysaccharides of a molecular weight typically in the range of 60000 to 130000 g / mol. Despite that pectins obtained from fruits offer advantages over cereal fibers for certain applications due to their better solubility, lower phytic acid content, and the presence of bioactive molecules, they are not often used as a source of dietary fiber. Instead, for food uses, they are most commonly used as gelling agents, thickening agents and stabilizers. Part of the reason is that they do gel and change the functional properties of foods. Also, due to their large size, they are not easy to fully solubilize, often leading to a gritty texture. This is especially the case at concentrations needed to provide a meaningful source of dietary fiber, for example sufficient to provide about 3 g per serving.

[0007] One method for improving the functionality of polysaccharides is to depolymerize them into shorter chain polymers such as oligosaccharides. Oligosaccharides are short chains of carbohydrates, usually having lengths in the range of 3 to 50 monomers. They are often soluble, more palatable, and easier to formulate than the original polysaccharides. Attempts have also been made to produce oligosaccharides from pectin. Because of the complex nature of pectins, these attempts have required pre-treatment to separate the various forms ofpectin from each other. Typical pre-treatment technologies include enzymatic, chemical, and physical treatments.

[0008] Enzymatic technologies are most common because of the specificity and selectivity of enzymes. However, due to the complexity of the pectin structure, several enzymes are needed. Methyl esters and acetyl groups on galacturonic acid residues are first removed using pectin methyl esterase and pectin acetyl esterase, respectively. Once they have been removed, an endo-polygalacturonase is then used to cleave the glycosidic bond of the a- (1— >4)-polygalacturonan in a random fashion. Exo-polygalacturonase can then be used to remove terminally (1-) linked Gal A residues from the non-reducing end of the homogalacturonan chains. The rhamnogal acturonan subunits can be separately degraded using rhamnogal acturonan hydrolase and rhamnogalacturonanlyase. These enzymes act on a-D- l ,4-Gal A-a-L- l ,2-Rha and a-L- l ,2-Rha -a-D- l ,4-Gal A linkages, respectively. Rhamnogal acturonan acetyl esterase can be used to remove acetyl groups and terminal rhamnosyl residues. The removal of side chains from rhamnogal acturonan-I can be achieved by a cocktail of various enzymes. Additional enzymes would be needed for any hemicellulose components of the fruit fibers, introducing additional complexity and cost.

[0009] Chemical hydrolysis of pectin has not been used extensively except for alkaline pretreatment which is generally used for the production of rhamnogal acturonan-I pectin. However, this approach has not generally achieved the desired degree of depolymerization. Physical pre-treatments like hydrothermal, dynamic high pressure microfluidization and irradiation have been tried. During hydrothermal pretreatment, pectin is partially hydrolyzed, and oligosaccharides can be effectively released from the biomass. Dynamic high pressure microfluidization, which is based on the principle of powerful shear, turbulence, impaction and cavitation, has been used on apple pectin. In general, these physical methods produce a wide range of mono-, di-, oligo- and polysaccharides which then require significant clean up and yield loss. The reproducibility of the technologies is also not clear.

[0010] A full description of the various technologies can be found in Babbar et al, 2016, Critical Reviews in Biotechnology (DOI: 10.3109 / 07388551.2014.996732). Also, a method for production oligosaccharides from carrot pectin using enzymes is described in patent application WO2019081523.

[0011] Recently new chemical methods for the depolymerization of polysaccharides using Fenton’s chemistry followed by cleavage using a base have been described in patentapplications WO2018236917 and WO2021097138. Additional depolymerization methods using Fenton’s chemistry are described in International patent application WO2023220318. The methods appear to be applicable to a range of polysaccharides. However, none of these applications describes depolymerization of dietary fibers extracted from complex polysaccharides which are rich in antioxidants, such as those obtained from many fruits. Further it is also unclear whether depolymerizing polysaccharides obtained from certain fruits will result in valorization of the polysaccharide.

[0012] Thus, there is a need for methods of valorizing polysaccharides obtained from fruit-based sources, and for oligosaccharides obtained from fruit-based sources.SUMMARY OF THE INVENTION

[0013] In one aspect, this invention provides a method for generating a mixture of polysaccharide cleavage products from a fruit-based starting material that is not pomegranate, the method comprising: (i) reacting one or more polysaccharide of a fruit-based starting material that is not pomegranate with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products. In preferred aspects, the fruit-based starting material is or comprises a pineapple-based starting material, an applebased starting material, or an orange-based starting material.

[0014] In this regard, the invention provides a method for depolymerizing polysaccharide containing material obtained from a fruit-based starting material to generate a mixture of polysaccharide cleavage products. In embodiments, fruit-based starting material is polysaccharide containing material, other than pomegranate, obtained from a fruit-based starting material which is optionally subjected to pre-processing prior to depolymerization as described herein. Polysaccharide containing material obtained from fruit-based starting material can be subjected to cleaning, drying or reduction of water content, pressing, cutting, chopping, grinding or other processes to adjust particle size, removal of juice, freezing, freeze-thawing, lyophilization, or any combination of the above prior to pre-treatment as described herein.

[0015] In an embodiment, the fruit-based starting material that is not pomegranate can comprise or be derived from any suitable fruit, including pineapple, apple, orange, lemon, lime, mango, or any combination thereof. In an embodiment, the fruit-based starting materialis or is derived from a polysaccharide source containing polyphenols. In an embodiment, the fruit-based starting material is or is derived from a polysaccharide source containing pectin.

[0016] In one aspect, this invention provides a method for generating a mixture of polysaccharide cleavage products from a pineapple-based starting material, the method comprising: (i) reacting one or more polysaccharide of a pineapple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0017] In another aspect, the invention provides a method for generating a mixture of polysaccharide cleavage products from an apple-based starting material, the method comprising: (i) reacting one or more polysaccharide of an apple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0018] In yet another aspect, the invention provides a method for generating a mixture of polysaccharide cleavage products from an orange-based starting material, the method comprising: (i) reacting one or more polysaccharide of an orange-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0019] In another aspect, the method for depolymerizing a polysaccharide of a fruit-based starting material further comprises pre-treating the polysaccharide prior to depolymerizing the polysaccharide. In an embodiment, pre-treatment involves extraction of polyphenols from the polysaccharide to provide a polyphenol-reduced fruit-based starting material containing polysaccharides. In an aspect, the polyphenols are extracted using water or an organic solvent. In embodiments, the organic solvent is an alkyl alcohol, optionally methanol, ethanol, n-propanol, or iso-propanol; a ketone, optionally acetone, or an alkyl ester; optionally ethyl acetate. In embodiments, the organic solvent for polyphenol removal is a mixture of chloroform and methanol. In embodiments, the organic solvent is a 1: 1 (v:v) mixture of chlorofornrmethanol. In embodiments, the organic solvent is acidified, for example by addition or an organic or mineral acid. In embodiments, the organic acid is a mono-, di-, or tricarboxylic acid. In embodiments, the tricarboxylic acid, citric acid, is used toacidify the organic solvent. In embodiments, the organic solvent contains 0.005 to 1% wt / v of the organic acid. In embodiments, the acidified organic solvent is acidified ethanol or acidified propanol. In embodiments, the acidified organic solvent is acidified with 0.01 to 0.1% wt / v citric acid. In embodiments, the acidified organic solvent is acidified with 0.01% to 0.02% wt / v citric acid. In embodiments, the acidified organic solvent is acidified with 0.05% to 0.15% wt / v citric acid. In embodiments, polyphenols are extracted with acidic ethanol or isopropanol containing 0.01% to 0.02% wt / v citric acid. In embodiments, polyphenols are extracted with acidic ethanol or isopropanol containing 0.05% -0.15% wt / v citric acid. In embodiments, polyphenols are extracted with 0.015% wt / v citric acid in ethanol or isopropanol. In embodiments, polyphenols are extracted with 0.1% wt / v citric acid in ethanol or isopropanol.

[0020] In embodiments, pre-treatment involves removal of protein from a polysaccharide of a fruit-based starting material. In embodiments, the proteins are removed using one or more proteases. In embodiments, proteins are removed by addition of two or more different proteases.

[0021] In embodiments, pre-treatment involves removal of one or more polysaccharides other than soluble fibers such as pectin. In embodiments, pre-treatment involves removal of starch and / or cellulose from a polysaccharide of a fruit-based starting material. In embodiments, starch is removed using one or more amylase or isoamylase. In embodiments, cellulose is removed using one or more cellulases. In embodiments, pre-treatment involves addition of one or more cellulases to the polysaccharide of the fruit-based starting material. In embodiments, pre-treatment involves addition of one or more glucanases to the polysaccharide of the fruit-based starting material. In embodiments, pre-treatment involves removal of monosaccharides and / or disaccharides. In embodiments, one or more disaccharidases are added to degrade disaccharides in the polysaccharide of the fruit-based starting material. In embodiments, the polysaccharide of the fruit-based starting material is treated with one or more of maltase, isomaltase, lactase, sucrase or combinations thereof to generate monosaccharides. In embodiments, monosaccharides present in or generated by enzyme treatment are removed from the polysaccharide of the fruit-based starting material. In embodiments, pre-treatment involves removal of lipids from the polysaccharide of the fruitbased starting material. In embodiments, pre-treatment involves removal of one or more of polyphenols, starch, cellulose, protein, lipid, monosaccharides, or disaccharides. Inembodiments, pre-treatment involves removal of polyphenols and one or more of starch, cellulose, protein, monosaccharides, or disaccharides.

[0022] In another aspect, pre-treatment involves extraction of polysaccharide from the fruit-based starting material, wherein the extracted polysaccharides provide a polysaccharide- enriched fruit-based starting material containing polysaccharides. In embodiments, pretreatment involves extraction of soluble fibers such as pectins and soluble hemicellulose polysaccharides and the extracted polysaccharides provide a soluble fiber-enriched fruitbased starting material containing polysaccharides, such as a pectin-enriched fruit-based starting material containing polysaccharides. In embodiments, polysaccharide extraction involves extraction using an acidic aqueous solution (e.g., water to which acid is added). In embodiments, polysaccharide extraction is extraction with water to which a mineral acid, e.g., sulfuric acid or hydrochloride acid is added. In embodiments, polysaccharide extraction is extraction with water to which an organic acid, e.g., acetic acid or citric acid is added. In embodiments, polysaccharide extraction employs acidic aqueous solution at pH of 1-3 (+ / - 0.5). In embodiments, polysaccharide extraction employs acidic aqueous solution at pH of 1- 2 (+ / -0.5). In embodiments, polysaccharide extraction employs acidic aqueous solution at pH of 1.0-1.5 (+ / -0.1). In embodiments, polysaccharide extraction employs acidic aqueous solution at pH of 1.2 (+ / -0.1). In embodiments, polysaccharide extraction employs acidic aqueous solution at pH of 1.0-1.5 (+ / -0.1) acidified with sulfuric or hydrochloric acid. In embodiments, polysaccharide extraction employs acidic aqueous solution at pH of 1.2 (+ / - 0.1) acidified with sulfuric or hydrochloric acid. In embodiments, extraction of polysaccharide separates polysaccharide from one or more of starch, cellulose, lignin, or protein. In embodiments, extraction of polysaccharide separates polysaccharide from one or more of starch, cellulose, lignin, lipids or protein. In embodiments, extraction of pectin separates pectin from one or more other polysaccharides, such as starch, cellulose or hemicellulose, and / or separates pectin from lignin, lipids or protein.

[0023] In another aspect, the method for depolymerizing a polysaccharide of a fruit-based starting material comprises or further comprises purifying the mixture of polysaccharide cleavage products to provide a purified mixture of polysaccharide cleavage products. In embodiments, the polysaccharide cleavage products are subjected to ethanol precipitation to remove ethanol-soluble components from the polysaccharide cleavage products. In embodiments, polysaccharide cleavage products are solubilized in appropriate solvent and thesolution is filtered to remove residual solids. In embodiments, salts and mono- and disaccharides are removed from the polysaccharide cleavage products by diafiltration.

[0024] The invention further provides mixtures of polysaccharide cleavage products having useful properties. In embodiments, the invention provides soluble fruit fiber useful in a variety of food, beverage, nutritional and pharmaceutical compositions. In embodiments, the invention provides soluble fruit fiber comprising a mixture of polysaccharide cleavage products, wherein a 20 g / L mixture of the polysaccharide cleavage product in water has turbidity of 40 nephelometric turbidity units (NTU) or less and / or a dynamic viscosity of 4 millipascal-second (mPa.s) or less at 25°C. In embodiments, the mixture of polysaccharide cleavage products contains 5% or more by weight of polysaccharide cleavage products having molecular weight less than 100 kDa. In embodiments, the mixture of polysaccharide cleavage products contains 40% or more by weight of polysaccharide cleavage products having molecular weight less than 100 kDa. In embodiments, the mixture of polysaccharide cleavage products of the soluble fruit fiber has a degree of polymerization (DP) of 3 to 100. In embodiments, a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 20 NTU or less, or a 20 g / L mixture of the soluble fruit fiber in water has dynamic viscosity of 2 millipascal-second (mPa.s) or less at 25°C, or both.

[0025] In embodiments, the mixture of polysaccharide cleavage products is formed by cleavage of a polysaccharide of a pineapple-based starting material, a pineapple-based starting material containing polysaccharides, a polyphenol-reduced pineapple-based starting material containing polysaccharides or a soluble fiber-enriched pineapple-based starting material containing polysaccharides.

[0026] In embodiments, the mixture of polysaccharide cleavage products is formed by cleavage of a polysaccharide of an apple-based starting material, an apple-based starting material containing polysaccharides, a polyphenol-reduced apple-based starting material containing polysaccharides or a soluble fiber-enriched apple-based starting material containing polysaccharides.

[0027] In embodiments, the mixture of polysaccharide cleavage products is formed by cleavage of a polysaccharide of an orange-based starting material, an orange-based starting material containing polysaccharides, a polyphenol-reduced orange-based starting material containing polysaccharides or a soluble fiber-enriched orange-based starting material containing polysaccharides.

[0028] In embodiments, the invention provides a composition comprising a mixture of polysaccharide cleavage products, wherein the composition is formed by:(i) reacting one or more polysaccharide of a fruit-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products. In embodiments, the fruit-based starting material comprises pomace, husk, seeds, rinds, peels, marcs, or any combination thereof. In a specific embodiment, the fruit-based starting material comprises peels.

[0029] In embodiments, the mixture of polysaccharide cleavage products contains 5% or more, by weight, of polysaccharide cleavage products with a molecular weight of less than 100 kDa. In embodiments, the mixture of polysaccharide cleavage products contains 10% or more, by weight, of polysaccharide cleavage products with a molecular weight of less than 100 kDa. In embodiments, the mixture of polysaccharide cleavage products contains 40% or more, by weight, of polysaccharide cleavage products with a molecular weight of less than 100 kDa. In embodiments, the mixture of polysaccharide cleavage products contains 60% or more, by weight, of polysaccharide cleavage products with a molecular weight of less than 100 kDa. In embodiments, each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products has a degree of polymerization (DP) of 3 to 100. In embodiments, each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products has a degree of polymerization (DP) of 3 to 50. In embodiments, the mixture of polysaccharide cleavage products of the composition in water at a concentration of 20 g / L has turbidity of 40 NTU or less and / or dynamic viscosity of 4 millipascal-second (mPa.s) or less at 25°C.

[0030] In embodiments of methods, soluble fruit fibers, and compositions, the mixture of polysaccharide cleavage products contains 40% or less by weight of polysaccharides with molecular weight of lOOkDa or more. In embodiments, the mixture of polysaccharide cleavage products generated contains 20% or less by weight of polysaccharides with molecular weight of lOOkDa or more. In embodiments, the mixture of polysaccharide cleavage products generated contains 10% or less by weight of polysaccharides with molecular weight of lOOkDa or more. In embodiments, the mixture of polysaccharide cleavage products generated contains 5% or less by weight of polysaccharides with molecular weight of lOOkDa or more. In embodiments, the mixture of polysaccharide cleavage productsgenerated contains less than 5% by weight of polysaccharide having molecular weight of 500 kDa or higher. In embodiments, the mixture of polysaccharide cleavage products generated contains less than 1% by weight of polysaccharide having molecular weight of 500 kDa or higher.

[0031] In an aspect, the polysaccharide of the fruit-based starting material is valorized by increasing the solubility of the fruit-based starting material. In embodiments, the mixture of polysaccharide cleavage products generated from the fruit-based starting material has a solubility in water or aqueous solution of 20 g / L or higher optionally with a turbidity below 20 NTU. In embodiments, the mixture of polysaccharide cleavage products has solubility in water or aqueous solution of 50 g / L or higher optionally with a turbidity below 40 NTU or below 20 NTU. In embodiments, the mixture of polysaccharide cleavage products has solubility in water or aqueous solution of 100 g / L or higher optionally with a turbidity below 40 NTU or below 20 NTU. In embodiments, the mixture of polysaccharide cleavage products has solubility in water or aqueous solution of 200 g / L or higher optionally with a turbidity below 40 NTU or below 20 NTU.

[0032] In an aspect, the polysaccharide of the fruit-based starting material is valorized by decreasing the viscosity of the fruit-based starting material. In embodiments, the mixture of polysaccharide cleavage products generated from the fruit-based starting material has a viscosity of less than 4 millipascal-second (mPa.s) at 25°C when dissolved in water at a concentration of at least 50 g / L (a 5% w / v aqueous solution). In embodiments, the mixture of polysaccharide cleavage products has a viscosity of less than 4 millipascal-second (mPa.s) at 25°C when dissolved in water at a concentration of 80 g / L. In embodiments, the viscosity of a 20 g / L water solution of the mixture of polysaccharide cleavage products is less than 4 mPa.s at 25°C. In embodiments, the viscosity of a 20 g / L water solution of the mixture of polysaccharide cleavage products is less than 2 mPa.s at 25°C. In embodiments, the mixture of polysaccharide cleavage products exhibits both low viscosity as described above and high solubility with low turbidity as described above.

[0033] In an aspect, the polysaccharide of the fruit-based starting material is valorized by increasing the prebiotic properties of the fruit-based starting material. In embodiments, the mixture of polysaccharide cleavage products has the ability, when consumed, alone or in combination with a food or beverage product, by an animal, to increase the relative abundance of beneficial bacteria in the gastrointestinal tract of the animal. In embodiments, the mixture of polysaccharide cleavage products increases the relative abundance of one ofmore of Anaerostipes, Bacleroides, Bifidobacterium, Blaulia, Faecalibacterium, Fusicatenibacter, and Prevotella in the intestinal microbiota of the animal. In embodiments, the mixture of polysaccharide cleavage products increases the relative abundance of one of more Bifidobacteria in the intestinal microbiota of the animal.

[0034] In an aspect, the polysaccharide of the fruit-based starting material is valorized by improving the organoleptic properties of the fruit-based starting material or of a food or beverage product containing the polysaccharide. For example, the mixture of polysaccharide cleavage products exhibits organoleptic properties suitable for its use in human beverages or liquid food products. In an embodiment, suitability in human beverages requires that beverage viscosity is not increased by the addition of the mixture of polysaccharide cleavage products to the point that a consumer will reject the product. In an embodiment, suitability in human beverages requires that beverage viscosity is not increased by the addition of the mixture of polysaccharide cleavage products to the point that a consumer will reject the product. In an embodiment, suitability in human beverages requires that a beverage does not acquire a gritty or grainy texture as measured by expert or consumer sensory analysis when the mixture of polysaccharide cleavage products is added. In an embodiment, suitability in human beverages requires that beverage clarity is not substantially decreased by the addition of the mixture of polysaccharide cleavage products as measured by turbidity or expert and consumer panels. In embodiments, the mixture of polysaccharide cleavage products exhibits reduced gritty or grainy texture alone or when combined in a beverage, an aqueous solution or a liquid or solid food product. In embodiments, an aqueous solution or slurry of the mixture of polysaccharide cleavage products exhibits reduced gritty or grainy texture. In embodiments, the mixture of polysaccharide cleavage products exhibits sweet taste. In some embodiments, a sweet taste is defined as having at least 10% of the sweetness of an equivalent amount of sucrose. For use in beverages, aqueous solutions or liquid or solid food products, the amount of the mixture of polysaccharide cleavage products added will vary with the application. In an embodiment, the amount of the mixture of polysaccharide cleavage products is added to a beverage, solution or liquid or solid food product ranges from O.lg / L to the 2000 g / L of beverage, solution or liquid or solid food product. In an embodiment, the amount of the mixture of polysaccharide cleavage products added to a beverage, solution or liquid or solid food product ranges from O.lg / L to the 200 g / L of beverage, solution or liquid or solid food product. In embodiments, the amount of the mixture of polysaccharide cleavage products added to a beverage, solution, or liquid or solid food product ranges from 0.5 g / L tothe solubility limit of the mixture of polysaccharide cleavage products in the beverage, solution, or liquid or solid food.

[0035] In additional embodiments of the methods, compositions, and soluble fruit fiber, the polysaccharide cleavage products comprise less than 10% by weight of residual monosaccharides, for example, less than 5% by weight of residual monosaccharide. In an embodiment, the residual monosaccharides comprise greater than 70% by weight of a combination of xylose, arabinose, galactose, and galacturonic acid. In another embodiment, the residual monosaccharides comprise greater than 70% by weight of a combination of galacturonic acid, xylose, and galactose. In another embodiment, the residual monosaccharides comprise greater than 70% by weight of a combination of arabinose, galactose, galacturonic acid, and xylose.

[0036] In embodiments, the optionally purified polysaccharide cleavage products contain glycosidic linkages comprising one or more of terminal arabinose, 3,5-linked arabinose, 5- linked arabinose, and 2,5-linked arabinose. In other embodiments, the optionally purified polysaccharide cleavage products contain glycosidic linkages comprising one or more of 4- linked galactose, terminal galactose, terminal xylose, and 3,4-linked galactose. In other embodiments, the optionally purified polysaccharide cleavage products contain glycosidic linkages comprising one or more of 4-linked galactose, terminal galactose, terminal arabinose, and 3,5-linked arabinose.

[0037] In an embodiment, the optionally purified polysaccharide cleavage products comprise monomeric saccharides of which greater than 70% by weight comprise galacturonic acid, glucose, and galactose. In a further embodiment, the monomeric saccharides further comprise rhamnose, xylose, and / or arabinose.

[0038] In an embodiment, galacturonic acid comprises 5% to 40% by weight of the monomeric saccharides, for example 6% to 35% by weight. In an embodiment, glucose comprises 5% to 45% by weight of the monomeric saccharides, for example 6% to 40% by weight. In an embodiment, galactose comprises 2% to 25% by weight of the monomeric saccharides, for example 2% to 20% by weight. In an embodiment, xylose comprises 2% to 50% by weight of the monomeric saccharides, for example 3% to 45% by weight. In an embodiment, arabinose comprises 1% to 40% by weight of the monomeric saccharides, for example 2% to 35% by weight.

[0039] In an embodiment, the optionally purified polysaccharide cleavage products contain glycosidic linkages comprising 4-linked glucose, 4-linked galactose, 6-linked galactose, 4-linked xylose, 3, 6-linked glucose, 3, 6-linked galactose, 4-linked rhamnose and 2, 3, 4-linked rhamnose.

[0040] In an embodiment, the optionally purified polysaccharide cleavage products comprise 43% by mass xylose, 26% by mass arabinose, 6% by mass galacturonic acid, 16% by mass galactose, and 6% by mass glucose.

[0041] In an embodiment, the optionally purified polysaccharide cleavage products comprise 4-linked xylose, terminal xylose, 2-linked xylose, terminal arabinose, 3,5-linked arabinose, 5-linked arabinose, 3% 2,5-linked arabinose, terminal galactose, and 4-linked glucose.

[0042] In an embodiment, the optionally purified polysaccharide cleavage products comprise 45% by mass xylose, 25% by mass arabinose, 6% by mass galacturonic acid, 14% by mass galactose, and 7% by mass glucose.

[0043] In an embodiment, the optionally purified polysaccharide cleavage products comprise 4-linked xylose, terminal xylose, 2-linked xylose, 2, 3, 4-linked xylose, 5-linked arabinose, 2,5-linked arabinose, terminal arabinose, 3,5-linked arabinose, terminal galactose, and 4-linked glucose.

[0044] In an embodiment, the optionally purified polysaccharide cleavage products comprise 6% by mass xylose, 35% by mass arabinose, 11% by mass galacturonic acid, 19% by mass galactose, 16% by mass glucose, 7% by mass mannose, and 2% by mass fructose.

[0045] In an embodiment, the optionally purified polysaccharide cleavage products contain glycosidic linkages comprising 4-linked xylose, terminal xylose, 2-linked xylose, terminal arabinose, 5-linked arabinose, 3,5-linked arabinose, terminal galactose, 4-linked galactose, 4-linked glucose, and terminal glucose.

[0046] In an embodiment, the optionally purified polysaccharide cleavage products comprise 3% by mass xylose, 2% by mass arabinose, 32% by mass galacturonic acid, 3% by mass galactose, 39% by mass glucose, and 20% by mass fructose.

[0047] In an embodiment, the optionally purified polysaccharide cleavage products comprise 3-linked glucose, 4-linked glucose, terminal glucose, 4-linked galactose, 3, 4-linked galactose, terminal galactose, 4-linked xylose, terminal xylose, and terminal arabinose.

[0048] In embodiments, the purified mixture of polysaccharide cleavage products contains less than 10% by weight polyphenols and preferably contains less than 5% by weight polyphenols. In embodiments, the purified mixture of polysaccharide cleavage products contains less than 10% by weight of proteins and preferably contains less than 5% by weight proteins. In embodiments, the purified mixture of polysaccharide cleavage products contains less than 10% by weight of mono- and / or disaccharides and preferably contains less than 5% by weight of mono- and / or disaccharides. In embodiments, the purified mixture of polysaccharide cleavage products contains less than 5% by weight ash and preferably contains less than 2% by weight ash. In embodiments, the purified mixture of polysaccharide cleavage products contains greater than 50% by weight or greater than 75% by weight of fiber. Preferably the purified mixture of polysaccharide cleavage products contains greater than 85% by weight of fiber. In embodiments, the purified mixture of polysaccharide cleavage products contains greater than 30% by weight of soluble fiber. In embodiments, the purified mixture of polysaccharide cleavage products contains greater than 50% by weight of soluble fiber. In embodiments, the purified mixture of polysaccharide cleavage products contains greater than 75% by weight of soluble fiber. In embodiments, the purified mixture of polysaccharide cleavage products contains greater than 85% by weight of soluble fiber.

[0049] In embodiments, the purified mixture of polysaccharide cleavage products comprises oligosaccharides, in which oligosaccharides having a degree of polymerization of 3 comprise 35% to 65% by weight of the oligosaccharides, for example 40% to 60% by weight of the oligosaccharides. In embodiments, the purified mixture of polysaccharide cleavage products comprises oligosaccharides, in which oligosaccharides having a degree of polymerization of 4 comprise 25% to 45% by weight of the oligosaccharides, for example 30% to 40% by weight of the oligosaccharides. In embodiments, the purified mixture of polysaccharide cleavage products comprises oligosaccharides, in which oligosaccharides having a degree of polymerization of 5 comprise 5% to 25% by weight of the oligosaccharides, for example 5% to 205% by weight of the oligosaccharides.

[0050] The invention is also directed to mixtures of polysaccharide cleavage products obtained from a fruit-based starting material, for example a pineapple-based starting material, prepared by the methods herein and having properties as described herein. In specific embodiments, the invention provides the mixture of polysaccharide cleavage productsdesignated as composition CLX136. In specific embodiments, the invention provides the mixture of polysaccharide cleavage products designated as composition CLX136S.

[0051] The invention is also directed to mixtures of polysaccharide cleavage products obtained from a fruit-based starting material, for example an orange-based starting material, prepared by the methods herein and having properties as described herein. In specific embodiments, the invention provides the mixture of polysaccharide cleavage products designated as composition CLX137C.

[0052] The invention is also directed to mixtures of polysaccharide cleavage products obtained from a fruit-based starting material, for example an apple-based starting material, prepared by the methods herein and having properties as described herein. In specific embodiments, the invention provides the mixture of polysaccharide cleavage products designated as composition CLX138.

[0053] The invention also provides prebiotic compositions and synbiotic compositions comprising the composition or soluble fruit fiber containing the mixture of polysaccharide cleavage products from fruit-based starting material. The prebiotic compositions stimulate growth of one or more beneficial bacteria. The synbiotic compositions comprise one or more beneficial bacteria.

[0054] The invention also provides a food, beverage, or nutritional or medicinal product comprising the compositions or soluble fruit fiber described herein. In embodiments, the medicinal products include medical foods or pharmaceutical products.

[0055] Other aspects and embodiments of the invention will be apparent to one of ordinary skill in the art on consideration of the detailed description, non-limiting examples and drawings provided.BRIEF DESCRIPTION OF THE DRAWINGS

[0056] FIG. 1 illustrates molecular weight distribution of composition CLX136, as measured by refractive index detection (RID).

[0057] FIG. 2 illustrates the molecular weight distribution of composition CLX136S, as measured by refractive index detection (RID).

[0058] FIG. 3 illustrates the molecular weight distribution of composition CLX137C, as measured by refractive index detection (RID).

[0059] FIG. 4 illustrates the molecular weight distribution of composition CLX138, as measured by refractive index detection (RID).

[0060] FIGs. 5A-5C illustrate the impact of CLX136 on the production of certain shortchain fatty acids (SCFAs) in ex-vivo human fecal samples. By the 24-hour timepoint, CLX136 promoted significant production of butyrate, acetate, and propionate compared to the untreated control. Selected results for butyrate production are shown in FIG. 5A where p = 0.0139, acetate production are shown in FIG. 5B where p = 1.77e-14; and propionate production are shown in FIG. 5C where p = 6.81e-9.

[0061] FIGs. 6A-6D illustrate the impact of CLX136 on the promotion of growth of butyrate-producing species in ex-vivo human fecal samples. By the 24-hour timepoint, CLX136 increased the relative abundance of Blautia (FIG. 6A), Anaerostipes (FIG. 6B), Faecalibacterium, (FIG. 6C) and Fusicatenibacter (FIG. 6D) in multiple fecal donors relative to the untreated control. In each of FIGs. 6A-6D, “UTRT” refers to the untreated control.

[0062] FIGs. 7A-7D illustrate the impact of CLX136 on the promotion of growth of beneficial bacteria in ex-vivo human fecal samples. By the 24-hour timepoint, CLX136 increased the relative abundance of beneficial bacterial taxa, including Bifidobacterium (FIG. 7A), Bacteroides (FIG. 7B), and Prevotella (FIG. 7C) species in multiple fecal donors relative to the untreated control, and prevented the expansion of the phylum Proteobacteria (FIG. 7D) relative to the untreated control. In each of FIGs. 7A-7D, “UTRT” refers to the untreated control.STATEMENTS REGARDING NOMENCLATURE

[0063] In general, the terms and phrases used in this specification have the meaning recognized in the art, which can be found by reference to standard texts, journal references and contexts known to those skilled in the art. For clarity, the following terms have the following meaning unless otherwise specified.

[0064] It will be understood by one of ordinary skill in the art that single numeric values or numeric values in a range, include slight variations or deviations from the stated value which may be used to achieve substantially the same results as the stated value. In cases, where a numeric value is one that is measured, it will be recognized that there is some level of uncertainty in the stated value due to experimental error, which can be determined by one of ordinary skill in the art. In circumstances where this definition cannot be applied to a givenstated value, is exceedingly difficult to apply, or wherein an uncertainty value is not specifically recited, then the numeric value has a reasonable deviation from the value, as known to a skilled person in the art. In embodiments, the reasonable deviation for a given value is + / -10% unless otherwise indicated.

[0065] “Ammonium bicarbonate” means solid ammonium bicarbonate, and / or an aqueous solution containing (i) ammonium and bicarbonate; (ii) ammonium, OH', and CO2, (iii) ammonia, H2O, and CO2, or (iv) any of the preceding and their equilibrium products.

[0066] “Ammonium hydroxide” means aqueous ammonium hydroxide and / or a solution containing (i) ammonia and H2O, (ii) ammonium and OH', (iii) ammonia and OH', or (iv) any of the preceding and their equilibrium products.

[0067] “Average molecular weight”, “average molecular mass”, “mean molecular weight”, “mean molecular mass”, or similar terms, refers to weight average molecular weight. Generally, unless otherwise specified, when a polysaccharide or a polysaccharide composition is described herein to have a specified average molecular weight (e.g., the mixture of polysaccharide cleavage products contains 40% or more by weight of polysaccharides with a molecular weight of less than 100 kDa), such values can be calculated with the aid of molecular weight analysis as described herein.

[0068] “Apple” refers to any part of the plant in the genus malus. "Apple" may refer to any of the cultivars of Malus domestica o Malus sieversii or other distinct species, varieties and hybrids. "Apple" may refer to fraction or extracts of the plant during harvest or food processing, non-limiting examples include apple skin, apple pulp, apple fiber, apple pomace, and apple seeds. "Apple" may refer to the solid material after roasting, fermentation, hot- water, enzymatic, chemical, alkaline, super critical fluid, sun drying, organic solvent, acidic, mechanical pressure or pressure-based extractions.

[0069] “Base” means a compound or collection of compounds that can accept hydrogen ions from a peroxyl oxidized carbohydrate, water, or non-aqueous solvent. Base can include Lewis bases, non-Arrhenius bases, strong-Arrhenius bases, weak-Arrhenius bases, other molecules that produce hydroxide ions through their decomposition, or other compounds that can accept hydrogen ions from a hydroperoxyl oxidized carbohydrate. Unless otherwise specified, base includes but does not specifically mean exclusively strong-Arrhenius bases (e g., Na+OH', K+0H', or Ca+2(OH')2).

[0070] “Biologically relevant increase” means a statistically significant increase as measured by parametric or non-parametric tests, generally in reference to the effects of a method comprising administering a polysaccharide composition or formulation thereof, relative to an otherwise identical method that does not include administering the polysaccharide composition or formulation thereof. In some aspects, a biologically relevant increase can be measured in feces, jejunum, cecum, ileum, stomach, large intestines, duodenum, mouth, respiratory tract, skin, urogenital tract, vaginal tract, or other microbial community. In some aspects, a biologically relevant increase is a 10% increase or a 5x increase or a lOx increase or a 50x increase or a lOOx or l,000x increase or 10,000x or more. In some aspects the biologically relevant increase can be in the absolute amount of a taxa or group of taxa, or amount of a given species. In some aspects, the biologically relevant increase can be the rate that a taxa, group of taxa, or other given species increases in the microbial community or in a subject (or location therein, such as a GI tract). In some aspects, the biologically relevant increase can be in the relative amount of a taxa, group of taxa, or other given species in a microbial community or in a subject (or location therein, such as a GI tract). In some aspects, an increase in abundance refers to the presence of one microbial taxa as compared to another microbial taxa, or one given species compared to another given species. “Biologically relevant decrease,” “biologically relevant change,” “biologically relevant amount,” and similar such terms can be similarly understood.

[0071] “Bronsted-Lowry base” means a compound or atom that can accept or bond to a hydrogen ion (e.g., methanol, formaldehyde, ammonia, etc.).

[0072] “Cleavage agent” or “cleavage reagent” means a single or a collection of strong Arrhenius bases, non-Arrhenius and / or weak-Arrhenius bases used to cleave polysaccharides after hydroperoxyl oxidation. In certain aspects, a cleavage agent or cleavage reagent breaks glycosidic bonds in the polysaccharide, which bonds may be present between any two saccharides of the polysaccharide. The cleavage reagent may also be, and preferably is, a peroxide quenching reagent, and in either case may be used in combination with an additional compatible peroxide-quenching agent that may or may not also be a cleavage agent. In some aspects, a cleavage reagent may be an enzyme, for example a glycosyl hydrolase, a lytic polysaccharide monooxygenase, a glycosyl transferase, transglycosidase, polysaccharide lyase, carbohydrate binding module, glycoysl transferase, carbohydrate esterase, a cocktail containing two or more of the aforementioned enzymes, or any enzyme that is carbohydrateactive. In some aspects, a cleavage reagent may be a solid-phase acid catalyst or a solid-phase base catalyst.

[0073] CLX136” means a mixture of polysaccharide cleavage products and / or an oligosaccharide containing composition in which 43% of the mass comprises xylose, 26% of the mass comprises arabinose, 6% of the mass comprises galacturonic acid, 16% of the mass comprises galactose, and 6% of the mass comprises glucose, as measured by hydrolytic monosaccharide compositional analysis, and as depicted in Table 1. In embodiments, CLX136 includes the subunits and weight percentages as listed. In embodiments, CLX136 includes the subunits and weight percentages within + / - 10% of the listed amounts. In embodiments, CLX136 includes the subunits and weight percentages within + / - 5% of the listed amounts. In embodiments, CLX136 includes subunits and weight percentages within + / - 1% of the listed amounts. In embodiments, the glycosidic linkage composition of CLX136 is as shown in Table 5, where the listed values are + / - 10%. In embodiments, the glycosidic linkage composition comprises 20% 4-linked xylose, 16% terminal arabinose, 13% 3,5-linked arabinose, 7% terminal galactose, 6% terminal xylose, 4% 2-linked xylose, 4% 4-linked glucose, 4% 5-linked arabinose, 3% 2,5-linked arabinose, and 5% other minor linkages shown in Table 5, again where each value is + / - 10%. The approximate molecular weight distribution of CLX136 comprises the values set forth in Table 6, as measured by refractive index detection (RID) (see also Figure 1), where each value is + / - 10%. CLX136 is generally derived from a pineapple-based starting material, but can also be derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP mono- and / or oligo-saccharides) that provide oligosaccharides that have the same (or substantially the same, e.g., values within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolytic monosaccharide composition, glycosidic linkage composition, and oligosaccharide analysis as CLX136. In embodiments, CLX136 is derived from pineapple fiber. In embodiments, CLX136 is prepared by the methods described in Example 1.

[0074] CLX136S” means a mixture of polysaccharide cleavage products and / or an oligosaccharide containing composition in which 45% of the mass comprises xylose, 25% of the mass comprises arabinose, 6% of the mass comprises galacturonic acid, 14% of the mass comprises galactose, and 7% of the mass comprises glucose, as measured by hydrolytic monosaccharide compositional analysis, and as depicted in Table 2. In embodiments, CLX136S includes the subunits and weight percentages as listed. In embodiments, CLX136Sincludes the subunits and weight percentages within + / - 10% of the listed amounts. In embodiments, CLX136S includes the subunits and weight percentages within + / - 5% of the listed amounts. In embodiments, CLX136S includes subunits and weight percentages within + / - 1% of the listed amounts. In embodiments, the glycosidic linkage composition of CLX136S is as shown in Table 5, where the listed values are + / - 10%. In embodiments, the glycosidic linkage composition of CLX136S comprises 23% 4-linked xylose, 15% terminal arabinose, 12% 3,5-linked arabinose, 6% terminal galactose, 7% terminal xylose, 4% 2- linked xylose, 3% 4-linked glucose, 4% 5-linked arabinose, 3% 2,5-linked arabinose, 3% 2, 3, 4-linked xylose, and 5% other minor linkages shown in Table 5, again where the listed values are + / - 10%. The approximate molecular weight distribution of CLX136S composition comprises the values set forth in Table 6, as measured by refractive index detection (RID) (see also Figure 2), where each value is + / - 10%. CLX136S is generally derived from a pineapple-based starting material, but can also be derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP mono- and / or oligosaccharides) that provide oligosaccharides that have the same (or substantially the same, e.g., values within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolytic monosaccharide composition, glycosidic linkage composition, and oligosaccharide analysis as CLX136S. In embodiments, CLX136S is derived from pineapple fiber. In embodiments, CLX136S is prepared by the methods described in Example 2.

[0075] CLX137C” means a mixture of polysaccharide cleavage products and / or an oligosaccharide containing composition in which 6% of the mass comprises xylose, 35% of the mass comprises arabinose, 11% of the mass comprises galacturonic acid, 19% of the mass comprises galactose, 16% of the mass comprises glucose, 7% of the mass comprises mannose, and 2% of the mass comprises fructose, as measured by hydrolytic monosaccharide compositional analysis, and as depicted in Table 3. In embodiments, CLX 137C includes the subunits and weight percentages as listed. In embodiments, CLX137C includes the subunits and weight percentages within + / - 10% of the listed amounts. In embodiments, CLX137C includes the subunits and weight percentages within + / - 5% of the listed amounts. In embodiments, CLX137C includes subunits and weight percentages within + / - 1% of the listed amounts. In embodiments, the glycosidic linkage composition of CLX137C is as shown in Table 5, where the listed values are + / - 10%. In embodiments, the glycosidic linkage composition of CLX137C comprises 3% 4-linked xylose, 22% terminal arabinose, 6% 3,5- linked arabinose, 6% terminal galactose, 4% terminal xylose, 3% 2-linked xylose, 11% 4-linked glucose, 3% 5-linked arabinose, 4% terminal glucose, 10% 4-linked galactose, and 13% other minor linkages shown in Table 5, again where the values are + / - 10%. The approximate molecular weight distribution of CLX137C composition comprises the values set forth in Table 6, as measured by refractive index detection (RID) (see also Figure 3), where each value is + / - 10%. CLX137C is generally derived from an orange-base starting material, but can also be derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP mono- and / or oligo-saccharides) that provide oligosaccharides that have the same (or substantially the same, e.g., values within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolytic monosaccharide composition, glycosidic linkage composition, and oligosaccharide analysis as CLX137C. In embodiments, CLX137C is derived from orange peel. In embodiments, CLX137C is prepared by the methods described in Example 3.

[0076] CLX138” means a mixture of polysaccharide cleavage products and / or an oligosaccharide containing composition in which 3% of the mass comprises xylose, 2% of the mass comprises arabinose, 32% of the mass comprises galacturonic acid, 3% of the mass comprises galactose, 39% of the mass comprises glucose, and 20% of the mass comprises fructose, as measured by hydrolytic monosaccharide compositional analysis, and as depicted in Table 4. In embodiments, CLX 138 includes the subunits and weight percentages as listed. In embodiments, CLX138 includes the subunits and weight percentages within + / - 10% of the listed amounts. In embodiments, CLX138 includes the subunits and weight percentages within + / - 5% of the listed amounts. In embodiments, CLX138 includes subunits and weight percentages within + / - 1% of the listed amounts. In embodiments, the glycosydic linkage composition of CLX138 is as shown in Table 5, where the listed values are + / - 10%. In embodiments, the glycosidic linkage composition comprises 12% 3-linked glucose, 28% 4- linked glucose, 23% terminal glucose, 6% 4-linked galactose, 2% 3, 4-linked galactose, 5% terminal galactose, 2% 4-linked xylose, 3% terminal xylose, 2% terminal arabinose, and 10% other minor linkages shown in Table 5, again where the values are + / - 10%. The approximate molecular weight distribution of CLX138 composition comprises the values set forth in Table 6, as measured by refractive index detection (RID) (see also Figure 4), where each value is + / - 10%. CLX138 is generally derived from an apple-based starting material, but can also be derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP mono- and / or oligo-saccharides) that provide oligosaccharides that have the same (or substantially the same, e.g., values within 10%, or within 15%, orwithin 20%, or within 25%, or within 30%) dynamic viscosity, hydrolytic monosaccharide composition, glycosidic linkage composition, and oligosaccharide analysis as CLX138. In embodiments, CLX138 is derived from apple pectin.

[0077] "Comprising" means including. Related terms such as "comprise" and "comprised" are to be interpreted in the same manner.

[0078] “Degree of polymerization” or “DP” refers to the total number of monosaccharide units that are part of a particular carbohydrate polymer. For example, a tetra oligosaccharide has a DP of 4. When used to describe a group of saccharides or polysaccharide cleavage products (e.g., a polysaccharide composition or a mixture of polysaccharide cleavage products), DP generally refers to the mean DP of the polysaccharides in the composition. In some aspects, the DP of a polysaccharide is referred to as “DP#”, where “#” corresponds to an integer representing the total number (or average number if used to describe a group of polysaccharides) of sugar monomer units (e.g., “DP3” means a degree of polymerization of 3). For saccharides, saccharide compositions, and polysaccharide cleavage products discussed herein having a DP between the range of 3-9, the recited DP includes a variance of up to ±2 monomer units of the recited value. For saccharides, saccharide compositions, and polysaccharide cleavage products having a DP of 10 or greater, the recited DP includes a variance of up to ±20% of the recited value, for example, an oligosaccharide having a DP of 20 may have a DP of 16, 17, 18, 19, 20, 21, 22, 23, or 24.

[0079] “Depolymerize” or “Depolymerization” and variants of this word refer generally to reacting a specific polysaccharide to reduce the number of monomer units therein, typically by cleavage of one or more glycosidic bonds. Such cleavage also results in a decrease in molecular weight of the polysaccharide. Such cleavage can result in generation of one or more polysaccharides of lower molecular weight, one or more oligosaccharides or one or more monosaccharides or combinations thereof. Depolymerization of a mixture of polysaccharides results in a mixture of polysaccharide cleavage products having a lower DP or lower average molecular weight than the original mixture. The result of such cleavage on mixtures of polysaccharides can also be described in terms of a weight % of polysaccharides therein having molecular weight greater than a selected values, e.g., a polysaccharide composition having less than 5% (or 2% or 1%) by weight of polysaccharides having molecular weight greater than 500 kDa (or 100 kDa or 50 kDa). In an embodiment, depolymerization can be affected by treatment with a Fenton’s reagent followed by cleavingthe reacted polysaccharide with a cleavage agent. As is apparent from examples herein, the extent of depolymerization of polysaccharides achieved varies with reaction conditions.

[0080] “Fenton’s reagent” means a reagent comprising a peroxide agent and a metal. In certain aspects, the peroxide agent is hydrogen peroxide. In certain aspects, the metal is Fe(II), Fe(III), Cu(I), Cu(II), Mn(II), Zn(II), Ni(II), and Co(II), alkaline earth metals Ca(II) and Mg(II), the lanthanide Ce(IV) or any combination thereof. The term, “high concentration Fenton’s reagent” and like terms refers to a Fenton’s reagent having a higher concentration of peroxide agent and / or metal relative to standard concentrations typically required to initiate deploymerization of a polysaccharide in the methods described herein. In an embodiment, the high concentration Fenton’s reagent comprises a higher concentration of peroxide agent, e.g., 0.1% v / v more, 0.2% v / v more, 0.5% v / v more, or 1% v / v more than the standard concentration typically required to depolymerize a polysaccharide. In an embodiment, the high concentration Fenton’s reagent comprises a higher concentration of a metal, e.g., 0.0005% w / v more, 0.001% w / v more, 0.0015% w / v more, or 0.002% w / v more than the standard concentration typically required to depolymerize a polysaccharide. Said higher concentration will vary depending on the starting material. For example, for a pineapplebased starting material, a high concentration Fenton’s reagent may comprise a final peroxide agent concentration of 6% v / v or more and / or a final metal species concentration of 0.018% w / v or more where a standard concentration Fenton’s regent comprises a final peroxide agent concentration of 5% v / v or more and / or a final metal species concentration of 0.016% w / v.

[0081] “Fiber” or “dietary fiber” is a carbohydrate that cannot be digested by a subject, particularly by an animal or human subject. Fiber cannot be broken down into mono- or disaccharides by the subject, and instead it passes through the body undigested by the subject. Fiber may, however, be digested by organisms in the microbiota of the subject. In embodiments, the mixture of polysaccharide cleavage products contains fiber. The amount of fiber present in a given composition can be determined by methods that are well known in the art. Fiber can be characterized as soluble or insoluble. “Insoluble fiber” is not soluble in water and generally is described as passing through the human gastrointestinal tract unchanged. “Soluble fiber” is generally described as attracting water or being soluble in water in the colon. Soluble fiber can also form a gel or gel-like substance in the colon. Soluble fiber can be degraded by bacteria in the colon. Even though fruit-based dietary fiber may contain soluble fiber, it is generally in the form of long-chain polysaccharides which generally makes it unsuitable for use in human foods and beverages. This invention providessoluble fruit fiber that exhibits enhanced water solubility with low turbidity and / or low viscosity in water solutions. More specifically, the invention provides soluble fruit fiber wherein a 20 g / L mixture of the polysaccharide cleavage product in water has turbidity of 40 NTU or less or dynamic viscosity of 4 millipascal-second (mPa.s) or less at 25°C, or both. The soluble fruit fiber as described herein is generally suitable for use in human foods and beverages as well as in various nutritional and pharmaceutical applications.

[0082] “Free monosaccharide compositional analysis” refers to the method described in Amicucci, Galermo et al. 2019, the invention of which is incorporated by reference herein for all purposes and specifically for a description of that method, with some modifications. The derivatization reaction to produce monosaccharides is performed at the optimized condition of 70°C for 30 minutes. Samples are run on an Agilent 1290 Infinity II ultra-high performance liquid chromatography (UHPLC) system coupled to an Agilent 6490A triple quadrupole (QqQ) mass spectrometer. Separation is carried out on an Agilent InfinityLab Poroshell HPH-C18 column (2.1 mm x 50 mm, 1.9 pm particle size) plus a guard column (5 mm) with the same solvent system described in the paper. With a constant flow rate of 1.2 mL / min, an isocratic gradient of 8.5% B is used for the first 4-minute elution period, followed by 15% B for 0.4 minutes. For the flush period, 97% B was held for 1 minute. The column thermostat is set at 35°C. For the mass spectrometry parameters, the only change from the method described in Amicucci, Galermo et al. 2019 is that the fragmentor voltage is set at 380V. For data analysis, the hydrolysis correction factor is not applied since the samples contain oligosaccharides instead of polysaccharides. In this analysis method, inherently free unpolymerized monosaccharides are calculated by quantifying the concentrations of 14 monosaccharides (glucose, galactose, fructose, xylose, arabinose, fucose, rhamnose, glucuronic acid, galacturonic acid, N-acetylglucosamine, N- acetylgalactosamine, mannose, allose, ribose) against their individual standard curves. For example, 30% free glucose, as measured by free monosaccharide compositional analysis, means containing 30 g of glucose per 100 g of the sum of all 14 monosaccharides described above.

[0083] “Fruit-based starting material containing polysaccharide” or “fruit-based starting material” refers to a whole fruit or a fraction, extract, or residue of a fruit, other than pomegranate, which contains polysaccharide. Fruit-based starting material is a polysaccharide source for the methods, compositions, and soluble fruit fibers described herein. The fruit-based starting material is a polysaccharide containing material derived fromany suitable fruit, other than pomegranate. In embodiments, the fruit-based starting material is, or is derived from, pomes, drupes, berries, melons, citrus fruits, pineapple, or a combination thereof. In embodiments, fruit-based starting material refers to fractions, extracts, or residues of fruit (other than pomegranate) during harvest or food processing, nonlimiting examples include, leaves, stems, seeds, peels, rinds, marcs, husk, pomace, flesh, or any combination thereof. In preferred embodiments, fruit-based starting material is a pineapple, an apple, or an orange, or fractions, extracts, or residues thereof. In these embodiments, the fruit-based starting material refers to a pineapple-based starting material, an apple-based starting material, and / or an orange based-starting material. In an embodiment, fruit-based starting material comprises two or more different fruits or fractions, extracts, or residues from two or more different fruits. In methods herein, a fruit-based starting material is treated to generate mixtures of polysaccharide cleavage products. More generally, the fruitbased starting material is treated to depolymerize at least a portion of the polysaccharides therein. The fruit-based starting material of this method can be any form of fruit fractions or extracts. In embodiments, fruit-based starting material comprises, consists of, or consists essentially of pectin. Other soluble fiber sources can also be present in the fruit-based starting material, such as hemicellulose fibers including xyloglucans, glucomannans and / or betaglucans. Fruit-based starting material can also, in addition to polysaccharide, contain, polyphenols, lignin, protein, or lipids, among others.

[0084] In embodiments, fruit-based starting material is or is derived from a fruit or fruit residue having polyphenols. In embodiments, fruit-based starting material is or is derived from a fruit having elevated levels of polyphenols relative to other fruits. Fruit-based starting material may be treated, for example, to reduce the level of polyphenols therein providing a “polyphenol-reduced fruit-based starting material.” In embodiments, the level of polyphenol in the “polyphenol-reduced fruit-based starting material” is reduced by 50% or more by weight compared to fruit -based starting material. In embodiments, the level of polyphenol in the “polyphenol-reduced fruit-based starting material” is reduced by 80% or more by weight compared to fruit-based starting material. In embodiments, the polyphenol level in polyphenol-reduced fruit-based starting material is 15% or less by weight. In embodiments, the polyphenol level in polyphenol-reduced fruit-based starting material is 10% or less by weight. In embodiments, the polyphenol level in polyphenol-reduced fruit-based starting material is 5% or less by weight. Use of the polyphenol-reduced fruit-based starting materialin Fenton-based depolymerization methods herein has been found to result in higher yields of polysaccharide cleavage products.

[0085] Fruit-based starting material can be treated to enhance the level of one or more polysaccharides therein. Fruit-based starting material can be treated to extract one or more polysaccharides and the extracted one or more polysaccharides can then be employed as a “polysaccharide-enriched fruit-based starting material.” The level of polysaccharides in the polysaccharide-enriched fruit-based starting material is generally higher than in fruit-based starting material containing polysaccharides. In embodiments, the level of polysaccharides is enhanced by 10% or more by weight compared to fruit-based starting material. In embodiments, the level of polysaccharides is enhanced by 25% or more by weight compared to fruit-based starting material. In embodiments, the level of polysaccharides is enhanced by 50% or more by weight compared to fruit-based starting material. In specific embodiments the polysaccharide that is extracted is pectin and a pectin-enriched fruit-based starting material is provided.

[0086] The amount of polysaccharide and its composition, as well as, the properties of the polysaccharides, including average molecular weight and DP and extent and kind of esterification, in the fruit-based starting material as well as the concentration of other components therein, will vary depending upon the source of the starting material, type of processing, components of the residue, among other variables. In embodiments, the fruitbased starting material contains at least 1% by weight (dry weight) polysaccharides and preferably contains 5%, 7% or 10% or more of polysaccharides. Polysaccharide obtained from certain fruit-based starting materials, such as an apple-based starting material, can contain pectin, starch, cellulose, galactomannan, xylan, arabinoxylan, xyloglucan and other types of polysaccharide. Polysaccharides in the fruit-based starting material can be esterified, for example by methyl or acetyl groups. The degree of esterification of polysaccharide will vary dependent, at least, upon the source of the fruit-based starting material and the components in the residues thereof.

[0087] Fruit-based starting material is optionally pre-treated, as described herein, prior to treatment with Fenton’s reagent. Pre-treatment can include, among others, removal or extraction of polyphenols; removal of starch, removal of cellulose, removal of lignin, removal of protein, removal of lipids, precipitation steps, and various filtration steps. Pretreatment, can include various extractions (e.g., with organic solvent, with acidic organic solvent, with aqueous acid), various enzyme treatments or combinations of such treatments(e.g., with one or more protease, with one or more endo- or exo-peptidase, with one or more amylase (including isoamylase), with one or more cellulase, with one or more glucanase, with one or more pectinase, with one or more xylanases, with one or more mannanases, or with one or more arabinanses, with one or more disaccharidases (e.g., maltase, isomaltase, lactase, or sucrose)). Pre-treatment can also include deesterification of polysaccharide (e.g., by chemical or enzymatic deesterification). In embodiments, pre-treatment includes a step of removal of polyphenols, particularly by extraction with organic solvent and more specifically by extraction with acidic alcohol. In embodiments, pre-treatment includes a step of extraction of polysaccharides, particularly pectin, from polysaccharide containing material. In embodiments, hemicellulose polysaccharides are also extracted. In embodiments, polysaccharide is extracted from polysaccharide containing material using extraction with an acidic aqueous solution. In embodiments, such polysaccharide extraction can function to separate polysaccharide from protein, lignin, and lipid. In embodiments, pre-treatment, includes a step of extraction of polyphenols and a step of extraction of polysaccharides from fruit-based starting material. In embodiments, pre-treatment does not include pre-treatment with a pectinase. When used with reference to a pre-treatment step the terms “remove” or “removing” refer to extraction and / or enzymatic degradation of a given component (e.g., polyphenols, starch, etc.) and include any measurable decrease in the given component. It will be appreciated that in embodiments, it is preferred to obtain a significant reduction in the component, for example a reduction of 10%, 25% or 50% or more of the component. In some embodiments, the component can be reduced to a non-detectible level. In some embodiments, the component is reduced to a level that such that yield of the depolymerization reaction is increased. In some embodiments, the level of polyphenol in the polysaccharide starting material is reduced to a level that such that yield of the depolymerization reaction is increased.

[0088] “Fruit residue” means by-products of processing of a fruit other than pomegranate fruit. In some aspects, the fruit residue is the fruit-based starting material for the methods, compositions, and soluble fruit fiber described herein. The residue can be generated by any processing of the fruit, such as roasting, fermentation, or various extraction processes so long as some polysaccharide, and in embodiments pectin, remains. In particular, fruit residue is generated as the by-product of juicing. Fruit residue can contain any portion of the fruit or a combination of such portions. Non-limiting examples include the husk which has a hard outer rind or exocarp or peel and a spongy mesocarp with attached sarcotestas membranescontaining arils, arils containing seeds, pomace, and the like. Pomace can be the residue of juicing or other processing of the arils of the fruit or of juicing any portion of the fruit, combination thereof, or the whole fruit. Fruit residue generally contains one or more polysaccharides unless treated to release or extract a given polysaccharide. In specific embodiments, fruit residue contains pectin. In specific embodiments, fruit residue contains hemicellulose polysaccharides. In specific embodiments, fruit residue contains polyphenols. In keeping with this embodiment, the fruit residue may comprise an elevated level of polyphenols relative to other fruit residues, such as 200 mg per 100 g of fresh mass of the residue or greater, 250 mg per 100 g of fresh mass of the residue or greater, 400 mg per 100 g of fresh mass of the residue or greater, or 500 mg per 100 g of fresh mass of the residue or greater. Fruit residue can also contain in addition to polysaccharides, polyphenols, lignin, protein, or lipids, among others. Fruit residue is optionally dried (e.g., air dried, oven dried or freeze-dried), chopped and / or processed into a powder and / or formed into pellets. Particles of fruit residue can be ground or otherwise processed to have a selected size-range.

[0089] Gastrointestinal tract” or “GI tract” means the passageway in the digestive system of a subject (e.g., animal, including humans) that includes all components from the esophagus to the anus (inclusive), as well as everything situated along the passageway including the stomach, intestines, and so forth. Generally, “gastrointestinal tract” is used interchangeably herein with the term “gut.”

[0090] “Glycosidic linkage composition”, “glycosidic linkage analysis”, “permethylated linkage composition analysis” or similar terms, refer to a method described in Galermo, Nandita et al. 2018, incorporated by reference herein in its entirety for all purposes and specifically for a description of such analysis, with some modifications. The permethylation reaction time is 30 minutes. Samples are run on an Agilent 1290 Infinity II UHPLC system coupled to an Agilent 6490A QqQ mass spectrometer. Separation is carried out on an Agilent InfinityLab Poroshell HPH-C18 column (2.1 mm * 100 mm, 1.9 pm particle size) plus a guard column (5 mm) with the same solvent system described in Galermo, Nandita et al. 2018. With a constant flow rate of 0.8 mL / min, an isocratic gradient of 14% B is used for the 16-minute elution period, followed by a 2-minute 99% B flush period. The column thermostat is set at 35°C. The glycosidic linkage composition is calculated by integrating the chromatographic peak area of all peaks with the following m / z values: 481.2, 495.2, 509.2, 523.3, 525.2, 537.3, 539.3, 553.3, 567.3, 581.3. For example, 20% 4-galactose, as measuredby the permethylated linkage composition analysis, refers to the peak area of 4-galactose being 20% of the sum of the peak area of all linkage peaks with the m / z values listed above.

[0091] “Hemicellulose fibers” or “hemicellulose polysaccharides” are saccharide polymers found in plant cell walls. Hemicellulose fibers are generally comprised of one or more of the following four fiber groups: 1) xylans, such as xylan and arabinoxylan; 2) mannans, such as glucomannan and galactomannan; 3) beta-glucans; and 4) xyloglucans.

[0092] “Hydrolytic monosaccharide compositional analysis” means the method described in Amicucci, Galermo et al. 2019, incorporated by reference in its entirety for all purposes, with the following modifications. The hydrolysis reaction to produce monosaccharides is performed at the optimized condition of 100°C for 2 hours. Samples are run on an Agilent 1290 Infinity II ultra-high performance liquid chromatography (UHPLC) system coupled to an Agilent 6490A triple quadrupole (QqQ) mass spectrometer. Separation is carried out on an Agilent InfinityLab Poroshell HPH-C18 column (2.1 mm x 50 mm, 1.9 pm particle size) plus a guard column (5 mm) with the same solvent system described in Amicucci, Galermo et al. 2019. With a constant flow rate of 1.2 mL / min, an isocratic gradient of 8.5% B is used for the first 4-minute elution period, followed by 15% B for 0.4 minutes. For the flush period, 97% B is held for 1 minute. The column thermostat is set at 35°C. For the mass spectrometry parameters, the only change from the method described in Amicucci, Galermo et al. 2019 is that the fragmentor voltage is set at 380V. For data analysis, the hydrolysis correction factor is not applied nor needed since the samples contain oligosaccharides instead of polysaccharides. In this analysis method, monosaccharide composition is calculated by quantifying the concentrations of 14 monosaccharides (glucose, galactose, fructose, xylose, arabinose, fucose, rhamnose, glucuronic acid, galacturonic acid, N-acetylglucosamine, N- acetylgalactosamine, mannose, allose, ribose) against their individual standard curves. For example, 30% glucose, as measured by hydrolytic monosaccharide compositional analysis, means containing 30 g of glucose per 100 g of the sum of all 14 monosaccharides described above.

[0093] “Increases abundance of’ or “increasing abundance of’ refers to a biologically relevant increase in the population of a certain bacterial taxa.

[0094] “Lewis base” means a compound or atom that can donate electron pairs (e.g., F’, benzene, H', pyridine, acetonitrile, acetone, urea, etc.).

[0095] “Linkage ratio”, “linkage peak area ratio”, “ratio of linkage” or other similar terms refer to any number of comparisons dependent upon the relationships observed in the glycosidic linkage composition analysis. Peak area for each linkage is calculated on a relative percent basis of the peak area in relationship to the summation of all other linkage peaks areas observed. Peak area ratios are calculated by dividing one contributing linkage by any other linkage of the same monosaccharide within the composition.

[0096] “Microbiota”, “microflora”, “microbiome” and “microbial community” mean a community of living microorganisms that typically inhabits a bodily organ or part, for example the gastro-intestinal organs of complex organisms, such as mammals and humans. In particular, the most dominant members of the gastrointestinal microbiota include microorganisms of the phyla of Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, Synergistetes, Verrucomicrobia, Fusobacteria, and Euryarchaeola: at genus level Anaerostipes, Bacteroides, Faecalibacterium, Fusicatenibacter, Bifidobacterium, Prevotella, Roseburia, Alistipes, Collinsella, Blautia, Coprococcus, Ruminococcus, Eubacterium and Dorea, at species level Bacteroides uniformis, Alistipes putredinis, Parabacteroides merdae, Ruminococcus bromii, Dorea longicatena, Bacteroides caccae, Bacteroides thetaiotaomicron, Eubacterium hallii, Ruminococcus torques, Faecalibacterium prausnitzii, Ruminococcus lactaris, Collinsella aerofaciens, Dorea formicigenerans, Bacteroides vulgatus and Roseburia intestinalis. The gastrointestinal microbiota includes the mucosa- associated microbiota, which are located in or attached to the mucous layer covering the epithelium of the gastrointestinal tract, and luminal-associated microbiota, which are found in the lumen of the gastrointestinal tract.

[0097] “Modulate”, “modulating”, “modulation” or other similar terms refer to the ability of a compound (e.g., oligosaccharide composition, or mixture of polysaccharide cleavage products) to alter the amount, degree, or rate of a biological function (including metabolite production), the progression of a disease, or amelioration of a condition. For example, modulating can refer to the ability of a compound to increase or decrease the abundance of a microorganism, increase or decrease production of a metabolite, or elicit a decrease in the inflammation, pain, incidence, or severity of a symptom associated with a particular condition or disease (e.g., associated with the gastrointestinal system). In embodiments, the modulation is a biologically relevant change.

[0098] “Molecular weight analysis” or “SEC -RID” or similar terms, refer to a method in which samples are prepared by reconstituting dried powders into a 10 mg / mL solution inHPLC grade water. Samples are analyzed on an Agilent Infinity II 1260 RID coupled to an Agilent Infinity II 1260 HPLC. Separation is performed on an Agilent AdvanceBio SEC column. Chromatographic solvents consisted of A: HPLC grade water and B: 95% acetonitrile in water (v / v). The RID is operated in positive signal polarity mode and at a 2.31 Hz peak width. Samples are integrated using the manual integration tool in ChemStation data analysis.

[0099] “Monosaccharide ratio”, “monosaccharide peak area ratio”, “ratio of monosaccharide” or similar terms refer to any number of the comparisons dependent upon the relationships observed in the hydrolytic monosaccharide compositional analysis. Absolute concentrations of each monosaccharide are calculated on a relative percent basis in relation to the summation of all other monosaccharides observed. Monosaccharide ratios are calculated by dividing one contributing monosaccharide by any other monosaccharide within the composition. Monosaccharide ratios are not intended to limit the composition to the listed monosaccharides. For example, a glucose:galactose ratio of 1: 1 means that there are roughly equal amounts of glucose subunits and galactose subunits in the composition, but the composition may also comprise mannose subunits, rhamnose subunits, or any other subunit.

[0100] “Nitrogen-based” means a compound that contains at least one nitrogen atom with four substituent groups that can contain any combination of lone pairs of electrons, hydrogens, or carbon atoms (e.g., ammonia, sodium amide, trimethylamine, di ethylamine, N,N-Diisopropylethylamine, urea, pyridine, ammonium hydroxide, ammonium bicarbonate, etc.). Exemplary nitrogen-based, peroxide-quenching, polysaccharide-cleavage agents are listed in Table A. A nitrogen-based reagent may have an unsubstituted or substituted ammonium group and can be present in neutral and / or ionic forms.

[0101] Table A. Exemplary polysaccharide (PS)-cleavage, and / or peroxide-quenching agents.

[0102] “NMR HSQC Analysis”, “1H-13C HSQC NMR”, “HSQC spectra” or other similar terms mean the data generated from two-dimensional spectral analysis of a sample via a Heteronuclear Single Quantum Coherence (HSQC) spin coupling of protons and bonded carbons present in the sample. HSQC experimentation depends on the solvation of samples in a deuterated solvent such as D6-DMSO or D2O. An HSQC spectrum contains a unique peak for each proton attached to the heteronuclear carbon atom being considered, allowing for identification of molecular structure of the analyzed sample. Each experiment is conducted with a Bruker AVANCE 600MHz NMR using heteronuclear single quantum coherence (HSQC) to illustrate the correlation between the 1H and 13C chemical shifts through 1JCH coupling. The resulting FIDs are processed using Bruker TopSpin 4.1.3 and the experimental chemical shifts are utilized to determine oligosaccharide structures and the anomeric characteristics of the glycosidic bonds with the aid of the CASPER program. Relative ratios between alpha and beta bonds are calculated through examination of the 2D 1H-13C HSQC via examination of signal strength in Hz. These values are then compared to determine the percent abundance of each linkage type among the same carbohydrate. NMR samples are dried via lyophilization, and the resulting material is then dissolved in 0.75mL of dimethyl sulfoxide-d6 (DMSO-d6) with a 0.03% (v / v) TMS internal standard at a concentration of 20mg / mL at a 4.5-6pH range.

[0103] “Non-Arrhenius base” means a compound or atom that can donate electrons (e.g., Lewis Bases), accept protons (e.g., Bronstead-Lowry Bases), or releases hydroxide ions through its decomposition (NH4HCO3), but does not qualify as an Arrhenius base.

[0104] “Oligosaccharide” means an oligomer of monosaccharides, in which the DP of the oligomer is between 2 and 30 monosaccharide units, such as between 3-30, 3-20, 3-15, 3-10, 3-8, 3-6, or 5-15, or any subrange thereof, monosaccharide units. An oligosaccharide can belinear, branched, primarily linear with pendant saccharide monomers, or any combination thereof. An oligosaccharide is an individual oligomer chain.

[0105] “Oligosaccharide composition”, “oligosaccharide pool” or “oligosaccharide mixture” means a mixture of two or more oligosaccharides, each of which can be the same or different from one another. Although efforts have been made to consistently use the terms “oligosaccharide” and “oligosaccharide composition” according to their preceding definitions, the intended meaning will be clear from context when such terms are used herein. In embodiments, “one or more oligosaccharides” refers to an oligosaccharide mixture when more than one oligosaccharide is present. In embodiments, “one or more oligosaccharides” refers to one oligosaccharide. In some embodiments, an oligosaccharide composition comprises one or more polysaccharides. In keeping with this aspect, an oligosaccharide composition may comprise up to 60-80% polysaccharides by mass, optionally less than 70% or less than 60% polysaccharides by mass (e.g., between 0.5% and 70% polysaccharides, between 0.5% and 60% polysaccharides, or between 0.5% and 50% polysaccharides). In embodiments, an oligosaccharide composition comprising up to 60-80% polysaccharides has a higher solubility, increased bioactivity, or a combination thereof, as compared to a composition comprising between 80-100% polysaccharides. Mixtures of polysaccharide cleavage products are examples of oligosaccharide compositions.

[0106] “Oligosaccharide analysis” or “oligosaccharide composition analysis” (or similar terms) refer to a HPLC-quadrupole time-of-flight (Q-TOF) method described in Amicucci, Nandita et al. 2020, incorporated by reference in its entirety for all purposes, with some modifications. For sample preparation, oligosaccharides are reduced by incubation with 2.0 M NaBH4 for 1 hour at 65 °C. The oligosaccharides are purified using C-18 cartridge 96- well plates: the plates are washed with 100% ACN, and the oligosaccharides are loaded and eluted with water. The oligosaccharides are subsequently purified using porous graphitized carbon (PGC) 96-well plates: PCG plates are washed with 80% acetonitrile and 0.1% (v / v) TFA in water, and the oligosaccharides from C-18 purification are loaded and washed with water. The oligosaccharides are eluted with 40% acetonitrile with 0.05% (v / v) TFA. Samples are completely dried by evaporative centrifugation and reconstituted for mass spectrometry analysis. Instrumentation is performed on an Agilent 1260 Infinity II HPLC coupled to an Agilent 6530 Q-TOF mass spectrometer. Using the same stationary (plus a 5 mm guard column) and mobile phase as described in Amicucci, Nandita et al. 2020, separation is carried out using the following gradient: 2-15% B, 0-20 minutes; 15-60 % B; 20-45 minutes. Thecolumn thermostat is set at 35°C. The fragmentor voltage is set at 75V. In this method, the “oligosaccharide weight %” (or “oligo wt%” or such terms) is calculated by dividing the chromatographic peak area of a particular oligosaccharide by the total peak area of all oligosaccharides identified in that sample during the defined chromatographic period. Generally, when an oligosaccharide composition is described to contain a specified weight percent of oligosaccharides on a dry basis having a degree of polymerization of a specified number (e.g., at least 50 wt. % oligosaccharides on a dry basis having a degree of polymerization of between 3 and 50 monosaccharide subunits), such values can be calculated with the aid of the oligosaccharide analysis described above; however, other methods can also aid this determination, such as size exclusion chromatography using a universal detector, or other methods known in the art.

[0107] “Orange” refers to any part of the plant in the genus Citrus. "Orange" may refer to Citrus maxima, Citrus reticulata, Citrus sinensis, Citrus aurantium, Citrus bergamia Risso, Citrus trifoliata or other distinct species, varieties and hybrids. "Orange" may refer to fraction or extracts of the plant during harvest or food processing, non-limiting examples include orange rind, orange pith orange pulp, orange fiber, orange pomace, or orange seeds. "Orange" may refer to the solid material after roasting, fermentation, hot-water, enzymatic, chemical, alkaline, super critical fluid, sun drying, organic solvent, acidic, mechanical pressure or pressure based extractions. "Orange" may refer to other non-citrus genus, which are colloquially known as sweet orange, bitter orange, Bergamot orange, Trifoliate orange or mandarin orange.

[0108] “Organoleptic” means a quality (such as taste, color, odor, and feel) of a substance that stimulates the sense organs. Organoleptic properties include among others taste (sweetness, sourness, bitterness, saltiness, umami or savory), mouthfeel or texture (the way a food or drink feels when put in the mouth and can include among others hardness / softness, chewy, gummy, slimy, gritty / grainy, crisp, and crunchy), smell or color. Organoleptic properties of a food or beverage are often assessed by expert taste testers or panels of consumers.

[0109] Other minor linkages” means the sum of linkages which are either not entirely annotated or constitute less than 2% of any samples. Therefore, the contributions of these linkages to the sample glycosidic linkage composition are summed into the “other minor linkages” category.

[0110] “Pectin” means a heteropolysaccharide found in the primary lamella, in the middle lamella, and in the cell walls of plants. Pectin is rich in galacturonic acid but may take different forms such as: 1) homogalacturonans which are linear chains of a-(l-4)-linked D- galacturonic acid; 2) branched galacturonans in which the D-galacturonic acid residues in the a-(l-4)-linked chains have branches, such as for example D-xylose branches from a backbone of D-galacturonic acid residues; 3) Rhamnogal acturonan I pectins (RG-I) which have a backbone of the repeating disaccharide: 4)-a-D-galacturonic acid-(l,2)-a-L-rhamnose- (1), often with sidechains of various neutral sugars branching off from many of the rhamnose residues; and 4) rhamnogal acturonan II pectins (RG-II) which are complex, highly branched polysaccharides in which the backbone is made primarily of D-galacturonic acid units. In embodiments, pectins comprise more than one of the forms. In embodiments, pectins comprise all the forms.

[0111] “Peeling reaction” or “peeling” means the sequential alkaline degradation of carbohydrates through a mechanism that releases monomeric units from the reducing end of the polymer.

[0112] “Peroxide agent” means a compound that contain oxygen-oxygen bonds that can produce, natively, with light, temperature, or catalyst (e.g., metals and enzymes), R-O. and / or R-O-O. species, where “R” refers to a hydrogen or carbon group that is attached to the rest of the molecule. In one aspect, a peroxide agent is hydrogen peroxide.

[0113] “Peroxide quenching reagent” means a compound or atom which is not a strong- Arrhenius base, and that can convert hydrogen peroxide, peroxyl radicals, and hydroperoxyl radicals to a less reactive or non-reactive state (e.g., ammonium hydroxide, ammonium bicarbonate, ammonia, etc.). In certain aspects, a peroxide quenching reagent converts hydrogen peroxide as well as radicals produced from hydrogen peroxide to less reactive species (e.g. water). In certain aspects, a peroxide quenching reagent may reduce the hydrogen peroxide concentration to zero, below 5 mg / L, below 10 mg / L, below 25 mg / L, or below 50 mg / L. In certain aspects, a peroxide quenching reagent may form water, hydroxide ions, or oxygen gas. In certain aspects, the peroxide quenching reagent may be an enzyme, for example, a catalase. The enzyme can be of microbial origin, from recombinant origin, or from animal origin, for example from bovine liver. In certain aspects, different enzymes may be mixed to quench the peroxide species.

[0114] “Pineapple” refers to any part of the plant in the genus Ananas. "Pineapple" may refer to any of the cultivars of Ananas comosus. or other distinct species, varieties and hybrids. "Pineapple" may refer to fraction or extracts of the plant during harvest or food processing, non-limiting examples include pineapple skin, pineapple core, pineapple pulp, pineapple fiber, or pineapple pomace. "Pineapple" may refer to the solid material after roasting, fermentation, hot-water, enzymatic, chemical, alkaline, super critical fluid, sun drying, organic solvent, acidic, mechanical pressure or pressure-based extractions.

[0115] “Polysaccharide” means a polymer of monosaccharide units having greater than 30 monosaccharide units, or a material comprising such a polymer. The polysaccharide can be linked to other non-carbohydrate moieties (e.g., glycoproteins, proteoglycans, glycopeptides, glycolipids, glycoconjugates, glycosides, or any combination thereof). The polysaccharide can be a linear polymer, branched polymer, primarily linear polymer with pendant saccharide monomers, or any combination thereof.

[0116] “Polysaccharide cleavage product” refers to a product formed from the chemical and / or enzymatic cleavage of a polysaccharide obtained from a fruit-based starting material other than pomegranate. In some aspects polysaccharide cleavage products comprises one or more polysaccharides. In some aspects, polysaccharide cleavage products comprises one or more oligosaccharides and one or more polysaccharides. In preferred embodiments of the methods herein, polysaccharide cleavage products are generated from a fruit-based starting material containing polysaccharides, such as a pineapple-based starting material, an applebased starting material, or an orange-based starting material. In some aspects, the mixture of polysaccharide cleavage products comprises 1% or more oligosaccharides by weight, such as 1% or more, 5% or more, 10% or more, or 15% or more oligosaccharides by weight. In some aspects, the mixture of polysaccharide cleavage products comprises 5% or more oligosaccharides by weight, such as 5% or more, 10% or more, 15% or more, or 20% or more oligosaccharides by weight. In some aspects, the mixture of polysaccharide cleavage products comprises 10% or more oligosaccharides by weight, such as 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more oligosaccharides by weight. In this regard, the mixture of polysaccharide cleavage products is an example of an oligosaccharide composition as defined herein.

[0117] “Polysaccharide composition”, “polysaccharide pool” or “polysaccharide mixture” means a mixture of two or more polysaccharides, each of which can be the same or different from one another. Although efforts have been made to consistently use the terms“polysaccharide” and “polysaccharide composition” according to their preceding definitions, the intended meaning will be clear from context when such terms are used herein. In embodiments, “one or more polysaccharides” refers to a polysaccharide mixture when more than one polysaccharide is present. In embodiments, “one or more polysaccharides” refers to one polysaccharide. In some aspects, polysaccharide composition refers to a composition comprising, or consisting of, polysaccharide cleavage products. In methods herein polysaccharides are obtained from a fruit-based starting material. In preferred embodiments of methods herein, polysaccharides are obtained from a pineapple-based starting material, an apple-based starting material, and / or an orange-based starting material.

[0118] “Polysaccharide source containing pectin” means a source of polysaccharides which includes pectin polysaccharides. The source of polysaccharides can also include other types of polysaccharides. The source of polysaccharides can be a residue, fraction, or extract obtained from a fruit. Suitable fruits include apples, pears, berries, citrus fruits such as oranges, lemons, pineapples, grapefruits, and the like. In preferred embodiments, the suitable fruits include apples, oranges, and pineapples.

[0119] “Polysaccharide source containing polyphenols” means a source of polysaccharides which includes polyphenols. The source of polysaccharides containing polyphenols can be a residue, fraction, or extract obtained from a fruit. Suitable fruits include apples, pears, berries such as blueberry, raspberry, blackberry, and raspberry, citrus fruits such as pineapples, and the like. In preferred embodiments, the suitable fruits include apples, oranges, and pineapples.

[0120] “Pomegranate” means any part of the fruit of the plant Punica granatum.

[0121] “Prebiotic” means a selectively fermented composition that results in specific changes in the composition and / or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health. In some aspects, a prebiotic is generally a non-digestible or partially-digestible (i.e., digestible by the subject / human / animal, and does not include digestion by microbes) food ingredient that beneficially affects a host when ingested by selectively stimulating the growth and / or the activity of one or a limited number of microbes in the gastrointestinal tract or other portion of the host.

[0122] “Purified” means a molecule or composition which is at least partially or substantially purified. A substantially purified molecule or composition has a purity of 95%, optionally for some applications 99%, optionally for some applications 99.9%, optionally forsome applications 99.99%, and optionally for some applications 99.999% pure. The terms “purified” and “isolated” are used interchangeably.

[0123] “Reaction mixture” means a mixture comprising reagents which may react chemically to form products which are distinct from the reagents.

[0124] The term “remove,” or “removing” relates to reducing or reduction of a component in a material.

[0125] "Reducing" or any variation of the term such as “reduction” means any measurable decrease to achieve a desired effect. In aspects, “reducing” refers to a biologically relevant decrease as defined herein.

[0126] “Retention factor” means the ratio obtained by dividing the retention time of a given peak observed in an oligosaccharide analysis (e.g., HPLC spectrum) by the first oligosaccharide peak (i.e., the lowest retention time) observed in the oligosaccharide analysis.

[0127] Short chain fatty acid” or “SCFA” includes one or more of butyrate, propionate, beta-hydroxybutyrate, lactate, or acetate.

[0128] “Specified reaction time” or “reaction time” means the time for a reaction to proceed toward an equilibrium state between reagents added and products produced by the reaction of the reagents. In certain aspects, specified reaction time allows sufficient time to reach an equilibrium. In certain other aspects, specified reaction time, while allowing time for the reaction to proceed toward equilibrium, does not provide the time needed to reach equilibrium.

[0129] “Strong- Arrhenius base” means a compound that completely dissociates in water to release one or more hydroxide ions into solution. Examples of a “strong- Arrhenius base” are KOH, NaOH, Ba(OH)2, CsOH, Sr(OH)2, Ca(OH)2, LiOH, and RbOH.

[0130] “Subject” or “patient” generally refer to a living organism suffering from or prone to a disease or condition that can be treated by administration of a compound or pharmaceutical composition, as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goat, sheep, cows, deer, and other nonmammalian animals. In some embodiment, a subject is an animal. In some embodiments, a subject is human. In those aspects where the subject is a human, the subject can be a pediatric or adult subject. In some embodiments, a subject is a mammal. In some embodiments, asubject is a mouse. In some embodiments, a subject is an experimental animal. In some embodiments, a subject is a rat. In some embodiments, a subject is a test animal.

[0131] “Substantially commensurate with initiation of peroxide-quenching” means the relationship between the timing of a cleavage reaction and the timing of a peroxide quenching reaction indicating that the initiation of the cleavage reaction and the initiation of the peroxide quenching reaction occur within a short time duration of each other (e.g. on the order of seconds, or on the order of minutes but not more than one day).

[0132] Subunit” (sometimes referred to herein as “unit”) means a species that is covalently bonded to or within an oligomer (e.g., oligosaccharide) or polymer (e.g., polysaccharide). Such species generally can include saccharides (e.g., glucose, galactose, mannose, etc.). For example, when an oligosaccharide composition comprises a glucose subunit, it means that the composition comprises a glucose molecule that is bound to or within an oligomer or polymer. Therefore, a composition that contains only free monomeric glucose would not contain a glucose subunit. Similarly, when an oligosaccharide composition comprises a sum of glucose, galactose, and mannose subunits in an amount of at least 70 wt. % based on total weight of saccharide subunits, this means that the mass of all of the glucose subunits, galactose subunits, and mannose subunits are summed, and the subunits of all saccharides are summed, and then the first sum is divided by the second sum. Additionally, when a polysaccharide composition comprises non-terminal galactose subunits, and at least 70 wt% of the non-terminal galactose subunits are specified to have at least one 4-linkage, this feature is calculated by summing the mass of all non-terminal galactose subunits having at least one 4-linkage (and this can include, for example, galactose subunits with 4,6-linkages and 4,3 -linkages), and then dividing by the total mass of non-terminal galactose subunits regardless of linkage type. The same concept is applicable to any feature where reference to “at least one X-linkage,” in which X is an integer (e.g., such as “a weight ratio of glucose subunits having at least one 4-linkage to glucose subunits having at least one 3-linkage is between 2: 1 to 4: 1” and other such features). Moreover, in such calculations the actual mass of the subunit is used (i.e., in bound form) rather than the mass of the unit as if it was hydrolyzed (which would add the mass of water). Other features can be calculated similarly. These features can be determined with the aid of the various analytical techniques, such as hydrolytic monosaccharide compositional analysis, oligosaccharide analysis, glycosidic linkage analysis, NMR HSQC Analysis, and so forth, as well as other techniques known in the art.

[0133] “Synthetic composition” means a composition which is artificially prepared and preferably means a composition containing at least one compound that is produced ex vivo chemically and / or biologically, e.g., by means of chemical reaction, enzymatic reaction, recombinantly, or any combination thereof. The synthetic composition typically comprises one or more compounds, including one or more of the polysaccharide cleavage products described herein. In some aspects, the polysaccharides and polysaccharide compositions can be formulated into a synthetic composition or administered as the polysaccharide alone or the polysaccharide cleavage product alone.

[0134] Treated polysaccharide” means a polysaccharide which has been contacted with at least one reagent capable of reacting with the polysaccharides (e.g. an enzyme or a Fenton’s reagent).

[0135] “Weak- Arrhenius base” means a compound that incompletely dissociates in water to release one or more hydroxide ions into solution, e.g. ammonium hydroxide, H2O, etc. As “weak-Arrhenius base” is used herein, there are no compounds which meet both the definitions used of strong-Arrhenius base and weak-Arrhenius base.

[0136] When the amount of a component is expressed in terms of weight or mole percent, it is intended that the amount is on a dry basis unless otherwise specified. “Dry basis” means in the absence of water or other solvent. For example, when a composition comprises 10 g of glucose, 40 g of xylose, and 50 g of water, it means the composition comprises 25% (mass% or wt.%) glucose on a dry basis, but the glucose is present in the composition at a concentration of 10% (mass% or wt.%).

[0137] Any viscosity measurement or property reported employs water as the solvent, unless specified otherwise.DETAILED DESCRIPTION OF THE INVENTION

[0138] In the following description, numerous specific details of the methods, compositions, and soluble fruit fibers of the invention are set forth to provide a thorough explanation of the precise nature of the invention. However, it will be apparent to those skilled in the art that the invention can be practiced without these specific details. Although the following description is divided into sections, it is contemplated that each section contains various aspects of the invention and as such invention from within each section and across two or more sections can be combined to form any aspect of the invention. When a table, spectrum, or other data is referred to as representing the features or properties possessed by aparticular composition, polysaccharide, or other compound or mixture, unless specified otherwise, the same analysis method and procedure used to obtain the table, spectrum, or other data is to be used to determine the properties of the particular composition, polysaccharide, or other compound or mixture.

[0139] It has been surprisingly found that a dietary fiber suitable for use in foods and beverages can be produced from fruit residues by (i) reacting polysaccharide of a fruit-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal, and (ii) cleaving the reacted polysaccharide with a cleavage agent to generate a mixture of polysaccharide cleavage products. The cleavage products have improved solubility, improved organoleptics and improved prebiotic properties, while retaining the basic monomer structure of the polysaccharide. The method thus valorizes a major component of fruit waste.

[0140] In one aspect, the mixture of polysaccharide cleavage products can be produced as broadly disclosed in WO 2018 / 236917, hereby incorporated by reference herein in its entirety for all purposes. This method comprises reacting a polysaccharide of a fruit-based starting material using a Fenton’s reagent composed of iron (Fe+, Fe2+) or other transition metal (including but not limited to, Cu1+, Co2+, etc.,) and hydrogen peroxide. The reaction can be allowed to proceed, for example, between 10 minutes and 24 hours, more specifically 15 minutes to 4 hours or 30 minutes to 3 hours. The transition metal or alkaline earth metal in the reaction mixture can, for example, be at a concentration of 10 pM to 5M, but generally at a concentration of more than 0.65 mM. If desired, prior to the Fenton reaction, the polysaccharides can be contacted with one or more polysaccharide degrading enzymes, such as an amylase, isoamylase, cellulase, maltase, glucanase, or a combination thereof. If desired, prior to the Fenton reaction, the polysaccharides can pre-treated as described herein, such as to extract polyphenols or extract polysaccharides. The reaction is subsequently quenched with base (e.g., an Arrhenius base or non-Arrhenius base, such as aqueous sodium hydroxide calcium hydroxide, potassium hydroxide, etc., or any combination thereof).

[0141] In another aspect, the mixture of polysaccharide cleavage products can be produced as broadly disclosed in in WO 2021 / 097138, hereby incorporated by reference in its entirety for all purposes. This method comprises a multi-step reaction that includes reacting the polysaccharide of the fruit-based starting material with a Fenton’s reagent and a then a peroxide-quenching / polysaccharide-cleavage step using either: a polysaccharide-cleavage agent that also functions as a peroxide-quenching agent; or using a polysaccharide-cleavageagent in combination with a compatible peroxide-quenching reagent that does not interfere with the polysaccharide-cleavage reaction. The polysaccharide-cleavage agent may be, for example, a weak-Arrhenius base or non- Arrhenius base. The polysaccharide-cleavage agent preferably also functions as a peroxide-quencher to quench (e.g., sufficiently reduce or eliminate) residual hydrogen peroxide and / or radicals thereof to minimize or eliminate off- target side reactions. In a specific embodiment, the method, for example, comprises reacting the polysaccharide of the fruit-based starting material with hydrogen peroxide and a suitable metal or metal ion (e.g., a transition metal, alkaline earth metal, or lanthanide, such as, for example, Fe(II), Fe(III), Cu(I), Cu(II), Ca(II), Mg(II), Mn(II), Zn(II), Ni(II), Ce(IV), Co(II) or other metal ions, or any combination thereof), followed by cleaving the glycosidic linkages in the hydroperoxyl-treated polysaccharides with a high-yield peroxide- quenching / cleavage agent such as ammonium bicarbonate, ammonium hydroxide, ammonia, urea, sodium amide, other ammonium-based reagent, a weak Arrhenius base, a nonArrhenius base, a Lewis base, a Bronsted-Lowry base, or any combination thereof. This generates high yields of polysaccharide cleavage products from the fruit-based starting material containing polysaccharide, while reducing or eliminating peeling and unwanted sidereactions.

[0142] The polysaccharide-cleavage reagent used in the methods can comprise at least one reagent selected from ammonium hydroxide, ammonia, ammonium bicarbonate, urea, etc., or a combination thereof. In some aspects, the polysaccharide-cleavage reagent may comprise the conjugate base of an alcohol or amine. In some aspects, the polysaccharide- cleavage reagent may comprise sodium methoxide, sodium ethoxide, sodium tertbutoxide, or other deprotonated alcohol. In some aspects, the polysaccharide-cleavage reagent may be or comprise one or more relatively “bulky bases” such as tert-butoxide, triethylamine, or other sterically hindered base. In some aspects, the use of such bulky cleavage reagents / bases results in selective cleavage of the accessible glycosidic bonds to provide oligosaccharide profiles unique / specific to the cleavage reagent / base. In some aspects the polysaccharide- cleavage reagent is not a base, / ?er se, but consists of, or comprises one or more reactive agent(s) that react to produce basic conditions and / or decomposition products. In all the methods described herein, the polysaccharide-cleavage reagent (cleavage initiator) can also be, and preferably is, a peroxide-quenching reagent, and in either case may be used in combination with an additional compatible peroxide-quenching agent that may or may not also be a polysaccharide-cleavage agent.

[0143] The use of weak Arrhenius bases and / or non-Arrhenius bases (e.g., ammonium- based peroxide-quenching / polysaccharide-cleavage reagents, etc.) as polysaccharide- cleavage agents not only provides for improved high-yield polysaccharide cleavage product production (relative to the strong Arrhenius bases), but also eliminates the need for costly and time-consuming post-reaction concentration, and desalting steps.

[0144] The cleavage initiator also can, and preferably does, function as a peroxidequencher to quench (sufficiently reduce or eliminate) residual peroxide and / or radicals thereof to reduce or eliminate peeling and unwanted side-reactions. Alternatively, the cleavage agent can be added to the reaction after, or along with addition of a compatible peroxide-quenching agent (that could also be a cleavage reagent). The peroxide- quenching / cleavage agent can be, and preferably is, selected from one or more nitrogen-based agents as described above. This not only provides high-yield cleavage and residual peroxide- quenching, but also provides for cleavage specificity tailoring (e.g., by replacing nitrogen bound hydrogen with larger moieties to sterically hinder or otherwise modify access by, or activity of the cleavage agent).

[0145] The transition metal or alkaline earth metal in the reaction mixture is at a concentration of at least about 10 pM, for example about 10 pm to about 20 mM. In some aspects, the concentration is at least about 0.65 mM (e.g. at least a value in the range of 0.5 to 0.7 mM). In some aspects, the transition metal or alkaline earth metal in the reaction mixture is at a concentration from 0.65 mM to 500 mM. In some aspects, the peroxide agent (e.g., hydrogen peroxide) in the reaction mixture is at a concentration of at least about 0.02 M (e.g. at least a value in the range of 0.015 to 0.025 M). In some aspects, the peroxide agent (e.g., hydrogen peroxide) in the reaction mixture is at a concentration of from 0.02 M to 1 M, or in some aspects up to 5 M. In some aspects, the peroxide agent (e.g., hydrogen peroxide) in the reaction mixture is at a concentration of from 1 M to 5 M. In some aspects, the cleavage reagent / base is or comprises ammonium hydroxide, ammonia, ammonium bicarbonate, a weak Arrhenius base, a non-Arrhenius base, a Lewis base, and / or a Bronsted-Lowry base. Moreover, combinations of two or more cleavage reagents / bases (e.g., such as the cleavage reagents / bases discussed herein) may be used. In some aspects, strong-Arrhenius bases (e.g., Na+OH', K+0H', or Ca+2(OH )2) can be used in combination with the cleavage reagents / bases discussed herein. In some aspects, ammonia gas can be in contact with the solution through bubbling or as an atmospheric component to act as a cleavage and / or quenching reagent. In some aspects, the cleavage reagent is at a concentration of at least about 0.1 M (+ / - 20%). Insome aspects, the cleavage reagent is at a concentration of from 0.1 M-5.0 M. In some aspects the cleavage reagent is present as a saturated solution or insoluble material. In some aspects the cleavage reagent brings the solution to pH 7.5, 8, 9, 10, 12, or higher. In all the methods described herein, the cleavage reagent (cleavage initiator) may also be, and preferably is a peroxide-quenching reagent, and in either case may be used in combination with an additional compatible peroxide-quenching agent that may or may not also be a cleavage agent.

[0146] In some aspects, the fruit-based starting material first undergo initial oxidative treatment with the hydrogen peroxide and a transition metal, alkaline earth metal, or lanthanide catalyst to render the glycosidic linkages more labile. Ammonium hydroxide, ammonium bicarbonate, ammonia, urea, etc., or other weak Arrhenius or non-Arrhenius base is then used for cleavage, which results in a variety of distinctive oligosaccharides (distinctive oligosaccharide profile), or smaller polysaccharides. In some aspects, peroxide-quenching and / or neutralization takes place immediately to reduce unwanted oxidation, or peeling, respectively. In some aspects the treated polysaccharide (e.g., the polysaccharide after treatment with a Fenton’s reagent) is allowed to react with the cleavage reagent at reduced, ambient, or room temperature to facilitate the production of oligosaccharides. In some aspects the cleavage reaction takes place at 4-100°C, 20-80°C, 30-60°C or about 40°C. In some aspects, cleavage and peroxide-quenching are immediate. In some aspects the cleavage step is conducted for 10-30 minutes, 20-60 minutes, 60-360 minutes. In some aspects the cleavage step is conducted for 2-6 hours, 3-12 hours, 6-24 hours or longer. In some aspects the cleavage step is conducted overnight. In all the methods described herein, the cleavage reagent (cleavage initiator) may also be, and preferably is a peroxide-quenching reagent, and in either case may be used in combination with an additional compatible peroxide-quenching agent that may or may not also be a cleavage agent.

[0147] In some aspects, the method of cleaving polysaccharides comprises multiple steps. For instance, the method can comprise: a) contacting one or more polysaccharide with a Fenton’s reagent, comprising a peroxide agent and metal ions to form a mixture; b) allowing the Fenton’s reagent to react with the polysaccharide for a specified reaction time; and c) after step b, adding a cleavage agent which may also be a peroxide quenching reagent to the mixture. In such aspect, the steps of contacting the polysaccharide with a Fenton’s reagent (step a) and allowing a specified reaction time to pass (step b) can be performed at the same or different pH wherein the pH is selected from within a range of pH 3 to 8, pH 4 to 7, pH 4.5to 6.5, and pH 5 to 6. The pH can be any possible value between the specified ranges of pH values. The step of adding a cleavage agent which may also be a peroxide quenching reagent (step c) can be performed at a pH selected from within a range of pH 6 to 11, pH 6.5 to 9.5, pH 7 to 9, and pH 7 to 8. The pH can be any possible value between the specified ranges of pH values. In such aspect, the step of contacting the polysaccharide with a Fenton’s reagent (step a) and passage of the specified reaction time (step b) can be performed at the same or different temperature wherein the temperature is selected from within a range of temperature between 10 and 70 degrees Celsius, between 20 and 60° Celsius, and between 25 and 55° Celsius. The temperature can be any possible value between the specified ranges of temperature values. The step of adding a cleavage agent which may also be a peroxide quenching reagent (step c) can be performed at a temperature selected from within a range of temperature between 10 and 70° Celsius, between 20 and 60° Celsius, and between 25 and 55° Celsius. The temperature can be any possible value between the specified ranges of temperature values.

[0148] In some aspects, if desired, the fruit-based starting material can optionally be treated with one or more polysaccharide-degrading enzyme(s) to reduce the average size or complexity of the polysaccharide therein before the resulting polysaccharides are treated with the Fenton’s reagent. Non-limiting examples of polysaccharide enzymes that can be used include for example, amylase, isoamylase, cellulase, maltase, glucanase, lactase, xylanase, arabinase, pectinase, mannanase, or a combination thereof.

[0149] In some aspects, the polysaccharides of the fruit-based starting material first undergo initial oxidative treatment with hydrogen peroxide and a transition metal or alkaline earth metal (e.g., iron(III) sulfate) catalyst to render the glycosidic linkages more labile. A weak-Arrhenius base or non-Arrhenius base is then used for base induced cleavage, which results in a variety of oligosaccharides. Immediate neutralization may take place to reduce any peeling reaction. This method can generate large amounts of biologically active oligosaccharides from a variety of fruit-based sources. The initial oxidative treatment can include hydrogen peroxide and a transition metal or an alkaline earth metal. Metals with different oxidation states, sizes, periodic groups, and coordination numbers have been tested and each of the different metals has shown activity in the reaction. The oxidative treatment is followed by a base treatment.

[0150] Prior to reacting the polysaccharide of the fruit-based starting material with the Fenton’s reagent, the polysaccharide can be purified from the starting material, i.e., the fruit-based polysaccharide containing material can be pre-treated. In embodiments, the polysaccharide can be enriched in the starting material by removing one or more of polyphenols, lignin, lipids, proteins, starch, salts, monosaccharides, and disaccharides. This can be done using various methods known for purification of polysaccharides from polysaccharide containing material. Polyphenols and lipids can be removed by any suitable extraction technology. Conventional batch extraction or continuous extraction, as well as extraction employing ultrasound or microwave assistance, as understood in the art, can be employed to enhance extraction efficiency. In embodiments, the content of polyphenols or lipids can be reduced by extraction with a suitable organic solvent (e.g., alcohols (e.g., methanol, ethanol, or isopropanol), ketones (e.g., acetone), esters (e.g., ethyl acetate) or a mixture of organic solvents (chloroform / methanol). Polyphenols or lipids can also be extracted with acidified organic solvent, e.g., acidified alcohol. In embodiments, supercritical fluid extraction, as known in the art, can be used to extract polyphenols as well as lipids.

[0151] In embodiments, the content of polyphenols and lipids can be reduced by extraction with an acidified solvent comprising a solvent and an acidifying agent. In embodiments, the solvent is an organic solvent rather than an aqueous solvent. In embodiments, the solvent can be any solvent suitable for processing foodstuffs, for example ethanol. The acidifying agent can be any suitable acidifying agent, for example a mineral or organic acid. The mineral acid can, for example, include sulfuric or hydrochloric acid. The organic acid can be a carboxylic acid, and more specifically can be a mono-, di- or tricarboxylic acid. In embodiments, the organic acid can be acetic acid, propionic acid or butyric acid. In embodiments, the organic acid can be malonic, succinic glutaric or adipic acid. In embodiments, the organic acid can be citric acid, lactic acid, malic acid, or propane- 1,2, 3 -tricarboxylic acid. In embodiments, the acidifying agent can be a salt of an organic acid.

[0152] In embodiments, the organic solvent can contain about 0.005% to about 1% wt / v of the acidifying agent, for example about 0.01% to about 1% wt / v of the acidifying agent or any subrange thereof. In embodiments, the amount of acidifying agent ranges from 0.01% to 0.5% wt / v. In embodiments, the amount of acidifying agent ranges from 0.01% to 0.2% wt / v. In embodiments, the amount of acidifying agent ranges from 0.01% to 0.05% wt / v. In embodiments, the amount of acidifying agent ranges from 0.01% to 0.02% wt / v. In embodiments, the amount of acidifying agent ranges from 0.05% to 0.5% wt / v. In embodiments, the amount of acidifying agent ranges from 0.05% to 0.2% wt / v. Inembodiments, the amount of acidifying agent ranges from 0.05% to 0.1% wt / v. In embodiments, the amount of acidifying agent ranges from 0.1% to 0.2% wt / v. In embodiments, the amount of acidifying agent ranges from 0.1% to 0.5% wt / v. In embodiments, the amount of acidifying agent ranges from 0.1% to 1% wt / v. In embodiments, the amount of acidifying agent ranges from 0.5% to 1% wt / v. In embodiments, the amount of acidifying agent is 0.013%-0.017 wt / v. In embodiments, the amount of acidifying agent ranges from 0.85 to 1% wt / v. The foregoing embodiments include all subranges of the listed ranges.

[0153] In an embodiment, the extraction can take place at room temperature (e.g., about 25°C) to about 90°C, for example at about 35°C to about 80°C. It will be appreciated that the temperature at which the extraction is conducted will depend upon the volatility of the solvent employed. In an embodiment, the extraction can take 1 hour to 5 hours, for example, about 2 hours to about 4 hours. In an embodiment, the ratio (v / w) of acidified solvent to starting material can be about 15: 1 to about 1 :1, or any subrange thereof, for example, about 10: 1 to about 4:1. After the extraction, the polyphenols and / or lipids can be removed with the liquid phase by filtration; for example, using centrifugation. Extraction can optionally be repeated multiple times. Extraction is optionally accompanied by one or more steps of washing the extracted solid with acidified solvent or solvent.

[0154] In one aspect, the polysaccharide can be enriched by removing the protein and starches from the fruit-based starting material. This can be achieved by utilizing any suitable protein and / or starch removal technology. An example of a suitable technology for removal is incubation with one or more enzymes followed by separation of peptides, other protein material, and / or monosaccharides, and / or disaccharides. For separation of the protein, the enzyme can be any suitable protease, for example the protease can be an endo- or exopeptidase or a mixture thereof as found in (Flavorzyme® peptidase), or a serine endopeptidase (subtilisinA) as found in Alcalase®, or the like. For separation of the starch, the enzyme can be any suitable amylase, including isoamylase. The polysaccharides can be separated from the peptides, other protein material and / or monosaccharides, and / or disaccharides by causing the polysaccharides to precipitate or solubilizing the peptides, other protein material and / or monosaccharides and / or disaccharides followed by filtration. In one example, the polysaccharides are separated by adding ethanol to precipitate polysaccharide followed by filtration such as centrifugation to separate precipitated polysaccharide. The process may be repeated as necessary. Ordinarily, salts, monosaccharides, and disaccharideswill also be removed in the liquid filtrate along with the peptides and other protein material. If desired, an acid extraction can also be used to remove peptides, salts, monosaccharides, and di saccharides.

[0155] In one aspect, polysaccharide can be extracted from fruit-based starting material containing polysaccharide using an acidic aqueous solution. This method is particularly useful for extracting pectin containing polysaccharide from the fruit-based starting materials. In embodiments, polysaccharide is extracted from fruit-based starting material with an aqueous solution having pH of 1 to 3 (+ / -0.5). In embodiments, the aqueous solution used for extraction has pH of 1 to 2 (+ / -0.5). In embodiments, the aqueous solution used for extraction has pH of 1.0 to 1.5 (+ / -0.1). In embodiments, the aqueous solution used for extraction has pH of 1.2 (+ / -0.1). In embodiments, any suitable acidifying agent can be used. In embodiments, the acidifying agent is a mineral acid or an organic acid. In embodiments, the mineral acid is sulfuric or hydrochloric acid. In embodiments, concentrated sulfuric acid or concentrated hydrochloric acid is added to water or an aqueous solution to obtain the desired pH. In embodiments, the organic acid can be a liquid or solid acid. A selected amount of the liquid or solid organic acid is added to water or an aqueous solution to obtain the desired pH. Useful organic acids include, among others, formic acid, acetic acid, propionic acid, butyric acid, malonic, succinic, glutaric, adipic acid, citric acid, lactic acid, malic acid, or propane- 1,2, 3 -tricarboxylic acid. In embodiments, preferred acidifying agents and solvents are those that are suitable for use in processing animal or human food products. In embodiments, the organic acidifying agent is acetic acid or citric acid.

[0156] In an embodiment, the extraction can take place at room temperature (e.g., about 25°C) to about 90°C, for example at about 35°C to about 80°C or at room temperature. Extraction with acidic aqueous solution can result in acid hydrolysis of polysaccharide. In embodiments, it is preferred to minimize hydrolysis of polysaccharide and maximize extraction of polysaccharide. It will be appreciated by one of ordinary skill in the art that the pH of the acidic aqueous solution employed as well as the temperature and length of extraction time can affect the amount of hydrolysis and extraction. It is preferred to adjust extraction conditions to minimize acid hydrolysis of polysaccharides, and particularly to minimize the hydrolysis of pectin. In an embodiment, the extraction can take 1 hour to 5 hours, for example 2 hours to 4 hours, or any subrange thereof. In an embodiment, the ratio (v / w) of acidified solvent to starting material can be 15: 1 to 1 : 1, or any subrange thereof, for example, 10: 1 to 4: 1. Polysaccharide, particularly pectin, is extracted from the fruit-basedstarting material into the aqueous phase. After extraction, the aqueous phase is separated from remaining solids by filtration; for example, using centrifugation. The aqueous phase containing polysaccharide is optionally extracted with a suitable organic solvent and or subjected to additional purification steps, for example the aqueous solution is optionally treated to remove mono- or disaccharides or salts, and particularly the aqueous phase can be treated by diafiltration. The aqueous phase after any further optional purification is neutralized and solvent can be removed by drying. Alternatively, polysaccharide, particularly pectin, can be precipitated from aqueous solution as is known in the art by addition of suitable organic solvent, more specifically an alkyl alcohol and yet more specifically ethanol. Precipitated polysaccharide is optionally washed one or more times with organic solvent, e.g., ethanol. Extraction can optionally be repeated multiple times.

[0157] In embodiments herein precipitation of polysaccharide from aqueous solution is referred to as ethanol precipitation. It will be apparent to one of ordinary skill in the art that other organic solvents can be employed to precipitate polysaccharides, such as isopropanol. In embodiments, the precipitating solvent is miscible with water. In embodiments, the precipitating organic solvent (e.g., ethanol or isopropanol) is added to the aqueous solution to achieve a 40%-80% v / v concentration of precipitation solvent to water. In embodiments, the v / v concentration of precipitating solvent to water ranges from 50%-70%. In embodiments, the v / v concentration of precipitating solvent to water ranges from 55%-65%. Optionally, the aqueous solution is subjected to multiple precipitations where the concentration of precipitation solvent added in different precipitations steps is the same or is different.

[0158] In embodiments, fruit-based starting material containing polysaccharide is pretreated prior to depolymerization (e.g., treatment with Fenton’s reagent) to remove starch, for example by treatment with amylase in aqueous solution. Optionally, one or more proteases are added as well to remove protein. Amylase-treated material or amylase / protease treated material is then subjected to ethanol precipitation to collect the insoluble fraction which contains purified polysaccharides from which starch or starch and protein has been removed. Optionally, one or more additional polysaccharide degrading enzymes (e.g., one or more cellulases) are added to the aqueous mixture. In an embodiment, polysaccharide from which starch (and protein and / or other polysaccharides, e.g., cellulose) has been removed is extracted with organic solvent or acidic organic solvent to remove polyphenols. Preferably, pectinase is not added to remove pectin. In embodiments, polysaccharide from which starch has been removed is treated in a separate step with one or more proteases to remove proteinand optionally in another separate step is treated to remove cellulose or other polysaccharide other than pectin. In embodiments, pre-treatment does not include treatment with one or more pectinases. In embodiments, one or more steps of ethanol precipitation follow removal of starch, optional removal of protein and optional removal of cellulose or polysaccharide other than pectin.

[0159] In embodiments, dried polysaccharide containing material obtained from a fruit is extracted with acidified organic solvent to remove polyphenols. In embodiments, the organic solvent is acidified with citric acid. In embodiments, the organic solvent is ethanol. In embodiments, the acidified organic solvent is ethanol acidified with citric acid. Polyphenol reduced polysaccharide material is then subjected to treatment with one or more proteases, one or more amylase or a combination thereof in aqueous medium. In embodiment, polysaccharide in the aqueous medium is subjected to ethanol precipitation to obtain pretreated polysaccharide containing material. The resulting pre-treated polysaccharide containing material can be used as the fruit-based starting material.

[0160] In an embodiment, dried polysaccharide containing material obtained from a fruit is treated with amylase in aqueous medium and thereafter subjected to ethanol precipitation. The resulting pre-treated polysaccharide containing material can be used as the fruit-based starting material.

[0161] In an embodiment, dried polysaccharide containing material obtained from a fruit is extracted with acidified organic solvent to remove polyphenols. In embodiments, the organic solvent is acidified with citric acid. In embodiments, the organic solvent is ethanol. In embodiments, the acidified organic solvent is ethanol acidified with citric acid. Polyphenol reduced polysaccharide containing material is subjected to acidic aqueous extraction (pH 1 to 3, preferably pH 1.2), followed by neutralization and treatment with one or more amylases, and optionally one or more proteases. This treatment is followed by ethanol precipitation to obtain solid purified polysaccharides which can be used as the fruit-based starting material.

[0162] In an embodiment, dried polysaccharide containing material obtained from a fruit is treated with one or more amylases in aqueous medium and thereafter subjected to ethanol precipitation to recover polysaccharide from which starch has been removed. This material is then subjected to extraction with organic solvent (e.g., chloroform / methanol / acetone extraction) to remove polyphenols. This pre-treated polysaccharide material can be used as the fruit-based starting material.

[0163] In an embodiment, dried polysaccharide containing material obtained from a fruit is extracted with acidic organic solvent, e.g., acidic ethanol, to remove polyphenols.Polyphenol reduced polysaccharide containing material is then subjected to acidic aqueous extraction (pH 1-3, preferably pH 1.2) to obtain extracted polysaccharide from which polyphenols have been removed. After neutralization extracted polysaccharide in aqueous medium is then treated with one or more amylases and the aqueous medium is separated form remaining solids, e.g., containing cellulose and protein. The separated aqueous medium is then subjected to ethanol precipitation to precipitate polysaccharide. This precipitated polysaccharide can be used as the fruit-based starting material.

[0164] In embodiments, polysaccharide containing material obtained from a fruit is first treated to extract polyphenols to extract polyphenols and is thereafter treated to remove starch, optionally to remove protein or other polysaccharides other than pectin. In embodiments, polysaccharide containing material obtained from a fruit is treated both to extract polyphenols and is treated to extract polysaccharides from the fruit-based starting material, wherein the order of treatment is not critical.

[0165] In additional aspects, the mixture of polysaccharide cleavage products can be subjected to further processing to purify them. This can be done using various methods known for purification of oligosaccharides.

[0166] In embodiments, the mixture of polysaccharide cleavage products can be purified by removing cellulose and other unreacted polysaccharide material. The cellulose and unreacted polysaccharide material can be removed using any suitable technology, for example, filtration such as ultrafiltration, or by precipitation, such as precipitation in ethanol.

[0167] In embodiments, the mixture of polysaccharide cleavage products can be purified by removing residual metals, such as iron or copper. In embodiments, the residual metals can be removed by suitable resins. In embodiments, the residual metals are removed by suitable nanofiltration membranes. In embodiments, useful nanofiltration membranes have molecular weight cutoffs of 400-600 daltons, 600-800 daltons, or 800-1000 daltons. In embodiments, the polysaccharide cleavage products can be purified by removing remaining hydrogen peroxide. In embodiments, residual hydrogen peroxide can be removed by addition of a quenching agent. In embodiments, hydrogen peroxide can be removed by enzymes, such as one or more catalase, for example Catazyme® catalase (Novozymes A / S, Denmark). In embodiments, the hydrogen peroxide is removed by suitable nanofiltration membranes. Inembodiments, suitable nanofiltration membranes have molecular weight cutoffs of 400-600 daltons, 600-800 daltons, or 800-1000 daltons.

[0168] In embodiments, the mixture of polysaccharide cleavage products can be purified by removing salts. In embodiments, salts are removed by applying a suitable resin material. Suitable resin materials may include anion-exchange, cation-exchange, mixed bed, decolorizing, other chelation resins. For example, suitable resins may include, but are not limited to, lonac NM-60, MBD-10 ULTRA, Thermax Tulsion MB, Cole-Parmer RR-1400, Amberlite MB20, DOWEX Monosphere MR-450, and LSF 973 resin. Two or more resins may be combined to create mixed-bed resins. In embodiments, the salts are removed by suitable nanofiltration membranes. In embodiments, the nanofiltration membranes have molecular weight cutoffs of 400-600 daltons, 600-800 daltons, or 800-1000 daltons. In embodiments, the mixture of polysaccharide cleavage products can be purified by removing residual monosaccharide or disaccharides. In embodiments, the residual monosaccharides and disaccharides are removed by suitable nano-filtration membranes. In embodiments, the nanofiltration membranes have molecular weight cutoffs of 400-600 daltons, 600-800 daltons, or 800-1000 daltons.

[0169] In embodiments, the mixture of polysaccharide cleavage products may be treated with carbon to remove undesired components. The carbon may be activated carbon, charcoal, graphitized carbon, porous graphitized carbon, or any carbon-based material that is added with the goal of purification.

[0170] In embodiments, the purified polysaccharide cleavage products contain less than 10% (less than 9%, less than 8%, less than 7%, or less than 6%) by weight of polyphenols. In preferred embodiments, purified polysaccharide cleavage products contain less than 5% (less than 4%, less than 3%, less than 2% or less than 1%) by weight of polyphenols. In embodiments, purified polysaccharide cleavage products contain less than 10% (less than 9%, less than 8%, less than 7%, or less than 6%) by weight of salt. In embodiments, purified polysaccharide cleavage products contain less than 10% (less than 9%, less than 8%, less than 7%, or less than 6%) by weight of starch. In embodiments, purified polysaccharide cleavage products contain less than 10% (less than 9%, less than 8%, less than 7%, or less than 6%) by weight of protein. In preferred embodiments, purified polysaccharide cleavage products contain less than 5% (less than 4%, less than 3%, less than 2% or less than 1%) by weight of protein. In embodiments, purified polysaccharide cleavage products contain less than 10% (less than 9%, less than 8%, less than 7%, or less than 6%) by weight of mono-and / ordi saccharides. In preferred embodiments, purified polysaccharide cleavage products contain less than 5% (less than 4%, less than 3%, less than 2% or less than 1%) by weight of mono- and / or disaccharides. In embodiments, purified polysaccharide cleavage products contain less than 5% (less than 4%, less than 3%, less than 2%) by weight of ash. In embodiments, purified polysaccharide cleavage products contain less than 2% (less than 1.75%, less than 1.5%, less than 1.25%) by weight of ash. In preferred embodiments, purified polysaccharide cleavage products contain less than 1% (less than 0.7%, less than 0.5%, or less than 0.2%) by weight of ash.

[0171] In embodiments, purified polysaccharide cleavage products contain 40% or more (or 50% or more, or 60% or more, or 70% or more, or 80% or more) by weight of fiber (e.g., dietary fiber). In preferred embodiments, purified polysaccharide cleavage products contain 85% or more (or 90% or more, or 95% or more, or 98% or more) by weight of fiber (e.g., dietary fiber).

[0172] In embodiments, one or more carbohydrate active enzymes can be used to modify the resulting mixture of polysaccharide cleavage products by either adding or removing monomeric units to make a new product.Source Polysaccharides and Source Materials Comprising Polysaccharides

[0173] Any suitable polysaccharide source can be used to prepare the polysaccharide cleavage products. For example, the polysaccharide source can be pineapple pomace obtained after juice or fruit processing of pineapples. Also, the polysaccharide source can be apple pomace obtained after juice or fruit processing of apples, or carrot pomace after juice or fruit processing of a carrots, or orange pomace after juice or fruit processing of oranges. The pomance comprises mainly parts of the husk, membranes and seeds. The polysaccharide source can also be in the form of press cake obtained by pressing residues after juice processing. Other suitable sources include lemon pomace or press cake, pear pomance and press cake, and the like. The polysaccharide source is not pomegranate or any part of the pomegranate fruit. In preferred embodiments, the polysaccharide source contains pectin.Polysaccharide cleavage products

[0174] The polysaccharide cleavage products have suitable features, structural characteristics, and other various properties. In some aspects, the polysaccharide cleavage products have the same, or substantially the same (i.e., within 10%, within 20%, within 30%of the values for) features as described for CLX136 and CLX136S. In some aspects, the polysaccharide cleavage products have the same, or substantially the same (i.e., within 10%, within 20%, within 30% of the values for) features as described for CLX137C. In some aspects, the polysaccharide cleavage products have the same, or substantially the same (i.e., within 10%, within 20%, within 30% of the values for) features as described for CLX136 and CLX136S.

[0175] In embodiments, the polysaccharide cleavage products can have a number of oligosaccharide / polysaccharide structures ranging in size from a DP of 3 to 500, or any subrange thereof. The exact profile of oligosaccharide structures will depend on the polysaccharide source and reaction conditions. The source material and the conditions can be altered to provide the desired profile.

[0176] In embodiments, the polysaccharide cleavage products can have an average DP in the range of 3 to 300, for example, 3 to 200, 3 to 100, 10 to 300, 10 to 200, 10 to 100 or any subrange thereof. The average DP may be adjusted as desired.

[0177] In embodiments, the polysaccharide cleavage products comprise a backbone containing galacturonic acid monomers, wherein each galacturonic acid monomer is optionally bonded in an alternating fashion to a rhamnose monomer, and wherein the total number of monomers in the synthetic oligosaccharide or polysaccharide ranges from 3 to 50. In embodiments, the backbone is branched with galactose, arabinose, or xylose monomers. In embodiments, the arabinose branches are further extended with additional arabinose monomers. In embodiments, the galactose branches are further extended with additional galactose monomers. In embodiments, the extended galactose monomers are further branched with arabinose monomers. Such synthetic oligosaccharides or polysaccharides can be obtained, for example, by depolymerizing a fruit-based starting material containing pectin according to the methods described herein. Pectin is known to contain both galacturonic acid and alternating galacturonic acid and rhamnose backbones with arabinose, galactose, and arabinogalactan branches. Arabinose branches contain 3- and 5-linked arabinose monomers. Galactose branches contain 4-linked galactose monomers. Arabinogalactan branches contain 4-linked galactose with terminal arabinose branches. Pectins may be obtained from whole fruits, fractions or extracts of fruits, fruit waste streams, and / or fruit juices.

[0178] In some aspects, the polysaccharide cleavage products have a solubility in water or aqueous solution of between 50 mg / ml and 2000 mg / ml, for example, between 50 mg / mland 2000 mg / ml, between 100 mg / ml and 2000 mg / ml, between 150 mg / ml and 2000 mg / ml, between 200 mg / ml and 2000 mg / ml, between 150 mg / ml and 1500 mg / ml, between 1000 mg / ml and 2000 mg / ml, between 500 mg / ml and 2000 mg / ml, between 50 mg / ml and 1000 mg / ml, between 100 mg / ml and 1000 mg / ml, between 150 mg / ml and 1000 mg / ml, between 200 mg / ml and 1000 mg / ml, between 150 mg / ml and 500 mg / ml, or between 200 mg / ml and 1000 mg / ml. In some embodiments, the polysaccharide cleavage products have a solubility of at least 200 mg / ml, optionally less than 1000 mg / ml. In further embodiments, the polysaccharide cleavage products have a solubility of at least 200 mg / ml, optionally less than 1000 mg / ml, with a turbidity value of less than 20 NTU, optionally greater than 0.5 NTU. In some embodiments, the polysaccharide cleavage products have a solubility of at least 200 mg / ml, of at least 500 mg / ml, of at least 1000 mg / ml or at least 2000 mg / ml.

[0179] In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 50 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 40 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 35 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 30 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 25 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 20 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 15 NTU. In embodiments, solutions of the polysaccharide cleavage products in water at 25 °C exhibit turbidity less than 10 NTU. In embodiments, the concentration of the polysaccharide cleavage products in water at 25 °C is 5 g / L or greater. In embodiments, the concentration of the polysaccharide cleavage products in water at 25 °C is 10 g / L or greater. In embodiments, the concentration of the polysaccharide cleavage products in water at 25 °C is 15 g / L or greater. In embodiments, the concentration of the polysaccharide cleavage products in water at 25 °C is 20 g / L or greater. In embodiments, the concentration of the polysaccharide cleavage products in water at 25 °C is 30 g / L or greater.

[0180] In embodiments, a solution of the polysaccharide cleavage products in water at 25 °C at a concentration of 5 g / L or greater has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU or less, or optionally 15 NTU or less. In embodiments, a solution of the polysaccharide cleavage products in water at 25 °C at a concentration of is 10 g / L or greater has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU orless, or optionally 15 NTU or less. In embodiments, a solution of the polysaccharide cleavage products in water at 25 °C at a concentration of 15 g / L or greater has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU or less, or optionally 15 NTU or less. In embodiments, a solution of the polysaccharide cleavage products in water at 25 °C at a concentration of 20 g / L or greater has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU or less, or optionally 15 NTU or less. In embodiments, the concentration of the polysaccharide cleavage products in water at 25 °C is 30 g / L or greater has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU or less, or optionally 15 NTU or less.

[0181] In some aspects, the polysaccharide cleavage products have an average molecular weight of less than 100 kDa, optionally greater than 0.5 kDa, 1 kDa, or 2 kDa. In some aspects, 50% or less (e.g., 40% or less, 30% or less, 20% or less, or 10% or less) of the polysaccharide cleavage products by weight have a molecular weight of about 100 kDa or more. In preferred aspects, 10% or less of the polysaccharide cleavage products by weight have a molecular weight of about 100 kDa or more. In keeping with this aspect, preferred polysaccharide cleavage products comprise between 5% and 10% (such as between 5% and 9%, between 5% and 8%, between 5% and 7%, between 6% and 10%, or any subranges thereof) polysaccharides by weight having a molecular weight of about 100 kDa or more.

[0182] In some aspects, the polysaccharide cleavage products have an average molecular weight ranging from 0.5-50 kDa. In some aspects, the polysaccharide cleavage products have an average molecular weight ranging from 1-50 kDa. In some aspects, the polysaccharide cleavage products have an average molecular weight ranging from 0.5-25 kDa. In some aspects, the polysaccharide cleavage products have an average molecular weight ranging from 1-25 kDa.Soluble Fruit Fiber

[0183] In an aspect, the invention provides soluble fruit fiber comprising, consisting essentially of or consisting of a mixture of polysaccharide cleavage products from a fruitbased starting material, as described herein. In preferred embodiments, the soluble fruit fiber comprises, consists essentially of or consists of a mixture of polysaccharide cleavage products from a pineapple-based starting material, an apple-based starting material, or an orange-based starting material. In embodiments, the soluble fruit fiber exhibits high solubility in water or aqueous solution with low turbidity. In embodiments, solutions of soluble fruitfiber in water or aqueous solution exhibit low viscosity. In embodiments, a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU or less or optionally 15 NTU or less. In embodiments, a 20 g / L mixture of the soluble fruit fiber in water has a dynamic viscosity of 4 millipascal-second (mPa.s) or less at 25°, optionally 2 mPa.s or less, optionally 1.5 mPa.s or less, or optionally ImPa.s or less. In embodiments, a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 40 NTU or less, optionally 30 NTU or less, optionally 20 NTU or less or optionally 15 NTU or less and has a dynamic viscosity of 4 millipascal-second (mPa.s) or less at 25°, optionally 2 mPa.s or less, optionally 1.5 mPa.s or less, or optionally ImPa.s or less.

[0184] In embodiments, the mixture of polysaccharide cleavage products of the soluble fruit fiber contains 40% or more, optionally 50% or more, optionally 60% or more, optionally 70% or more, optionally 80% or more, optionally 90% or more by weight of polysaccharide cleavage products having molecular weight less than 100 kDa. In embodiments, each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products of the soluble fruit fiber has a degree of polymerization (DP) of 3 to 100, optionally of 3 to 75, optionally of 3-50, optionally of 3-25 or optionally of 23-20.

[0185] In embodiments, the mixture of polysaccharide cleavage products of the soluble fruit fiber is formed by cleavage of a polysaccharide of a fruit-based starting material. In embodiments, the fruit-based starting material comprises, consists essentially of or consists of pomace, husk, peels, seeds, or a combination thereof. In a specific embodiment, the fruitbased starting material, consists essentially of or consists of peels.

[0186] In embodiments, polysaccharides of the fruit-based starting material are depolymerized by any chemical or physical method that will achieve desired level of depolymerization, for example, the specified degree of polymerization or the specifies range of molecular weights. In embodiments, the depolymerization method is a chemical and / or enzymatic method. In embodiments, the fruit-based starting material is first treated to remove polyphenols prior to depolymerization. In embodiments, the mixture of polysaccharide cleavage products is formed by:(i) reacting one or more polysaccharides in a fruit-based starting material containing polysaccharide with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products.Uses for Soluble Fruit Fiber and Oligosaccharide Compositions

[0187] The oligosaccharide compositions (e.g., polysaccharide cleavage products) and soluble fruit fiber herein have a variety of beneficial uses. In some aspects, the polysaccharide cleavage products and / or the soluble fruit fibers are useful as synbiotics, prebiotics, immune modulators, digestion aids, food additives, pharmaceutical excipients, or analytical standards. In some aspects, the polysaccharide cleavage products and / or the soluble fruit fibers are combined with other ingredients to produce foodstuffs and supplements including infant formula, geriatric supplements, baking flours, and snack foods. The polysaccharide cleavage products and / or the soluble fruit fibers can also be used as pharmaceutical products.

[0188] In embodiments, the polysaccharide cleavage products and / or the soluble fruit fibers can be used as a prebiotic to selectively stimulate growth of one or more probiotic bacteria. In some aspects, the polysaccharide cleavage product and / or the soluble fruit fiber compositions can be administered as a prebiotic formulation (i.e., without bacteria) or as a synbiotic formulation (i.e., with one or more desirable bacteria such as bifidobacteria as described herein). In general, any food or beverage that can be consumed by humans or animals, or otherwise suitably administered, may be used to make formulations containing the prebiotic and probiotic compositions. Exemplary foods include those with a semi-liquid consistency to allow easy and uniform dispersal of the prebiotic and probiotic compositions described herein. However, other consistencies (e.g., powders, liquids, etc.) can also be used without limitation. Accordingly, such food items include, without limitation, dairy -based products such as cheese, cottage cheese, yogurt, and ice cream. Processed fruits and vegetables, including those targeted for infants / toddlers, such as apple sauce or strained peas and carrots, are also suitable for use in combination with the polysaccharide cleavage products and / or the soluble fruit fibers of the present invention. Both infant cereals such as rice- or oat-based cereals and adult cereals such as Cream of Wheat™, etc., are also suitable for use in combination with the polysaccharide cleavage products and / or the soluble fruit fibers. The polysaccharide cleavage products and / or the soluble fruit fibers can also be used in medical foods, for example, such as Pedialyte™, Ensure™, etc. In addition to foodstargeted for human consumption, animal feeds may also be supplemented with the prebiotic and probiotic compositions.

[0189] In embodiments, the polysaccharide cleavage products and / or the soluble fruit fibers can be used to supplement a beverage. Examples of such beverages include, without limitation, infant formula, follow-on formula, toddler’s beverage, milk, fermented milk, fruit juice, fruit-based drinks, waters, and sports drinks. Many infant and toddler formulas are known in the art and are commercially available, including, for example, Carnation Good Start™ (Nestle Nutrition Division; Glendale, Calif.) and Nutrish AB™ produced by Mayfield Dairy Farms (Athens, Tenn.). Other examples of infant or baby formula include those disclosed in U.S. Patent No. 5,902,617. Other beneficial formulations of the compositions include the supplementation of animal milks, such as cow's milk. In embodiments, the polysaccharide cleavage products and / or soluble fruit fibers may also be used to help the absorption of other nutrients and minerals.

[0190] In some aspects, the polysaccharide cleavage products as described herein, can be used to stimulate microbes of any sort. Examples of microbes that can be stimulated by the polysaccharide cleavage products include, for example, soil microbes (e.g., mycorrhizal fungi and bacteria and other microbes used as soil inoculants such as Azosprillum sp.), oral bacterial (e.g., Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguis, and S. oralis) and skin bacteria (e.g., Propionibacterium acnes, also ammonia oxidizing bacteria, including but not limited to Nitrosomonas, Nitrosococcus, Nitrosospira, Nitrosocvstis, Nitrosolobus, and Nitrosovibrio.

[0191] In some aspects, the polysaccharide cleavage product and / or soluble fruit fiber compositions can be used as bulking-agents. In some aspects, the polysaccharide cleavage product and / or soluble fruit fiber compositions can be used as bulking-agents in reduced sugar food applications. In some aspects these polysaccharide cleavage products and / or soluble fruit fibers can be used as bulking-agents that do not affect flavor, odor, rheological, and textural properties.

[0192] In another aspect, the polysaccharide cleavage product and / or soluble fruit fiber compositions are administered to a subject for promotion of resistance to bacterial or yeast infections, e.g., Candidiasis or diseases induced by sulfate reducing bacteria. The polysaccharide cleavage products and / or the soluble fruit fibers described herein, can be used to stimulate yeast.Formulations and Administration

[0193] In some aspects, the polysaccharide cleavage product and / or the soluble fruit fiber can be formulated into a synthetic composition or administered alone. The synthetic composition can be in the form of a nutritional composition, a pharmaceutical composition, a prebiotic composition, a probiotic composition, and / or a synbiotic composition.

[0194] In some aspects, the prebiotic, probiotic, and synbiotic compositions can be formulated into pills or tablets or encapsulated in capsules, such as gelatin capsules. Tablet forms can optionally include, for example, one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphates, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid, and other excipients, colorants, fillers, binders, diluents, buffering agents, moistening agents, preservatives, flavoring agents, dyes, disintegrating agents, and pharmaceutically compatible carriers. Lozenge or candy forms can comprise the compositions in a flavor, e.g., sucrose, as well as pastilles comprising the compositions in an inert base, such as gelatin and glycerin or sucrose and acacia emulsions, gels, and the like containing, in addition to the active ingredient, carriers known in the art. The prebiotic or probiotic containing formulations may also contain conventional food supplement fillers and extenders such as, for example, rice flour.

[0195] In some aspects, the prebiotic, probiotic, and synbiotic containing compositions will comprise or further comprise a non-human protein, non-human lipid, non-human carbohydrate, or other non-human component. For example, in some aspects, the compositions may comprise a bovine (or other non-human) milk protein, a soy protein, a rice protein, beta-lactoglobulin, whey, soybean oil or starch. In some aspects, the polysaccharide cleavage products and / or soluble fruit fibers are combined with polysaccharides. In some aspects, the polysaccharide cleavage products and / or soluble fruit fibers are combined with the fruit-based starting material.

[0196] The dosages of the prebiotic and probiotic polysaccharide cleavage product and / or the soluble fruit fiber containing compositions will vary depending upon the requirements of the individual, and / or will take into account factors such as age (infant versus adult), weight, and reasons for loss of beneficial gut bacteria (e.g., antibiotic therapy, chemotherapy, radiation therapy, disease, or age). The administration regimen, and amount administered to, or consumed by an individual, in the context of the present invention should preferably be sufficient to establish colonization of the gut with beneficial bacteria over time. Theadministration regimen and / or the size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects that may accompany the administration of the provided prebiotic and probiotic polysaccharide cleavage product and / or the soluble fruit fiber containing compositions. In some administration aspects, the dosage range will be effective as a food supplement and for reestablishing beneficial bacteria in the intestinal tract. In some administration aspects, the dosage of a composition of the present invention ranges from about 1 micrograms / L to about 25 grams / L of polysaccharide cleavage product and / or soluble fruit fibers. In some aspects, the dosage of a composition is about 100 micrograms / L to about 15 grams / L of polysaccharide cleavage product and / or soluble fruit fibers. In some aspects, the dosage of an polysaccharide cleavage product and / or soluble fruit fiber composition is about 1- 10 g / L, 5-15 g / L, 10-50 g / L, or as high as 200 g / L. In some aspects, the dosage is 50-70 g / day. In some aspects, the dosage is 10 g / day. In some aspects, the dosage is between 1 and 10 g / day. In some aspects, the dosage is over 100 g / day. In some aspects, the dosage is 0.25-3 g / day. Exemplary Bifidobacterium dosages include, but are not limited to, about 104to about 1012colony forming units (CFU) per dose. A further advantageous range is about 106to about 1010CFU. Other bacterium can also be dosed at similar concentrations, but are not limited to, about 104to about 1012colony forming units (CFU) per dose or about 106to about 1010CFU.

[0197] The disclosed prebiotic and probiotic polysaccharide cleavage product and / or the soluble fruit fiber containing formulations can be administered to any subject / individual in need thereof. In some aspects, the individual is an infant or toddler. For example, in some aspects, the individual is less than, e.g., 3 months, 6 months, 9 months, one year, two years or three years old. In some aspects, the individual is between 3-18 years old. In some aspects, the individual is an adult (e.g., 18 years or older). In some aspects, the individual is over 50, 55, 60, 65, 70, or 75 years old.

[0198] Exemplary Bifidobacterium that can be included in the synbiotic compositions of the invention include, but are not limited to, Bifidobacterium longum subsp. infantis, B. longum subsp. longum, Bifidobacterium breve, Bifidobacterium adolescentis, and B. pseudocatenulatum. The Bifidobacterium used will depend in part on the target consumer.

[0199] It will be appreciated that it may be advantageous for some applications to include other Bifidogenic factors in the formulations described herein. Such additional components may include, but are not limited to, fructoligosaccharides such as Raffinose (Rhone-Poulenc, Cranbury, New Jersey), inulin (Imperial Holly Corp., Sugar Land, Texas), and Nutraflora(Golden Technologies, Westminister, Colorado), as well as lactose, xylooligosaccharides, soyoligosaccharides, lactulose / lactitol and galactooligosaccharides among others. In some applications, other beneficial bacteria, such as Lactobacillus, Rumminococcus, Akkermansia, Bacteroides, Faecalibacterium can be included in the formulations.

[0200] The nutritional composition can be a food, a beverage, a rehydration solution, a medical food or food for special medical purposes, a nutritional supplement and the like. The nutritional composition can contain sources of protein, lipids and / or digestible carbohydrates and can be in solid, powdered or liquid forms. The synthetic composition can be designed to be the sole source of nutrition, or as a food or nutritional supplement which forms part of the diet.

[0201] Suitable protein sources include milk proteins, soy protein, rice protein, pea protein and oat protein, or mixtures thereof. Milk proteins can be in the form of milk protein concentrates, milk protein isolates, whey protein or casein, or mixtures of both. The protein can be whole protein or hydrolyzed protein, either partially hydrolyzed or extensively hydrolyzed. Hydrolyzed protein offers the advantage of easier digestion which can be important for humans with inflamed or compromised GI tracts. The protein can also be provided in the form of free amino acids. The protein can comprise about 5 % to about 30% of the energy of the nutritional composition, normally about 10 % to 20%.

[0202] The protein source can be a source of glutamine, threonine, cysteine, serine, proline, or a combination of these amino acids. The glutamine source can be a glutamine dipeptide and / or a glutamine enriched protein. Glutamine can be included due to the use of glutamine by enterocytes as an energy source. Threonine, serine and proline are important amino acids for the production of mucin. Mucin coats the gastrointestinal tract and can improve intestinal barrier function and mucosal healing. Cysteine is a major precursor of glutathione, which is key for the antioxidant defenses of the body.

[0203] Suitable digestible carbohydrates include maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup, com syrup solids, high fructose com syrup, rice-derived carbohydrates, pea-derived carbohydrates, potato-derived carbohydrates, tapioca, sucrose, glucose, fructose, sucrose, lactose, honey, sugar alcohols (e.g. maltitol, erythritol, sorbitol), or mixtures thereof. Preferably the composition is reduced in or free from added lactose or other FODMAP carbohydrates. Generally digestible carbohydrates provideabout 35 % to about 55 % of the energy of the nutritional composition. A particularly suitable digestible carbohydrate is a low dextrose equivalent (DE) maltodextrin.

[0204] Suitable lipids include medium chain triglycerides (MCT) and long chain triglycerides (LCT). Preferably the lipid is a mixture of MCTs and LCTs. For example, MCTs can comprise about 30 % to about 70 % by weight of the lipids, more specifically about 50 % to about 60 % by weight. MCTs offer the advantage of easier digestion which can be important for humans with inflamed or compromised GI tracts. Generally, the lipids provide about 35 % to about 50 % of the energy of the nutritional composition. The lipids can contain essential fatty acids (omega-3 and omega-6 fatty acids). Preferably these polyunsaturated fatty acids provide less than about 30 % of total energy of the lipid source.

[0205] Suitable sources of long chain triglycerides are rapeseed oil, sunflower seed oil, palm oil, soy oil, milk fat, corn oil, high oleic oils, and soy lecithin. Fractionated coconut oils are a suitable source of medium chain triglycerides. The lipid profile of the nutritional composition is preferably designed to have a polyunsaturated fatty acid omega-6 (n-6) to omega-3 (n-3) ratio of about 4: 1 to about 10: 1. For example, the n-6 to n-3 fatty acid ratio can be about 6: 1 to about 9: 1. The nutritional composition may also include vitamins and minerals. If the nutritional composition is intended to be a sole source of nutrition, it preferably includes a complete vitamin and mineral profile. Examples of vitamins include vitamins A, B-complex (such as Bl, B2, B6 and Bl 2), C, D, E and K, niacin and acid vitamins such as pantothenic acid, folic acid and biotin. Examples of minerals include calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium and boron.

[0206] The nutritional composition can also include a carotenoid such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoid included can vary from about 0.001 pg / ml to about 10 pg / ml. Lutein can be included in an amount of from about 0.001 pg / ml to about 10 pg / ml, preferably from about 0.044 pg / ml to about 5 pg / ml of lutein. Lycopene can be included in an amount from about 0.001 pg / ml to about 10 pg / ml, preferably about 0.0185 pg / ml to about 5 pg / ml of lycopene. Beta-carotene can comprise from about 0.001 pg / ml to about 10 mg / ml, for example about 0.034 pg / ml to about 5 pg / ml of beta-carotene.

[0207] The nutritional composition preferably also contains reduced concentrations of sodium; for example, from about 300 mg / 1 to about 400 mg / 1. The remaining electrolytes canbe present in concentrations set to meet needs without providing an undue renal solute burden on kidney function. For example, potassium is preferably present in a range of about 1180 to about 1300 mg / 1; and chloride is preferably present in a range of about 680 to about 800 mg / 1.

[0208] The nutritional composition can also contain various other conventional ingredients such as preservatives, emulsifying agents, thickening agents, buffers, fiber and prebiotics (e.g. fructooligosaccharides, galactooligosaccharides), probiotics (e.g. B. animalis subsp. lactis BB-12, B. lactis HN019, B. lactis Bi07, B. infantis ATCC 15697, L. rhamnosus GG, L. rhamnosus HN001, L. acidophilus LA-5, L. acidophilus NCFM, L.fermentum CECT5716, B. longum BB536, B. longum AH1205, B. longum AH1206, B. breve M-16V, L. reuteri ATCC 55730, L. reuteri ATCC PTA-6485, L. reuteri DSM 17938), anti oxi dant / anti- inflammatory compounds including tocopherols, carotenoids, ascorbate / vitamin C, ascorbyl palmitate, polyphenols, glutathione, and superoxide dismutase (melon), other bioactive factors (e.g. growth hormones, cytokines, TFG-P), colorants, flavors, and stabilizers, lubricants, and so forth.

[0209] The nutritional composition can be formulated as a soluble powder, a liquid concentrate, or a ready -to-use formulation. The composition can be fed to a human in need via a nasogastric tube or orally. Various flavors, fibers and other additives can also be present.

[0210] The nutritional compositions can be prepared by any commonly used manufacturing techniques for preparing nutritional compositions in solid or liquid form. For example, the composition can be prepared by combining various feed solutions. A protein-in- fat feed solution can be prepared by heating and mixing the lipid source and then adding an emulsifier (e.g. lecithin), fat soluble vitamins, and at least a portion of the protein source while heating and stirring. A carbohydrate feed solution is then prepared by adding minerals, trace and ultra-trace minerals, thickening or suspending agents to water while heating and stirring. The resulting solution is held for 10 minutes with continued heat and agitation before adding carbohydrates (e.g. the [..] oligosaccharide and digestible carbohydrate sources). The resulting feed solutions are then blended together while heating and agitating and the pH adjusted to 6.6-7.0, after which the composition is subjected to high-temperature short-time processing during which the composition is heat treated, emulsified and homogenized, and then allowed to cool. Water soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range if necessary, flavors are added, and water is added to achieve the desired total solid level.

[0211] The resulting feed solutions are then blended together while heating and agitating and the pH adjusted to 6.6-7.0, after which the composition is subjected to high-temperature short-time processing during which the composition is heat treated, emulsified and homogenized, and then allowed to cool. Water soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range if necessary, flavors are added, and water is added to achieve the desired total solid level.

[0212] For a liquid product, the resulting solution can then be aseptically packed to form an aseptically packaged nutritional composition. In this form, the nutritional composition can be in ready-to-feed or concentrated liquid form. Alternatively, the composition can be spray- dried and processed and packaged as a reconstitutable powder.

[0213] When the nutritional product is a ready-to-feed nutritional liquid, it may be preferred that the total concentration of polysaccharide cleavage products in the liquid, by weight of the liquid, is from about 0.1 % to about 1.5 %, including from about 0.2 % to about 1.0 %, for example from about 0.3 % to about 0.7 %. When the nutritional product is a concentrated nutritional liquid, it may be preferred that the total concentration of polysaccharide cleavage products in the liquid, by weight of the liquid, is from about 0.2 % to about 3.0 %, including from about 0.4 % to about 2.0 %, for example from about 0.6 % to about 1.5 %.

[0214] The nutritional composition can also be in a unit dosage form. The unit dosage form can contain an acceptable food-grade carrier, e.g. phosphate buffered saline solution, mixtures of ethanol in water, water and emulsions such as an oil / water or water / oil emulsion, as well as various wetting agents or excipients. The unit dosage form can also contain other materials that do not produce an adverse, allergic or otherwise unwanted reaction when administered to a subject. The carriers and other materials can include solvents, dispersants, coatings, absorption promoting agents, controlled release agents, and one or more inert excipients, such as starches, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrating agents. Preferably carriers and other materials are low in FODMAPs or contain no FODMAPs.

[0215] The unit dosage form can be administered orally, e.g. as a tablet, capsule, or pellet containing a predetermined amount of the mixture, or as a powder or granules containing a predetermined concentration of the mixture or a gel, paste, solution, suspension, emulsion, syrup, bolus, electuary, or slurry, in an aqueous or non-aqueous liquid, containing apredetermined concentration of the mixture. An orally administered composition can include one or more binders, lubricants, inert diluents, flavoring agents, and humectants. An orally administered composition such as a tablet can optionally be coated and can be formulated to provide sustained, delayed or controlled release of the polysaccharide cleavage product and / or soluble fruit fiber.

[0216] The unit dosage form can also be administered by naso-gastric tube or direct infusion into the GI tract or stomach.

[0217] The unit dosage form can also include therapeutic agents such as antibiotics, probiotics, analgesics, and anti-inflammatory agents.

[0218] The proper dosage of the nutritional composition can be determined in a conventional manner, based upon factors such as the subject’s condition, immune status, body weight and age. In general, the dosage of the nutritional composition is such that the amount of polysaccharide cleavage product and / or soluble fruit fiber delivered is in the range from about 0.5 g to about 15 g per day, in certain embodiments from about 1 g to about 10 g per day, for example about 2 g to about 7.5 g per day. Appropriate dose regimes can be determined by methods known to those skilled in the art.

[0219] Pharmaceutical compositions herein comprise a named active ingredient in an amount effective for achieving the desired biological activity for a given form of administration to a given patient and optionally contain a pharmaceutically acceptable carrier. Pharmaceutical compositions can include an amount (for example, a unit dosage) of one or more of the disclosed compounds together with one or more non-toxic pharmaceutically acceptable additives, including carriers, diluents, and / or adjuvants, and optionally other biologically active ingredients. Such pharmaceutical compositions can be prepared by standard pharmaceutical formulation techniques such as those disclosed in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. (19th Edition).

[0220] The pharmaceutical composition can contain a pharmaceutically acceptable carrier, e.g. phosphate buffered saline solution, mixtures of ethanol in water, water and emulsions such as an oil / water or water / oil emulsion, as well as various wetting agents or excipients. Pharmaceutically acceptable carriers are those carriers that are compatible with the other ingredients in the formulation and are biologically acceptable. Carriers can be solid or liquid. It is currently contemplated that preferred carrier are liquid carriers. Carriers can include one or more substances that can also act as solubilizers, suspending agents, fillers,glidants, compression aids, binders, tablet-disintegrating agents, or encapsulating materials. Liquid carriers can be used in preparing solutions, suspensions, emulsions, syrups and elixirs. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water (of appropriate purity, e.g., pyrogen-free, sterile, etc.), an organic solvent, a mixture of both, or a pharmaceutically acceptable oil or fat. The liquid carrier can contain other suitable pharmaceutical additives such as, for example, solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators. Compositions for oral administration can be in either liquid or solid form.

[0221] Suitable examples of liquid carriers for oral and parenteral administration include water of appropriate purity, aqueous solutions (particularly containing additives, e.g. cellulose derivatives, sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols e.g. glycols) and their derivatives, and oils. Sterile liquid carriers are used in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant. Liquid pharmaceutical compositions that are sterile solutions or suspensions can be administered by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. Compositions for oral administration can be in either liquid or solid form. The carrier can also be in the form of creams and ointments, pastes, and gels. The creams and ointments can be viscous liquid or semisolid emulsions of either the oil-in-water or water-in-oil type.

[0222] The pharmaceutical composition can also contain other materials that do not produce an adverse, allergic or otherwise unwanted reaction when administered to humans. The carriers and other materials can include solvents, dispersants, coatings, absorption promoting agents, controlled release agents, and one or more inert excipients, such as starches, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrating agents. Preferably carriers and other materials are low in FODMAPs or contain no FODMAPs.

[0223] The pharmaceutical compositions can be administered orally, e.g. as a tablet, capsule, or pellet containing a predetermined amount, or as a powder or granules containing a predetermined concentration or a gel, paste, solution, suspension, emulsion, syrup, bolus, electuary, or slurry, in an aqueous or non-aqueous liquid, containing a predeterminedconcentration. Orally administered compositions can include binders, lubricants, inert diluents, flavoring agents, and humectants. Orally administered compositions such as tablets can optionally be coated and can be formulated to provide sustained, delayed or controlled release of the mixture therein.

[0224] The pharmaceutical compositions can also be administered by rectal suppository, aerosol tube, naso-gastric tube or direct infusion into the GI tract or stomach.

[0225] The pharmaceutical compositions can also include therapeutic agents such as antibiotics, probiotics, analgesics, and anti-inflammatory agents. The proper dosage of these compositions for a human can be determined in a conventional manner, based upon factors such condition, immune status, body weight and age. In general, the dosage of the nutritional composition is such that the amount of polysaccharide cleavage product and / or soluble fruit fiber delivered is in the range from about 0.5 g to about 15 g per day, in certain embodiments from about 1 g to about 10 g per day, for example from about 2 g to about 7.5 g per day. Appropriate dose regimes can be determined by conventional methods.

[0226] The amount of polysaccharide cleavage product and / or soluble fruit fiber required to be administered for the treatment of chronic gastrointestinal conditions associated with an impaired intestinal barrier function, reducing the risk of occurrence of a chronic gastrointestinal condition associated with an impaired intestinal barrier function, treating or reducing the risk of occurrence of a chronic metabolic condition, treating or reducing the risk of occurrence of a chronic kidney condition, treating or reducing the risk of occurrence of an atopic allergy, and / or treating or reducing the risk of occurrence of a chronic medical condition associated with dysfunction in gut brain interactions, will vary depending upon factors such as the risk and severity of the underlying condition, any other medical conditions or diseases, age, the form of the composition, and other medications being administered. Further the amount may vary depending upon whether the polysaccharide cleavage product and / or soluble fruit fiber is being used to deliver a direct effect (when the dose may be higher) or whether the polysaccharide cleavage product and / or soluble fruit fibers are being used as a secondary prevention / maintenance (when the dose may be lower). However, the required amount can be readily set by a medical practitioner and would generally be in the range from about 0.5 g to about 15 g per day, in certain embodiments from about 1 g to about 10 g per day, for example from about 2 g to about 7.5 g per day. An appropriate dose can be determined based on several factors, including, for example, body weight and / or condition, the severity of the underlying condition being treated or prevented, other ailments and / ordiseases, the incidence and / or severity of side effects and the manner of administration. Appropriate dose ranges may be determined by methods known to those skilled in the art. During an initial treatment phase, the dosing can be higher (for example 3 g to 15 g per day, preferably 4 g to 7.5 g per day). During a maintenance phase, the dosing can be reduced (for example, 1 g to 10 g per day, preferably 2 g to 7.5 g per day, more preferably about 2 g to about 5 g per day)).

[0227] Aspects of the Invention: Various aspects are contemplated herein, several of which are set forth in the paragraphs below. It is explicitly contemplated that any aspect or portion thereof can be combined to form an aspect. In addition, it is explicitly contemplated that any aspect (e.g., Aspect A13) that references an aspect (e.g., Aspect Al) for which there are sub-aspects having the same top level number (e.g., Aspect Ala, Alb, Ale, and so forth) necessarily includes reference to those sub-aspects Ala, Alb, Ale, and so forth. In other words, if Aspect Al 3 refers to Aspect Al, and there are Aspects Ala and Alb present, then Aspect A13 refers to Aspects Ala or Alb. Furthermore, although the aspects below are subdivided into aspects A, B, C, D, and so forth, it is explicitly contemplated that aspects in each of subdivisions A, B, C, D, etc. can be combined in any manner. Moreover, the term “any preceding aspect” means any aspect that appears prior to the aspect that contains such phrase (in other words, the sentence “Aspect B 13: The method of any one of aspects Bl -Bl 2, or any preceding aspect, ...” means that any aspect prior to aspect B 13 is referenced, including aspects Bl-B 12 and all of the “A” aspects). For example, it is contemplated that, optionally, any method or composition of any the below aspects may be useful with or combined with any other aspect provided below. Further, for example, it is contemplated that any embodiment described elsewhere herein, including above this paragraph, may optionally be combined with any of the below listed aspects. Within the aspects described herein, including those aspects set forth below, any “comprising” term (and grammatical variations thereof) can be replaced by “collectively comprising” (and grammatical variations thereof). In this regard, any structural feature of one or more oligosaccharides, or an oligosaccharide composition, can be described in terms of “comprising” or “collectively comprising” (and grammatical variations thereof). In some instances in the aspects below, or elsewhere herein, two open ended ranges are disclosed to be combinable into a range. For example, “at least X” is disclosed to be combinable with “less than Y” to form a range, in which X and Y are numeric values. For the purposes of forming ranges herein, it is explicitly contemplated that“at least X” combined with “less than Y” forms a range of X-Y inclusive of value X and value Y.

[0228] Aspect Al : A method for generating a mixture of polysaccharide cleavage products from a fruit-based starting material, the method comprising:(i) reacting one or more polysaccharide of a fruit-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0229] Aspect B 1 : A method for generating a mixture of polysaccharide cleavage products from a pineapple-based starting material, the method comprising:(i) reacting one or more polysaccharide of a pineapple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0230] Aspect Cl : A method for generating a mixture of polysaccharide cleavage products from an apple-based starting material, the method comprising:(i) reacting one or more polysaccharide of an apple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0231] Aspect D 1 : A method for generating a mixture of polysaccharide cleavage products from an orange-based starting material, the method comprising:(i) reacting one or more polysaccharide of an orange-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

[0232] Aspect El : The method of any preceding aspect, further comprising pretreating the starting material prior to reacting the one or more polysaccharide therein.

[0233] Aspect E2: The method of aspect El, or any preceding aspect, wherein the pretreating comprises extracting polyphenols from the starting material with an extractionsolvent to generate a polyphenol-reduced starting material prior to reacting the one or more polysaccharide of the polyphenol-reduced starting material.

[0234] Aspect E3 : The method of aspect E2, or any preceding aspect, wherein the polyphenols are extracted with water, methanol, ethanol, acetone or ethyl acetate.

[0235] Aspect E4: The method of aspect E2, or any preceding aspect, wherein the extraction solvent is acidified alcohol.

[0236] Aspect E5 : The method of aspect E2, or any preceding aspect, wherein the extraction solvent is acidified ethanol or isopropanol.

[0237] Aspect E6: The method of aspect E4 or E5, or any preceding aspect, wherein the alcohol is acidified by addition of an organic acid.

[0238] Aspect E7: The method of aspect E4 or E5, or any preceding aspect, wherein the alcohol is acidified by addition of a carboxylic acid that is optionally a di- or tri-carboxylic acid.

[0239] Aspect E8: The method of aspect E4 or E5, or any preceding aspect, wherein the alcohol is acidified by addition of a citric acid.

[0240] Aspect E9: The method of aspect E2, or any preceding aspect, wherein the extracting polyphenols employs ultrasound or microwave assistance.

[0241] Aspect E10: The method of aspect E2, or any preceding aspect, wherein the extracting polyphenols comprises using supercritical fluid extraction.

[0242] Aspect El l : The method of any one of aspects E2-E10, or any preceding aspect, wherein the pretreating comprises or further comprises extracting pectin from the starting material or the reduced-polyphenol starting material to generate a pectin-enriched starting material prior to reacting the one or more polysaccharide of the pectin-enriched starting material.

[0243] Aspect El la: The method of any one of aspects E2-E11, or any preceding aspect, wherein the pretreating comprises or further comprises extracting hemicellulose fibers from the starting material, the polyphenol-reduced starting material, or the pectin-enriched starting material to generate a hemicellulose-enriched starting material prior to reacting the one or more polysaccharide of the hemicellulose-enriched starting material.

[0244] Aspect E12: The method of aspect El 1, or any preceding aspect, wherein the extracting pectin comprises extraction with an acidic aqueous solution.

[0245] Aspect E12a: The method of aspect E12, or any preceding aspect, wherein the acidic aqueous solution comprises acidified alcohol, optionally acidified ethanol, optionally acidified isopropanol, or any combination thereof.

[0246] Aspect El 3: The method of any one of aspects El -El 2, or any preceding aspect, wherein the pretreating comprises or further comprises removing one or more polysaccharides other than pectin from the starting material.

[0247] Aspect E13a: The method of any one of aspects E1-E12, or any preceding aspect, wherein the pretreating comprises or further comprises removing one or more polysaccharides other than hemicellulose fibers from the starting material.

[0248] Aspect E13b: The method of any one of aspects E1-E12, or any preceding aspect, wherein the pretreating comprises or further comprises removing one or more polysaccharides other than pectin and hemicellulose fibers from the starting material.

[0249] Aspect E14: The method of aspect El 3, or any preceding aspect, wherein the one or more polysaccharides other than pectin are removed by extraction or enzyme treatment.

[0250] Aspect E14a: The method of aspect E13a, or any preceding aspect, wherein the one or more polysaccharides other than hemicellulose fibers are removed by extraction or enzyme treatment.

[0251] Aspect E14b: The method of aspect E13a, or any preceding aspect, wherein the one or more polysaccharides other than pectin and hemicellulose fibers are removed by extraction or enzyme treatment.

[0252] Aspect El 5: The method of aspect El 3, or any preceding aspect, wherein the one or more polysaccharides other than pectin is starch.

[0253] Aspect El 5a: The method of aspect El 5, or any preceding aspect, wherein the starch is removed by treatment with one or more amylases.

[0254] Aspect El 6: The method of any one of aspects El -El 5, or any preceding aspect, wherein the pretreating comprises or further comprises removal of protein from the starting material prior to reacting the one or more polysaccharide therein.

[0255] Aspect El 7: The method of aspect El 6, or any preceding aspect, wherein the protein is removed using one or more proteases.

[0256] Aspect El 8: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a solubility of greater than 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) in 1.2% (w / w) solution.

[0257] Aspect El 9: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a solubility of greater than 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) in 2.3% (w / w) solution.

[0258] Aspect E20: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a solubility of greater than 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) in 9.1% (w / w) solution.

[0259] Aspect E21 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a solubility of greater than 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) in 16.7% (w / w) solution.

[0260] Aspect E22: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a viscosity of less than 2 mPa s at 25°C (e.g., less than 2 mPa s at 25°C, less than 1.75 mPa s at 25°C, less than 1.5 mPa s at 25°C, less than 1.25 mPa s at 25°C, or less than 1 mPa s at 25°C) when dissolved in water at a concentration of 20 g / L.

[0261] Aspect E23 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a viscosity of less than 4 mPa s at 25°C (e.g., less than 4 mPa s at 25°C, less than 3.5 mPa s at 25°C, less than 3 mPa s at 25°C, less than 2.5 mPa s at 25°C, or less than 2 mPa s at 25°C) when dissolved in water at a concentration of 20 g / L.

[0262] Aspect E24: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a turbidity of less than 200 NTU (e.g., less than 200 NTU, less than 175 NTU, less than 150 NTU, less than 125 NTU, less than 100 NTU, less than 50 NTU, less than 40 NTU, or less than 20 NTU) at a concentration of 2.3% (w / w) or less.

[0263] Aspect E25: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products has a turbidity of less than 2500 NTU (e.g., less than 2500 NTU, less than 2000 NTU, less than 1500 NTU, less than 1000 NTU, less than 750 NTU,less than 500 NTU, less than 250 NTU, or less than 100 NTU) at a concentration of 16.7% (w / w) or less.

[0264] Aspect E26: The method of any preceding aspect, further comprising purifying the mixture of polysaccharide cleavage products to provide a purified mixture of polysaccharide cleavage products.

[0265] Aspect E27 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products are subjected to filtration or ethanol precipitation followed by filtration to remove ethanol insoluble material from the mixture of polysaccharide cleavage products.

[0266] Aspect E28: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products are treated to remove protein.

[0267] Aspect E28: The method of aspect E28, or any preceding aspect, wherein the mixture of polysaccharide cleavage products are treated with one or more proteases to remove protein, optionally two or more proteases to remove protein.

[0268] Aspect E29: The method of any preceding aspect, wherein salts and / or mono- and / or di-saccharides are removed from the mixture of polysaccharide cleavage products.

[0269] Aspect E30: The method of aspect 34, or any preceding aspect, wherein the salts and / or mono- and di-saccharides are removed from the mixture of polysaccharide cleavage products by nanofiltration and / or treatment with ion exchange resins.

[0270] Aspect E31 : The method of aspect Al, or any preceding aspect, wherein the fruitbased starting material is a citrus fruit-based starting material.

[0271] Aspect E32: The method of any preceding aspect, wherein the fruit-based starting material is or comprises pomace, husks, peels, rinds, marcs, seeds, skin, or any combination thereof.

[0272] Aspect E33: The method of any preceding aspect, wherein the fruit-based starting material comprises peels.

[0273] Aspect E34: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises less than 10% (e.g., less than 10%, less than 7.5%, less than 5%, less than 2.5%, or less than 1%) by weight of residual monosaccharides.

[0274] Aspect E35: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises less than 15% (e.g., less than 15%, less than 10%, less than 5%, less than 2.5%, or less than 1%) by weight of ash.

[0275] Aspect E35a: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises less than 10% (e.g., less than 10%, less than 7.5%, less than 5%, less than 2.5%, or less than 1%) by weight of ash.

[0276] Aspect E36: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products contains 40% or less (e.g., 40% or less, 35% or less, 30% or less, 25% or less, 20% or less, or 15% or less) by weight of polysaccharides with molecular weight of lOOkDa or more.

[0277] Aspect E37: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products contains 60% or less (e.g., 60% or less, 50% or less, 40% or less, 25% or less, 20% or less, or 15% or less), by weight, of polysaccharides with a molecular weight of 100 kDa or more.

[0278] Aspect E38: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products contains 20% or less (e.g., 200% or less, 15% or less, 10% or less, or 5% or less) by weight of polysaccharides with molecular weight of 100 kDa or more.

[0279] Aspect E39: The method of any preceding aspect, wherein the polysaccharide cleavage products of the mixture of polysaccharide cleavage products have an average degree of polymerization (DP) of 3 to 100.

[0280] Aspect E40: The method of any preceding aspect, wherein the polysaccharide cleavage products of the mixture of polysaccharide cleavage products have an average degree of polymerization (DP) of 3 to 50.

[0281] Aspect E41 : The method of any preceding aspect in which the mixture of polysaccharide cleavage products has solubility in water of 20 g / L or higher, optionally less than 500 g / L, optionally less than 250 g / L, optionally less than 100 g / L, at 25°C.

[0282] Aspect E42: The method of any preceding aspect in which the mixture of polysaccharide cleavage products has solubility in water of 50 g / L or higher, optionally less than 1000 g / L, optionally less than 500 g / L, optionally less than 250 g / L, optionally less than 100 g / L, at 25°C.

[0283] Aspect E43 : The method of any preceding aspect in which the mixture of polysaccharide cleavage products has solubility in water of 100 g / L or higher, optionally less than 2000 g / L, optionally less than 1000 g / L, optionally less than 500 g / L, at 25°C.

[0284] Aspect E44: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products exhibits the ability, when consumed by an animal, to increase the relative abundance of beneficial bacteria in the gastrointestinal tract of the animal.

[0285] Aspect E45: The method of aspect E44, or any preceding aspect, in which the relative abundance of one of more of Anaerostipes, Bacteroides, Bifidobacterium, Blaulia, Faecali bacterium, Fusicatenibacter , and Prevotella is increased when the mixture of polysaccharide cleavage products is consumed by an animal.

[0286] Aspect E46: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products are fermented by the microbiome of an animal to produce short chain fatty acids.

[0287] Aspect E47 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products exhibits the ability, when consumed by an animal, to increase the levels of butyrate, acetate, propionate, or any combination thereof, in the gut of the animal.

[0288] Aspect E48: The method of any preceding aspect in which the organoleptic properties of the mixture of polysaccharide cleavage products in a food or beverage product are suitable for consumption of the food or beverage product by a human.

[0289] Aspect E49: The method of any preceding aspect in which the organoleptic properties of the mixture of polysaccharide cleavage products in a liquid food or beverage product are suitable for consumption of the liquid food or beverage product by a human.

[0290] Aspect E50: The method of any preceding aspect, wherein the addition of the mixture of polysaccharide cleavage products to a food or beverage product is such that consumers will not reject the product.

[0291] Aspect E51 : The method of any preceding aspect, wherein, on addition of the mixture of polysaccharide cleavage products to a food or beverage product, the food or beverage product does not acquire a gritty or grainy texture as measured by expert or consumer sensory analysis.

[0292] Aspect E52: The method of any preceding aspect, wherein the clarity of a food or beverage product is not substantially decreased on addition of the mixture of polysaccharide cleavage products as measured by turbidity or expert and consumer sensory analysis.

[0293] Aspect E53 : The method of aspect E52, or any preceding aspect, wherein the food or beverage product is a beverage to which the mixture of polysaccharide cleavage products is added.

[0294] Aspect E54: The method of any preceding aspect, wherein a 20 g / L mixture of the mixture of polysaccharide cleavage products in water has turbidity of 40 NTU or less, optionally 35 NTU or less, optionally 30 NTU or less, optionally 25 NTU or less, optionally 20 NTU or less, optionally 15 NTU or less, optionally 10 NTU or less.

[0295] Aspect E55: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises one or more subunits selected from the group consisting of: xylose, arabinose, galactose, and galacturonic acid.

[0296] Aspect E56: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 30% to 60% by weight xylose subunits, 15% to 40% by weight arabinose subunits, 10% to 20% by weight galactose subunits, and 1% to 15% by weight galacturonic acid subunits, as measured by hydrolytic monosaccharide compositional analysis.

[0297] Aspect E57: The method of any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: terminal arabinose, 3, 5 -linked arabinose, 5 -linked arabinose, and 2, 5 -linked arabinose, as measured by glycosidic linkage analysis.

[0298] Aspect E58: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 10% to 25% of peak area of terminal arabinose, 5% to 20% of peak area of 3,5-linked arabinose, 1% to 10% of peak area of 5-linked arabinose, and 1% to 10% of peak area of 2, 5-linked arabinose, as measured by glycosidic linkage analysis.

[0299] Aspect E59: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 1% to 15% of peak area of terminal galactose, as measure by glycosidic linkage analysis.

[0300] Aspect E60: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises one or more subunits selected from the group consisting of: galacturonic acid, xylose, and galactose.

[0301] Aspect E61 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 45% to 70% by weight galacturonic acid subunits, 2% to 10% by weight xylose subunits, 2% to 10% by weight galactose subunits, and 2% to 10% by weight galactose subunits, as measured by hydrolytic monosaccharide compositional analysis.

[0302] Aspect E62: The method of any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal xylose, and 3, 4-linked galactose, as measured by glycosidic linkage analysis.

[0303] Aspect E63 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 3% to 15% of peak area of 4-linked galactose, 2% to 15% of peak area of terminal galactose, 1% to 10% of peak area of terminal xylose, and 1% to 10% of peak area of 3, 4-linked galactose, as measured by glycosidic linkage analysis.

[0304] Aspect E64: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises one or more subunits selected from the group consisting of: arabinose, galactose, galacturonic acid, and xylose.

[0305] Aspect E65: The method of any preceding aspect, wherein the mixture of soluble fruit fibers comprises 25% to 50% by weight arabinose subunits, 10% to 30% by weight galactose subunits, 5% to 40% by weight galacturonic acid subunits, and 3% to 15% by weight xylose subunits, as measured by hydrolytic monosaccharide compositional analysis.

[0306] Aspect E66: The method of any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal arabinose, 3,5-linked arabinose, as measured by glycosidic linkage analysis.

[0307] Aspect E67 : The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 5% to 20% of peak area of 4-linked galactose,2% to 12% of peak area of terminal galactose, 10% to 30% of peak area of terminal arabinose, and 3% to 15% of peak area of 3,5-linked arabinose, as measured by glycosidic linkage analysis.

[0308] Aspect E68: The method of any preceding aspect, wherein the mixture of polysaccharide cleavage products contains 70% or less (e.g., 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, or 10% or less) by weight of polysaccharides with molecular weight of lOOkDa or more.

[0309] Aspect Fl : Soluble fruit fiber comprising a mixture of polysaccharide cleavage products from a fruit-based starting material.

[0310] Aspect Fla: Soluble fruit fiber comprising a mixture of polysaccharide cleavage products from a fruit-based starting material, wherein a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 40 NTU or less, optionally 35 NTU or less, optionally 30 NTU or less, optionally 25 NTU or less, optionally 20 NTU or less, optionally 15 NTU or less, optionally 10 NTU or less.

[0311] Aspect Fib: Soluble fruit fiber comprising a mixture of polysaccharide cleavage products from a fruit-based starting material, wherein a 20 g / L mixture of the soluble fruit fiber in water has a dynamic viscosity of 4 millipascal-second (mPa.s) or less, optionally 3.5 mPa.s or less, optionally 3 mPa.s or less, optionally 2.5 mPa.s or less, optionally 2 mPa.s or less, optionally 1.5 mPa.s or less, at 25°C.

[0312] Aspect Fl c: Soluble fruit fiber comprising a mixture of polysaccharide cleavage products from a fruit-based starting material, wherein a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 40 NTU or less, optionally 35 NTU or less, optionally 30 NTU or less, optionally 25 NTU or less, optionally 20 NTU or less, optionally 15 NTU or less, optionally 10 NTU or less and a dynamic viscosity of 4 millipascal-second (mPa.s) or less, optionally 3.5 mPa.s or less, optionally 3 mPa.s or less, optionally 2.5 mPa.s or less, optionally 2 mPa.s or less, optionally 1.5 mPa.s or less, at 25°C.

[0313] Aspect F2: The soluble fruit fiber of aspect Fl, or any preceding aspect, wherein the mixture of polysaccharide cleavage products contains 40% or more (e.g., 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more) by weight of polysaccharide cleavage products having molecular weight less than 100 kDa.

[0314] Aspect F3: The soluble fruit fiber of aspect Fl or F2, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage product has a degree of polymerization (DP) of 3 to 100.

[0315] Aspect F3a: The soluble fruit fiber of aspect Fl or F2, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage product has a degree of polymerization (DP) of 3 to 50.

[0316] Aspect F4: The soluble fruit fiber of any one of aspects F1-F3, or any preceding aspect, wherein a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 20 NTU or less, optionally 15 NTU or less, optionally 10 NTU or less.

[0317] Aspect F5: The soluble fruit fiber of any one of aspects F1-F4, or any preceding aspect, wherein a 20 g / L mixture of the soluble fruit fiber in water has dynamic viscosity of 2 millipascal-second (mPa.s) or less, optionally 1.5 mPa.s or less, optionally 1 mPa.s or less, at 25°C.

[0318] Aspect F6: The soluble fruit fiber of any one of aspects F1-F5, or any preceding aspect, wherein the mixture of soluble fruit fiber is formed by cleavage of a polysaccharide obtained from a pineapple-based starting material.

[0319] Aspect F7: The soluble fruit fiber of any one of aspects F1-F5, or any preceding aspect, where the mixture of soluble fruit fiber is formed by cleavage of a polysaccharide obtained from an apple-based starting material.

[0320] Aspect F8: The soluble fruit fiber of any one of aspects F1-F5, or any preceding aspect, where the mixture of soluble fruit fiber is formed by cleavage of a polysaccharide obtained from a citrus-based starting material.

[0321] Aspect F9: The soluble fruit fiber of any one of aspects F1-F5, or any preceding aspect, where the mixture of soluble fruit fiber is formed by cleavage of a polysaccharide obtained from an orange-based starting material.

[0322] Aspect F10: The soluble fruit fiber of any one of aspects F1-F5, or any preceding aspect, wherein the fruit-based starting material is or comprises pomace, husk, peels, rinds, marcs, seeds, or a combination thereof.

[0323] Aspect Fl 1 : The soluble fruit fiber of aspect F10, or any preceding aspect, wherein the fruit-based starting material comprises peels.

[0324] Aspect G1 : A composition comprising a mixture of polysaccharide cleavage products, wherein the composition is formed by:(i) reacting one or more polysaccharide of a fruit-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products; wherein the mixture of polysaccharide cleavage products contains 5% or more, such as 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more, by weight of polysaccharides with a molecular weight of less than 100 kDa.

[0325] Aspect Hl : A composition comprising a mixture of polysaccharide cleavage products, wherein the composition is formed by:(i) reacting one or more polysaccharide of a pineapple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products; wherein the mixture of polysaccharide cleavage products contains 5% or more, such as 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more, by weight of polysaccharides with a molecular weight of less than 100 kDa.

[0326] Aspect II : A composition comprising a mixture of polysaccharide cleavage products, wherein the composition is formed by:(i) reacting one or more polysaccharide of an apple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products;wherein the mixture of polysaccharide cleavage products contains 5% or more, such as 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more, by weight of polysaccharides with a molecular weight of less than 100 kDa.

[0327] Aspect JI : A composition comprising a mixture of polysaccharide cleavage products, wherein the composition is formed by:(i) reacting one or more polysaccharide of an orange-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products; wherein the mixture of polysaccharide cleavage products contains 5% or more, such as 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 95% or more, by weight of polysaccharides with a molecular weight of less than 100 kDa.

[0328] Aspect KI : The composition of any one of aspects Gl-Jl, or any preceding aspect, wherein the starting material is or comprises pomace, husk, peels, rinds, marcs, seeds, or a combination thereof.

[0329] Aspect K2: The composition of aspect KI, or any preceding aspect, wherein the starting material comprises peels.

[0330] Aspect K3: The composition of any one of aspects G1-K2, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products has a degree of polymerization (DP) of 3 to 100.

[0331] Aspect K4: The composition of any one of aspects G1-K3, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products has a degree of polymerization (DP) of 3 to 50.

[0332] Aspect K5: The composition of any one of aspects G1-K4, or any preceding aspect, wherein a 20 g / L mixture of the mixture of polysaccharide cleavage products in water has turbidity of 40 NTU or less, optionally 35 NTU or less, optionally 30 NTU or less, optionally 25 NTU or less, optionally 20 NTU or less, optionally 15 NTU or less, optionally 10 NTU or less.

[0333] Aspect K6: The composition of any one of aspects G1-K5, or any preceding aspect, wherein a 20 g / L mixture of the mixture of polysaccharide cleavage products in water has a dynamic viscosity of 4 millipascal-second (mPa.s) or less, optionally 3.5 mPa.s or less, optionally 3 mPa.s or less, optionally 2.5 mPa.s or less, optionally 2 mPa.s or less, optionally 1.5 mPa.s or less, at 25°C.

[0334] Aspect K7: The composition or soluble fruit fiber of any one of aspects F1-K6, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises one or more subunits selected from the group consisting of: xylose, arabinose, galactose, and galacturonic acid.

[0335] Aspect K8: The composition or soluble fruit fiber of any one of aspects F1-K7, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 30% to 60% by weight xylose subunits.

[0336] Aspect K9: The composition or soluble fruit fiber of any one of aspects F1-K8, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 15% to 40% by weight arabinose subunits.

[0337] Aspect K10: The composition or soluble fruit fiber of any one of aspects F1-K9, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 10% to 20% by weight galactose subunits.

[0338] Aspect KI 1 : The composition or soluble fruit fiber of any one of aspects F1-K10, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 1% to 15% by weight galacturonic acid subunits.

[0339] Aspect K12: The composition or soluble fruit fiber of any one of aspects Fl -KI 1, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 30% to 60% by weight xylose subunits, 15% to 40% by weight arabinose subunits, 10% to 20% by weight galactose subunits, and 1% to 15% by weight galacturonic acid subunits, as measured by hydrolytic monosaccharide compositional analysis.

[0340] Aspect K13: The composition or soluble fruit fiber of any one of aspects F1-K12, or any preceding aspect, wherein at least 25% (e.g., at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, or at least 75%) of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 3,5-linked arabinose, 5-linked arabinose, and 2,5-linked arabinose, as measured by glycosidic linkage analysis.

[0341] Aspect K14: The composition or soluble fruit fiber of any one of aspects F1-K13, or any preceding aspect, wherein at least 50% (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95%) of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 3,5-linked arabinose, 5-linked arabinose, and 2,5-linked arabinose, as measured by glycosidic linkage analysis.

[0342] Aspect KI 5: The composition or soluble fruit fiber of any one of aspects Fl -KI 4, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 3,5-linked arabinose, 5-linked arabinose, and 2,5-linked arabinose, as measured by glycosidic linkage analysis.

[0343] Aspect K16: The composition or soluble fruit fiber of any one of aspects F1-K15, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 10% to 25% of peak area of terminal arabinose, 5% to 20% of peak area of 3,5-linked arabinose, 1% to 10% of peak area of 5-linked arabinose, and 1% to 10% of peak area of 2,5- linked arabinose, as measured by glycosidic linkage analysis.

[0344] Aspect KI 7: The composition or soluble fruit fiber of any one of aspects Fl -KI 6, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 1% to 15% of peak area of terminal galactose, as measure by glycosidic linkage analysis.

[0345] Aspect KI 8: The composition or soluble fruit fiber of any one of aspects Fl -KI 7, or any preceding aspect, wherein the mixture of polysaccharide cleavage products is or comprises CLX136.

[0346] Aspect KI 9: The composition or soluble fruit fiber of any one of aspects Fl -KI 8, or any preceding aspect, wherein the mixture of polysaccharide cleavage products is or comprises CLX136S.

[0347] Aspect K20: The composition or soluble fruit fiber of any one of aspects F1-K6, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises one or more subunits selected from the group consisting of: galacturonic acid, xylose, and galactose.

[0348] Aspect K21 : The composition or soluble fruit fiber of aspect K20, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 45% to 70% by weight galacturonic acid subunits.

[0349] Aspect K22: The composition or soluble fruit fiber of aspect K20 or K21, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 2% to 10% by weight xylose subunits.

[0350] Aspect K23 : The composition or soluble fruit fiber of any one of aspects K20- K22, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 2% to 10% by weight galactose.

[0351] Aspect K24: The composition or soluble fruit fiber of any one of aspects K20- K23, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 45% to 70% by weight galacturonic acid subunits, 2% to 10% by weight xylose subunits, 2% to 10% by weight galactose subunits, and 2% to 10% by weight galactose subunits, as measured by hydrolytic monosaccharide compositional analysis.

[0352] Aspect K25: The composition or soluble fruit fiber of any one of aspects K20- K24, or any preceding aspect, wherein at least 25% (e.g., at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, or at least 75%) of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal xylose, and 3, 4-linked galactose, as measured by glycosidic linkage analysis.

[0353] Aspect K26: The composition or soluble fruit fiber of any one of aspects K20- K25, or any preceding aspect, wherein at least 50% (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95%) of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal xylose, and 3, 4-linked galactose, as measured by glycosidic linkage analysis.

[0354] Aspect K27: The composition or soluble fruit fiber of any one of aspects K20- K26, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal xylose, and 3, 4-linked galactose, as measured by glycosidic linkage analysis.

[0355] Aspect K28: The composition or soluble fruit fiber of any one of aspects K20- K27, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 3% to 15% of peak area of 4-linked galactose, 2% to 15% of peak area of terminalgalactose, 1% to 10% of peak area of terminal xylose, and 1% to 10% of peak area of 3,4- linked galactose, as measured by glycosidic linkage analysis.

[0356] Aspect K29: The composition or soluble fruit fiber of any one of aspects K20- K28, or any preceding aspect, wherein the mixture of polysaccharide cleavage products is or comprises CLX138.

[0357] Aspect K30: The composition or soluble fruit fiber of any one of aspects F1-K6, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises one or more subunits selected from the group consisting of: arabinose, galactose, galacturonic acid, and xylose.

[0358] Aspect K31 : The composition or soluble fruit fiber of aspect K30, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 25% to 50% by weight arabinose subunits.

[0359] Aspect K32: The composition or soluble fruit fiber of aspect K30 or K31, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 10% to 30% by weight galactose subunits.

[0360] Aspect K33 : The composition or soluble fruit fiber of any one of aspects K30- K32, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 5% to 40% by weight galacturonic acid subunits.

[0361] Aspect K34: The composition or soluble fruit fiber of any one of aspects K30- K33, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 3% to 15% by weight xylose subunits.

[0362] Aspect K35: The composition or soluble fruit fiber of any one of aspects K30- K34, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 25% to 50% by weight arabinose subunits, 10% to 30% by weight galactose subunits, 5% to 40% by weight galacturonic acid subunits, and 3% to 15% by weight xylose subunits, as measured by hydrolytic monosaccharide compositional analysis.

[0363] Aspect K36: The composition or soluble fruit fiber of any one of aspects K30- K35, or any preceding aspect, wherein at least 25% (e.g., at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, or at least 75%) of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidiclinkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal arabinose, 3,5-linked arabinose, as measured by glycosidic linkage analysis.

[0364] Aspect K37: The composition or soluble fruit fiber of any one of aspects K30- K36, or any preceding aspect, wherein at least 50% (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95%) of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal arabinose, 3,5-linked arabinose, as measured by glycosidic linkage analysis.

[0365] Aspect K38: The composition or soluble fruit fiber of any one of aspects K30- K37, or any preceding aspect, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 4-linked galactose, terminal galactose, terminal arabinose, 3,5-linked arabinose, as measured by glycosidic linkage analysis.

[0366] Aspect K39: The composition or soluble fruit fiber of any one of aspects K30- K38, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises 5% to 20% of peak area of 4-linked galactose, 2% to 12% of peak area of terminal galactose, 10% to 30% of peak area of terminal arabinose, and 3% to 15% of peak area of 3,5-linked arabinose, as measured by glycosidic linkage analysis.

[0367] Aspect K40: The composition or soluble fruit fiber of any one of aspects K30- K39, or any preceding aspect, wherein the mixture of polysaccharide cleavage products is or comprises CLX137C.

[0368] Aspect K41 : The composition or soluble fruit fiber of any one of aspects F1-K40, or any preceding aspect, wherein the composition or soluble fruit fiber comprises 15% or less (e.g., 15% or less, 10% or less, 7.5% or less, 5% or less, or 2% or less) by weight of ash.

[0369] Aspect K41a: The composition or soluble fruit fiber of any one of aspects F1-K40, or any preceding aspect, wherein the composition or soluble fruit fiber comprises 5% or less (e.g., 5% or less, 4% or less, 3% or less, 2.5% or less, or 2% or less) by weight of ash.

[0370] Aspect K42: The composition or soluble fruit fiber of any one of aspects F1-K41, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises less than 10% (e.g., 10% or less, 7.5% or less, 5% or less, or 2% or less) by weight of mono- and di saccharides.

[0371] Aspect K43: The composition or soluble fruit fiber of any one of aspects F1-K42, or any preceding aspect, wherein the mixture of polysaccharide cleavage products is purified.

[0372] Aspect K44: The composition or soluble fruit fiber of any one of aspects F1-K43, or any preceding aspect, wherein the mixture of polysaccharide cleavage products comprises less than 10% (e.g., 10% or less, 7.5% or less, 5% or less, or 2% or less) by weight of residual monosaccharides.

[0373] Aspect LI : A prebiotic composition comprising the composition or soluble fruit fiber of any one of aspects F1-K44, or any preceding aspect, wherein the prebiotic composition stimulates growth of one or more beneficial bacteria.

[0374] Aspect L2: The prebiotic composition of aspect LI, or any preceding aspect, wherein the one or more beneficial bacteria is selected from the group consisting of: Anaerostipes, Bacteroides, Bifidobacterium, Blaulia, Faecalibacterium, Fusicatenibacter, and Prevotella.

[0375] Aspect Ml : A synbiotic composition comprising the composition or soluble fruit fiber of any one of aspects Fl -K44, or any preceding aspect, and one or more beneficial bacteria.

[0376] Aspect M2: The synbiotic composition of aspect Ml, or any preceding aspect, wherein the one or more beneficial bacteria are selected from the group consisting of: Anaerostipes, Bacteroides, Bifidobacterium, Blautia, Faecalibacterium, Fusicatenibacter, and Prevotella.

[0377] Aspect N1 : A food, beverage, or nutritional or medicinal product comprising the composition or soluble fruit fiber of any one of aspects F1-K44, or any preceding aspect.

[0378] Aspect Nla: A food product comprising the composition or soluble fruit fiber of any one of aspects F1-K44, or any preceding aspect, wherein the food product is or comprises one or more of a dairy -based product, a processed fruit, a processed vegetable, or a cerealbased product.

[0379] Aspect Nib: A beverage product comprising the composition or soluble fruit fiber of any one of aspects F1-K44, or any preceding aspect, wherein the beverage product is or comprises one or more of infant formula, follow-on formula, toddler’s beverages, milk, fermented milk, fruit juice, fruit-based drinks, waters, or sports drinks.

[0380] Aspect Nlc: A nutritional product comprising the composition or soluble fruit fiber of any one of aspects F1-K44, or any preceding aspect.

[0381] Aspect 01 : The nutritional product of aspect Nl, or any preceding aspect, further comprising a source of a protein, a lipid, a digestible carbohydrate, or any combination thereof.

[0382] The invention can be further understood by the following non-limiting examples.THE EXAMPLESEXAMPLE 1: Bench Scale Production of Oligosaccharide Compositions Derived from a Pineapple-Based Starting Material (CLX136)

[0383] Pineapple pomace is homogenized at a concentration of 10% w / v in 200 proof ethanol with 1% w / v citric acid and 5% v / v methanol and stirred for 3 hours at 60°C. The mixture is centrifuged at 4000g for 15 minutes at 20°C and the pellet dried under vacuum at 45°C for 15 hours. This process may be used to remove and / or reduce polyphenol content of pineapple pomace.

[0384] The dried material is then homogenized in water at a concentration of 5% w / v and adjusted to pH 5.5 with 0.25 M NaOH at 55°C. Cellulase (Celluclast - Novozymes) at a concentration of 0.3% v / v is added and the mixture is stirred at 55°C for 48 hours to remove cellulose. Polysaccharides are then precipitated in 60% v / v ethanol and centrifuged at 4700 RPM for 15 minutes at 20°C. The resulting pellet is lyophilized to obtain a fine beige powder.

[0385] The pellet of purified pineapple polysaccharides is dissolved in water at a final concentration of 5% w / v with 50mM ammonium acetate buffer adjusted to pH 5.5 with acetic acid. To this mixture 30% v / v hydrogen peroxide is added to a final concentration of 5% v / v. Copper (II) sulfate is then added to a final concentration of 0.016% w / v. The mixture is stirred at 55°C for 2 hours. The reaction mixture is then cooled to 20°C and the pH is adjusted to -pH 9 with the addition of 3.85% v / v 35% w / v ammonium hydroxide to initiate cleavage of the polysaccharides. The cleavage is allowed to proceed at 45°C for 2 hours while stirring. On completion, the mixture is clarified with a glass fiber filter, with any solid material being discarded. The filtrate is deionized in MB 10 mixed bed resin and lyophilized to produce a white powder made up of pineapple oligosaccharides (composition CLX136).

[0386] Composition CLX136 is subjected to hydrolytic monosaccharide compositional analysis to determine the monosaccharide composition of the pineapple oligosaccharides.The results are shown in Table 1. CLX136 comprises about 43% by mass of xylose, about 26% by mass of arabinose, about 6% by mass of galacturonic acid, about 16% by mass of galactose, and about 6% by mass of glucose.Table 1. Monosaccharide composition of CLX136 prior to removal of mono- and disaccharides. Hex refers to hexose sugars, Pent refers to pentose sugars, and HexA refers to hexur onic acid sugars.

[0387] Composition CLX136 is also subjected to glycosidic linkage analysis as described in Galermo, Nandita et al. 2018. The glycosidic linkage composition of CLX136 comprises 20% 4-linked xylose, 16% terminal arabinose, 13% 3,5-linked arabinose, 7% terminal galactose, 6% terminal xylose, 4% 2-linked xylose, 4% 4-linked glucose, 4% 5-linked arabinose, 3% 2, 5-linked arabinose, and 10% other minor linkages as shown in Table 5 in Example 5.EXAMPLE 2; Bench Scale Production of Oligosaccharide Compositions Derived from a Pineapple-Based Starting Material (CLX136S)

[0388] Pineapple pomace is homogenized at a concentration of 10% w / v in 200 proof ethanol with 1% w / v citric acid and 5% v / v methanol and stirred for 3 hours at 60°C. The mixture is then centrifuged at 4000g for 15 minutes at 20°C and the resulting pellet is dried under vacuum at 45°C for 15 hours. This process may be used to remove and / or reduce polyphenol content of pineapple pomace.

[0389] The dried pellet is homogenized in water at a concentration of 5%w / v and then the pH is adjusted to 5.5 using 0.25 M NaOH at 55°C. Celluclast (cellulase - Novozymes) at a concentration of 0.3%v / v is then added and the mixture is allowed to stir at 55°C for 48 hours to remove cellulose. Polysaccharides are then precipitated in 60% v / v ethanol and centrifuged at 4700 RPM for 15 minutes at 20°C. The resulting pellet is dried by lyophilization to obtain a fine beige powder.

[0390] The pellet of purified pineapple polysaccharides is dissolved in water at a final concentration of 5% w / v with 50mM ammonium acetate buffer adjusted to pH 5.5 with acetic acid. To this mixture 30% v / v hydrogen peroxide is added to a final concentration of 6% v / v and copper (II) sulfate is added to a final concentration of 0.018% w / v. The mixture is stirredat 55°C for 2 hours. The reaction mixture is then cooled to 20°C and the pH is adjusted to -pH 9 with the addition of 3.85% v / v 35% w / v ammonium hydroxide to initiate base cleavage. The base cleavage is allowed to proceed at 45°C for 2 hours while stirring. On completion, the mixture is clarified using a glass fiber filter, with the solid material being discarded. The filter effluent is deionized using a MB 10 mixed bed resin and dried by lyophilization to obtain a white powder made up of pineapple oligosaccharides (composition CLX136S).Table 2: Monosaccharide composition of CLX136S prior to removal of mono- and disaccharides. Hex refers to hexose sugars, Pent refers to pentose sugars, and HexA refers to hexur onic acid sugars.

[0391] Composition CLX136S is also subjected to glycosidic linkage analysis as described in Galermo, Nandita et al. 2018. The glycosidic linkage composition of CLX136S comprises 23% 4-linked xylose, 15% terminal arabinose, 12% 3,5-linked arabinose, 6% terminal galactose, 7% terminal xylose, 4% 2-linked xylose, 3% 4-linked glucose, 4% 5- linked arabinose, 3% 2,5-linked arabinose, 3% 2, 3, 4-linked xylose, and 5% other minor linkages as shown in Table 5 in Example 5.EXAMPLE 3: Bench Scale Production of Oligosaccharide Compositions Derived from an Orange-Based Starting Material (CLX137C)

[0392] Purified orange polysaccharides are dissolved in water at a final concentration of 10% w / v with 50mM ammonium acetate buffer adjusted to pH 5.5 with acetic acid. To this mixture, 30% v / v hydrogen peroxide is added to a final concentration of 5% v / v. Then copper (II) sulfate is added to a final concentration of 0.0159% w / v. The mixture is allowed to stir at 55°C for 2 hours. Once the time has elapsed, the reaction mixture is cooled to 15°C and the pH is adjusted to -pH 9 with the addition of 4.71% v / v 35% w / v ammonium hydroxide. The base cleavage is allowed to proceed at 45°C for 2 hours while stirring.

[0393] On completion, the mixture is clarified via centrifugation at 4700 RPM for 15 minutes at 8°C, with the solid material being discarded, then clarified further with a glass fiber filter. The filter effluent is deionized with an MB 10 mixed bed resin and dried via lyophilization resulting in a white powder.Table 3: Monosaccharide composition of CLX137C prior to removal of mono- and disaccharides. Hex refers to hexose sugars, Pent refers to pentose sugars, and HexA refers to hexur onic acid sugars.

[0394] Composition CLX137C is also subjected to glycosidic linkage analysis as described in Galermo, Nandita et al. 2018. The glycosidic linkage composition of CLX137C comprises 3% 4-linked xylose, 22% terminal arabinose, 6% 3,5-linked arabinose, 6% terminal galactose, 4% terminal xylose, 3% 2-linked xylose, 11% 4-linked glucose, 3% 5- linked arabinose, 4% terminal glucose, 10% 4-linked galactose, andl3% other minor linkagesas shown in Table 5 in Example 5.EXAMPLE 4: Bench Scale Production of Oligosaccharide Compositions Derived from an Apple-Based Starting Material (CLX138)

[0395] Purified Apple Pectin Powder polysaccharides from Monterey Bay Herb Co are dissolved in water at a final concentration of 10.0% w / v with 50mM ammonium acetate buffer adjusted to pH 5.5 with acetic acid. To this mixture, 30% v / v hydrogen peroxide is added to a final concentration of 6% v / v. Then iron (II) sulfate is added to a final concentration of 0.032% w / v. The mixture is allowed to stir at 55°C for 2 hours. Once the time has elapsed, the reaction mixture is cooled to 20°C and the pH is adjusted to -pH 9 with the addition of 9.91% v / v 35% w / v ammonium hydroxide. The base cleavage is allowed to proceed for 3 hours at 45°C for 2 hours while stirring.

[0396] On completion, the mixture is clarified with a glass fiber filter, with the solid material being discarded. The filter effluent is deionized with an MB 10 mixed bed resin and dried via lyophilization resulting in a white powder.Table 4: Monosaccharide composition of CLX138 prior to removal of mono- and disaccharides. Hex refers to hexose sugars, Pent refers to pentose sugars, and HexA refers to hexur onic acid sugars.

[0397] Composition CLX138 is also subjected to glycosidic linkage analysis as described in Galermo, Nandita et al. 2018. The glycosidic linkage composition of CLX138 comprises 12% 3-linked glucose, 28% 4-linked glucose, 23% terminal glucose, 6% 4-linked galactose, 2% 3, 4-linked galactose, 5% terminal galactose, 2% 4-linked xylose, 3% terminal xylose, 2% terminal arabinose, and 10% other minor linkages as shown in Table 5 in Example 5.EXAMPLE 5: Determination of Glycosidic Linkages and Molecular Weight Distribution

[0398] Compositions CLX136, CLX136S, CLX137C and CLX138 are subjected to glycosidic linkage analysis as described in Galermo, Nandita et al. 2018. Data are presented in units of peak area%. “Other” refers to linkages making up less than 1%. The notationrepresents a linkage that exists in an amount less than 1% (which can be 0%) of the total oligosaccharide weight. If linkage is not fully described, it is denoted by the monosaccharide, when known, or the type of monosaccharide, either pentose or hexose, followed by multiple or a single “x” denoting the number of branch points and finally the retention time, in parentheses, in the units of minutes.Table 5: Glycosidic linkage analysis of CLX136, CLX136S, CLX137C and CLX138.

[0399] Compositions CLX136, CLX136S, CLX137C and CLX138 are subjected to refractive index detection to determine their molecular weight distribution. Exemplary resulting molecular weight distributions are depicted in Table 6 below, and exemplary resulting refractive index graphs are depicted in FIGs. 1-4, with FIG. 1 showing results for CLX136, FIG. 2 showing results for CLX136S, FIG. 3 showing results for CLX137C, and FIG. 4 showing results for CLX138.Table 6: Molecular weight distribution of CLX136, CLX136S, CLX137C and CLX138 as determined by refractive index detection. Data are presented in units of peak area%.EXAMPLE 6: Effect of CLX136 on Ex-vivo Human Fecal Samples

[0400] Fecal samples are collected from healthy donors and stored at -80°C until processing. Aliquots of slurry from the fecal samples are prepared mixing three parts of fecal samples, one part of glycerol and one part of PBS. Slurries are stored at -80°C.

[0401] Static fecal fermentations are conducted in a deep 96-well format, under anaerobic conditions (Anaerobic Chamber Vinyl Type B), using a mix of gas (carbon dioxide 5%, hydrogen 5%, nitrogen balance), using inoculum from either individual donors or a pool of feces (2% of the fermentation mix).

[0402] The oligosaccharide composition, CLX136, is tested at a concentration of 0.6% w / v. Fermentation media is optimized to support diverse microbial taxa and control pH within the range of the colon physiological conditions, containing mineral and vitamin solution, CaC12 (10 mg / ml) and basic fermentation medium as described by MacFarlane GT et al (1989).

[0403] A mix of background sugars (xylan, amylopectin, potato starch, and pectin) are included in the basic fermentation media in low concentration to sustain microbial networks and minimize changes due to lack of nutrients which would confound the experimental results. Multiple samples are taken at different time points (Oh, 6h, lOh and 24h) for short chain fatty acid (SCFA) analysis and metabolomics analysis. After 24 hours of fermentation, gDNA is extracted from each well and sent for 16S rRNA sequencing.

[0404] Supernatants of fecal bacterial growths are derivatized with 3- nitrophenylhydrazine (3-NPH) for LC-MS / QTOF analysis. In brief, 20pL of supernatant is added to 20 pL of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1-EDC HC1) in 5% pyridine. Then 40 pL of 200mM 3-NPH in 80% acetonitrile with 50mM HC1 is added and briefly vortexed prior to incubating for 30 minutes at 40°C. After incubation, samples are diluted with 400 pL of 10% acetonitrile and vortexed. The dilutions are centrifuged prior to LC-MS / QTOF analysis.

[0405] Short chain fatty acids are analyzed with a 1290 Infinity II LC (Agilent Technologies, Santa Clara, CA) equipped with a reverse phase column (Zorbax Eclipse C18 2.1 x50mm; Agilent Technologies, Santa Clara, CA) and 6530 LC-MS QTOF (Agilent Technologies, Santa Clara, CA). LC separation is performed with 5%acetonitrile with 0.1% formic acid (solvent A) and 95% acetonitrile with 0.1% formic acid (solvent B). A separation gradient is as follows: 5% to 15%B for 2 minutes, then 15% to 30%B for 3 minutes, 30- 100%B in 0.1 minute, hold at 100%B for 1.9 minutes, return to 5% B in 0.1 minute, and equilibrate at 5% B for 1.9minutes. The MS conditions are set to positive mode with a scan range set at m / z 50-1100 at 1 spectra / sec scan rate. Peak area is quantitated using Agilent Quantitative Analysis software and areas are normalized to internal standards and compared to an external standard curve for quantitation.

[0406] The microbial short chain and organic acids are monitored during the fecal fermentation at time points 6-hour, 10-hour, and 24-hour and are compared to a sample containing only background sugars. Short chain and organic acids (SCFAs) make up the key intermediate and products of carbohydrate fermentation, many of which are known to be beneficial for human health. By the 24-hour timepoint, CLX136 promoted significant production of butyrate, acetate, and propionate compared to the untreated control. Selected results are shown in FIGs. 5A-5C (FIG. 5A: Butyrate; FIG. 5B: Acetate; FIG. 5C: Propionate).

[0407] By the 24-hour timepoint, CLX136 promoted the expansion of butyrate-producing species, including Blautia (FIG. 6A), Anaerostipes (FIG. 6B), Faecalibacterium, (FIG. 6C) and Fusicatenibacter (FIG. 6D) in multiple fecal donors relative to the untreated control. CLX136 also supported the expansion of the potentially beneficial bacterial taxa Bifidobacterium (FIG. 7A), Bacteroides (FIG. 7B), and Prevotella (FIG. 7C) species and prevented the expansion of the phylum Proteobacteria (FIG. 7D), which contains numerous enteric pathogens. Selected results are shown in FIGs. 7A-7D.

[0408] The results suggest that CLX136 oligosaccharides is a good candidate for applications involving improvement of conditions related to gut-brain interactions. These oligosaccharides can increase the production of SCFA, which play an important role in maintenance of intestinal gut health, assisting regeneration and maintaining optimum gut barrier impermeability, providing energy to colonic cells, regulation of immune system, prevention of inflammatory chronic diseases, such us obesity, insulin resistance, type 2 diabetes, and reduction of risk of colorectal cancer. Additionally, both Bifidobacterium andBacteroides species have been associated with modulation of the gut-brain axis through GABA production and regulation of enteroendocrine cells, respectively.EXAMPLE 7: Physical and Chemical Properties of CLX136

[0409] 296.7mg, 593.5mg, 2.5004g, and 5.0002g of CLX136 were weighed out and dissolved into four separate vials and 25 ml of water was added to make 1.2%, 2.3%, 9.1%, and a 16.7% w / w solutions respectively. Each vial was vortexed to aid in solubility. The vials were then sonicated for lOmins to ensure there were no bubbles in the mixture.Measurements of pH, EC, turbidity, and brix were measured (Table 7).Table 7: Initial physical characteristics of CLX136.

[0410] To test how these physical parameters may be impacted by processing, the samples were allowed to rest for 30 minutes at room temperature before being moved to an incubator set at 30° C for thirty more minutes. Again, pH, EC, turbidity, and brix were measured (Table 8).Table 8: Post-heat processing physical characteristics of CLX136.

[0411] All initial pH values were between 4.2 and 4.4, which was only slightly altered by incubation. EC, turbidity, and Brix values were not dramatically altered before and after incubation and showed a linear increase with increasing concentration.

[0412] Finally, to quantitatively assess solubility at each concentration, the solutions were centrifuged, the soluble fraction was separated from the pellet and freeze dried. The mass of each dried, soluble pellet was compared to the initial mass to determine % Solubility. 100%of the 1.2% solution was soluble, 94% of the 2.3% solution was soluble, 100% of the 9.1% solution was soluble, and 95% of the 16.7% solution was soluble. This result indicates, that even at high concentrations, >95% of the formulation is soluble.REFERENCES

[0413] All references listed below, or anywhere else throughout this description, are hereby incorporated by reference in their entireties for all purposes:

[0414] Alvarez, R., Araya, H., et al. (2016). “Evaluation of Polyphenol Content and Antioxidant Capacity of Fruits and Vegetables Using a Modified Enzymatyic Extraction.” Food Technol. Biotechnol. 54(4): 462-467.

[0415] Amicucci, M. J., Galermo, A.G., et al. (2019). "A rapid-throughput adaptable method for determining the monosaccharide composition of polysaccharides." International Journal of Mass Spectrometry 438: 22-28.

[0416] Babbar et al., 2016, “Pectic oligosaccharides from agricultural by-products: production, characterization and health benefits”, Critical Reviews in Biotechnology 30(4).

[0417] Guarner, F. and Malagelada, J.R. (2003). “Gut flora in health and disease.” The Lancet 361(9356): 512-519.

[0418] Galermo, A. G., Nandita, E., et al. (2018). “Liquid chromatography-tandem mass spectrometry approach for determining glycosidic linkages.” Analytical Chemistry' 90(21): 13073-13080.STATEMENTS REGARDING INCORPORATION BY REFERENCE AND VARIATIONS

[0419] All references throughout this application, for example patent documents and patent application publications, and non-patent literature documents or other source material, are hereby incorporated by reference in their entireties, as though individually incorporated by reference, to the extent each reference is at least partially not inconsistent with the invention in this application (for example, a reference that is partially inconsistent is incorporated by reference except for the partially inconsistent portion of the reference). All patents and publications mentioned in the specification are indicative of the levels of skill ofthose skilled in the art to which the invention pertains. References cited are incorporated by reference in their entirety to indicate the state of the art, in some cases as of their filing date, and it is intended that this information can be employed, if needed, to exclude (e.g., to disclaim) specific embodiments that are in the prior art. For example, when a compound is claimed, it should be understood that compounds known in the prior art, including certain compounds disclosed in the references disclosed (particularly in referenced patent documents), are not intended to be included in the claim.

[0420] This application incorporates each of the following by reference its entirety for all purposes, to the extent not inconsistent with the present invention: PCT Patent Application PCT / US23 / 21949, filed May 11, 2023 (WO2023 / 220318), PCT Patent Application No. PCT / US22 / 29065, filed May 12, 2022 (WO 2022 / 241163 Al), PCT Patent Application No. PCT / US2018 / 038350, filed June 19, 2018 (WO 2018 / 236917 Al), PCT Patent Application No. PCT / US2020 / 035748, filed June 2, 2020 (WO 2020 / 247389 Al), and PCT Patent Application No. PCT / US2020 / 060297, filed November 12, 2020 (WO 2021 / 097138 Al).

[0421] The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although this invention has been specifically disclosed by preferred embodiments, exemplary embodiments and optional features, modification and variation of the concepts disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the claims. The specific embodiments provided are examples of useful embodiments of the invention and it will be apparent to one skilled in the art that the invention may be carried out using a large number of variations of the devices, device components, methods steps set forth in the description.

[0422] As used in this specification and in the appended claims, the singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise. Also, the terms "a" (or "an"), "one or more" and "at least one" can be used interchangeably. It is also to be noted that the terms "comprising", "including", and "having" can be used interchangeably. The expression “of any of claims XX- YY” (wherein XX and YY refer to claim numbers) is intended to provide a multiple dependent claim in the alternative form, andin some embodiments is interchangeable with the expression “as in any one of claims XX- YY.”

[0423] When a group of substituents is disclosed, it is understood that all individual members of that group and all subgroups, are disclosed separately. When a Markush group or other grouping is used, all individual members of the group and all combinations and subcombinations possible of the group are intended to be individually included in the invention. Specific names of compounds are intended to be exemplary, as it is known that one of ordinary skill in the art can name the same compounds differently.

[0424] Every device, system, formulation, combination of components, or method described or exemplified can be used to practice the invention, unless otherwise stated.

[0425] Whenever a range is given in the specification, for example, a temperature range or composition range, the range is inclusive of the listed end points and all intermediate ranges and subranges, as well as all individual values included in the ranges given are intended to be included in the invention.

[0426] One of ordinary skill in the art will appreciate that starting materials, biological materials, reagents, synthetic methods, purification methods, analytical methods, assay methods, and biological methods other than those specifically exemplified can be employed in the practice of the invention without resort to undue experimentation. All functional equivalents known in the art, of any such materials and methods are intended to be included in this invention. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims.

Claims

WE CLAIM:

1. A method for generating a mixture of polysaccharide cleavage products from a fruitbased starting material, the method comprising:(i) reacting one or more polysaccharide of a fruit-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

2. A method for generating a mixture of polysaccharide cleavage products from a pineapple-based starting material, the method comprising:(i) reacting one or more polysaccharide of a pineapple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

3. A method for generating a mixture of polysaccharide cleavage products from an apple-based starting material, the method comprising:(i) reacting one or more polysaccharide of an apple-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

4. A method for generating a mixture of polysaccharide cleavage products from an orange-based starting material, the method comprising:(i) reacting one or more polysaccharide of an orange-based starting material with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products, and (ii) cleaving the reaction products with a cleavage agent to generate a mixture of polysaccharide cleavage products.

5. The method of any preceding claim, further comprising pretreating the starting material prior to reacting the one or more polysaccharide therein.

6. The method of claim 5, wherein the pretreating comprises extracting polyphenols from the starting material with an extraction solvent to generate a polyphenol-reduced starting material prior to reacting the one or more polysaccharide of the polyphenol- reduced starting material.

7. The method of claim 6, wherein the pretreating comprises or further comprises extracting pectin from the starting material or the polyphenol-reduced starting material to generate a pectin-enriched starting material prior to reacting the one or more polysaccharide of the pectin-enriched starting material.

8. The method of any one of claims 5-7, wherein the pretreating comprises or further comprises extracting hemicellulose fibers from the starting material, the polyphenol- reduced starting material, or the pectin-enriched starting material to generate a hemicellulose-enriched starting material prior to reacting the one or more polysaccharide of the hemicellulose-enriched starting material.

9. The method of any preceding claim, wherein salts and / or mono- and / or di-saccharides are removed from the mixture of polysaccharide cleavage products.

10. Soluble fruit fiber comprising a mixture of polysaccharide cleavage products from a fruit-based starting material, wherein a 20 g / L mixture of the soluble fruit fiber in water has turbidity of 40 NTU or less and / or a dynamic viscosity of 4 millipascal- second (mPa.s) or less at 25°C.

11. The soluble fruit fiber of claim 10, wherein the mixture of polysaccharide cleavage products contains 40% or more by weight of polysaccharide cleavage products having molecular weight less than 100 kDa.

12. The soluble fruit fiber of claim 10 or 11, wherein each of the polysaccharide cleavage products of the mixture of polysaccharide cleavage product has a degree of polymerization (DP) of 3 to 100, optionally a DP of 3 to 50.

13. The soluble fruit fiber of any one of claims 10-12, wherein the fruit-based starting material is or comprises a pineapple-based starting material, an apple-based starting material, a citrus-based starting material, or any combination thereof.

14. A composition comprising a mixture of polysaccharide cleavage products, wherein the composition is formed by:(i) reacting one or more polysaccharide of a fruit-based starting material, optionally a pineapple-based starting material, optionally an apple-based starting material, or optionally an orange-based starting material, with a Fenton’s reagent comprising a peroxide agent and a metal to generate reaction products; and(ii) cleaving the reaction products with a cleavage agent to generate the mixture of polysaccharide cleavage products; wherein the mixture of polysaccharide cleavage products contains 5% or more by weight of polysaccharides with a molecular weight of less than 100 kDa.

15. The composition or soluble fruit fiber of any one of claims 10-14, wherein the mixture of polysaccharide cleavage products comprises 30% to 60% by weight xylose subunits, 15% to 40% by weight arabinose subunits, 10% to 20% by weight galactose subunits, and 1% to 15% by weight galacturonic acid subunits, as measured by hydrolytic monosaccharide compositional analysis.

16. The composition or soluble fruit fiber of any one of claims 10-15, wherein at least 25% of the polysaccharide cleavage products of the mixture of polysaccharide cleavage products comprises at least one glycosidic linkage selected from the group consisting of: 3, 5 -linked arabinose, 5 -linked arabinose, and 2, 5 -linked arabinose, as measured by glycosidic linkage analysis.

17. The composition or soluble fruit fiber of any one of claims 10-14, wherein the mixture of polysaccharide cleavage products comprises 45% to 70% by weight galacturonic acid subunits, 2% to 10% by weight xylose subunits, 2% to 10% by weight galactose subunits, and 2% to 10% by weight galactose subunits, as measured by hydrolytic monosaccharide compositional analysis.

18. The composition or soluble fruit fiber of any one of claims 10-14, wherein the mixture of polysaccharide cleavage products is or comprises CLX136, CLX136S, CLX138, or CLX137C.

19. A prebiotic composition comprising the composition or soluble fruit fiber of any one of claims 10-18, wherein the prebiotic composition stimulates growth of one or more beneficial bacteria.

20. A food, beverage, or medicinal product comprising the composition or soluble fruit fiber of any one of claims 10-18.

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