Methods of treating biomass to produce oligosaccharides and related compositions

Abstract

The present application relates to methods and related compositions for treating biomass to produce oligosaccharides. Provided herein are methods of forming ingredients for human consumption. The method can include separating one or more soluble polysaccharides from the biomass, producing one or more oligosaccharides from the biomass, and combining the one or more separated soluble polysaccharides with the produced oligosaccharides to form a component. Methods of pre-treating biomass are also provided. The method may include applying a physical pretreatment to the biomass, applying a mild pretreatment to the physically pretreated biomass, and applying a strong pretreatment to the mildly pretreated biomass. Ingredients for human consumption are also provided.

Description

[0001] This application is a divisional application of the application with the filing date of August 14, 2020, application number 202080072717.0, and invention title "Methods and Related Compositions for Processing Biomass to Produce Oligosaccharides".

[0002] Cross-reference

[0003] This application claims the benefit of UK Patent Application No. 1911762.1 filed on August 16, 2019, UK Patent Application No. 1911764.7 filed on August 16, 2019, and UK Patent Application No. 2002315.6 filed on February 19, 2020, the entire contents of which are incorporated herein by reference. BACKGROUND OF THE INVENTION

[0004] Sugary foods and beverages are an important part of cultures and lifestyles around the world, but the sugars they contain are associated with obesity, diabetes, poor dental health, and disruptive behavior in people. For these reasons, consumer preferences have shifted away from sugary foods, and governments are increasingly implementing regulations to encourage reduced sugar consumption.

[0005] Therefore, for decades, the industry has been searching for suitable low-calorie sweeteners to replace sugars in foods and beverages. Unfortunately, many sugar substitutes are produced from non-natural resources and often have a slightly bitter or other unpleasant taste associated with their sweetness, which is unappealing to consumers. Additionally, while many sweeteners can mimic the sweetness of sugars in foods and beverages, few can mimic the broad roles that sugars play in foods, such as increasing volume, modulating texture, providing structure, acting as preservatives, and regulating color and flavor through caramelization and Maillard reactions. Moreover, many of the incremental sweeteners that can mimic these physical properties of sugars have gastrointestinal tolerance issues, which limit their use to levels far below those required to replace sugars in a standard Western diet.

[0006] Dietary fiber is an important part of a healthy diet and helps maintain the health of the digestive system and proper regulation of the gut microbiota. This fiber includes carbohydrates of different chain lengths and types. In addition to being naturally present in a wide variety of foods, fiber can also be produced separately and added to other foods during production. SUMMARY OF THE INVENTION

[0007] Other aspects and advantages of the present disclosure will become apparent to those skilled in the art from the following detailed description, which illustrates and describes only illustrative embodiments of the present disclosure. As will be appreciated, the present disclosure is capable of other different embodiments and several details thereof can be modified in various obvious aspects, all without departing from the present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature and not restrictive.

[0008] In some embodiments, methods for producing ingredients for human consumption are described herein. A method for producing an ingredient for human consumption can include: (a) separating one or more soluble polysaccharides from biomass; (b) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides; (c) separating one or more oligosaccharides; and (d) combining a portion of the one or more soluble polysaccharides from step (a) with a portion of the one or more oligosaccharides from step (c) to form the ingredient.

[0009] In some cases, the method can include purifying the separated one or more soluble polysaccharides.

[0010] In some cases, the method can include purifying the separated one or more oligosaccharides.

[0011] In some cases, the method can include treating the biomass to dissolve one or more soluble polysaccharides.

[0012] In some cases, the method can include purifying the separated one or more soluble polysaccharides.

[0013] In some cases, the treatment includes thermochemical treatment.

[0014] In some cases, the thermochemical treatment includes at least one of hot water treatment or hot alkali treatment.

[0015] In some cases, the hot alkali treatment uses an alkali with a pH of 10 to 14.

[0016] In some cases, the hot alkali treatment uses at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, or ammonia water.

[0017] In some cases, the treatment can be carried out at a temperature of 30 °C to 180 °C.

[0018] In some cases, the treatment can be carried out for 10 minutes to 24 hours.

[0019] In some cases, one or more soluble polysaccharides and / or one or more oligosaccharides are dried before step (d).

[0020] In some cases, one or more soluble polysaccharides and / or one or more oligosaccharides are dried after step (d).

[0021] In some cases, the component is soluble in water.

[0022] In some cases, the solubility of the component in water can be at least 80 g of the component per 100 g of water at 50 °C.

[0023] In some cases, the method includes combining the component with a liquid to form a liquid component.

[0024] In some cases, the viscosity of the liquid component can be similar to the viscosity of corn syrup.

[0025] In some cases, the viscosity of the liquid component can be similar to the viscosity of high fructose corn syrup.

[0026] In some cases, the calories per gram of the liquid component are lower than those of corn syrup or high fructose corn syrup.

[0027] In some cases, the glycemic index of the liquid component is lower than that of corn syrup or high fructose corn syrup.

[0028] In some cases, the liquid contains water.

[0029] In some cases, the liquid component contains at least 20% dry weight of at least one oligosaccharide and at least 2% dry weight of at least one polysaccharide.

[0030] In some cases, the viscosity of the liquid component is 5 cps to 100,000 cps, 8,000 cps to 100,000 cps, 10,000 cps to 50,000 cps, or 15,000 cps to 25,000 cps.

[0031] In some cases, the liquid component contains at least 2% dry weight of xylan.

[0032] In some cases, the liquid component contains at least 2% dry weight of mannan.

[0033] In some cases, the liquid component contains at least 2% dry weight of cellulose derivatives.

[0034] In some cases, the polysaccharide concentration of the liquid component is 0.1% to 50% w / v.

[0035] In some cases, the ratio of the amount of polysaccharide to oligosaccharide in the liquid component is 1:100 to 1:1.

[0036] In some cases, the one or more soluble polysaccharides include at least one of mannan, xylan, mixed-linkage glucan, lignocellulose, hemicellulose, cellulose derivatives, chitosan, or xyloglucan.

[0037] In some cases, the cellulose derivatives include at least one of cellulose acetate, hydroxyethyl cellulose, or hydroxypropyl cellulose.

[0038] In some cases, the biomass includes at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass, or softwood biomass.

[0039] In some cases, the one or more oligosaccharides include at least one of the following: i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2 to 12; iii) arabinoxylooligosaccharides with a DP of 3 to 15; iv) mannanooligosaccharides with a DP of 2 to 12; v) mixed-linkage glucan oligosaccharides with a DP of 2 to 5; vi) xyloglucan oligosaccharides with a DP of 4 to 12; or vii) chitosan oligosaccharides with a DP of 2 to 12.

[0040] In some cases, the composition includes at least two of the oligosaccharides listed in (i) to (vii).

[0041] In some cases, the composition includes at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other.

[0042] In some embodiments, compositions for human consumption are described herein. Compositions for human consumption may include: soluble polysaccharides; and oligosaccharides, which may include at least one of the following: (i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; (ii) xylooligosaccharides with a DP of 2 to 12; (iii) mannanooligosaccharides with a DP of 2 to 12; (iv) arabinoxylooligosaccharides with a DP of 3 to 15; (v) mixed-linkage glucan oligosaccharides with a DP of 2 to 5; or (vi) chitosan oligosaccharides with a DP of 2 to 12, wherein the composition contains less than 5% dry weight of insoluble polysaccharides.

[0043] In some cases, the composition may be substantially free of insoluble polysaccharides.

[0044] In some cases, the composition may be soluble in water.

[0045] In some cases, the solubility of the composition in water may be at least 80 g of the composition per 100 g of water at 50 °C.

[0046] In some cases, the composition further includes a liquid, thereby forming a liquid composition.

[0047] In some cases, the liquid may be water.

[0048] In some cases, the liquid composition comprises at least one oligosaccharide at at least 20% dry weight and at least one polysaccharide at at least 2% dry weight.

[0049] In some cases, the viscosity of the liquid composition is from 5 cps to 100,000 cps, from 8,000 cps to 100,000 cps, from 10,000 cps to 50,000 cps or from 15,000 cps to 25,000 cps.

[0050] In some cases, the liquid composition comprises at least 2% dry weight of xylan.

[0051] In some cases, the liquid composition comprises at least 2% dry weight of mannan.

[0052] In some cases, the liquid composition comprises at least 2% dry weight of a cellulose derivative.

[0053] In some cases, the polysaccharide concentration of the liquid composition is from 0.1% to 50% w / v.

[0054] In some cases, the ratio of the amounts of polysaccharide and oligosaccharide comprised in the liquid composition is from 1:100 to 1:1.

[0055] In some cases, the one or more soluble polysaccharides comprise at least one of mannan, xylan, mixed-link glucan, lignocellulose, hemicellulose, cellulose derivative, chitosan or xyloglucan.

[0056] In some cases, the cellulose derivative comprises at least one of cellulose acetate, hydroxyethyl cellulose or hydroxymethyl cellulose.

[0057] In some cases, the biomass comprises at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass or softwood biomass.

[0058] In some cases, the one or more oligosaccharides comprise at least one of the following: i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2 to 12; iii) arabinoxylooligosaccharides with a DP of 3 to 15; iv) mannanooligosaccharides with a DP of 2 to 12; v) mixed-link glucan oligosaccharides with a DP of 2 to 5; vi) xyloglucan oligosaccharides with a DP of 4 to 12; or vii) chitooligosaccharides with a DP of 2 to 12.

[0059] In some cases, the composition comprises at least two of the oligosaccharides listed in (i) to (vii).

[0060] In some cases, the components comprise at least two oligosaccharides in a ratio to each other of from 1:9 to 1:1.

[0061] In some embodiments, methods for producing ingredients for human consumption are described herein. The methods for producing ingredients for human consumption may include: (a) applying a physical pretreatment to biomass to reduce the average size of the biomass; (b) applying a mild pretreatment to the physically pretreated biomass, the mild pretreatment may include: (i) incubating the physically pretreated biomass in an aqueous solution to dissolve monosaccharides and / or disaccharides from the physically pretreated biomass; and (ii) removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution; (c) applying a strong pretreatment to the mildly pretreated biomass to improve the digestibility of the biomass; (d) contacting one or more polysaccharide-cleaving enzymes with the strongly pretreated biomass in a solution or suspension to form one or more oligosaccharides; and (e) enriching the solution or suspension to increase the concentration of the one or more oligosaccharides to form the ingredient.

[0062] In some cases, the mild pretreatment may be an incubation cycle.

[0063] In some cases, the strong pretreatment may be a thermochemical treatment, and the thermochemical treatment may include incubating the mildly pretreated biomass in one of an acidic solution or a basic solution.

[0064] In some cases, the method may further include removing at least 25% or 50% of the dissolved monosaccharides and / or disaccharides from the incubation solution in step (b)(ii).

[0065] In some cases, the strongly pretreated biomass composition after step (c) contains less than 10% w / w monosaccharides.

[0066] In some cases, the method may further include purifying one or more oligosaccharides from the solution or suspension.

[0067] In some cases, the strongly pretreated biomass composition after step (c) contains less than 20% w / w monosaccharides.

[0068] In some cases, the method may further include repeating step (b).

[0069] In some cases, step (b) may be carried out two, three, four, or five times.

[0070] In some cases, the method may further include repeating step (c).

[0071] In some cases, step (c) may be carried out two, three, four, or five times.

[0072] In some cases, the method may further include concentrating a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

[0073] In some cases, the method may further include discarding a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

[0074] In some cases, a portion of the dissolved monosaccharides and / or disaccharides removed in step (b) may not be combined with a portion of one or more oligosaccharides in step (e) to form the composition.

[0075] In some cases, the composition contains less than 15% dry weight of monosaccharides.

[0076] In some cases, the composition contains less than 50% dry weight of disaccharides.

[0077] In some cases, the composition may be substantially free of monosaccharides.

[0078] In some cases, the composition may be substantially free of disaccharides.

[0079] In some cases, the one or more oligosaccharides include at least one of the following: i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2 to 12; iii) arabinoxylooligosaccharides with a DP of 3 to 15; iv) mannanooligosaccharides with a DP of 2 to 12; v) mixed-link glucan oligosaccharides with a DP of 2 to 5; vi) xyloglucan oligosaccharides with a DP of 4 to 12; or vii) chitooligosaccharides with a DP of 2 to 12.

[0080] In some cases, the composition contains at least two of the oligosaccharides listed in (i) to (vii).

[0081] In some cases, the composition contains at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other.

[0082] In some cases, the composition contains less than 50% total dry weight w / w of sucrose, maltose, lactose, glucose, fructose, or galactose, which accounts for the total dry weight w / w of all the oligosaccharides of i - vii.

[0083] In some cases, the monosaccharides and / or disaccharides include at least one of sucrose, maltose, lactose, glucose, fructose, or galactose.

[0084] In some cases, in addition to the monosaccharides and / or disaccharides, step (b) also dissolves one or more organic acids.

[0085] In some cases, the one or more organic acids include at least one of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid, or acetic acid.

[0086] In some cases, the total weight of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid, and acetic acid can be greater than 10% of the total weight of sucrose, maltose, lactose, glucose, fructose, and galactose in the portion dissolved and removed in step b.

[0087] In some cases, the physical pretreatment in step (a) includes at least one of cutting, chopping, grinding, ball milling, polishing, trimming, or mixing the biomass.

[0088] In some cases, the mild pretreatment in step (b) is carried out in an aqueous solution containing water.

[0089] In some cases, the mild pretreatment in step (b) is carried out at a temperature of 5°C to 150°C.

[0090] In some cases, the mild pretreatment in step (b) can be carried out for 15 minutes to 1 hour.

[0091] In some cases, the strong pretreatment in step (c) includes heating the mildly pretreated biomass in an acidic solution or an alkaline solution.

[0092] In some cases, the heating can be carried out at a temperature of 50°C to 150°C.

[0093] In some cases, the heating can be carried out for 30 minutes to 4 hours.

[0094] In some cases, step (c) includes treating the mildly pretreated biomass in an alkaline solution with a pH of 8 to 11.

[0095] In some cases, the alkaline solution contains at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, ammonia water, ammonium sulfate, or ammonium hydroxide.

[0096] In some cases, step (c) includes treating the mildly pretreated biomass in an acidic solution with a pH of 4 to 6.

[0097] In some cases, the acidic solution contains at least one of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, or oxalic acid.

[0098] In some cases, the biomass includes at least one of sugarcane, corn stover, corn cob, wheat bran, wheat straw, hardwood, or softwood.

[0099] In some cases, the biomass includes at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed-link glucan, mannan, or lignocellulose.

[0100] In some cases, the one or more polysaccharide-cleaving enzymes comprise at least one of cellulase, xylanase, xyloglucanase, endoglucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase (LPMO).

[0101] In some cases, the one or more polysaccharide-cleaving enzymes comprise at least one of AA9, AA10, AA11, AA13, AA14, or AA15.

[0102] In some cases, the one or more polysaccharide-cleaving enzymes can be prepared from filamentous fungi such as Trichoderma reesei.

[0103] In some cases, the one or more polysaccharide-cleaving enzymes can be operably linked to a catalytic module.

[0104] In some cases, the one or more polysaccharide-cleaving enzymes can be operably linked to a non-catalytic module.

[0105] In some cases, the non-catalytic module can be a carbohydrate-binding module.

[0106] In some cases, a water-soluble composition for human consumption can comprise at least one of the following oligosaccharides: i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2 to 12; iii) arabinoxylooligosaccharides with a DP of 3 to 15; iv) mannanooligosaccharides with a DP of 2 to 12; v) mixed-link glucan oligosaccharides with a DP of 2 to 5; vi) xyloglucan oligosaccharides with a DP of 4 to 12; or vii) chitooligosaccharides with a DP of 2 to 12; and at least one of the following monosaccharides or disaccharides: sucrose, maltose, lactose, fructose, or galactose, wherein the total dry weight of the monosaccharide or disaccharide is less than 10% of the total dry weight of the oligosaccharides with a DP of 2 to 12.

[0107] In some cases, the composition can comprise at least two of the oligosaccharides listed in (i) to (vii). In some cases, the composition can further comprise at least one of the following organic acids: oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid, or acetic acid. In some cases, the composition can comprise at least two of the organic acids. In some cases, the composition can comprise at least two of the oligosaccharides listed in (i) to (vii) in a ratio of 1:9 to 1:1 relative to each other.

[0108] In some cases, the composition may comprise at least one monosaccharide or disaccharide selected from: glucose, fructose or sucrose; and at least one organic acid selected from: oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid or acetic acid, wherein the total weight of the organic acid is greater than 10% of the total weight of the monosaccharide or disaccharide.

[0109] In some cases, a method of producing an ingredient for human consumption may include:

[0110] (a) applying a physical pretreatment to the biomass to reduce the average size of the biomass; (b) applying a mild pretreatment to the physically pretreated biomass, the mild pretreatment comprising: (i) incubating the physically pretreated biomass in an aqueous solution to dissolve monosaccharides and / or disaccharides from the physically pretreated biomass; and (ii) removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution; (c) applying a strong pretreatment to the mildly pretreated biomass to dissolve polysaccharides and increase the digestibility of the plant biomass; (d) separating one or more dissolved polysaccharides from the biomass; (e) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides; (f) separating one or more oligosaccharides; and (g) combining a portion of one or more soluble polysaccharides from step (d) with a portion of one or more oligosaccharides from step (f) to form the ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0111] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the invention will be obtained by reference to the following detailed description and the drawings, which illustrate exemplary embodiments in which the principles of the invention are utilized, in the drawings (also referred to herein as "FIGURES"):

[0112] Figure 1 Depicts the results of thin layer chromatography (TLC) analysis showing the monosaccharide and disaccharide content of the indicated biomass after 1 - 5 washing cycles (incubation pretreatment).

[0113] Figure 2 Depicts the results of TLC analysis comparing the levels of monosaccharides and disaccharides in the enzyme hydrolysis product mixture when washing (+) and not washing (-) (incubation pretreatment) was performed.

[0114] Figure 3 Depicts the results of preparing a glaze with the compositions of the present disclosure and comparative compositions.

[0115] Figure 4 Depicts a baked product using the liquid ingredient of the present disclosure.

[0116] Figure 5 Depicts the high performance anion exchange chromatography (HPAEC) analysis of sample 4.

[0117] Figure 6 Depicts a cereal bar product produced using the liquid components of the present disclosure.

[0118] Figure 7 Depicts the HPAEC chromatogram of sugars after washing corncobs.

[0119] Figure 8 Is a simplified flowchart describing a method for processing biomass according to some embodiments of the present disclosure.

[0120] Figure 9 Is a simplified flowchart depicting another method for processing biomass according to some embodiments of the present disclosure.

[0121] Figure 10A Is a diagram depicting a method for processing biomass.

[0122] Figure 10B Describes the measurement of sugars in multiple samples.

[0123] Figure 10C Depicts the measurement of organic acids in multiple samples.

[0124] Figure 10D Depicts according to Figure 10A Visual observation of a sample processed by the method.

[0125] Figure 11A Illustrates a comparison between cereal bars made using soluble polysaccharides and cereal bars made using insoluble polysaccharides.

[0126] Figure 11B Depicts the Figure 11A Hardness and stickiness of a cereal bar using penetration testing.

[0127] Figure 11C Depicts the Figure 11A Hardness of a cereal bar using cutting testing. Detailed Description

[0128] Introduction

[0129] The present disclosure provides a method for generating one or more components. The method may include separating one or more soluble polysaccharides from biomass or feedstock (such as plant biomass). Then, the remaining biomass may be contacted with one or more enzymes or treated with one or more enzymes to form one or more oligosaccharides, and the one or more oligosaccharides may be enriched or separated. In addition, at least a portion of the one or more soluble polysaccharides separated from the biomass may be combined with a portion of the one or more enriched or separated oligosaccharides to form the component.

[0130] The texture of the ingredient can be continuous and smooth. Additionally, the ingredient can have certain properties such that it can be used as a sweetener and / or sugar substitute. The properties of the ingredient can include sweetness, smooth texture, desirable mouthfeel, binding ability, sizing ability or ability to form a paste, moistness, viscosity, bulking ability, and / or caramelization ability. Compared to corn syrup or high fructose corn syrup, the ingredient can also have lower calories, a reduced glycemic index, a reduced glycemic load, increased fiber, and / or reduced sugar.

[0131] The present disclosure also provides methods for producing ingredients for incorporation into foods, nutraceuticals, and / or cosmetics, where the methods can include one or more pretreatment steps on the biomass. For example, the method can include a first pretreatment step, a second pretreatment step, a third pretreatment step, or additional pretreatment steps on the biomass. The pretreatment steps can be carried out in a specified order.

[0132] The biomass for producing one or more oligosaccharides can be plant biomass. Examples of plant biomass include, but are not limited to: sugarcane, corn stover, corn cobs, wheat bran, wheat straw, hardwood, or softwood. In some cases, the biomass can contain cellulose, chitin, chitosan, xylan, xyloglucan, mixed-linkage glucan, mannan, or lignocellulose.

[0133] The biomass can be digested into one or more oligosaccharides. The biomass digestion can be carried out enzymatically. The enzymatic digestion can be performed after one or more pretreatment steps. One or more pretreatment steps can be carried out to reduce the size of the biomass and / or increase the surface area of the biomass available for digestion. One or more pretreatment steps can include one or more washing steps, solubilization steps, or predigestion treatments. In some cases, one or more pretreatment steps can be carried out to reduce the monosaccharides and / or disaccharides present in the biomass. In some cases, one or more pretreatment steps can be carried out to recover a soluble polysaccharide fraction from the biomass.

[0134] During one or more pre-treatment steps, monosaccharides and / or disaccharides can be removed from the starting material (e.g., biomass). In other words, monosaccharides and / or disaccharides can be removed from biomass during one or more pre-treatment steps. Thus, after completion of one or more pre-treatment steps, little or no monosaccharides and / or disaccharides are produced. That is to say, the pre-treated biomass may contain little or no monosaccharides and / or disaccharides. This can improve the efficiency of the method for producing ingredients incorporated into foods, nutraceuticals, and / or cosmetics as provided herein. For example, since a portion of the monosaccharides and / or disaccharides have been removed from the biomass during one or more pre-treatment steps, less ingredient purification (e.g., to remove monosaccharides and / or disaccharides) may be required. Specifically, fewer or less stringent filtration steps may be required during purification to produce the ingredients disclosed herein.

[0135] The first pre-treatment step (Pre-treatment step 1) can include physically treating the biomass (e.g., cutting the biomass). The second pre-treatment step (Pre-treatment step 2 or mild pre-treatment) can include subjecting the physically treated biomass to a incubation cycle or a washing cycle. The incubation cycle can include incubating the physically treated biomass (from Pre-treatment step 1) in an aqueous solution to dissolve monosaccharides and / or disaccharides from the physically treated biomass. The incubation cycle can also include removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution. In some cases, the aqueous solution can include water. In some other cases, the incubation cycle can be carried out at about 25 °C for a period of about 30 minutes to about 1.5 hours. Pre-treatment step 2 can be a mild pre-treatment step. For example, the conditions of Pre-treatment step 2 (e.g., solution, temperature, time, etc.) can be milder than the conditions of Pre-treatment step 3 described in further detail below.

[0136] The third pretreatment step (pretreatment step 3 or strong pretreatment) can include treating the incubated biomass from pretreatment step 2 in one of an acidic solution or an alkaline solution. Pretreatment step 3 can improve the digestibility of the biomass (e.g., by enzymes). Pretreatment step 3 can also improve the access of enzymes to the biomass. In various cases, pretreatment step 3 can occur in an alkaline solution (e.g., 1% w / v NaOH solution) at a temperature above room temperature (e.g., about 90 °C to about 110 °C). Additionally, pretreatment step 3 can be carried out by maintaining at the desired temperature for about 30 minutes to 1 hour. For example, the effective temperature - 90 °C in this example - is maintained for 1 hour (which can be changed according to the desired characteristics). The solution in the third pretreatment step can be retained. In other words, when the treated biomass moves from the pretreatment step to the steps after the pretreatment step described in further detail below, the solution can be not discarded. In some cases, pretreatment step 3 can include thermochemical treatment. That is, pretreatment step 3 can be carried out in an acidic or alkaline solution and / or can be carried out at a temperature above room temperature.

[0137] In various cases, after one or more pretreatment steps, the method for producing a composition for human consumption can include contacting one or more polysaccharide-cleaving enzymes with the biomass from pretreatment step 3 in a solution or suspension to form one or more oligosaccharides. The method can also include enriching the solution or suspension to increase the concentration of one or more oligosaccharides to form the composition (e.g., a composition for human consumption).

[0138] Compared with some other methods, the pretreatment steps can improve the efficiency of the method. Since less downstream processing may be required, the efficiency can be improved. In some cases, downstream processing can include processes for removing monosaccharides and / or disaccharides from the pretreated biomass. In some embodiments, it may be difficult to remove disaccharides alone, e.g., from an intermediate solution or fraction produced by an enzymatic digestion method. Therefore, removing disaccharides during the pretreatment step may be more effective.

[0139] The steps of downstream processing can include ion exchange chromatography, ultrafiltration, microfiltration, nanofiltration, etc. Part of the role of the nanofiltration step can be to remove excess monosaccharides from the oligosaccharide mixture. The nanofiltration can be carried out multiple times to reach the desired monosaccharide level. When there are fewer monosaccharides in the pretreated biomass (e.g., when monosaccharides have been removed by washing or incubation), the number of said nanofiltration steps can be reduced. Additionally, ultrafiltration generally cannot distinguish between the desired disaccharides (e.g., cellobiose) and the unwanted disaccharides. Therefore, removing unwanted disaccharides during pretreatment and producing the desired disaccharides during the enzymatic treatment step can also reduce the number of steps involved or required during downstream processing.

[0140] Although various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, changes, and substitutions can be made by those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed.

[0141] As used in the specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "sample" also includes multiple samples, including mixtures thereof.

[0142] As used herein, the term "about" may mean within 1 or more standard deviations. Alternatively, "about" may mean a range of up to 10%, up to 5%, or up to 1% of a given value. For example, about may mean up to ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of a given value.

[0143] As used herein, "food" and "foodstuff" generally refer to any article for consumption, which can be consumed by humans or any other animal. It can be food, feed, beverage, or an ingredient for producing any of the foregoing.

[0144] As used herein, "nutraceutical" generally refers to any composition introduced into a human or other animal by ingestion, injection, absorption, or any other method for the purpose of providing nutrition to the human or other animal. The use of such nutraceuticals can take the form of beverages with added dietary fiber, probiotic additives, pills, or other capsules, or any other suitable use.

[0145] As used herein, "cosmetic" generally refers to any composition intended for use on a human or other animal to enhance its aesthetic appeal or prevent future loss of aesthetic appeal, as well as any other composition generally referred to as a cosmetic. Aesthetic appeal is not limited to visual aesthetics but also applies to structural or any other aspect of appeal. Cosmetics can be mascara, foundation, lip gloss, eyeshadow cream, eyeliner cream, primer, lipstick, blush, nail polish, bronzer, or any other makeup; shampoo, conditioner, styling mousse, styling gel, hairspray, hair dye, hair wax, or any other hair product; moisturizer, exfoliant, sunscreen, cleanser, toothpaste, cream, lotion, ointment, or any other composition that effectively modifies the teeth, skin, hair, or other parts of the body in an aesthetic manner. Additionally, cosmetics can be compositions used as components of face masks, brushes, curling irons, other styling devices, other solid structures, or any other suitable composition.

[0146] As used herein, "ingredient" generally refers to any composition suitable for incorporation into a food, cosmetic, or nutritional product, which may include those used directly as the product itself. It can be a dry ingredient or a liquid ingredient, unless it is specifically referred to as "dry" or "liquid". This includes compositions that can be considered intermediates during the methods of the present disclosure, such as compositions formed after combining one or more oligosaccharides and one or more soluble polysaccharides before any further purification, optimization, drying, dissolution, or any other such step, and including the final compositions obtained from the methods.

[0147] As used herein, "polysaccharide" generally refers to any sugar polymer having a length greater than about 20 residues. Polysaccharides can be highly branched, mildly branched, or unbranched. Polysaccharides can contain any combination of any type of glycosidic bond, any number of, for example, α or β bonds, and any combination of monomer types (such as glucose, glucosamine, mannose, xylose, galactose, fucose, fructose, glucuronic acid, arabinose, or their derivatives), such as any combination of the above monomers modified with an acetyl group or other group. Polysaccharides can be cellulose or hemicellulose polymers, and contemplated hemicellulose polymers include xylan, glucuronoxylan, arabinoxylan, glucomannan, and xyloglucan. In some embodiments, the cellulose polymer can be cellulose.

[0148] As used herein, "lignocellulose" generally refers to aggregates containing polysaccharides that are plant cell wall materials or derived from plant cell wall materials. For example, they can contain one or more of the following polysaccharides associated together: cellulose, xylan, mannan, and mixed-linkage glucan.

[0149] As used herein, "highly branched", "mildly branched", and "unbranched" generally refer to the number of side chains per main chain in a sugar. A highly branched sugar has an average of 4 to 10 side chains per 10 main chain residues, a mildly branched sugar has an average of 1 to 3 side chains per 10 main chain residues, and an unbranched sugar has only one main chain and no side chains. The average is calculated by dividing the number of side chains in the sugar by the number of main chain residues.

[0150] As used herein, "sugar" generally refers to any polysaccharide and / or oligosaccharide, such as monosaccharides and / or disaccharides.

[0151] As used herein, "oligosaccharide" generally refers to a sugar polymer having a chain length less than or equal to about 20 sugar residues. Oligosaccharides can be highly branched, mildly branched, or unbranched; and can contain any combination of glycosidic bonds, any number of α or β bonds, and any combination of monomer types (such as glucose, glucosamine, mannose, xylose, galactose, fucose, fructose, glucuronic acid, arabinose, or their derivatives). Suitable derivatives include the above monomers containing an acetyl group or other group.

[0152] As used herein, "monosaccharide" and "disaccharide" generally refer to sugar compounds composed of one or two residues, respectively. Monosaccharides are compounds such as glucose, glucosamine, xylose, galactose, fucose, fructose, glucuronic acid, arabinose, galacturonic acid; or their epimers or other derivatives. Suitable derivatives include acetyl or other groups. Disaccharides are compounds composed of two monosaccharides linked by any glycosidic bond.

[0153] As used herein, "fructooligosaccharide" generally refers to an oligosaccharide composed of one or more glucose residues linked by β-1,4-glycosidic bonds, and can be chemically related thereto by oxidation, reduction, esterification, epimerization or another chemical modification.

[0154] As used herein, "xylooligosaccharide" generally refers to an oligosaccharide composed mainly of xylose residues (usually linked by β-1,4-glycosidic bonds), and may also contain glucuronic acid residues and / or arabinose residues and / or acetyl groups and / or any other modifications, and can be chemically related thereto by oxidation, reduction, esterification, epimerization, further glycosylation or another chemical modification.

[0155] As used herein, "arabinoxylan oligosaccharide" generally refers to an oligosaccharide composed of xylose residues (usually linked by β-(1→4)-bonds substituted by arabinose side chains, usually linked by (1→2)-bonds or (1→3)-bonds), and can be chemically related thereto by oxidation, reduction, esterification, epimerization, further glycosylation or another chemical modification.

[0156] As used herein, "mixed-linkage glucan-oligosaccharide" generally refers to an oligosaccharide composed of one or more glucose residues linked by at least one β-1,3-glycosidic bond and at least one β-1,4-glycosidic bond, and can be chemically related thereto by oxidation, reduction, esterification, epimerization or another chemical modification.

[0157] As used herein, "mannooligosaccharide" generally refers to an oligosaccharide composed of one or more mannose residues and optionally containing one or more glucose and / or galactose residues, and can be chemically related thereto by oxidation, reduction, esterification, epimerization or another chemical modification;

[0158] As used herein, "chitooligosaccharide" generally refers to an oligosaccharide composed of one or more glucosamine and / or N-acetyl-glucosamine residues, and can be chemically related thereto by oxidation, reduction, esterification, epimerization or another chemical modification.

[0159] As used herein, "cellulose" generally refers to a polysaccharide composed of glucose residues linked by β-1,4-glycosidic bonds and its derivatives. As used herein, "xylan" generally refers to a polysaccharide composed of a backbone of xylose residues and may also contain glucuronic acid residues and / or arabinose residues and / or acetyl groups and / or any other modifications. As used herein, "mixed-linkage glucan" generally refers to a polysaccharide composed of glucose residues linked by β-1,3-glycosidic bonds and β-1,4-glycosidic bonds. As used herein, "mannan" generally refers to a polysaccharide composed of greater than 40% mannose residues and optionally containing glucose and / or galactose residues. As used herein, "chitin" or "chitosan" generally refers to a polysaccharide composed of glucosamine and / or N-acetyl-glucosamine residues. The polysaccharides of cellulose, xylan, mixed-linkage glucan, mannan, chitin or chitosan may include chemical variants that have been modified by oxidation, reduction, esterification, epimerization or another chemical modification.

[0160] As used herein, "soluble", "solubility" and their grammatical variants generally refer to solubility in an aqueous solution (e.g., water). As used herein, "dissolve" generally refers to incorporating a solid into an aqueous solution or liquid to form a solution.

[0161] As used herein, "suspension" generally refers to a composition comprising at least two immiscible phases (e.g., a solid phase and a liquid phase), wherein the weight of the solid phase can be from about 0.5% to about 30%, from about 1% to about 20%, from about 2% to about 15% or from about 3% to about 10%, by weight percentage of the composition. The suspension may contain a suitable solvent, which may be water.

[0162] As used herein, "viscosity" generally refers to a quantity representing the magnitude of internal friction in a fluid, measured by the force resisting uniform flow per unit area. Viscosity can be measured by a variety of methods, but unless otherwise stated, the values given herein refer to those obtained by using a standard test method using a Brookfield HDB VE rotational viscometer, operating in accordance with the manufacturer's instructions for the range, and collecting a 400 mL sample in a tall-form beaker to ensure that no container effects occur.

[0163] As used herein, "dissolved" generally refers to a solid incorporated into a liquid to form a solution.

[0164] I. Pretreatment

[0165] Physical Pretreatment

[0166] Mechanical or physical pretreatment can be performed on the biomass to digest the biomass into one or more oligosaccharides. The mechanical and / or physical pretreatment can be the first pretreatment step in the biomass digestion process. Alternatively, the mechanical or physical pretreatment step can be performed after another pretreatment step. For example, the mechanical or physical pretreatment step can be performed after another pretreatment step (such as a washing pretreatment step).

[0167] The biomass can be mechanically or physically pretreated to reduce the size of the biomass. Examples of the mechanical or physical pretreatment step include, but are not limited to: cutting, chopping, grinding, ball milling, milling, trimming, mixing, and / or steam explosion of the biomass. More than one physical pretreatment can be performed on the biomass.

[0168] Dissolution step

[0169] The biomass can be subjected to a dissolution step. The dissolution step can be a mild pretreatment step or part of a mild pretreatment step, and the dissolution step dissolves the polysaccharide fraction or the monosaccharide and / or disaccharide fraction from the biomass. The dissolution pretreatment step can be a washing step, an incubation step, a thermochemical step, or a chemical treatment step. The dissolution pretreatment can be performed before the physical or mechanical pretreatment step. The dissolution pretreatment can be performed after the physical or mechanical pretreatment step.

[0170] In some embodiments, a dissolution step can be performed to remove a portion of the soluble polysaccharides. The soluble polysaccharides can be added to one or more oligosaccharides, for example, after purification of the soluble polysaccharides. The soluble polysaccharide fraction can be used to produce food types with desired taste, texture, quality, viscosity, and odor. The presence of the soluble polysaccharides can also contribute to the production of a better quality product that does not produce precipitates (or particulate matter) in the food.

[0171] The dissolution step can be a chemical or thermochemical treatment of the biomass. The chemical or thermochemical treatment can include one or more aqueous solutions. The aqueous solution can contain one or more salts, acids, bases, or ions. The aqueous solution can include one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, ammonia water, dilute sulfuric acid, dilute acetic acid, dilute hydrochloric acid, or dilute phosphoric acid.

[0172] The aqueous solution can be an alkaline solution with a pH of 10 to 14. The aqueous solution can be an alkaline solution with a pH of 10 to 11, 10 to 12, 10 to 13, 10 to 14, 11 to 12, 11 to 13, 11 to 14, 12 to 13, 12 to 14, or 13 to 14. The aqueous solution can be an acidic solution with a pH of 2 to 6. The aqueous solution can be an acidic solution with a pH of 2 to 3, 2 to 4, 2 to 5, 2 to 6, 3 to 4, 3 to 5, 3 to 6, 4 to 5, 4 to 6, or 5 to 6.

[0173] The dissolution step can be carried out at a temperature of from 30°C to 180°C. The dissolution step can be carried out at a temperature of at least 30°C. The dissolution step can be carried out at a temperature of at most 180°C. The dissolution step can be carried out at the following temperatures: 30°C to 60°C, 30°C to 90°C, 30°C to 120°C, 30°C to 150°C, 30°C to 180°C, 60°C to 90°C, 60°C to 120°C, 60°C to 150°C, 60°C to 180°C, 90°C to 120°C, 90°C to 150°C, 90°C to 180°C, 120°C to 150°C, 120°C to 180°C or 150°C to 180°C. The dissolution step can be carried out at a temperature of at least 30°C, 60°C, 90°C, 120°C, 150°C or 180°C.

[0174] The duration of the dissolution step can be varied depending on the biomass used, the desired soluble polysaccharide fraction and / or the complexity of the desired soluble polysaccharide. The dissolution step can be carried out for about 10 minutes to about 24 hours. The dissolution step can be carried out for at least 10 minutes. The dissolution step can be carried out for at most 60 minutes. The dissolution step can be carried out for 10 minutes to 30 minutes, 10 minutes to 60 minutes or 30 minutes to 60 minutes. The dissolution step can be carried out for at least 10 minutes, 30 minutes, or 60 minutes. The dissolution step can be carried out for 1 hour to 24 hours. The dissolution step can be carried out for at least 1 hour. The dissolution step can be carried out for at most 24 hours. The dissolution step can be carried out for 1 hour to 4 hours, 1 hour to 8 hours, 1 hour to 12 hours, 1 hour to 16 hours, 1 hour to 20 hours, 1 hour to 24 hours, 4 hours to 8 hours, 4 hours to 12 hours, 4 hours to 16 hours, 4 hours to 20 hours, 4 hours to 24 hours, 8 hours to 12 hours, 8 hours to 16 hours, 8 hours to 20 hours, 8 hours to 24 hours, 12 hours to 16 hours, 12 hours to 20 hours, 12 hours to 24 hours, 16 hours to 20 hours, 16 hours to 24 hours, or 20 hours to 24 hours. The dissolution step can be carried out for at least 1 hour, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours or 24 hours.

[0175] Soluble polysaccharides can be removed from the solution after the dissolution step. A portion of the soluble polysaccharides can be removed from the soluble fraction. In some cases, at least 5% of the soluble polysaccharides can be removed and / or purified. In some cases, up to 100% of the soluble polysaccharides can be removed and / or purified. In some cases, 5% to 10%, 5% to 20%, 5% to 30%, 5% to 40%, 5% to 60%, 5% to 80%, 5% to 90%, 5% to 100%, 10% to 20%, 10% to 30%, 10% to 40%, 10% to 60%, 10% to 80%, 10% to 90%, 10% to 100%, 20% to 30%, 20% to 40%, 20% to 60%, 20% to 80%, 20% to 100%, 30% to 40%, 30% to 60%, 30% to 80%, 30% to 100%, 40% to 60%, 40% to 80%, 40% to 100%, 60% to 80% or 60% to 100% of the soluble polysaccharides can be removed and / or purified. In some cases, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80% or 100% of the soluble polysaccharides can be removed and / or purified.

[0176] Mild pretreatment

[0177] The biomass can be subjected to a mild pretreatment step. The mild pretreatment step can dissolve the polysaccharide fraction or the monosaccharide and / or disaccharide fraction from the biomass. In some cases, the mild pretreatment step can include at least a part or all of the dissolution step. In other words, the dissolution step can be a part or a sub-step of the mild pretreatment step. The mild pretreatment step can be a washing step, an incubation step, a thermochemical step or a chemical treatment step. The mild pretreatment can be performed before the physical or mechanical pretreatment step. The mild pretreatment can be performed after the physical or mechanical pretreatment step. The mild pretreatment can be performed simultaneously with the physical or mechanical pretreatment step. The mild pretreatment step can be performed one or more times. The mild pretreatment can be carried out 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.

[0178] The mild pretreatment step can be an incubation step or a washing step. The biomass (physically treated or untreated) can be incubated in an aqueous solution. The aqueous solution can be water, or the aqueous solution can contain salts, acids, bases, ions, alcohols, and / or other chemicals. The pH of the aqueous solution can be from 6.2 to 8.5. The pH of the aqueous solution can be at least 6.2. The pH of the aqueous solution can be at most 8.5. The pH value of the aqueous solution can be 6.2 to 6.5, 6.2 to 7, 6.2 to 7.2, 6.2 to 7.5, 6.2 to 7.7, 6.2 to 8, 6.2 to 8.2, 6.2 to 8.5, 6.5 to 7, 6.5 to 7.2, 6.5 to 7.5, 6.5 to 7.7, 6.5 to 8, 6.5 to 8.2, 6.5 to 8.5, 7 to 7.2, 7 to 7.5, 7 to 7.7, 7 to 8, 7 to 8.2, 7 to 8.5, 7.2 to 7.5, 7.2 to 7.7, 7.2 to 8, 7.2 to 8.2, 7.2 to 8.5, 7.5 to 7.7, 7.5 to 8, 7.5 to 8.2, 7.5 to 8.5, 7.7 to 8, 7.7 to 8.2, 7.7 to 8.5, 8 to 8.2, 8 to 8.5, or 8.2 to 8.5. The pH of the aqueous solution can be 6.2, 6.5, 7, 7.2, 7.5, 7.7, 8, 8.2, or 8.5.

[0179] The pH of the aqueous solution can be from 9 to 12. The pH of the aqueous solution can be at least 9. The pH of the aqueous solution can be at most 12. The pH of the aqueous solution can be 9 to 9.5, 9 to 10, 9 to 10.5, 9 to 11, 9 to 11.5, 9 to 12, 9.5 to 10, 9.5 to 10.5, 9.5 to 11, 9.5 to 11.5, 9.5 to 12, 10 to 10.5, 10 to 11, 10 to 11.5, 10 to 12, 10.5 to 11, 10.5 to 11.5, 10.5 to 12, 11 to 11.5, 11 to 12, or 11.5 to 12. The pH of the aqueous solution can be 9, 9.5, 10, 10.5, 11, 11.5, or 12.

[0180] The incubation step can be carried out for 15 minutes to 60 minutes. The incubation step can be carried out for at least 15 minutes. The incubation step can be carried out for at most 60 minutes. The incubation step can be carried out for 15 minutes to 30 minutes, 15 minutes to 45 minutes, 15 minutes to 60 minutes, 30 minutes to 45 minutes, 30 minutes to 60 minutes, or 45 minutes to 60 minutes. The incubation step can be carried out for at least 15 minutes, 30 minutes, 45 minutes, or 60 minutes. The incubation step can be carried out for 1 hour to 24 hours. The incubation step can be carried out for at least 1 hour. The incubation step can be carried out for at most 24 hours. The incubation step can be carried out for 1 hour to 4 hours, 1 hour to 8 hours, 1 hour to 12 hours, 1 hour to 16 hours, 1 hour to 20 hours, 1 hour to 24 hours, 4 hours to 8 hours, 4 hours to 12 hours, 4 hours to 16 hours, 4 hours to 20 hours, 4 hours to 24 hours, 8 hours to 12 hours, 8 hours to 16 hours, 8 hours to 20 hours, 8 hours to 24 hours, 12 hours to 16 hours, 12 hours to 20 hours, 12 hours to 24 hours, 16 hours to 20 hours, 16 hours to 24 hours, or 20 hours to 24 hours. The incubation step can be carried out for at least 1 hour, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours.

[0181] After the mild pretreatment step, dissolved polysaccharides, monosaccharides, and / or disaccharides can be removed from the aqueous solution. In some cases, monosaccharides and / or disaccharides can be removed from the aqueous solution after the mild pretreatment step.

[0182] Soluble polysaccharides can be removed from the solution after the mild pretreatment step. At least a portion of the dissolved polysaccharides can be removed from the dissolved fraction. In some cases, at least 5% of the dissolved polysaccharides can be removed and / or purified. In some cases, up to 100% of the dissolved polysaccharides can be removed and / or purified. In some cases, 5% to 10%, 5% to 20%, 5% to 30%, 5% to 40%, 5% to 60%, 5% to 80%, 5% to 100%, 10% to 20%, 10% to 30%, 10% to 40%, 10% to 60%, 10% to 80%, 10% to 100%, 20% to 30%, 20% to 40%, 20% to 60%, 20% to 80%, 20% to 100%, 30% to 40%, 30% to 60%, 30% to 80%, 30% to 100%, 40% to 60%, 40% to 80%, 40% to 100%, 60% to 80%, or 60% to 100% of the dissolved polysaccharides can be removed. In some cases, after the mild pretreatment step, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80%, or 100% of the dissolved polysaccharides can be removed and / or purified.

[0183] After a mild pretreatment step, monosaccharides and / or disaccharides can be removed from the solution. A portion of the dissolved monosaccharides and / or disaccharides can be removed from the aqueous solution after an incubation step. In some cases, at least 5% of the dissolved monosaccharides and / or disaccharides can be removed. In some cases, up to 100% of the dissolved monosaccharides and / or disaccharides can be removed. In some cases, 5% to 10%, 5% to 20%, 5% to 30%, 5% to 40%, 5% to 60%, 5% to 80%, 5% to 100%, 10% to 20%, 10% to 30%, 10% to 40%, 10% to 60%, 10% to 80%, 10% to 100%, 20% to 30%, 20% to 40%, 20% to 60%, 20% to 80%, 20% to 100%, 30% to 40%, 30% to 60%, 30% to 80%, 30% to 100%, 40% to 60%, 40% to 80%, 40% to 100% or 60% to 100% of the dissolved monosaccharides and / or disaccharides can be removed. In some cases, after a mild pretreatment step, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80% or 100% of the dissolved monosaccharides and / or disaccharides can be removed and / or purified. After the incubation step, the monosaccharide and / or disaccharide fraction can be discarded. In certain cases, the portion of the dissolved monosaccharides and / or disaccharides removed in this step may not be combined with the portion of one or more oligosaccharides produced in the biomass treatment.

[0184] After a mild pretreatment step, an aqueous solution can be removed from the biomass. After a mild pretreatment step, a portion of the aqueous solution can be removed from the biomass. After a mild pretreatment step, 5% to 100% of the aqueous solution can be removed from the biomass. After a mild pretreatment step, at least 5% of the aqueous solution can be removed from the biomass. After a mild pretreatment step, at most 98% of the aqueous solution can be removed from the biomass. After a mild pretreatment step, 5% to 10%, 5% to 20%, 5% to 40%, 5% to 50%, 5% to 60%, 5% to 80%, 5% to 90%, 5% to 98%, 10% to 20%, 10% to 40%, 10% to 50%, 10% to 60%, 10% to 80%, 10% to 90%, 10% to 98%, 20% to 40%, 20% to 50%, 20% to 60%, 20% to 80%, 20% to 90%, 20% to 98%, 40% to 50%, 40% to 60%, 40% to 80%, 40% to 90%, 40% to 98%, 50% to 60%, 50% to 80%, 50% to 90%, 50% to 98%, 60% to 80%, 60% to 90%, 60% to 98%, 80% to 90%, 80% to 98% or 90% to 98% of the aqueous solution can be removed from the biomass. After a mild pretreatment step, at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90% or 98% of the aqueous solution can be removed from the biomass. A portion of the aqueous solution can be removed using pressure filtration, centrifugation, sedimentation, filtration, and / or any other suitable method.

[0185] Severe pretreatment

[0186] The biomass can be subjected to a severe pretreatment step. The severe pretreatment step can dissolve the polysaccharide fraction or the monosaccharide fraction and / or the disaccharide fraction from the biomass. The severe pretreatment step can be performed to make the biomass more readily digestible by enzymes. The severe pretreatment can help break the hydrogen bonds in the biomass. The severe pretreatment step can be a washing step, a thermochemical step, or a chemical treatment step. The severe pretreatment can be performed before a physical or mechanical pretreatment step. The severe pretreatment step can be performed after a physical or mechanical pretreatment step. The severe pretreatment step can be performed before a mild pretreatment step. The severe pretreatment step can be performed after a mild pretreatment step. The severe pretreatment step can be performed one or more times. The severe pretreatment can be performed 2, 3, 4, 5, 6, 7, 8, 9, or 10 times.

[0187] The strong pretreatment step can be a thermochemical treatment. The chemical or thermochemical treatment can include one or more aqueous solutions. The aqueous solution can contain one or more salts, acids, bases or ions. The aqueous solution can be an alkaline solution, which contains one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide or ammonia water. The aqueous solution can be an acidic solution, which contains at least one of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid or oxalic acid. In some embodiments, after completing one or more strong pretreatment steps, the aqueous solution can be retained. In other words, the aqueous solution may not be discarded or abandoned.

[0188] The thermochemical pretreatment can be carried out at a pH of about 2 to 6.5. The thermochemical pretreatment can be carried out at a pH of at least 2. The thermochemical pretreatment can be carried out at a pH of at most 6.5. The thermochemical pretreatment can be carried out at the following pH values: 2 to 2.5, 2 to 3, 2 to 3.5, 2 to 4, 2 to 4.5, 2 to 5, 2 to 5.5, 2 to 6, 2 to 6.5, 2.5 to 3, 2.5 to 3.5, 2.5 to 4, 2.5 to 4.5, 2.5 to 5, 2.5 to 5.5, 2.5 to 6, 2.5 to 6.5, 3 to 3.5, 3 to 4, 3 to 4.5, 3 to 5, 3 to 5.5, 3 to 6, 3 to 6.5, 3.5 to 4, 3.5 to 4.5, 3.5 to 5, 3.5 to 5.5, 3.5 to 6, 3.5 to 6.5, 4 to 4.5, 4 to 5, 4 to 5.5, 4 to 6, 4 to 6.5, 4.5 to 5, 4.5 to 5.5, 4.5 to 6, 4.5 to 6.5, 5 to 5.5, 5 to 6, 5 to 6.5, 5.5 to 6, 5.5 to 6.5 or 6 to 6.5. The thermochemical pretreatment can be carried out at a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6 or 6.5.

[0189] The thermochemical pretreatment can be carried out at a pH of from 7.5 to 12. The thermochemical pretreatment can be carried out at a pH of at least 7.5. The thermochemical pretreatment can be carried out at a pH of at most 12. The thermochemical pretreatment can be carried out at the following pH values: 7.5 to 8, 7.5 to 8.5, 7.5 to 9, 7.5 to 9.5, 7.5 to 10, 7.5 to 10.5, 7.5 to 11, 7.5 to 11.5, 7.5 to 12, 8 to 8.5, 8 to 9, 8 to 9.5, 8 to 10, 8 to 10.5, 8 to 11, 8 to 11.5, 8 to 12, 8.5 to 9, 8.5 to 9.5, 8.5 to 10, 8.5 to 10.5, 8.5 to 11, 8.5 to 11.5, 8.5 to 12, 9 to 9.5, 9 to 10, 9 to 10.5, 9 to 11, 9 to 11.5, 9 to 12, 9.5 to 10, 9.5 to 10.5, 9.5 to 11, 9.5 to 11.5, 9.5 to 12, 10 to 10.5, 10 to 11, 10 to 11.5, 10 to 12, 10.5 to 11, 10.5 to 11.5, 10.5 to 12, 11 to 11.5, 11 to 12 or 11.5 to 12. The thermochemical pretreatment can be carried out at a pH of about 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5 or 12.

[0190] The thermochemical pretreatment can be carried out at a temperature of from 50 °C to 150 °C. The thermochemical pretreatment can be carried out at a temperature of at least 50 °C. The thermochemical pretreatment can be carried out at a temperature of at most 150 °C. The thermochemical pretreatment can be carried out at the following temperatures: 50 °C to 75 °C, 50 °C to 80 °C, 50 °C to 90 °C, 50 °C to 100 °C, 50 °C to 120 °C, 50 °C to 130 °C, 50 °C to 150 °C, 75 °C to 80 °C, 75 °C to 90 °C, 75 °C to 100 °C, 75 °C to 120 °C, 75 °C to 130 °C, 75 °C to 150 °C, 80 °C to 90 °C, 80 °C to 100 °C, 80 °C to 120 °C, 80 °C to 130 °C, 80 °C to 150 °C, 90 °C to 100 °C, 90 °C to 120 °C, 90 °C to 130 °C, 90 °C to 150 °C, 100 °C to 120 °C, 100 °C to 130 °C, 100 °C to 150 °C, 120 °C to 130 °C, 120 °C to 150 °C or 130 °C to 150 °C. The thermochemical pretreatment can be carried out at a temperature of at least 50 °C, 75 °C, 80 °C, 90 °C, 100 °C, 120 °C, 130 °C or 150 °C.

[0191] The thermochemical treatment can be carried out for 0.5 hours to 4 hours. The thermochemical treatment can be carried out for at least 0.5 hours. The thermochemical treatment can be carried out for at most 4 hours. The thermochemical treatment can be carried out for 0.5 hours to 0.75 hours, 0.5 hours to 1 hour, 0.5 hours to 1.5 hours, 0.5 hours to 2 hours, 0.5 hours to 2.5 hours, 0.5 hours to 3 hours, 0.5 hours to 3.5 hours, 0.5 hours to 4 hours, 0.75 hours to 1 hour, 0.75 hours to 1.5 hours, 0.75 hours to 2 hours, 0.75 hours to 2.5 hours, 0.75 hours to 3 hours, 0.75 hours to 3.5 hours, 0.75 hours to 4 hours, 1 hour to 1.5 hours, 1 hour to 2 hours, 1 hour to 2.5 hours, 1 hour to 3 hours, 1 hour to 3.5 hours, 1 hour to 4 hours, 1.5 hours to 2 hours, 1.5 hours to 2.5 hours, 1.5 hours to 3 hours, 1.5 hours to 3.5 hours, 1.5 hours to 4 hours, 2 hours to 2.5 hours, 2 hours to 3 hours, 2 hours to 3.5 hours, 2 hours to 4 hours, 2.5 hours to 3 hours, 2.5 hours to 3.5 hours, 2.5 hours to 4 hours, 3 hours to 3.5 hours, 3 hours to 4 hours or 3.5 hours to 4 hours. The thermochemical treatment can be carried out for at least 0.5 hours, 0.75 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours or 4 hours.

[0192] The thermochemically treated biomass may contain from less than 1% w / w to 30% w / w of monosaccharides. The thermochemically treated biomass may contain from less than 1% w / w to 2% w / w, 1% w / w to 5% w / w, 1% w / w to 10% w / w, 1% w / w to 15% w / w, 1% w / w to 20% w / w, 1% w / w to 25% w / w, 1% w / w to 30% w / w, 2% w / w to 5% w / w, 2% w / w to 10% w / w, 2% w / w to 15% w / w, 2% w / w to 20% w / w, 2% w / w to 25% w / w, 2% w / w to 30% w / w, 5% w / w to 10% w / w, 5% w / w to 15% w / w, 5% w / w to 20% w / w, 5% w / w to 25% w / w, 5% w / w to 30% w / w, 10% w / w to 15% w / w, 10% w / w to 20% w / w, 10% w / w to 25% w / w, 10% w / w to 30% w / w, 15% w / w to 20% w / w, 15% w / w to 25% w / w, 15% w / w to 30% w / w, 20% w / w to 25% w / w, 20% w / w to 30% w / w or 25% w / w to 30% w / w of monosaccharides. The thermochemically treated biomass may contain less than 1% w / w, 2% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w or 30% w / w of monosaccharides.

[0193] The thermochemically treated biomass can contain from 5% w / w to 50% w / w of disaccharides. The thermochemically treated biomass can contain from 5% w / w to 10% w / w, 5% w / w to 15% w / w, 5% w / w to 20% w / w, 5% w / w to 25% w / w, 5% w / w to 30% w / w, 5% w / w to 35% w / w, 5% w / w to 40% w / w, 5% w / w to 50% w / w, 10% w / w to 15% w / w, 10% w / w to 20% w / w, 10% w / w to 25% w / w, 10% w / w to 30% w / w, 10% w / w to 35% w / w, 10% w / w to 40% w / w, 10% w / w to 50% w / w, 15% w / w to 20% w / w, 15% w / w to 25% w / w, 15% w / w to 30% w / w, 15% w / w to 35% w / w, 15% w / w to 40% w / w, 15% w / w to 50% w / w, 20% w / w to 25% w / w, 20% w / w to 30% w / w, 20% w / w to 35% w / w, 20% w / w to 40% w / w, 20% w / w to 50% w / w, 25% w / w to 30% w / w, 25% w / w to 35% w / w, 25% w / w to 40% w / w, 25% w / w to 50% w / w, 30% w / w to 35% w / w, 30% w / w to 40% w / w, 30% w / w to 50% w / w, 35% w / w to 40% w / w, 35% w / w to 50% w / w or 40% w / w to 50% w / w of disaccharides. The thermochemically treated biomass can contain less than 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w or 50% w / w of disaccharides.

[0194] II. Enzymatic treatment and downstream processing

[0195] The methods of the present disclosure can further include contacting the thermochemically treated biomass with one or more polysaccharide-cleaving enzymes in solution or suspension to form one or more oligosaccharides. Additionally, these methods can include enriching the solution or suspension to increase the concentration of one or more oligosaccharides to form the composition. One or more oligosaccharides can be purified from the solution or suspension provided herein.

[0196] One or more steps of the method of forming or making a composition can be enzymatic reactions, where one or more enzymes are placed in a suitable reaction vessel together with one or more feedstocks or biomass (e.g., plant biomass) and a suitable solvent, and the one or more feedstocks or biomass can be soluble or insoluble in water. As used herein, unless otherwise specified, the term "plant biomass" can be replaced with the term "feedstock" or "biomass" (e.g., biomass not derived from plants).

[0197] A variety of enzymes can be suitable for the enzymatic reaction. Any enzyme that produces oligosaccharides when acting on a polysaccharide-containing feedstock is suitable. For example, the enzymatic reaction can include cellulase, endoglucanase, cellobiohydrolase, lytic polysaccharide monooxygenase (LPMO), lichenase, xyloglucan endoglucanase (XEG), mannanase, chitinase, xylanase, and / or one or more suitable enzymes.

[0198] In various cases, the enzymatic reaction can include a cellulolytic preparation from a species such as Trichoderma reesei, which can be purified and / or pretreated and / or supplemented with one or more additional enzymes, such as the addition of β-glucanase, β-xylanase, and cellobiohydrolase; β-glucanase, β-xylanase, LPMO, and cellobiohydrolase; LPMO and xylanase; or LPMO, xylanase, and lichenase. The various enzymes can be provided to the enzymatic reaction in the form of purified enzymes, semi-purified mixtures from some natural sources or laboratory-grown cultures, microbial strains engineered to produce the enzymes, or in any other suitable form. The fusion of these enzymes with other enzymes or with non-enzyme modules such as carbohydrate-binding modules (CBMs) is also contemplated. For example, LPMO fused with CBM, xylanase fused with CBM, or xylanase fused with LPMO can be used.

[0199] Aerobic conditions can be used for one or more enzymatic reactions. Aerobic conditions can include the addition of oxygen, which can be provided by aerating the substrate mixture with an oxygen-containing gas such as air. Aeration can be carried out by introducing oxygen-containing bubbles into the aqueous substrate mixture through various systems such as air ejectors, aeration frits, membrane systems, or internal circulation air-lift reactors. In some cases, the molecular oxygen concentration in the enzymatic reaction can be from about 4 mg / L to about 14 mg / L.

[0200] Another exemplary enzyme is lichenase, which can be selected from the GH5, GH7, GH8, GH9, GH12, GH16, GH17, or GH26 families. For example, a GH16 enzyme such as a GH16 enzyme derived from Bacillus subtilis can be used. The enzyme can act on, for example, mixed-linkage glucans (which are glucans that are a mixture of β-1,3 and β-1,4 linkages), and can cleave them at the β-1,4 glycosidic bond. In cases where the lichenase acts on mixed-linkage glucans, the resulting β-glucans can mainly fall within the size range of 3 to about 7 residues, and thus they are particularly suitable for the food, cosmetic, and nutritional industries. Mixed-linkage glucans are abundant in members of the grass and horsetail families. Thus, grassy raw materials such as rice straw usually have a high level of mixed-linkage glucans and can be effectively acted upon by lichenase. The lichenase can include a GH5 lichenase derived from Bacillus subtilis.

[0201] Another alternative enzyme is xylanase, which can act on, for example, raw materials containing a xylan backbone. The xylanase can be, for example, glucuronoxylanase, arabinoxylanase, or glucuronarabinoxylanase. The enzyme can act on a variety of polymers having a xylan backbone, such as glucuronoxylan, arabinoxylan, and glucuronarabinoxylan. These polymers are usually abundant in raw materials from various plant sources. For example, both hardwoods and softwoods can contain suitable polysaccharides, where hardwoods usually contain glucuronoxylan, while softwoods usually contain arabinoglucuronoxylan. The xylanase can include GH5 xylanases from Ruminiclostridium thermocellum and Gonapodya prolifera, and GH30 xylanases from Dickeya chrysanthemi, Bacillus subtilis, Bacteroides ovatus, and Trichoderma reesei.

[0202] Another alternative enzyme is mannanase, which can act on, for example, raw materials containing a mannan backbone. Mannanase can be, for example, mannanase, glucomannanase, galactomannanase or galactoglucomannanase. The enzyme can be active against various polymers having a mannan backbone, such as mannan, glucomannan, galactomannan or galactoglucomannan. These polymers are usually abundant in raw materials from various plant sources. For example, both hardwoods and softwoods can contain suitable polysaccharides. Suitable mannanases can include GH5 mannanases from Trichoderma reesei and Aspergillus niger, and GH26 mannanases from Aspergillus niger.

[0203] Other enzymes can include xyloglucanase and xyloglucan endoglucanase (XEG), which are produced by many organisms, including plant pathogenic microorganisms. Xyloglucanase and XEG can be capable of acting on xyloglucan, which is a hemicellulose β-1,4 glucan chain abundant in the primary cell wall of higher plants, which is modified with xylose and some xylose residues are also modified with other residues (such as galactose). When the appropriate xyloglucanase or XEG acts on xyloglucan, the product can contain xyloglucan oligosaccharides having a backbone length useful in the food, cosmetic and nutritional industries. Suitable xyloglucanases can include the GH5 xyloglucanase from Bacteroides ovatus and the GH74 xyloglucanase from Trichoderma reesei.

[0204] The enzymatic reaction can be carried out in solution and / or suspension. The enzymatic reaction can be carried out in a suitable reaction vessel. In some cases, the enzymatic reaction can be carried out at a temperature or temperature profile suitable for a particular combination of enzyme and raw material, and the reaction can be allowed to proceed for a certain amount of time (e.g., a predetermined amount of time) until the product has reached the desired concentration or until some other requirement is met.

[0205] To ensure optimal contact between the enzyme and the raw material, the reaction mixture can be stirred continuously or intermittently. Stirring can take the form of: (i) rhythmic movement of the entire reaction vessel, (ii) a blower or other stirring device, (iii) bubble injection, or (iv) any other suitable stirring method.

[0206] The enzymatic reaction can be microbial fermentation. The temperature and reaction time can be suitable for the growth of the microorganism used. The microorganism can be genetically altered to produce an enzyme suitable for the production of the oligosaccharide composition. The microorganism can be a bacterium, such as Escherichia coli, or a fungus, such as Saccharomyces cerevisiae or Trichoderma reesei.

[0207] In some embodiments, an expression vector suitable for modifying the subject microorganism can be used such that it produces an enzyme or enzyme mixture as described elsewhere herein. If desired, the expression vector can be a plasmid or any other nucleic acid capable of inducing enzyme production. In some cases, the expression vector can contain one or more of the following regulatory sequences to control the expression of the exogenous enzyme: regulatory sequences of heat shock genes, regulatory sequences of virulence genes, regulatory sequences of sporulation genes, or any other suitable regulatory sequences.

[0208] The enzymatic reaction can be carried out at a temperature or temperature profile suitable for the enzyme and substrate used. For example, the enzymatic reaction can be carried out at a constant temperature of 10 °C to 100 °C, 20 °C to 80 °C, or 40 °C to 60 °C. In some cases, if the enzymatic reaction takes the form of microbial fermentation, the temperature can be suitable therefor. For example, the enzymatic reaction can include the growth of Escherichia coli, and / or the temperature can be substantially constant and about 37 °C.

[0209] The pH of the solution or suspension can affect the activity of the enzyme. Control of the pH can help ensure that the enzymatic reaction proceeds at a suitable rate. The enzymatic reaction can be carried out within a pH range of 2 to 10, 3 to 8, or 4 to 6.

[0210] The enzymatic reaction can be allowed to continue for a certain period of time, then quenched and the product separated or otherwise collected. The period can be from 1 minute to 6 days, 0.5 days to 5 days, or 16 hours to 96 hours. Alternatively, the reaction can be allowed to proceed until no further catalysis occurs.

[0211] One or more feedstocks added to the enzymatic reaction can contain a polysaccharide. Such a polysaccharide can have been produced by a separate reaction carried out simultaneously or substantially simultaneously in the reaction vessel. The polysaccharide present in the enzymatic reaction can be partially cleaved by the enzyme into useful oligosaccharides, leaving partially cleaved or uncleaved polysaccharides, which can include, but are not limited to, cellulose, xylan (such as glucuronoxylan, arabinoxylan, or glucuronarabinoxylan), mannan (such as glucomannan, galactomannan, or galactoglucomannan), mixed-linkage glucan, xyloglucan, chitin, chitosan, or lignocellulose.

[0212] The enzymatic reaction can be allowed to continue until 5% to 75%, 5% to 70%, 5% to 65%, 5% to 55%, or 10% to 50% of the feedstock containing undigested polysaccharide remains. It can be monitored or checked by reducing-end assays (such as the anthrone assay) and / or chromatography (such as thin-layer chromatography and / or high-performance anion-exchange chromatography).

[0213] Any substance containing suitable polysaccharides can form part of the raw material. Since the food, cosmetic, and nutritional industries commonly use a wide variety of oligosaccharides, the polysaccharides suitable for participating in enzymatic reactions are not particularly limited. Raw materials applicable to generating oligosaccharide profiles can include, for example, cellulose, lignocellulose, chitin, chitosan, xylans (such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan), and / or mannans (such as glucomannan, galactomannan, or galactoglucomannan). However, any raw material that can function appropriately is envisioned. The raw materials can include sugarcane, corn stover, corn cobs, wheat bran, wheat straw, hardwood, softwood, or any other suitable biomass or plant biomass.

[0214] There is also no particular limitation on the raw materials containing such polysaccharides because most plant materials are rich in such polymers. Thus, the raw materials can include plant biomass such as grains, husks, pods, seed coats, and / or other seed materials; seaweeds; corn stover, straw, bagasse, miscanthus, sorghum bagasse, switchgrass, bamboo, and / or other monocotyledonous plant tissues; water hyacinth, leaf tissues, roots, and / or other plant materials; hardwood, hardwood chips, hardwood pulp, softwood, softwood chips, softwood pulp, paper, pulp, cardboard, and / or other lignaceous raw materials; crab shells, squid biomass, shrimp shells, and / or other marine biomass, and / or any combination of suitable raw materials. The raw materials can include wheat straw or wood. Since any given natural raw material may contain a mixture of different polysaccharides, sometimes a mixture of different enzymes will be beneficial. Such a mixture can contain one or more of any suitable enzymes as discussed herein. For example, such a mixture can contain LPMO and endoglucanase, xylanase and lichenase, cellobiohydrolase and mannanase, or endoglucanase and cellobiohydrolase. In some embodiments, the enzyme conjugates can be present in a molar ratio of, for example, 1:100 to 100:1. Additionally, since many suitable raw materials are recalcitrant, pretreatment of the raw materials is envisioned.

[0215] After the enzymatic reaction has proceeded to the desired point, one or more oligosaccharides and one or more polysaccharides can be separated from the enzymatic reaction mixture. Depending on the composition of the biomass used and the specificity of the enzymes used, this process can be carried out in a variety of ways. Since the reaction mixture typically contains a mixture of soluble oligosaccharides and insoluble polysaccharides, the reaction mixture can be filtered to remove the insoluble material and the resulting soluble oligosaccharides can be prepared for further processing.

[0216] Oligosaccharides can also be separated from polysaccharides in a variety of ways. Oligosaccharides can be separated based on solubility so that only compositions of soluble saccharides are extracted for further processing and / or separated by chromatography to produce compositions with a narrower band of oligosaccharide chain lengths. For example, the separation can be based on precipitation, size exclusion chromatography, ion exchange chromatography, or filtration, ultrafiltration, microfiltration, and nanofiltration. In the case of separation based on solubility, the distribution of saccharides present in the separated composition will generally depend on the original enzymatic reaction, since different polysaccharides generally have different rates of decreasing solubility with length.

[0217] It is also contemplated to further process all or part of the oligosaccharides produced prior to incorporation into food, cosmetics, or nutraceuticals to produce further products. This further processing can include any chemical, physical, or enzymatic steps, such as reduction, for example reductive amination when appropriate; oxidation, caramelization, modification with Schiff bases or by the Maillard reaction or by any combination of these steps, and can provide different products with achieved or improved properties for the desired purpose. For example, caramelization characteristics, calorific value, flavor, and color can be modified. The oligosaccharides can also be purified, for example by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration.

[0218] It is also contemplated to further process all or part of the polysaccharide fraction produced prior to incorporation into food, cosmetics, or nutraceuticals to produce a product with improved properties. Such further processing can include any chemical, physical, or enzymatic steps, such as alkylation or acid treatment. The polysaccharides can also be purified, for example by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration.

[0219] In some cases, after modification and / or purification of the oligosaccharide fraction and the polysaccharide fraction, all or part of the fractions can then be recombined in a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, for example, 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5. The specific ratio can depend on the desired properties of the final composition and the modifications and purifications that have been applied to the fractions. It may not be necessary to recombine all of the oligosaccharides and polysaccharides separated from the enzymatic reaction.

[0220] The fractions can be recombined in a variety of ways, such as by mixing a solution containing all or part of the oligosaccharide fraction and a solution and / or suspension containing all or part of the polysaccharide fraction, which can be further dried, lyophilized, or otherwise concentrated. The fractions can also be recombined by mixing the dry form containing all or part of the oligosaccharide fraction produced by drying, lyophilizing, or otherwise concentrating with the dry form containing all or part of the polysaccharide fraction produced by drying, lyophilizing, or otherwise concentrating.

[0221] The oligosaccharide component of the final composition can comprise one or more oligosaccharides of any type. For example, the oligosaccharide component can include cellooligosaccharide, xylo-oligosaccharide, mixed-linkage glucan oligosaccharide, mannan oligosaccharide, xyloglucan oligosaccharide, chitosan oligosaccharide, arabinoxylan oligosaccharide, or derivatives of any of the foregoing oligosaccharides.

[0222] Any such dry or liquid composition can be considered an ingredient suitable for incorporation into food, cosmetics, or nutraceuticals at any stage of the process. This includes compositions that can be considered intermediates during the method, such as compositions formed after recombining the oligosaccharide and polysaccharide fractions, prior to any further purification, optimization, drying, dissolution, or any other such steps, and including the final composition obtained from the method.

[0223] As described herein, the dry composition can be formed by drying and / or lyophilization. The dry composition can be dissolved in solutions of various liquids including water, syrups, pastes, solvents, alcohols, etc. to form a liquid composition ingredient suitable for incorporation into food, cosmetics, or nutraceuticals. The liquid composition is particularly useful for foods that require a smooth texture, such as confections, chocolates, and yogurts.

[0224] In some embodiments, after modification and / or purification of the oligosaccharide fraction and the polysaccharide fraction, all or part of the fractions can then be recombined in a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, such as 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5. The specific ratio may depend on the desired properties of the final ingredient and the modifications and purifications that have been applied to the fractions.

[0225] After obtaining a composition of the oligosaccharide product suitable for the intended application, further processing and / or separation can be carried out. Deriving foods, cosmetics, or nutraceuticals from the composition can provide a wide range of potential uses. The ingredients as described herein can be used in applications that conventionally use oligosaccharides, sugars, bulking sweeteners, low-intensity sweeteners, or other related food ingredients.

[0226] The polysaccharide-cleaving enzyme can be one of the following: cellulase, xylanase, xyloglucanase, endoglucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase (LPMO), e.g., selected from AA9, AA10, AA11, AA13, AA14, and AA15. The polysaccharide-cleaving enzyme can be prepared from the fungus Trichoderma reesei, and / or the enzymatic reaction can proceed until 5 - 75%, 5 - 65%, or 5 - 50% of the undigested polysaccharide-containing starting material remains.

[0227] The polysaccharide-cleaving enzyme can be effectively linked to a catalytic or non-catalytic module, for example, where the polysaccharide-cleaving enzyme can be effectively linked to a non-catalytic module. And the non-catalytic module is a carbohydrate-binding module.

[0228] In various embodiments, after separation of one or more oligosaccharides and one or more polysaccharides, the one or more oligosaccharides and one or more polysaccharides can be: purified; and / or subjected to chemical, physical, or enzymatic treatment, such as reduction, oxidation, caramelization, or Maillard reaction; and / or can be reconstituted by mixing a dry oligosaccharide powder with a dry polysaccharide powder.

[0229] In some embodiments, the composition can comprise three or more different molecular weight oligosaccharides, where the method can include forming the three or more different molecular weight oligosaccharides by an enzymatic reaction, and the enzymatic reaction comprises the step of contacting one or more polysaccharide-cleaving enzymes with one or more starting materials in solution or suspension.

[0230] III. Food, cosmetic, or nutritional ingredient

[0231] Composition

[0232] The polysaccharide component of the composition can comprise one or more polysaccharides of any type. For example, the polysaccharide can include cellulose, lignocellulose, xylan, mixed-linkage glucan, mannan, xyloglucan, chitin, chitosan, or derivatives of any of the foregoing polysaccharides.

[0233] The composition or ingredient can contain various oligosaccharides. The composition can include different amounts of oligosaccharides, e.g., depending on the desired properties of the composition. In some cases, the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of cellooligosaccharides with a degree of polymerization of 2 to 6, and / or the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of xylooligosaccharides with a degree of polymerization of 2 to 12, and / or the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of mixed-linkage glucan oligosaccharides with a degree of polymerization of 2 to 5, and / or the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of mannan oligosaccharides with a degree of polymerization of 2 to 12, and / or the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of xyloglucan oligosaccharides with a degree of polymerization of 4 to 12, and / or the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of chitooligosaccharides with a degree of polymerization of 2 to 12, and / or the composition can contain at least 20% by dry weight, such as at least 30% by dry weight, of arabinoxylooligosaccharides with a degree of polymerization of 3 to 15. In certain embodiments, it is understood that the composition can contain up to 100% by dry weight of the above oligosaccharides, and thus the above embodiments (where the oligosaccharides are present at least 20% by dry weight) do not include all seven types of oligosaccharides.

[0234] In various embodiments, the composition or ingredient can contain about 5% to about 50% w / w of cellooligosaccharides with a degree of polymerization of 2 to 6. In certain embodiments, the composition or ingredient can contain about 5% to about 50%, about 10% to about 40%, about 15% to about 35% w / w of cellooligosaccharides with a degree of polymerization of 2 to 6. The composition or ingredient can contain at least 5%, 8%, 10%, 15%, 20% or 25% w / w of cellooligosaccharides with a degree of polymerization of 2 to 6. In some embodiments, the composition or ingredient can contain about 20% to about 90% w / w of cellooligosaccharides with a degree of polymerization of 2 to 6. In certain embodiments, the composition or ingredient can contain about 5% to about 95%, about 10% to about 92.5%, about 30% to about 80%, about 40% to about 70%, or about 50% to about 60% w / w of cellooligosaccharides with a degree of polymerization of 2 to 6.

[0235] In various embodiments, the composition or ingredient may comprise from about 20% to about 90% w / w xylo-oligosaccharides having a degree of polymerization of 2 to 5. In certain embodiments, the composition or ingredient may comprise from about 5% to about 95%, from about 10% to about 92.5%, from about 30% to about 80%, from about 40% to about 70%, or from about 50% to about 60% w / w xylo-oligosaccharides having a degree of polymerization of 2 to 5. For example, the composition may comprise at least 30% w / w xylo-oligosaccharides having a degree of polymerization of 2 to 5. The composition or ingredient may comprise at least 5%, 8%, 10%, 15%, 20%, or 25% w / w xylo-oligosaccharides having a degree of polymerization of 2 to 5.

[0236] In certain embodiments, the composition or ingredient may comprise from about 0.1% to about 15% w / w arabino-xylo-oligosaccharides having a degree of polymerization of 3 to 12. The composition or ingredient may comprise at least 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or 15% w / w arabino-xylo-oligosaccharides having a degree of polymerization of 3 to 12. In various embodiments, the composition or ingredient may comprise from about 0.5% to about 25% w / w arabino-xylo-oligosaccharides having a degree of polymerization of 3 to 15. The composition or ingredient may comprise at least 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, or 35% w / w arabino-xylo-oligosaccharides having a degree of polymerization of 3 to 15.

[0237] In some embodiments, provided herein is the use of an oligosaccharide mixture in the formation of a food, cosmetic, or nutritional product, wherein the oligosaccharide mixture comprises two oligosaccharides selected from the following list:

[0238] i) cello-oligosaccharides having a degree of polymerization of 2 to 6;

[0239] ii) xylo-oligosaccharides having a degree of polymerization of 2 to 12;

[0240] iii) mixed-linkage glucan oligosaccharides having a degree of polymerization of 2 to 5;

[0241] iv) manno-oligosaccharides having a degree of polymerization of 2 to 12;

[0242] v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 10;

[0243] vi) chito-oligosaccharides having a degree of polymerization of 2 to 12; and / or

[0244] vii) arabino-xylo-oligosaccharides having a degree of polymerization of 3 to 15, wherein the two oligosaccharides may be present in a ratio of 1:9 to 9:1, 1:4 to 4:1, or 2:3 to 3:2 relative to each other.

[0245] In some cases, arabinoxylan oligosaccharides can contain at least 0.1% arabinose residues. Arabinoxylan oligosaccharides can contain at least 0.1%, 0.2%, 0.5%, 1%, 5% or 10% arabinose residues.

[0246] The amount of each oligosaccharide can vary according to the desired properties of the resulting food, cosmetic or nutritional product. For example, two oligosaccharides can be present in a ratio of 1:9 to 1:1, 1:2 to 1:1, or 2:3 to 1:1 relative to each other.

[0247] The oligosaccharide mixture can further contain a third oligosaccharide. The oligosaccharide mixture can contain a third oligosaccharide and a fourth oligosaccharide. The oligosaccharide mixture can contain a third oligosaccharide, a fourth oligosaccharide and a fifth oligosaccharide. The oligosaccharide mixture can further contain a third oligosaccharide, a fourth oligosaccharide, a fifth oligosaccharide and a sixth oligosaccharide. The oligosaccharide mixture can further contain a third oligosaccharide, a fourth oligosaccharide, a fifth oligosaccharide, a sixth oligosaccharide and a seventh oligosaccharide. These oligosaccharides can be selected from the same list as at least two of the oligosaccharides provided above.

[0248] An oligosaccharide mixture of at least two oligosaccharides can contain xylooligosaccharides, such as a combination of xylooligosaccharides and xylooligosaccharides. Optional compositions can contain a combination of xylooligosaccharides and mannan oligosaccharides. In some embodiments, the oligosaccharide mixture can include xylooligosaccharides, xylooligosaccharides and arabinoxylan oligosaccharides in combination with each other.

[0249] An oligosaccharide mixture of at least two oligosaccharides can additionally contain a polysaccharide, such as a cellulose polysaccharide, such as cellulose; or a polysaccharide derivative, such as a cellulose derivative, such as carboxymethyl cellulose; or a polysaccharide aggregate, such as a part of lignocellulosic biomass. In some cases, the ratio in the combination can be 1:100 to 1:1 polysaccharide / polysaccharide derivative / polysaccharide aggregate:oligosaccharide, such as 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5. Thus, the ratio between the first oligosaccharide, the second oligosaccharide and the polysaccharide can be 2:2:1 to 30:30:1, such as about 3:3:1.

[0250] Combination of oligosaccharides

[0251] The composition can contain a mixture of one or more oligosaccharides. The oligosaccharide mixture can contain two forms or types of oligosaccharides, for example, xylooligosaccharides and xylooligosaccharides. The oligosaccharide mixture can contain three forms of oligosaccharides, for example, xylooligosaccharides, mannan oligosaccharides and xylooligosaccharides. The oligosaccharide mixture can contain four forms of oligosaccharides, for example, xylooligosaccharides, mannan oligosaccharides, mixed-link glucan oligosaccharides and chitosan oligosaccharides.

[0252] The oligosaccharide mixture may contain two forms of oligosaccharides, for example, a first oligosaccharide and a second oligosaccharide. The oligosaccharide mixture may contain about 5% w / w of the first oligosaccharide and about 95% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 10% w / w of the first oligosaccharide and about 90% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 15% w / w of the first oligosaccharide and about 85% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 20% w / w of the first oligosaccharide and about 80% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 25% w / w of the first oligosaccharide and about 75% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 30% w / w of the first oligosaccharide and about 70% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 35% w / w of the first oligosaccharide and about 65% w / w of the second oligosaccharide. The oligosaccharide mixture may contain about 40% w / w of the first oligosaccharide and about 50% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 45% w / w of the first oligosaccharide and 55% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 50% w / w of the first oligosaccharide and 50% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 55% w / w of the first oligosaccharide and 45% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 60% w / w of the first oligosaccharide and 30% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 65% w / w of the first oligosaccharide and 35% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 70% w / w of the first oligosaccharide and 30% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 75% w / w of the first oligosaccharide and 25% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 80% w / w of the first oligosaccharide and 20% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 85% w / w of the first oligosaccharide and 15% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 90% w / w of the first oligosaccharide and 10% w / w of the second oligosaccharide. The oligosaccharide mixture may contain 95% w / w of the first oligosaccharide and 5% w / w of the second oligosaccharide. In some cases, the first oligosaccharide may be cellooligosaccharide, and the second oligosaccharide may be xylooligosaccharide. In some cases, the first oligosaccharide may be cellooligosaccharide, and the second oligosaccharide may be mannan oligosaccharide. In some embodiments, the first oligosaccharide may be xylooligosaccharide, and the second oligosaccharide may be mannan oligosaccharide. Other combinations of the first oligosaccharide and the second oligosaccharide are also within the scope of the present disclosure.

[0253] The oligosaccharide mixture may comprise three forms of oligosaccharides, such as a first oligosaccharide, a second oligosaccharide, and a third oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide, 40% w / w of the second oligosaccharide, and 40% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 30% w / w of the first oligosaccharide, 30% w / w of the second oligosaccharide, and 40% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 10% w / w of the first oligosaccharide, 10% w / w of the second oligosaccharide, and 80% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide, 20% w / w of the second oligosaccharide, and 60% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide, 30% w / w of the second oligosaccharide, and 50% w / w of the third oligosaccharide. In some examples, the first oligosaccharide may be mannan oligosaccharide, the second oligosaccharide may be xylooligosaccharide, and the third oligosaccharide may be cellooligosaccharide. In some examples, the first oligosaccharide may be xyloglucan-oligosaccharide, the second oligosaccharide may be xylooligosaccharide, and the third oligosaccharide may be cellooligosaccharide. Other combinations of the first oligosaccharide, the second oligosaccharide, and the third oligosaccharide are also within the scope of the present disclosure.

[0254] The oligosaccharide mixture may comprise two or more oligosaccharides, a first oligosaccharide and a second oligosaccharide different from the first oligosaccharide. For example, the first oligosaccharide may be xylooligosaccharide or cellooligosaccharide or mannan oligosaccharide or other oligosaccharides provided herein, and the second oligosaccharide may be xylooligosaccharide or cellooligosaccharide or mannan oligosaccharide or other oligosaccharides not used as the first oligosaccharide. In other words, the first oligosaccharide may be different from the second oligosaccharide (e.g., the first oligosaccharide may be a different type of oligosaccharide from the second oligosaccharide). The ratio of the first oligosaccharide to the second oligosaccharide in the mixture may be about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, or 1:9.

[0255] The ratio of the first oligosaccharide to the second oligosaccharide in the mixture may be about 2:1, 2:3, 2:5, 2:7, or 2:9. The oligosaccharide may be cellooligosaccharide, mannan oligosaccharide, xylooligosaccharide, xyloglucan-oligosaccharide, mixed-linkage oligosaccharide, chitosan oligosaccharide, arabinoxylan oligosaccharide, or other oligosaccharides provided herein, wherein the first oligosaccharide is selected to be different from the second oligosaccharide. In other words, the first oligosaccharide may be a different type of oligosaccharide from the second oligosaccharide.

[0256] The ratio of the first oligosaccharide to the second oligosaccharide in the mixture may be about 3:1, 3:2, 3:4, 3:5, 3:7, or 3:8. The oligosaccharide may be cellooligosaccharide, mannan oligosaccharide, xylooligosaccharide, xyloglucan-oligosaccharide, mixed-linkage oligosaccharide, chitosan oligosaccharide, arabinoxylan oligosaccharide, or other oligosaccharides provided herein, wherein the first oligosaccharide is selected to be different from the second oligosaccharide.

[0257] In an oligosaccharide mixture containing two or more oligosaccharides, the ratio of the first oligosaccharide to the second oligosaccharide can be from 1:9 to 9:1, 1:4 to 4:1, 1:3 to 3:1, or 2:3 to 3:2. The oligosaccharides can be the xylooligosaccharides, mannan oligosaccharides, xylooligosaccharides, xyloglucan-oligosaccharides, mixed-linkage oligosaccharides, chitosan oligosaccharides, arabinoxylan oligosaccharides or other oligosaccharides provided herein, wherein the first oligosaccharide is selected as an oligosaccharide different from the second oligosaccharide.

[0258] In some cases, the composition or ingredient can include at least 1% w / w, 2% w / w, 3% w / w, 4% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w or more of cellobiose, xylobiose, mannotriose (e.g., Man-β-1,4-Man), Glc-β-1,4-Man, Man-β-1,4-Glc, laminaribiose, gentiobiose, sophorose, maltose, lactose or sucrose. In certain cases, the composition or ingredient can include at least 1% w / w, 2% w / w, 3% w / w, 4% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w or more of cellotriose, xylotriose, monoarabinosylated xylobiose, monoglucuronidated xylobiose, maltotriose, mannotriose (e.g., Man-β-1,4-Man-β-1,4-Man, Glc-β-1,4-Man-β-1,4-Man, Man-β-1,4-Glc-β-1,4-Man, Man-β-1,4-Man-β-1,4-Glc, Man-β-1,4-Glc-β-1,4-Glc, Glc-β-1,4-Man-β-1,4-Glc, Glc-β-1,4-Glc-β-1,4-Man, Glc-β-1,3-Glc-β-1,4-Glc or Glc-β-1,4-Glc-β-1,3-Glc).In certain cases, the composition or ingredient can include at least 1% w / w, 2% w / w, 3% w / w, 4% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w or more of xylo - tetrose, cello - tetrose, mono - arabinosylated xylotriose, mono - glucuronosylated xylotriose, di - arabinosylated xylobiose, di - glucuronosylated xylobiose, maltotetrose, mannotetrose (e.g., Man-β-1,4-Man-β-1,4-Man-β-1,4-Man, Glc-β-1,4-Man-β-1,4-Man-β-1,4-Man, Man-β-1,4-Glc-β-1,4-Man-β-1,4-Man, Man-β-1,4-Man-β-1,4-Glc-β-1,4-Man, Man-β-1,4-Man-β-1,4-Man-β-1,4-Glc, Glc-β-1,4-Glc-β-1,4-Man-β-1,4-Man, Man-β-1,4-Glc-β-1,4-Glc-β-1,4-Man, Man-β-1,4-Man-β-1,4-Glc-β-1,4-Glc, Glc-β-1,4-Man-β-1,4-Glc-β-1,4-Man, Glc-β-1,4-Man-β-1,4-Man-β-1,4-Glc, Man-β-1,4-Glc-β-1,4-Man-β-1,4-Glc, Glc-β-1,3-Glc-β-1,4-Glc-1,4-Glc, Glc-β-1,4-Glc-β-1,3-Glc-1,4-Glc, Glc-β-1,4-Glc-β-1,4-Glc-1,3-Glc, or Glc-β-1,3-Glc-β-1,4-Glc-1,3-Glc. In certain cases, the composition or ingredient can include at least 0.01% w / w, 0.05% w / w, 0.1% w / w, 0.5% w / w, 1% w / w, 2% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w or more of xylopentaose, cellopentaose, mono - arabinosylated xylo - tetrose, mono - glucuronosylated xylo - tetrose, di - arabinosylated xylotriose, di - glucuronosylated xylotriose, maltopentaose, mannotetraose (e.g., Man-β-1,4-Man-β-1,4-Man-β-1,4-Man-β-1,4-Man), mixed - linkage glucan - derived pentasaccharide or mannan - derived pentasaccharide.

[0259] The composition or ingredient may comprise 1% to 50%, 5% to 40%, 10% to 30%, or 15% to 25% w / w of cellobiose. The composition or ingredient may comprise 2.5% to 90%, 5% to 80%, 10% to 70%, or 20% to 60% w / w of xylobiose. The composition or ingredient may comprise 2.5% to 75%, 5% to 50%, 10% to 40%, or 20% to 30% w / w of xylotriose.

[0260] Oligosaccharide composition with different degrees of polymerization

[0261] The average degree of polymerization of the oligosaccharides in the composition may be 1 to 50, 1.5 to 25, 2 to 15, 2.1 to 10, 2.1 to 7, or 2.2 to 5.

[0262] In the xylooligosaccharide mixture, the concentration of xylooligosaccharides with a degree of polymerization of 2 may be about 2% to about 80% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 2 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25%, or 30% w / w. In some cases, the concentration of xylooligosaccharides with a degree of polymerization of 2 may be higher, for example, up to 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or 80% w / w.

[0263] In the xylooligosaccharide mixture, the concentration of xylooligosaccharides with a degree of polymerization of 3 may be about 2% to about 20% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, or 20% w / w.

[0264] In the xylooligosaccharide mixture, the concentration of xylooligosaccharides with a degree of polymerization of 4 may be about 5% to about 20% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18%, or 20% w / w.

[0265] In the xylooligosaccharide mixture, the concentration of xylooligosaccharides with a degree of polymerization of 5 may be about 5% to about 20% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18%, or 20% w / w.

[0266] In the xylooligosaccharide mixture, the concentration of xylooligosaccharides with a degree of polymerization of 6 may be about 5% to about 25% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20%, or 25% w / w.

[0267] In the xylooligosaccharide mixture, the concentration of xylooligosaccharides with a degree of polymerization of 7 may be about 2% to about 20% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 7 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17%, or 20% w / w.

[0268] The concentration of xylooligosaccharides with a degree of polymerization of 8 in the xylooligosaccharide mixture can be from about 1% to about 15% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0269] The concentration of xylooligosaccharides with a degree of polymerization of 9 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0270] The concentration of xylooligosaccharides with a degree of polymerization of 10 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0271] The concentration of xylooligosaccharides with a degree of polymerization of 11 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0272] The concentration of xylooligosaccharides with a degree of polymerization of 12 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0273] The concentration of cellooligosaccharides with a degree of polymerization of 2 in the cellooligosaccharide mixture can be from about 2% to about 80% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w. In some cases, the concentration of cellooligosaccharides with a degree of polymerization of 2 may be higher, for example, at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% w / w.

[0274] The concentration of cellooligosaccharides with a degree of polymerization of 3 in the cellooligosaccharide mixture can be from about 2% to about 20% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0275] The concentration of cellooligosaccharides with a degree of polymerization of 4 in the cellooligosaccharide mixture can be from about 5% to about 20% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0276] In the fiber oligosaccharide mixture, the concentration of fiber oligosaccharide with a degree of polymerization of 5 can be from about 5% to about 20% w / w. The concentration of fiber oligosaccharide with a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0277] In the fiber oligosaccharide mixture, the concentration of fiber oligosaccharide with a degree of polymerization of 6 can be from about 5% to about 25% w / w. The concentration of fiber oligosaccharide with a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0278] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 2 can be from about 2% to about 30% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w.

[0279] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 3 can be from about 2% to about 20% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0280] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 4 can be from about 5% to about 20% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0281] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 5 can be from about 5% to about 20% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0282] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 6 can be from about 5% to about 25% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0283] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 7 can be from about 2% to about 20% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0284] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 8 can be about 1% to about 15% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0285] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 9 can be about 2% to about 15% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0286] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 10 can be about 2% to about 15% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0287] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 11 can be about 2% to about 15% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0288] In the mannan oligosaccharide mixture, the concentration of mannan oligosaccharide with a degree of polymerization of 12 can be about 2% to about 15% w / w. The concentration of mannan oligosaccharide with a degree of polymerization of 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0289] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 4 can be about 5% to about 20% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0290] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 5 can be about 5% to about 20% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0291] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 6 can be about 5% to about 25% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0292] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 7 can be about 2% to about 20% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0293] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 8 can be about 1% to about 15% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0294] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 9 can be about 2% to about 15% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0295] In the xyloglucan oligosaccharide mixture, the concentration of xyloglucan oligosaccharide with a degree of polymerization of 10 can be about 2% to about 15% w / w. The concentration of xyloglucan oligosaccharide with a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0296] In the mixed-linkage glucan oligosaccharide mixture, the concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 2 can be about 2% to about 30% w / w. The concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w.

[0297] In the mixed-linkage glucan oligosaccharide mixture, the concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 3 can be about 2% to about 20% w / w. The concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0298] In the mixed-linkage glucan oligosaccharide mixture, the concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 4 can be about 5% to about 20% w / w. The concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0299] In the mixed-linkage glucan oligosaccharide mixture, the concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 5 can be about 5% to about 20% w / w. The concentration of mixed-linkage glucan oligosaccharide with a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0300] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 2 can be from about 2% to about 30% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w.

[0301] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 3 can be from about 2% to about 20% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0302] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 4 can be from about 5% to about 20% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0303] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 5 can be from about 5% to about 20% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0304] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 6 can be from about 5% to about 25% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0305] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 7 can be from about 2% to about 20% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0306] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 8 can be from about 1% to about 15% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0307] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 9 can be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0308] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 10 can be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0309] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 11 can be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0310] In the chitosan oligosaccharide mixture, the concentration of chitosan oligosaccharide with a degree of polymerization of 12 can be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0311] In the arabinoxylan oligosaccharide mixture, the concentration of arabinoxylan oligosaccharide with a degree of polymerization of 3 can be from about 2% to about 20% w / w. The concentration of arabinoxylan oligosaccharide with a degree of polymerization of 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0312] In the arabinoxylan oligosaccharide mixture, the concentration of arabinoxylan oligosaccharide with a degree of polymerization of 4 can be from about 5% to about 20% w / w. The concentration of arabinoxylan oligosaccharide with a degree of polymerization of 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0313] In the arabinoxylan oligosaccharide mixture, the concentration of arabinoxylan oligosaccharide with a degree of polymerization of 5 can be from about 5% to about 20% w / w. The concentration of arabinoxylan oligosaccharide with a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0314] In the arabinoxylan oligosaccharide mixture, the concentration of arabinoxylan oligosaccharide with a degree of polymerization of 6 can be from about 5% to about 25% w / w. The concentration of arabinoxylan oligosaccharide with a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0315] In the arabinoxylan oligosaccharide mixture, the concentration of arabinoxylan oligosaccharide with a degree of polymerization of 7 can be from about 2% to about 20% w / w. The concentration of arabinoxylan oligosaccharide with a degree of polymerization of 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0316] In the arabinoxylan oligosaccharide mixture, the concentration of arabinoxylan oligosaccharide with a degree of polymerization of 8 can be from about 1% to about 15% w / w. The concentration of arabinoxylan oligosaccharide with a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0317] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 9 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0318] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 10 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0319] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 11 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0320] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 12 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0321] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 13 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 13 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0322] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 14 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 14 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0323] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 15 in the arabinoxylan oligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 15 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0324] The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 3 to 12 in the arabinoxylan oligosaccharide mixture can be from about 0.1% to about 15% w / w. The concentration of arabinoxylan oligosaccharides with a degree of polymerization of 3 to 12 can be at least 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0325] Compositions having a combination of monosaccharides, polysaccharides, and / or oligosaccharides

[0326] In some embodiments, the composition or ingredient (e.g., an ingredient for human consumption) is water-soluble. The solubility of the ingredient in water can be at least 80 g of the ingredient per 100 g of water at 50 °C.

[0327] The ingredient can be combined with a liquid to form a liquid ingredient. In some cases, the viscosity of the liquid ingredient can be comparable or similar to the viscosity of corn syrup. In some other cases, the viscosity of the liquid ingredient can be comparable or similar to the viscosity of high fructose corn syrup. For example, the viscosity of the liquid ingredient can be 5 cps to 100,000 cps, 8,000 cps to 100,000 cps, 10,000 cps to 50,000 cps, or 15,000 cps to 25,000 cps. Additionally, the calories per gram of the liquid ingredient are lower than those of corn syrup or high fructose corn syrup. The glycemic index of the liquid ingredient is lower than that of corn syrup or high fructose corn syrup.

[0328] In some embodiments, the liquid can include water or any other suitable liquid. The liquid ingredient can include at least 5%, 10%, 20%, 30%, 40%, or 50% dry weight of at least one oligosaccharide. Additionally, the liquid ingredient can contain at least 0.2%, 0.5%, 1%, 2%, 3%, 5%, or 10% dry weight of at least one polysaccharide. For example, the liquid ingredient can contain at least 20% dry weight of at least one oligosaccharide and at least 2% dry weight of at least one polysaccharide. Other combinations of at least one oligosaccharide and at least one polysaccharide are also within the scope of the present disclosure.

[0329] The liquid ingredient contains at least 0.2%, 0.5%, 1%, 2%, 3%, 5%, or 10% dry weight of xylan. The liquid ingredient can contain at least 0.2%, 0.5%, 1%, 2%, 3%, 5%, or 10% dry weight of mannan. The liquid ingredient can contain at least 0.2%, 0.5%, 1%, 2%, 3%, 5%, or 10% dry weight of a cellulose derivative.

[0330] In various cases, the polysaccharide concentration of the liquid component can be 0.1% to 50%, 0.1% to 40%, 0.1% to 30%, 0.1% to 20%, 0.1% to 10%, 0.5% to 50% or 1% to 50% w / v. For example, the polysaccharide concentration of the liquid component can be 0.1% to 50% w / v. The liquid component can contain a certain amount of polysaccharides and oligosaccharides, and the ratio of polysaccharides to oligosaccharides is 1:200 to 1:1, 1:150 to 1:1, 1:125 to 1:1, 1:100 to 1:1, 1:90 to 1:1, 1:80 to 1:1, 1:70 to 1:1, 1:60 to 1:1, 1:50 to 1:1, 1:25 to 1:1, or 1:10 to 1:1. For example, the liquid component can contain polysaccharides and oligosaccharides in a ratio of 1:100 to 1:1.

[0331] One or more soluble polysaccharides can include one or more of mannan, xylan, mixed-linkage glucan, lignocellulose, hemicellulose, cellulose derivatives, chitosan, xyloglucan, or any other suitable soluble polysaccharides. The cellulose derivatives can include at least one of cellulose acetate, hydroxyethyl cellulose, hydroxypropyl cellulose, or any other suitable cellulose derivatives.

[0332] The biomass can include at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass, softwood biomass, or any other suitable biomass.

[0333] In some cases, the composition for human consumption can include soluble polysaccharides and oligosaccharides, and the oligosaccharides include at least one of the following: (i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; (ii) xylooligosaccharides with a DP of 2 to 12; (iii) mannanooligosaccharides with a DP of 2 to 12; (iv) arabinoxylan oligosaccharides with a DP of 3 to 15; (v) mixed-linkage glucan oligosaccharides with a DP of 2 to 5; or (vi) chitosan oligosaccharides with a DP of 2 to 12. The composition can include less than 5% dry weight of soluble polysaccharides. In some cases, the composition can include less than 1%, 2%, 5%, 7.5%, 10%, or 20% dry weight of soluble polysaccharides. In some embodiments, the composition can be free or substantially free of insoluble polysaccharides.

[0334] The composition can comprise a combination of polysaccharides and oligosaccharides. In some embodiments, the composition can comprise a combination of oligosaccharides and soluble polysaccharides. The source of the polysaccharide (or soluble polysaccharide) in such compositions can comprise cellulose, such as biomass, e.g., the undigested components of partially digested biomass, e.g., the undigested biomass from the same reaction as the reaction that produces the oligosaccharides. The polysaccharides in the undigested biomass can include lignin, polyphenols, cellulose, lignocellulose, or any other suitable polysaccharide described herein. A polysaccharide (e.g., a soluble polysaccharide) can be added to the oligosaccharide mixture to improve the gastrointestinal tolerance of the oligosaccharide mixture. Consumption of oligosaccharides can cause gastrointestinal discomfort, including diarrhea, discomfort, and flatulence. Compared to commercially available sugar compositions or sugar compositions that primarily comprise monosaccharides and / or disaccharides, the compositions described herein can have improved gastrointestinal tolerance, e.g., less or no discomfort, flatulence, diarrhea, or gastrointestinal distress. For example, compared to if or when a subject ingests a commercially available sugar composition or a sugar composition that primarily comprises monosaccharides and / or disaccharides, a subject who ingests one or more of the compositions provided herein can have improved gastrointestinal tolerance, e.g., less or no discomfort, flatulence, diarrhea, or gastrointestinal distress.

[0335] The concentration of undigested biomass in the composition can be from 1% to 50% w / w. The concentration of undigested biomass in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of undigested biomass in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of undigested biomass in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of undigested biomass in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0336] The concentration of the soluble polysaccharide in the composition can be from 1% to 50% w / w. The concentration of the soluble polysaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of the soluble polysaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of the soluble polysaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of the soluble polysaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0337] The concentration of xylo-oligosaccharide in the composition can be 1% to 50% w / w. The concentration of xylo-oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of xylo-oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of xylo-oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of xylo-oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0338] The concentration of the xylo-oligosaccharide in the composition can be from 1% to 50% w / w. The concentration of the xylo-oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of the xylo-oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of the xylo-oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of the xylo-oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0339] In some embodiments, the composition can comprise at least 5% w / w of xylo-oligosaccharide and at least 5% w / w of a second oligosaccharide (e.g., at least 5% w / w of xylo-oligosaccharide, mannan-oligosaccharide, mixed-linkage glucan oligosaccharide, xyloglucan oligosaccharide, chitosan oligosaccharide, arabinoxylan oligosaccharide, or any other suitable oligosaccharide).

[0340] The concentration of mannan oligosaccharide in the composition can be from 1% to 50% w / w. The concentration of mannan oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of mannan oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of mannan oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of mannan oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0341] The concentration of chitosan oligosaccharide in the composition can be from 1% to 50% w / w. The concentration of chitosan oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of chitosan oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of chitosan oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of chitosan oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0342] The concentration of xyloglucan oligosaccharides in the composition can be from 1% to 50% w / w. The concentration of xyloglucan oligosaccharides in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of xyloglucan oligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of xyloglucan oligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of xyloglucan oligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0343] The concentration of the mixed-linkage glucan oligosaccharide in the composition can be from 1% to 50% w / w. The concentration of the mixed-linkage glucan oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of the mixed-linkage glucan oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of the mixed-linkage glucan oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of the mixed-linkage glucan oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0344] The concentration of arabinoxylan oligosaccharide in the composition can be from 1% to 50% w / w. The concentration of arabinoxylan oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50% or 45% to 50% w / w. The concentration of arabinoxylan oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of arabinoxylan oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of arabinoxylan oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0345] The composition can comprise one or more polysaccharides (e.g., one or more soluble polysaccharides) and one or more oligosaccharides. The composition can comprise a polysaccharide and one type of oligosaccharide. The composition can comprise a polysaccharide or multiple polysaccharides and two forms of oligosaccharides. The composition can comprise a polysaccharide or multiple polysaccharides and three forms of oligosaccharides. The composition can comprise a polysaccharide or multiple polysaccharides and four forms of oligosaccharides. The composition can comprise a polysaccharide or multiple polysaccharides and five forms of oligosaccharides. The oligosaccharide can be xylooligosaccharide, cellooligosaccharide, mannan oligosaccharide, mixed-link glucan oligosaccharide, xyloglucan oligosaccharide, chitosan oligosaccharide, arabinoxylan oligosaccharide, or any other suitable oligosaccharide described herein.

[0346] The composition may comprise from about 1% to 50% w / w of a polysaccharide (such as in the form of undigested biomass or extracted soluble polysaccharide), and from about 5% to about 95% w / w of an oligosaccharide. The composition of the polysaccharide can be at least about 1%, 2%, 2.5%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The oligosaccharide in such a mixture can be present at greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% w / w. The oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition can comprise 5% w / w of undigested biomass and 50% w / w of an oligosaccharide mixture. In another case, as described elsewhere herein, the composition can comprise 2.5% w / w of soluble polysaccharide and 50% w / w of an oligosaccharide mixture.

[0347] The composition may comprise from about 5% w / w of a polysaccharide (such as in the form of undigested biomass), and from about 5% to about 95% w / w of an oligosaccharide. The oligosaccharide in such a mixture can be present at greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% w / w. The oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition can comprise 5% w / w of undigested biomass and 50% w / w of an oligosaccharide mixture. In another case, as described elsewhere herein, the composition can comprise 5% w / w of soluble polysaccharide and 50% w / w of an oligosaccharide mixture.

[0348] The composition may comprise from about 7% w / w of a polysaccharide (such as in the form of undigested biomass), and from about 5% to about 93% w / w of an oligosaccharide. The oligosaccharide can form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 93% w / w of such a mixture. The oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition can comprise 7% w / w of undigested biomass and 50% w / w of an oligosaccharide mixture. In another case, as described elsewhere herein, the composition can comprise 7% w / w of soluble polysaccharide and 50% w / w of an oligosaccharide mixture.

[0349] The composition may comprise about 10% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 90% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or 90% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 10% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 10% w / w of a polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0350] The composition may comprise about 12% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 95% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or 88% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 12% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 12% w / w of a soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0351] The composition may comprise about 15% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 85% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80% or 85% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 15% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 15% w / w of a soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0352] The composition may comprise about 20% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 80% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 20% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 20% w / w of soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0353] The composition may comprise about 25% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 75% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 25% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 25% w / w of soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0354] The composition may comprise about 30% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 70% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 30% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 30% w / w of soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0355] The composition may comprise about 40% w / w of a polysaccharide (such as in the type of undigested biomass), and about 5% to about 60% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% or 60% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 40% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 40% w / w of a soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0356] The composition may comprise about 50% w / w of a polysaccharide (such as of the undigested biomass type) and about 5% to about 50% w / w of an oligosaccharide. The oligosaccharide may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w of such a mixture. The oligosaccharide may be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, the composition may comprise 50% w / w of undigested biomass and 50% w / w of a mixture of oligosaccharides. In another case, as described elsewhere herein, the composition may comprise 50% w / w of a soluble polysaccharide and 50% w / w of a mixture of oligosaccharides.

[0357] In some embodiments, the composition or ingredient may comprise less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w of a monosaccharide. For example, the composition may comprise less than 20% w / w of a monosaccharide. The composition may include 10% to 40%, 15% to 30%, 18% to 25% or about 20% w / w of a monosaccharide. In some embodiments, the composition or ingredient may comprise less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w of glucose. For example, the composition may comprise less than 10% w / w of glucose. The composition may include 10% to 40%, 15% to 30%, 18% to 25% or about 20% w / w of glucose. In some embodiments, the composition or ingredient may comprise less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w of xylose. For example, the composition may comprise less than 10% w / w of xylose. The composition may include 10% to 40%, 15% to 30%, 18% to 25% or about 20% w / w of xylose.

[0358] In certain cases, the ratio of glucose residues to xylose residues (e.g., glucose:xylose) within the composition or ingredient may be from 1:1 to 1:9, 1:1 to 1:7, 1:1 to 1:5, 1:1 to 1:3 or 1:1 to 1:2.

[0359] In certain embodiments, the composition may comprise less than 30%, 40%, 50%, 60%, 65%, 70%, 75% or 80% w / w disaccharide. For example, the composition may comprise less than 70% w / w disaccharide. The composition may include 10% to 95%, 15% to 90%, 20% to 80%, 30% to 70% or 40% to 60% w / w disaccharide. The composition may comprise 5% to 95%, 10% to 92.5%, 15% to 90%, 20% to 70%, 30% to 60% or 40% to 50% disaccharide. In various embodiments, the composition may comprise at least 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15% or 20% w / w trisaccharide. For example, the composition may comprise at least 5% w / w trisaccharide. In various embodiments, the composition may comprise at least 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15% or 20% w / w trisaccharide. For example, the composition may comprise at least 5% w / w trisaccharide. The composition may comprise 1% to 75%, 2.5% to 60%, 5% to 50%, 10% to 40% or 20% to 30% trisaccharide. In some cases, the composition may comprise at least 0.1%, 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15% or 20% w / w tetrasaccharide. For example, the composition may comprise at least 1% w / w tetrasaccharide. In various cases, the composition may comprise at least 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.5%, 0.5%, 1%, 2.5%, 5%, 7.5% or w / w pentasaccharide. For example, the composition may comprise at least 0.1% w / w pentasaccharide.

[0360] Use of the composition as an ingredient

[0361] In some embodiments, the composition is an ingredient (e.g., in a food product). In certain embodiments, the ingredient comprises at least 50%, 60%, 70%, 80%, 90%, 95%, 99% or 99.5% sugar present on a dry weight basis. The ingredient may consist essentially of sugar. For example, the ingredient may have less than 0.5%, 0.3% or 0.1% of other substances on a dry weight basis.

[0362] As described elsewhere herein, the ingredient may comprise a mixture of oligosaccharides. The ingredient may comprise at least two oligosaccharides. For example, the ingredient may comprise three oligosaccharides. The ingredient may comprise four oligosaccharides. The ingredient may comprise five oligosaccharides. The ingredient may comprise six oligosaccharides. The ingredient may comprise seven oligosaccharides.

[0363] In some embodiments, the ingredient comprises xylooligosaccharides, such as a combination of xylooligosaccharides and xylooligosaccharides. An alternative ingredient may comprise a combination of xylooligosaccharides and mannan oligosaccharides.

[0364] The ingredient can be used to prepare the final product. The ingredient can also be treated in some physical or chemical ways before or during incorporation into food, cosmetics, or nutraceuticals. The ingredient can be incorporated directly into the product, or the ingredient can be incorporated into, for example, dough, cake mixtures, chocolate mixtures, or other food precursors; into cosmetic base compositions; or into nutraceuticals and treated, for example, by steaming or in other ways that can cause chemical modification, texture change, color change, or other modifications.

[0365] Food, cosmetics, or nutraceuticals can be produced from the ingredients described herein. For example, in the food industry, sugar preparations produced by current methods can be used as sweeteners, bulking agents, added dietary fiber, or humectants. The ingredient can be used as a sugar substitute. The ingredient can be incorporated into cakes, bread, or other baked goods, or into chocolate or other confections, such as toffee, fudge, meringue, jam, jelly, or caramel; or into beverages, for example, to provide good flavor or color characteristics or to increase the dietary fiber content. In some cases, the ingredient can be incorporated into animal feed, for example, as a separate ingredient or by directly using the enzyme reaction mixture as a raw material.

[0366] In the cosmetics industry, sugars can be used as ingredients because they can improve texture and moisture retention, act as ultraviolet-absorbing molecules, maintain gel or cream structure, and / or act as bulking agents. The compositions described herein can be incorporated into nutraceutical compositions because the dietary fiber they provide can promote digestive health, well-regulate the gut microbiota, and provide other benefits to health. In such cases, they can also be used as ingredients in probiotic beverages or other prebiotic or probiotic preparations.

[0367] The compositions or ingredients described herein can be used to alter one or more properties of the final product. Such properties include, but are not limited to, sweetness, texture, mouthfeel, adhesiveness, glazing, smoothness, wetness, viscosity, color, hygroscopicity, flavor, bulking, water retention, caramelization, surface texture, crystallization, structural properties, and solubility.

[0368] In some cases, compared to the same properties provided by a sugar mixture mainly containing monosaccharides and / or disaccharides, the compositions and / or ingredients described herein can provide comparable or better properties for the final product. The control composition can be sugars commonly used in consumables, such as monosaccharide compositions, such as glucose, fructose, etc.; disaccharide compositions, such as sucrose, or artificial sugar compositions. The control composition can be table sugar, corn syrup, high-fructose corn syrup, or any other suitable composition. As used herein, the term "comparable" generally means that the two compositions can have up to 100%, up to 95%, up to 90%, or up to 80% identity. For example, comparable can mean that the composition has up to 90% identity with the control composition.

[0369] In some cases, the compositions described herein can be used as sweetener compositions. The sweetener compositions can be used alone or as an ingredient in a final product. Compared to an equal amount of a control composition, the compositions described herein can provide about the same level of sweetness or a higher level of sweetness, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a sweetener in a final product. In some cases, the sweetness of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90%, or 100% higher than that of an equal amount of the control composition.

[0370] Compared to an equal amount of a control composition, the compositions described herein can provide a comparable flavor profile or a better flavor profile, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a flavor enhancer in a final product. In some cases, the flavor of the composition can exceed that of an equal amount of the control composition by 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90%, or 100%.

[0371] Compared to an equal amount of a control composition, the compositions described herein can provide a comparable texture profile or a better texture profile, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a texture enhancer in a final product.

[0372] Compared to an equal amount of a control composition, the compositions described herein can provide a comparable binding profile or a better binding profile, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a binding enhancer in a final product.

[0373] Compared to an equal amount of a control composition, the compositions described herein can provide a comparable gloss profile or a better gloss profile, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a gloss enhancer in a final product.

[0374] Compared to an equal amount of a control composition, the compositions described herein can provide a comparable humidity or better humidity, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition to provide humidity in a final product.

[0375] Compared with the same amount of a control composition, the compositions described herein can provide a comparable color profile or a better color profile, wherein the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a color enhancer in the final product.

[0376] Compared with the same amount of a control composition, the compositions described herein can provide a comparable dissolution profile or a better dissolution profile, wherein the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a dissolution enhancer in the final product. In some cases, the dissolution of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than that of the same amount of the control composition.

[0377] Compared with the same amount of a control composition, the compositions described herein can provide a comparable taste or a better taste, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0378] Compared with the same amount of a control composition, the compositions described herein can provide a comparable viscosity or a better viscosity, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0379] Compared with the same amount of a control composition, the compositions described herein can provide a comparable hygroscopicity or a better hygroscopicity, wherein the control composition mainly comprises monosaccharides and / or disaccharides. In some cases, the hygroscopicity of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than that of the same amount of the control composition.

[0380] Compared with the same amount of a control composition, the compositions described herein can provide a comparable water retention or a better water retention, wherein the control composition mainly comprises monosaccharides and / or disaccharides. In some cases, the water retention of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than that of the same amount of the control composition.

[0381] Compared with the same amount of a control composition, the compositions described herein can provide a lower calorie composition, wherein the control composition mainly comprises monosaccharides and / or disaccharides. In some cases, the calorie content of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% lower than that of the same amount of the control composition.

[0382] Compared to an equal amount of a control composition, the compositions described herein can provide a lower glycemic index, wherein the control composition mainly comprises monosaccharides and / or disaccharides. In some cases, the glycemic index of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% lower than that of an equal amount of the control composition.

[0383] Compared to an equal amount of a control composition, the compositions described herein can provide comparable or better leavening, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0384] Compared to an equal amount of a control composition, the compositions described herein can provide comparable or better caramelization, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0385] Compared to an equal amount of a control composition, the compositions described herein can provide comparable or better surface texture, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0386] Compared to an equal amount of a control composition, the compositions described herein can provide comparable or better crystallization, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0387] Compared to an equal amount of a control composition, the compositions described herein can provide comparable structural properties, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0388] Compared to an equal amount of a control composition, the compositions described herein can provide less aftertaste, wherein the control composition mainly comprises monosaccharides and / or disaccharides.

[0389] Compared to oligosaccharides used alone, different oligosaccharide compositions can have improved dissolution profiles, hygroscopicity profiles, and taste profiles.

[0390] The compositions or ingredients described herein can be used to increase the fiber content of final products such as foods or nutraceuticals. Compared to an equal amount of a control composition, the composition can provide a higher level of fiber in the final product, wherein the control composition mainly comprises monosaccharides and / or disaccharides. In some cases, the composition can improve the fiber content of the final product without negatively or substantially negatively affecting any other properties, such as taste, sweetness, mouthfeel, texture, adhesiveness, or any other property described herein. In some cases, the fiber content of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than that of an equal amount of the control composition.

[0391] The ingredients can be used to change the properties of a final product, such as a food or a nutritional or cosmetic product. To change the properties of the final product, the final product can additionally contain a polysaccharide, such as a cellulose polysaccharide, such as cellulose; or a polysaccharide derivative, such as a cellulose derivative, such as carboxymethyl cellulose; or a polysaccharide aggregate, such as a part of lignocellulosic biomass. In some cases, the final product can contain from greater than 0% to 40% by dry weight of the polysaccharide, polysaccharide derivative or polysaccharide aggregate, for example, from greater than 1% to 30% by dry weight of the polysaccharide, polysaccharide derivative or polysaccharide aggregate, for example from greater than 5% to 25% by dry weight of the polysaccharide, polysaccharide derivative or polysaccharide aggregate, for example from greater than 10% to 20% by dry weight of the polysaccharide, polysaccharide derivative or polysaccharide aggregate.

[0392] The concentration of the composition containing the polysaccharide and oligosaccharide mixture in the final product can be any value between 0.1% and 40% w / w. The concentration of the composition containing the polysaccharide and oligosaccharide mixture in the final product can be about 0.1% to about 0.5%, about 0.1% to about 1%, about 0.1% to about 5%, about 0.1% to about 10%, about 0.1% to about 15%, about 0.1% to about 20%, about 0.1% to about 25%, about 0.1% to about 30%, about 0.1% to about 35%, about 0.1% to about 40%, about 0.5% to about 1%, about 0.5% to about 5%, about 0.5% to about 10%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 0.5% to about 30%, about 0.5% to about 35%, about 0.5% to about 40%, about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 1% to about 35%, about 1% to about 40%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 5% to about 40%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 10% to about 40%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 15% to about 40%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 20% to about 40%, about 25% to about 30%, about 25% to about 35%, about 25% to about 40%, about 30% to about 35%, about 30% to about 40% or about 35% to about 40% w / w. The concentration of the composition containing the polysaccharide and oligosaccharide mixture in the final product can be about 0.1%, about 0.5%, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35% or about 40% w / w. The concentration of the composition containing the polysaccharide and oligosaccharide mixture in the final product can be at least 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30% or 35% w / w. The concentration of the composition containing the polysaccharide and oligosaccharide mixture in the final product can be at most 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35% or 40% w / w.

[0393] In some cases, the oligosaccharide mixture (e.g., cellooligosaccharide and xylooligosaccharide) can form at least 20%, 30%, 40%, 50%, 60% or 70% w / w of the edible composition or ingredient. For example, a combination of 50% w / w cellooligosaccharide and xylooligosaccharide can form an edible composition or ingredient.

[0394] In some cases, the composition may include maize cob extract (MCE). MCE can be a mixture of oligosaccharides mainly composed of arabinoxyloligosaccharides, xylooligosaccharides, cellooligosaccharides, and cellulose. In certain cases, the oligosaccharides can be indigestible or substantially indigestible. Arabinoxyloligosaccharides can be oligomers in which the xylose backbone is linked by β-(1→4)-bonds substituted by arabinose side chains. Arabinoxyloligosaccharides can be indigestible. Arabinoxyloligosaccharides can be produced by hydrolyzing arabinoxylan (a polysaccharide of β-(1→4)-bonded xylose units substituted by arabinose side chains). In addition, the degree of polymerization (DP) of arabinoxyloligosaccharides can be from 3 to 15.

[0395] In various cases, xylooligosaccharides can be oligomers in which the xylose backbone is linked by β-(1→4)-bonds. Xylooligosaccharides can be indigestible. Xylooligosaccharides can be produced by hydrolyzing arabinoxylan. In addition, the DP of xylooligosaccharides can be from 2 to 8.

[0396] In certain cases, cellooligosaccharides can be oligomers in which the glucose backbone is linked by β-(1→4)-bonds. Cellooligosaccharides can be indigestible. Cellooligosaccharides can be produced by the hydrolysis of cellulose (a polysaccharide of β-(1→4)-bonded glucose units). In addition, the DP of cellooligosaccharides can be from 2 to 4, with most having a DP of 2.

[0397] IV. Exemplary Embodiments

[0398] Exemplary method for extracting soluble polysaccharides for subsequent combination with the generated oligosaccharides

[0399] Figure 8 is a simplified flowchart of an embodiment showing a method for extracting soluble polysaccharides for subsequent combination with the generated oligosaccharides to form a composition.

[0400] In the method shown, extracting or removing at least a portion of the soluble polysaccharide 140 before the enzyme treatment 150 can ensure that at least a portion of the soluble polysaccharide is retained or preserved for combination 144 with the generated oligosaccharides to form the composition 105. In some other methods (not shown), the soluble polysaccharide can be digested by one or more enzymes because the soluble polysaccharide is not extracted before the enzyme treatment. The composition 105 of the method shown can be a sweetener or sugar substitute, which can remain substantially soluble or completely soluble. Thus, the composition 105 can be delivered as a syrupy product (e.g., a viscous liquid). In some cases, the composition 105 can be a substitute or partial substitute for: corn syrup, high fructose corn syrup, maple syrup, honey, molasses, golden syrup, treacle, glucose syrup, high fructose syrup, agave syrup, date syrup, brown rice syrup, coconut syrup, corn syrup, or other suitable liquid sweeteners in syrupy foods.

[0401] As depicted, biomass 107 (e.g., corncobs or any other suitable biomass) can be physically pretreated 110 (e.g., by chopping or any other suitable method of physically pretreating the biomass). The physically treated biomass 112 can then undergo or be subjected to a thermochemical pretreatment 130 (e.g., heating 15% w / v chopped corncobs in 1% w / v NaOH for 1 hour). In some embodiments, the thermochemical pretreatment 130 can be followed by a neutralization step (not shown), and then soluble compounds or materials 140 can be extracted from the physically pretreated biomass 112. The extraction of the soluble compounds or materials 140 can include removing the liquid portion (e.g., supernatant) of the physically pretreated biomass 112. The liquid portion can include soluble compounds 146 from the physically pretreated biomass 112. The soluble compounds 146 can include soluble polysaccharides. In certain embodiments, then 15% of the liquid portion (including soluble compounds or materials 146) can be extracted or removed from the thermochemically pretreated biomass 112. The extraction portion including the soluble compounds or materials 146 (e.g., soluble polysaccharides) can then undergo one or more purification steps 142b (e.g., ultrafiltration) to enrich the soluble polysaccharides 147.

[0402] In addition, the liquid portion not extracted in step 140 can include soluble compounds or materials 146 and insoluble compounds or materials 148. For example, in the case of extracting 15% of the liquid portion as described above, the remaining portion of the physically pretreated biomass 112 (including 85% of the liquid portion) can include soluble and insoluble polysaccharides. The solution including the soluble compounds or materials 146 and the insoluble compounds or materials 148 can then undergo or be subjected to an enzymatic treatment 150 as disclosed herein. For example, one or more polysaccharide-cleaving enzymes can be added to the solution including the soluble compounds or materials 146 and the insoluble compounds or materials 148 to 0.5% w / v and incubated at 50 °C for 24 hours. Then, the enzymatically treated biomass 151 can be processed (e.g., filtered) 152 to remove at least a portion of the undigested biomass. In certain cases, the removed undigested biomass can be disposed of or discarded. Then, the digested biomass 141 can undergo or be subjected to purification 142a (e.g., ion exchange chromatography, nanofiltration, microfiltration, ultrafiltration, or any other suitable purification method) to enrich the oligosaccharides 154 described herein. The extracted, separated, and / or purified oligosaccharides 154 and the extracted, separated, and / or purified soluble polysaccharides 147 can be combined, mixed, and / or spray dried to form the composition 105.

[0403] Exemplary methods for pretreating biomass to remove monosaccharides and / or disaccharides

[0404] Figure 9It is a simplified flowchart showing an embodiment of a method for pretreating biomass to remove monosaccharides and / or disaccharides prior to enzymatic treatment.

[0405] Figure 9 Embodiments of may include components or steps that are in some respects similar to Figure 8 components or steps of embodiments of. For example, Figure 9 embodiments of include a physical pretreatment step 110 that may be similar to Figure 8 the physical pretreatment 210 of. It should be understood that the illustrated embodiments may have similar features. Accordingly, similar features are denoted by similar reference numerals, with a leading digit added to increment each reference numeral by 100. For example, the physical pretreatment is denoted as "110" in Figure 8 ; a similar physical pretreatment is denoted as "210" in Figure 9 . Accordingly, the related disclosure regarding similar identified features may not be repeated hereinafter. Additionally, Figure 9 specific features of the methods and related components or steps shown may not be shown in the figures or identified by reference numerals, or specifically discussed in the subsequent written description. However, such features may clearly be the same or substantially the same as features depicted and / or described with respect to other embodiments. Accordingly, the related description of these features equally applies to Figure 9 the methods and related components or steps of. Any suitable combination of the features described with respect to Figure 8 the methods shown and their variants may be used for Figure 9 the methods and components or steps of, and vice versa. This disclosure pattern equally applies to other embodiments depicted in the subsequent figures and / or described hereinafter.

[0406] As shown, the mild pretreatment 220 (e.g., the washing or incubation cycles provided herein) of the physically pretreated biomass 212 may include removing 224 soluble compounds 246. In some cases, the soluble compounds 246 may include monosaccharides and / or disaccharides. Then, the removed soluble monosaccharides and / or disaccharides may be discarded and / or disposed of. Accordingly, the mild pretreatment 220 may be carried out or performed to remove soluble monosaccharides and / or disaccharides from the biomass 207.

[0407] The biomass 207 can be physically pretreated 210 (e.g., chopped), and then the physically pretreated biomass 212 can be subjected to or undergo a mild pretreatment 220, such as washing or incubation (e.g., for 30 minutes in water at 25 °C). Then, the soluble sugars 246 (e.g., soluble monosaccharides and / or disaccharides) can be removed from the solution comprising the mildly pretreated biomass 226. Then, the mildly pretreated biomass 226 can be subjected to or undergo an intense pretreatment 230. In some cases, the intense pretreatment 230 can be a thermochemical pretreatment. For example, the mildly pretreated biomass 226 can be treated in 1% w / v NaOH at 100 °C for 60 minutes. In some embodiments, after the intense pretreatment 230, a neutralization step (not shown) can be performed prior to the enzymatic treatment 250. Then, as discussed herein, the intensively pretreated biomass 232 can be treated with an enzyme 250. Additionally, the enzymatically treated biomass 251 can then be subjected to downstream processing 260 to produce the component 205.

[0408] Other exemplary embodiments for extracting soluble polysaccharides for combination with oligosaccharides

[0409] In some cases, the present disclosure relates to new methods for processing plant biomass materials to produce food, cosmetic, or nutraceutical ingredients.

[0410] Sugary foods and beverages are an important part of the culture and lifestyle habits worldwide, but the sugars they contain are associated with people's obesity, diabetes, poor dental health, and disruptive behavior. For these reasons, consumer preferences have shifted away from sugary foods, and governments are increasingly implementing regulations to encourage the consumption of less sugar.

[0411] Therefore, for decades, the industry has been searching for suitable low-calorie sweeteners to replace the sugars in foods and beverages. Unfortunately, many sugar substitutes are produced from non-natural resources and often come with a slightly bitter or other unpleasant taste, neither of which is appealing to consumers. Additionally, while many sweeteners can mimic the sweetness of sugar in foods and beverages, few can mimic the broad roles of sugar in foods and beverages, such as increasing volume, modulating texture, providing structure, acting as a preservative, and regulating color and flavor through caramelization and the Maillard reaction.

[0412] Dietary fiber is an important part of a healthy diet and helps maintain digestive system health and good regulation of the gut microbiota. This fiber contains sugars of different chain lengths and types. In addition to being naturally present in a wide variety of foods, fiber can also be produced separately and added to other foods during the manufacturing process.

[0413] Biomass is a good source of sugars and can be used to replace sugars and add fiber to foods. However, there is still a need to optimize methods for obtaining these sugars from biomass and processing these sugars into compositions that can be used as ingredients in foods, cosmetics, or nutraceuticals.

[0414] First, sugars typically need to be produced by controlled decomposition. Different amounts and sizes of sugars in the composition can affect its nutritional value, as well as other properties such as hygroscopicity, which in turn affects properties such as the texture of products made using the composition. It is also desirable for the composition to include polysaccharides, as polysaccharides can improve gastrointestinal tolerance. However, due to the rate at which some particularly desirable polysaccharides decompose during the enzymatic reactions of previously known methods, these desirable polysaccharides are difficult to isolate and then incorporated into the composition.

[0415] In addition, foods that require a smooth texture (such as confections, chocolates, and yogurts) typically require the composition to be soluble to achieve a smooth texture. Compositions containing insoluble polymeric materials can produce a granular texture. However, due to the insolubility of certain polysaccharides, it can be difficult to prepare a composition containing completely soluble polysaccharides, particularly when prepared from a single piece of biomass in a one-pot process. Generally, soluble polysaccharides decompose more quickly than insoluble polysaccharides, so after exposing plant biomass to enzymes, as in previously known methods, the soluble polysaccharides are completely decomposed and only the insoluble polysaccharides remain.

[0416] Surprisingly, methods have been identified herein that can allow for the isolation of polysaccharides and their incorporation into soluble food, cosmetic, or nutraceutical ingredients containing oligosaccharides, thereby maintaining the benefit of increasing the gastrointestinal tolerance of the ingredient and allowing its use in foods with a smooth texture. The polysaccharides can be isolated from the same plant biomass, while other desired sugars can be obtained by providing an efficient and streamlined production method.

[0417] Accordingly, in a first aspect of the present disclosure, there is provided a method for producing a food, cosmetic, or nutraceutical ingredient, the method comprising the steps of:

[0418] a) providing plant biomass comprising one or more soluble polysaccharides and one or more insoluble polysaccharides;

[0419] b) treating the plant biomass to dissolve the one or more soluble polysaccharides;

[0420] c) removing a portion of the dissolved one or more soluble polysaccharides;

[0421] d) reacting the remaining plant biomass with one or more enzymes to form one or more oligosaccharides;

[0422] e) removing the one or more oligosaccharides; and

[0423] f) Combine a portion of the dissolved one or more soluble polysaccharides from step (c) with one or more oligosaccharides from step (e) to form a composition.

[0424] Accordingly, there is also provided a food, cosmetic or nutritional composition obtainable by the methods of the present disclosure.

[0425] In another aspect of the present disclosure, there is provided a food, cosmetic or nutritional liquid composition comprising at least one oligosaccharide selected from:

[0426] i) xylooligosaccharides having a degree of polymerization of 2 to 6;

[0427] ii) xylooligosaccharides having a degree of polymerization of 2 to 12;

[0428] iii) mannan oligosaccharides having a degree of polymerization of 2 to 12;

[0429] iv) mixed-linkage glucan oligosaccharides having a degree of polymerization of 2 to 5;

[0430] v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 12; and

[0431] vi) chitosan oligosaccharides having a degree of polymerization of 2 to 12;

[0432] and at least one polysaccharide selected from:

[0433] i) xylan;

[0434] ii) mannan;

[0435] iii) cellulose derivatives;

[0436] iv) mixed-linkage glucan;

[0437] v) xyloglucan; and

[0438] vi) chitosan;

[0439] wherein the liquid composition comprises at least 20% dry weight of at least one oligosaccharide and at least 2% dry weight of at least one polysaccharide, and wherein the viscosity of the liquid composition is from 5 to 100,000 cps.

[0440] Preparing food, cosmetic or nutritional compositions in the manner provided herein can allow for the efficient use of biomass by incorporating oligomeric and polymeric materials from the same biomass source to prepare soluble compositions. In addition, these methods can allow for purification, derivatization or other modifications, as well as control of the oligomeric and polymeric ratios, which can improve the functional, nutritional and tolerability properties of the compositions.

[0441] Any material containing suitable polysaccharides can be plant biomass. Since the food, cosmetic, and nutritional industries use a variety of oligosaccharides, there is no particular limitation on the polysaccharides applicable to this method. The plant biomass applicable to produce the oligosaccharide profiles of the present disclosure can include, for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan), xyloglucan, mixed-linkage glucan, and / or mannan (such as glucomannan, galactomannan, or galactoglucomannan). However, any plant biomass that can act appropriately is envisioned. One or more soluble polysaccharides contained in the plant biomass can include any of the following: mannan, mixed-linkage glucan, lignocellulose, hemicellulose, certain cellulose derivatives (such as cellulose acetate, hydroxyethyl cellulose, and hydroxymethyl cellulose), and chitosan. In some embodiments, the plant biomass contains hemicellulose. In certain embodiments, the hemicellulose contains xylan and / or mannan.

[0442] Thus, the plant biomass can be grains, husks, pods, seed coats, and / or other seed materials; seaweeds; corn stover, straw, bagasse, miscanthus, sorghum bagasse, switchgrass, bamboo, and / or other monocotyledonous tissues; water hyacinths, leaf tissues, roots, and / or other plant materials; and / or any combination of suitable plant biomass. In some cases, the plant biomass comprises or is suitably composed of the following: sugarcane biomass (such as bagasse), corn biomass (such as corn cobs or corn stover), wheat biomass (such as wheat straw or wheat bran), hardwood or softwood. In certain cases, the plant biomass includes corn cobs, bagasse, wheat straw, or straw.

[0443] In various cases, in step (b), the "treatment" is a thermochemical treatment of the plant biomass. As used herein, "thermochemical" generally refers to heating the plant biomass in a chemical substance above room temperature (room temperature can be about 20 °C to 22 °C), such as heating in a solution containing water, an alkali, or an ionic solvent. The thermochemical step can physically and chemically modify the chemical composition of the plant biomass. For example, free hydroxide ions from water or an alkali can break the hydrogen bonds between sugars, thereby dissolving certain types of sugars (e.g., hemicellulose) and making it easier for enzymes in subsequent steps to break down the sugars. These broken hydrogen bonds may be between monomers of the same sugar chain, which contributes to the tertiary structure of the chain. The broken hydrogen bonds may also be between monomers of different sugar chains, which contributes to the quaternary structure of more than one chain. Subsequently, the treatment can result in one or more polysaccharides being soluble (i.e., polysaccharides particularly susceptible to breakage by hydroxide ions, especially, for example, hemicellulose) to dissolve into the chemical substance used. One or more insoluble polysaccharides (such as cellulose) do not dissolve into the chemical substance.

[0444] The heating in process step (b) can be carried out within a certain temperature range, suitably from 30°C to 180°C, 50°C to 150°C, or 70°C to 120°C. Higher temperatures can contribute to a faster dissolution of the soluble polysaccharides. However, excessively high temperatures may be more difficult to achieve in an efficient and cost-effective manner and can chemically modify the biomass components, including sugars (e.g., in an undesirable way).

[0445] The heating can be carried out within a certain time scale range, especially when a large amount of biomass can be exposed to heating for a longer period of time, which can be adjusted accordingly. For example, the heating of plant biomass can be for 1 minute to 72 hours, 10 minutes to 24 hours, 20 minutes to 12 hours, or 25 minutes to 8 hours.

[0446] In some cases, the thermochemical treatment can include heating the plant biomass in water, i.e., at a neutral pH of about 7.

[0447] In certain cases, the thermochemical treatment can include heating the plant biomass in an alkaline solution with a pH of 8 to 14, 9 to 14, or 10 to 14. The solution can contain any one of the bases selected from the following, or suitably consist of any one of the following bases: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and ammonia water. In various cases, the base can be sodium hydroxide. Combinations of the listed bases are also envisaged.

[0448] Multiple different consecutive thermochemical treatment steps are also envisaged. For example, there can be two consecutive thermochemical treatments, three consecutive thermochemical treatments, or four or more consecutive thermochemical treatments. In some cases, the biomass can be subjected to thermochemical treatment in a neutral aqueous solution and then in an alkaline aqueous solution.

[0449] After the treatment step, step (c) can include removing a portion of the dissolved one or more soluble polysaccharides. The purpose of this step can be to separate and remove the soluble polysaccharides from the plant biomass so that they are not decomposed and thus lost in subsequent enzymatic reactions. This can enable the use of these polysaccharides when forming components in subsequent steps. Depending on the amount required in the final component, all of the dissolved polysaccharides or a portion of the dissolved polysaccharides can be removed. The soluble polysaccharides can be removed using simple steps, such as filtering the chemical in which the soluble polysaccharides are dissolved.

[0450] Step d) comprises reacting the remaining plant biomass, which may be in the form of a solution and / or a suspension, with one or more enzymes to form one or more oligosaccharides. Soluble and insoluble polysaccharides present in the remaining plant biomass solution and / or suspension may be partially or completely cleaved into oligosaccharides (such as useful oligosaccharides) by one or more enzymes, possibly leaving partially cleaved or uncleaved polysaccharides, which may include cellulose, xylan (such as glucuronoxylan, arabinoxylan or glucuronoarabinoxylan), mannan (such as glucomannan, galactomannan or galactoglucomannan), mixed-link glucan, xyloglucan, chitin, chitosan or lignocellulose.

[0451] The enzymatic reaction can be carried out in a suitable reaction vessel in solution and / or suspension. The enzymatic reaction can be carried out at a temperature or temperature profile suitable for the particular combination of enzyme and plant biomass, and the reaction can be allowed to proceed for a period of time until the product reaches the desired concentration or until some other requirement has been met.

[0452] To ensure optimal contact between the enzyme and the plant biomass, the reaction mixture can be agitated continuously or intermittently. Agitation can take the form of rhythmic movement of the entire reaction vessel, a blower or other agitation device, bubble injection or any other agitation method.

[0453] The enzymatic reaction can be microbial fermentation. The temperature and reaction time can be suitable for the growth of the microorganism used. The microbial organism can be genetically altered to produce an enzyme suitable for producing the oligosaccharides of the present disclosure. The microorganism can be, for example, a bacterium (such as Escherichia coli) or a fungus (such as Saccharomyces cerevisiae, Aspergillus niger or Trichoderma reesei).

[0454] Further embodied in the present disclosure is an expression vector suitable for modifying the subject microorganism to cause the microorganism to produce the enzyme or enzyme mixture of the present disclosure. If desired, the expression vector can be a plasmid or any other nucleic acid capable of inducing enzyme production, and the expression vector can contain one or more of the following regulatory sequences to control the expression of the exogenous enzyme: regulatory sequences of heat shock genes, regulatory sequences of virulence genes, and regulatory sequences of sporulation genes.

[0455] The enzymatic reaction can be carried out at a temperature or temperature profile suitable for the enzyme and substrate used. For example, the enzymatic reaction can be carried out at a constant temperature in the range of about 10°C to about 100°C, about 20°C to about 70°C or about 30°C to about 60°C. If the enzymatic reaction takes the form of microbial fermentation, the temperature may be suitable for this, for example, the enzymatic reaction can include the growth of Escherichia coli and / or the temperature can be constant and about 37°C.

[0456] The pH value of the solution or suspension can affect the activity of the enzyme. Controlling the pH can ensure that the enzymatic reaction proceeds at a suitable rate. The enzymatic reaction of the present disclosure can be carried out at a pH within the range of about 2 to about 10, about 3 to about 8, or about 4 to about 6.

[0457] The enzymatic reaction can be allowed to proceed for a period of time and then quenched, and the product can be separated or otherwise collected. The period of time can be from about 1 minute to about 6 days, about 0.5 days to about 5 days, or about 16 hours to about 96 hours. Alternatively, the reaction can be allowed to proceed until no further catalysis occurs.

[0458] The enzymatic reaction can be allowed to continue until the remaining undigested polysaccharide-containing plant biomass is less than 75%, less than 70%, less than 65%, less than 55%, or less than 50%. This can be monitored or checked by reducing end assays (such as the anthrone assay) and / or chromatography (such as thin layer chromatography and high performance anion exchange chromatography). The reaction can be carried out until all polysaccharides are converted to oligosaccharides.

[0459] There are many enzymes suitable for the enzymatic reaction of the present method. For example, "lytic polysaccharide monooxygenase" and "LPMO", the lytic polysaccharide monooxygenase and LPMO refer to a class of enzymes that can use a copper-containing moiety and use an oxygen source (such as dioxygen molecule, peroxide or any other oxygen source); and oxidatively cleave polysaccharides with a suitable reducing agent. Thus, when using LPMO, the enzymatic reaction can be carried out under aerobic conditions. The suitable reducing agent is not particularly limited, but examples include ascorbic acid, gallic acid, cysteine, NADH, NADPH, pyrogallol, dithiothreitol, cyanoborohydride, borohydride, photosynthetic pigments, lignin, ligninol, and a combination of cellobiose and cellobiose dehydrogenase. A variety of photosynthetic pigments can be used. In some embodiments, thylakoids and purified fractions or chlorophyllin can be used, and light can be provided. LPMO can be selected from the following families: AA9, AA10, AA11, AA13, AA14, and AA15. In various cases, LPMO can be PaLPMO9E (SEQ ID NO:1), which is an AA9 LPMO originally isolated from the ascomycete fungus (Podospora anserina), or LPMO can be an AA9 LPMO from Trichoderma reesei (SEQ ID NO:23).

[0460] An aerobic condition can include the addition of oxygen, which can be provided by aerating the substrate mixture with an oxygen-containing gas such as air. Aeration can be carried out by introducing oxygen-containing bubbles into the aqueous substrate mixture through various systems such as an air injector, an aeration frit, a membrane system, or an internal circulation air-lift reactor. The concentration of molecular oxygen in the enzymatic reaction can be from about 4 mg / L to about 14 mg / L.

[0461] Another enzyme that can be used in the method is "cellulase", which has hydrolytic activity towards cellulose, for example, endo-1,4-β-glucanase, cellobiohydrolase, and / or β-glucosidase activity. The enzyme is capable of cleaving glycosidic bonds in one or more forms of cellulose, including the cellulose found in plant biomass. During this process, cellulose produces products including glucose and cellooligosaccharides. In certain cases, the β-glucanase can include enzymes selected from the GH5, GH7, and GH12 enzymes, such as those derived from Aspergillus niger (SEQ ID NOs: 12, 13, and 14) and Trichoderma reesei (SEQ ID NOs: 24 and 25).

[0462] Another enzyme is "cellobiohydrolase", which has hydrolytic activity towards cellulose and mainly produces cellobiose as a product. Cellobiose is a disaccharide and is a type of cellooligosaccharide. Such enzymes are capable of cleaving glycosidic bonds in one or more forms of cellulose, including the cellulose found in plant biomass. In various cases, the cellobiohydrolase can be from the GH6 and GH7 families, such as the Cel6A or Cel7A enzymes from Trichoderma reesei (SEQ ID NOs: 10 and 11, respectively).

[0463] Another enzyme is "β-glucosidase", which has hydrolytic activity towards cellulose and mainly produces glucose as a product. Such enzymes are capable of cleaving glycosidic bonds in one or more forms of cellulose, including the cellulose found in plant biomass. In some embodiments, the β-glucosidase can include GH3 β-glucosidase, such as an enzyme derived from Trichoderma reesei (SEQ ID NO: 22).

[0464] Another class of enzymes is lichenase, which can be selected from the GH5, GH7, GH8, GH9, GH12, GH16, GH17 or GH26 families. In some embodiments, the lichenase can be a GH16 enzyme, such as a GH16 enzyme derived from Bacillus subtilis (SEQ ID NO:2). This enzyme is capable of acting on, for example, mixed-link glucans (which are glucans that are a mixture of β-1,3 bonds and β-1,4 bonds), and can cleave them at the β-1,4 glycosidic bonds. In cases where the lichenase acts on mixed-link glucans, the resulting β-glucans can mainly fall within the size range of about 3 to about 7 residues, and thus they are particularly useful in the food, cosmetic and nutritional industries. Mixed-link glucans are abundant in members of the grass and horsetail families, and thus, grassy biomass such as straw has a high level of mixed-link glucans and can be effectively acted upon by lichenase.

[0465] Another class of enzymes is xylanase, which acts on, for example, plant biomass containing a xylan backbone. The xylanase can be, for example, glucuronoxylanase, arabinoxylanase or glucuronarabinoxylanase. The enzyme can be active against a variety of polymers having a xylan backbone, such as glucuronoxylan, arabinoxylan and glucuronarabinoxylan. These polymers are abundant in various plant biomass, for example, both hardwoods and softwoods can contain suitable polysaccharides, where hardwoods typically contain glucuronoxylan and softwoods typically contain arabinoglucuronoxylan. In some cases, the xylanase can include GH5 xylanases derived from Clostridium thermocellum (SEQ ID NO:3) and Saprolegnia ferax (SEQ ID NO:4), and GH30 xylanases derived from Erwinia chrysanthemi (SEQ ID NO:5), Bacillus subtilis (SEQ ID NO:6), Bacteroides ovatus (SEQ ID NO:7) and Trichoderma reesei (SEQ ID NO:15).

[0466] Other enzymes useful in the present disclosure can include xyloglucanase and xyloglucan endo-glucanase (XEG), which are produced by a variety of organisms, including plant pathogenic microorganisms. Xyloglucanase and xyloglucan endo-glucanase are capable of acting on xyloglucan, which is a hemicellulose β-1,4 glucan chain abundant in the primary cell walls of higher plants, which is modified with xylose and some xylose residues are further modified with other residues such as galactose. When a suitable xyloglucanase or XEG acts on xyloglucan, the product contains xyloglucooligosaccharides having a backbone length that can be used in the food, cosmetic and nutritional industries. In some cases, the xyloglucanase can include a GH5 xyloglucanase derived from Bacteroides ovatus (SEQ ID NO:8) and a GH74 xyloglucanase derived from Trichoderma reesei.

[0467] Since any given natural plant biomass may contain a mixture of different polysaccharides, sometimes a mixture of different enzymes may be beneficial. Such a mixture can contain one or more of any other enzyme. For example, such a mixture can contain LPMO and endoglucanase, xylanase and lichenase, cellobiohydrolase and mannanase, or endoglucanase and cellobiohydrolase, where the enzyme partners are present in a molar ratio of, for example, 1:100 to 100:1.

[0468] In some cases, one or more enzymes can be a mixture of different enzymes, such as a crude enzyme preparation or a semi-crude enzyme preparation. As used herein, the term "crude enzyme preparation" generally refers to a soluble preparation extracted from a microbial fermentate that undergoes minimal processing after extraction, for example, typically the preparation may only be filtered to remove insoluble components. As used herein, the term "semi-crude enzyme preparation" generally refers to a soluble preparation extracted from a microbial fermentate that undergoes some processing after extraction, for example, the preparation may be filtered to remove insoluble components, increase the enzyme concentration, and / or nanofiltration to remove small molecular weight compounds.

[0469] In some cases, the crude enzyme preparation or the semi-crude enzyme preparation can be from bacteria or fungi. In some embodiments, the crude enzyme preparation or the semi-crude enzyme preparation can be from fungi, such as filamentous cellulolytic fungi, such as from species of Trichoderma or Aspergillus. In certain embodiments, the enzyme can be a crude enzyme preparation or a semi-crude enzyme preparation from Trichoderma reesei strain.

[0470] In step (e), one or more oligosaccharides formed in step (d) are removed, which can be carried out in a variety of ways. It can be separated based on solubility such that only a composition of soluble sugars is extracted for further processing, and / or by chromatographic separation to produce a composition with a narrower band of oligosaccharide chain lengths. The separation can be, for example, based on precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration. In the case of separation based on solubility, the sugar profile present in the separated composition can depend on the initial enzymatic reaction, since the solubility of different sugars decreases at different rates with length.

[0471] Also contemplated within the scope of the present disclosure is the further processing of all or part of the one or more removed oligosaccharides to produce further products, which are then combined with one or more dissolved polysaccharides to form a composition. The further processing may include any chemical, physical or enzymatic steps, such as reduction, for example reductive amination carried out where appropriate; oxidation, caramelization, modification with Schiff bases or by the Maillard reaction or by any combination of these steps, and can provide different products with improved properties for the desired purpose. For example, caramelization characteristics, calorific value, flavor and color can be modified. The oligosaccharides can also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0472] Also contemplated within the scope of the present disclosure is the further processing of all or part of the one or more dissolved soluble polysaccharides to produce a product with improved properties, which is then combined with one or more removed oligosaccharides to form a composition. The further processing may include any chemical, physical or enzymatic steps, such as alkylation or acid treatment. The polysaccharides can also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0473] In some cases, after modification and / or purification of the oligosaccharides and polysaccharides, all or part of the oligosaccharides and polysaccharides are combined, as in step (f), and the polysaccharide:oligosaccharide ratio can be from 1:100 to 1:1, 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4 or 1:60 to 1:5. The specific ratio may depend on the desired properties of the final composition and the modifications and purifications that have been applied to the sugars. In some embodiments, it is not necessary to recombine all of the removed oligosaccharides and polysaccharides.

[0474] In step (f), the combination can be carried out in a variety of ways. For example, by mixing a solution containing all or part of one or more soluble polysaccharides and a solution and / or suspension containing all or part of the soluble polysaccharides, which can be further spray dried, freeze dried or otherwise concentrated. The soluble polysaccharides and the removed oligosaccharides can also be mixed by combining a dry form containing all or part of one or more removed oligosaccharides produced by spray drying, freeze drying or otherwise concentrating (after removing them in step (e)) with a dry form containing all or part of one or more removed polysaccharides produced by spray drying, freeze drying or otherwise condensing (after removing them in step (c)). Alternatively, either (i) one or more soluble polysaccharides or (ii) one of the removed oligosaccharides can be in dry form while the other is in solution form when they are combined.

[0475] When the ingredient is in a dry form, the method may further include the step (g) of mixing and dissolving the ingredient in a liquid to form a liquid ingredient. In various cases, the liquid may be an aqueous solution, such as water.

[0476] The ingredient formed in step (f) and the liquid ingredient formed in step (g) according to the second aspect of the present disclosure may contain various oligosaccharides and contain different amounts depending on the desired properties. In some cases, the ingredient and the liquid ingredient may contain at least 20% dry weight or at least 30% dry weight of cellooligosaccharides with a degree of polymerization of 2 to 6, the ingredient may contain at least 20% dry weight or at least 30% dry weight of xylooligosaccharides with a degree of polymerization of 2 to 12, the ingredient may contain at least 20% dry weight or at least 30% dry weight of mixed-linkage glucan oligosaccharides with a degree of polymerization of 2 to 5, the ingredient may contain at least 20% dry weight or at least 30% dry weight of mannan oligosaccharides with a degree of polymerization of 2 to 12, the ingredient may contain at least 20% dry weight or at least 30% dry weight of xyloglucan oligosaccharides with a degree of polymerization of 4 to 12, and / or the ingredient may contain at least 20% dry weight or at least 30% dry weight of chitosan oligosaccharides with a degree of polymerization of 2 to 12. In some cases, the ingredient may contain up to 100% dry weight of the above oligosaccharides and the polysaccharides described herein, so the above embodiments in which the oligosaccharides are present at least 20% dry weight do not include five or six types of oligosaccharides.

[0477] The ingredient and the liquid ingredient may contain at least 50%, 60%, 70%, 80%, 90%, 95%, 99% or 99.5% dry weight of sugars present. The ingredient and the liquid ingredient may consist essentially of sugars. For example, the ingredient may have other substances less than 0.5% dry weight or less than 0.3% dry weight, such as 0.1% dry weight.

[0478] In some cases, the viscosity of the liquid component can be 5 to 100,000 cps, 10 to 80,000 cps, 20 to 60,000 cps, 30 to 40,000 cps, 40 to 20,000 cps, or 50 to 10,000 cps. When the liquid component is to be included in a beverage, a low syrup viscosity may be required, such as about 20 to 300 cps, 50 to 200 cps, or 100 to 150 cps. Higher viscosity values may be required in applications such as chocolate manufacturing, so when the liquid component is in syrup, its viscosity can be about 8,000 to 100,000, about 10,000 to 50,000 cps, or about 15,000 to 25,000 cps. The viscosity values are consistent with tests conducted using a Brookfield HDBVE rotational viscometer with a standard test procedure (where 400 mL of the sample is taken in a tall-form beaker to ensure no container effect). The instrument is operated according to the manufacturer's instructions regarding the range (using a rotational viscometer with spindle code 61, spindle speed 100 rpm, at 22 °C).

[0479] In some cases, the flow rate of the liquid component can be 100 to 350 seconds, 150 to 300 seconds, or 200 to 250 seconds. As used herein, "flow rate" generally refers to the volume of fluid passing per unit time. Unless otherwise specified, the flow rate values specified herein are determined by the following procedure: timing the flow rate of 5 mL of the liquid component from a vertical upright syringe (BD Plastipak 300613) filled with 20 mL of the test liquid under gravity at room temperature.

[0480] In various cases, the oligosaccharide concentration of the liquid component can be 1 to 200% w / v, 10 to 150% w / v, 20 to 140% w / v, 30 to 130% w / v, 40 to 120% w / v, 50 to 115% w / v, or 60 to 110% w / v.

[0481] In some cases, the polysaccharide concentration of the liquid component can be 0.1 to 50% w / v, 0.2 to 40% w / v, 0.3 to 30% w / v, 0.5 to 20% w / v, or 1 to 20% w / v.

[0482] In certain cases, the total concentration of oligosaccharides and polysaccharides in the liquid component can be 1 to 200% w / v, 10 to 160% w / v, 20 to 150% w / v, 30 to 140% w / v, 40 to 130% w / v, 50 to 120% w / v, or 60 to 110% w / v. In various cases, the higher the concentration of oligosaccharides and polysaccharides in the liquid, the thicker and more viscous the liquid can become. The liquid component can be a homogeneous solution.

[0483] In another aspect, the composition and the liquid composition can comprise at least two oligosaccharides. The amount of each oligosaccharide can vary depending on the desired properties of the resulting food, cosmetic or nutritional product. The two oligosaccharides can be present in a ratio of 1:9 to 9:1 or 1:2 to 2:1. Additionally, the composition and the liquid composition can comprise three oligosaccharides, they can comprise four oligosaccharides, they can comprise five oligosaccharides, or they can comprise six oligosaccharides.

[0484] The composition and the liquid composition can comprise xylooligosaccharides, such as a combination of xylooligosaccharides and xylooligosaccharides. Alternatively, the composition and the liquid composition can comprise a combination of xylooligosaccharides and mannan oligosaccharides.

[0485] One or more soluble polysaccharides in the composition can be particularly soluble in water or base. For example, the soluble polysaccharides used in the present disclosure can include hemicelluloses, such as xylan, mannan, mixed-linkage glucan, and certain cellulose derivatives (such as cellulose acetate, hydroxyethyl cellulose, and hydroxymethyl cellulose), as well as chitosan. In some embodiments, one or more soluble polysaccharides can comprise hemicelluloses. In certain embodiments, the hemicelluloses can comprise xylan and / or mannan.

[0486] In some cases, the composition and the liquid composition can comprise at least 2% dry weight or at least 3% dry weight of xylan. In various cases, the composition and the liquid composition can comprise at least 2% dry weight or at least 3% dry weight of mannan. In certain cases, the composition and the liquid composition can comprise at least 2% dry weight or at least 3% dry weight of cellulose derivatives. In some cases, the composition and the liquid composition can comprise at least 2% dry weight or at least 3% dry weight of mixed-linkage glucan. In various cases, the composition and the liquid composition can comprise at least 2% dry weight or at least 3% dry weight of xyloglucan. In certain cases, the composition and the liquid composition can comprise at least 2% dry weight or at least 3% dry weight of chitosan.

[0487] In certain cases, the composition and the liquid composition can comprise 2 to 40% dry weight of one or more soluble polysaccharides (including polysaccharide derivatives), 3 to 30% dry weight of one or more soluble polysaccharides, 5 to 25% dry weight of one or more soluble polysaccharides, or 8 to 20% dry weight of one or more soluble polysaccharides.

[0488] In some cases, the polysaccharide:oligosaccharide ratio of the composition and the liquid composition is 1:100 to 1:1, 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5.

[0489] The produced ingredients and liquid ingredients can be used in the applications of oligosaccharides, sugars, bulking sweeteners, low-intensity sweeteners or other related food ingredients for conventional use. For example, as sweeteners, bulking agents, added dietary fibers or humectants. Of particular note is the use of reduced sucrose in foods. It can be incorporated into cakes, bread or other baked goods; chocolates or other confections, such as toffees, fudges, meringues, jams, jellies or caramels; or incorporated into beverages, for example to provide good flavor or color characteristics or to increase the dietary fiber content. Alternatively, the ingredient can be incorporated into animal feed, for example as a separate ingredient or by directly using the enzymatic reaction mixture as a raw material.

[0490] The compositions or ingredients as described herein can be used to modify one or more properties of the final product. Such properties include but are not limited to sweetness, texture, mouthfeel, adhesiveness, glossiness, smoothness, moistness, viscosity, color, hygroscopicity, flavor, fluffiness, water retention, caramelization, surface texture, crystallization, structural properties, calorie reduction, glycemic index reduction, blood glucose load reduction, fiber increase, sugar reduction and solubility. These can be possible improvements over the currently used different types of sugars, sugar substitutes and / or other such compounds.

[0491] In the cosmetic industry, the ingredient can improve texture and moisture retention, act as an ultraviolet absorbing molecule, maintain the gel or cream structure, and / or act as a bulking agent. In addition, the ingredient and the liquid ingredient can be used in nutritional compositions because it has been shown that the dietary fiber they provide can promote digestive health, well regulate the gut microbiota and other benefits to health. In this case, the ingredients provided herein can also be used as ingredients in probiotic beverages or other prebiotic or probiotic preparations.

[0492] The detailed description is further supplemented with reference to the embodiments numbered below. 1) A method for producing a food, cosmetic or nutritional ingredient, said food, cosmetic or nutritional ingredient comprising one or more oligosaccharides and one or more soluble polysaccharides, said method comprising the following steps: (a) providing plant biomass comprising one or more soluble polysaccharides and one or more insoluble polysaccharides; (b) treating said plant biomass to dissolve said one or more soluble polysaccharides; (c) removing a portion of the dissolved one or more soluble polysaccharides; (d) reacting the remaining plant biomass with one or more enzymes to form one or more oligosaccharides; (e) removing said one or more oligosaccharides; and (f) combining a portion of the dissolved one or more soluble polysaccharides from step (c) with the one or more oligosaccharides from step (e) to form an ingredient. 2) The method according to embodiment numbered 1, wherein the treatment in step (b) is a thermochemical treatment. 3) The method according to embodiment numbered 2, wherein said thermochemical treatment is a hot water treatment or a hot alkali treatment. 4) The method according to embodiment numbered 3, wherein the alkali treatment uses an alkali with a pH of 10 to 14. 5) The method according to embodiment numbered 3 or 4, wherein the alkali treatment uses sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide and ammonia water. 6) The method according to any of the preceding numbered embodiments, wherein the treatment in step (b) is carried out at a temperature of 30°C to 180°C. 7) The method according to any of the preceding numbered embodiments, wherein the treatment in step (b) is carried out for 10 minutes to 24 hours. 8) The method according to any of the preceding numbered embodiments, wherein after removing the one or more oligosaccharides, the one or more oligosaccharides and / or the dissolved one or more soluble polysaccharides are subjected to chemical, physical or enzymatic treatment, such as reduction, oxidation, caramelization or Maillard reaction. 9) The method according to any of the preceding numbered embodiments, wherein the dissolved one or more soluble polysaccharides and / or the one or more oligosaccharides are dried and then combined together in step (f). 10) The method according to embodiment numbered 9, wherein the method further comprises the following step: (g) mixing and dissolving the ingredient in a liquid to form a liquid ingredient, wherein the viscosity of the liquid ingredient is 5 to 100,000 cps. 11) The method according to embodiment numbered 10, wherein the concentration of oligosaccharides and polysaccharides in the liquid ingredient is 1 to 200% w / v. 12) The method according to any of the preceding numbered embodiments, wherein the one or more soluble polysaccharides comprise at least one selected from the following: mannan, mixed-link glucan, lignocellulose, hemicellulose, certain cellulose derivatives (such as cellulose acetate, hydroxyethyl cellulose and hydroxymethyl cellulose) and chitosan. 13) The method according to embodiment numbered 12, wherein the hemicellulose comprises xylan and / or mannan.14) A method according to any of the preceding numbered embodiments, wherein the plant biomass comprises sugarcane biomass, corn biomass, wheat biomass, hardwood or softwood. 15) A food, cosmetic or nutritional ingredient obtained by the method according to any of the preceding numbered embodiments. 16) A food, cosmetic or nutritional liquid ingredient comprising at least one oligosaccharide selected from the group consisting of: i) cellooligosaccharides having a degree of polymerization of 2 to 6; ii) xylooligosaccharides having a degree of polymerization of 2 to 12; iii) mannanooligosaccharides having a degree of polymerization of 2 to 12; iv) mixed-link glucan oligosaccharides having a degree of polymerization of 2 to 5; v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 12; and vi) chitosan oligosaccharides having a degree of polymerization of 2 to 12; and at least one polysaccharide selected from the group consisting of: i) xylan; ii) mannan; iii) cellulose derivatives; iv) mixed-link glucan; v) xyloglucan; and vi) chitosan; wherein the liquid ingredient comprises at least 20% dry weight of at least one oligosaccharide and at least 2% dry weight of at least one polysaccharide, and wherein the viscosity of the liquid ingredient is 5 to 100,000 cps, 8,000 to 100,000 cps, 10,000 to 50,000 cps or 15,000 to 25,000 cps. 17) The liquid ingredient according to numbered embodiment 16, wherein the liquid ingredient comprises at least two of the oligosaccharides listed in (i) to (vi). 18) The liquid ingredient according to numbered embodiment 16 or 17, wherein the liquid ingredient comprises at least 20% dry weight of cellooligosaccharides having a degree of polymerization of 2 to 6. 19) The liquid ingredient according to any one of numbered embodiments 16 to 18, wherein the liquid ingredient comprises at least 20% dry weight of xylooligosaccharides having a degree of polymerization of 2 to 12. 20) The liquid ingredient according to any one of numbered embodiments 16 to 19, wherein the liquid ingredient comprises at least 20% dry weight of mannanooligosaccharides having a degree of polymerization of 2 to 12. 21) The liquid ingredient according to any one of numbered embodiments 16 to 20, wherein the liquid ingredient comprises at least 2% dry weight of xylan. 22) The liquid ingredient according to any one of numbered embodiments 16 to 21, wherein the liquid ingredient comprises at least 2% dry weight of mannan. 23) The liquid ingredient according to any one of numbered embodiments 16 to 22, wherein the liquid ingredient comprises at least 2% dry weight of cellulose derivatives. 24) The liquid ingredient according to any one of numbered embodiments 16 to 23, wherein the polysaccharide concentration of the liquid ingredient is 0.1 to 50% (w / v). 25) The liquid ingredient according to any one of numbered embodiments 16 to 24, wherein the oligosaccharide concentration of the liquid ingredient is 1 to 200% (w / v). 26) The liquid ingredient according to any one of numbered embodiments 16 to 25, wherein the polysaccharide and oligosaccharide concentration of the liquid ingredient is 1 - 200% (w / v).27) The liquid composition according to any one of numbered embodiments 16 to 26, wherein the ratio of the amounts of polysaccharides and oligosaccharides contained in the liquid composition is from 1:100 to 1:1. 28) The liquid composition according to any one of numbered embodiments 16 to 27, wherein the ratio of two oligosaccharides contained in the liquid composition is from 1:9 to 9:1. 29) Use of the liquid composition according to any one of numbered embodiments 16 to 28 in food, cosmetics or nutritional products.

[0493] Other exemplary embodiments of pretreating biomass to remove monosaccharides and / or disaccharides

[0494] In some cases, the present disclosure relates to new methods of physically and thermochemically treating plant biomass materials to produce ingredients for food, cosmetics or nutritional products.

[0495] Sugary foods and beverages are an important part of the culture and lifestyle around the world, but the sugars they contain are associated with people's obesity, diabetes, poor dental health and disruptive behavior. For these reasons, consumer preferences have shifted away from sugary foods, and governments are increasingly implementing regulations to encourage the consumption of less sugar.

[0496] Therefore, for decades, the industry has been looking for suitable low-calorie sweeteners to replace sugar in food and beverages. Unfortunately, many sugar substitutes are produced from non-natural resources and often have a slightly bitter or other unpleasant taste accompanying their sweetness, and these two tastes are not appealing to consumers. In addition, while many sweeteners can mimic the sweetness of sugar in food and beverages, few sweeteners can mimic the broad role of sugar in food, such as increasing volume, regulating texture, providing structure, acting as a preservative, and regulating color and flavor through caramelization and Maillard reactions.

[0497] Dietary fiber is an important part of a healthy diet and helps maintain digestive system health and good regulation of the gut microbiota. This fiber contains sugars of different chain lengths and types. In addition to being naturally present in a wide variety of foods, fiber can also be produced separately and added to other foods during manufacturing.

[0498] Biomass is a good source of sugars and can be used to replace sugar and add fiber to food. Compositions made from raw materials have been provided. However, there is still a need to optimize methods for obtaining these sugars from biomass and processing these sugars into compositions that can be used as ingredients for food, cosmetics or nutritional products on a large scale and a commercial scale. The enzymatic decomposition of large amounts of plant biomass can take a relatively long time. In addition, sugars usually need to be produced by controlled decomposition. For this purpose, enzymatic decomposition is required because the product size can be precisely controlled to ensure that no and / or very little monosaccharide is produced.

[0499] The methods provided herein can produce food, cosmetic, or nutritional ingredients from plant biomass starting materials economically and efficiently, which are faster than previously used methods, produce purer final products, and can be used on a large scale and a commercial scale. The method can be carried out by: performing a pre-washing step to remove endogenous monosaccharides and / or disaccharides from the biomass, and employing a thermochemical pretreatment step to ensure controlled decomposition and release of the sugars required for manufacturing the ingredients. In summary, these steps can ensure that little or no monosaccharides are produced and / or generated during the pre-enzymatic processing. This can maximize efficiency and limit the amount of post-reaction purification required.

[0500] Accordingly, in another aspect of the present disclosure, there is provided a method for producing a food, cosmetic, or nutritional ingredient, the ingredient comprising one or more oligosaccharides, wherein the method comprises the following steps:

[0501] a) physical pretreatment of plant biomass comprising monosaccharides and / or disaccharides;

[0502] b) a washing cycle (also referred to herein as an incubation cycle), which comprises the steps of: (i) washing (e.g., incubating) the plant biomass to dissolve at least a portion of the monosaccharides and / or disaccharides, and (ii) removing at least a portion of the monosaccharides and / or disaccharides;

[0503] c) thermochemical pretreatment of the plant biomass;

[0504] d) forming one or more oligosaccharides by an enzymatic reaction, the enzymatic reaction comprising the step of contacting one or more polysaccharide-cleaving enzymes with the plant biomass in solution or suspension;

[0505] e) separating (also referred to herein as enriching or separating) one or more oligosaccharides from the enzymatic reaction mixture and forming an ingredient using the one or more oligosaccharides.

[0506] Preparing food, cosmetic, or nutritional ingredients in the manner provided herein can allow for: efficient use of biomass by combining oligomeric and polymeric materials from the same biomass source, purification, derivatization, or other modification, and controlling the ratio of oligomeric and polymeric materials, which can improve the functional, nutritional, and tolerability properties of the ingredients.

[0507] Steps (a), (b), and (c) of the method of the present disclosure are all steps of performing "pretreatment" on the plant biomass starting material. As used herein, "pretreatment" generally refers to steps performed on plant biomass and then contacting the polysaccharide-cleaving enzyme with the plant biomass.

[0508] Step (a) is a physical pretreatment of plant biomass, the purpose of which can be to physically break down the plant biomass to prepare it for subsequent steps. The physical step can help to accelerate the overall process as it can increase the available surface area of the plant biomass, enabling the chemicals used in subsequent steps to be active on, for example, more plant biomass simultaneously. The physical pretreatment step can include cutting, chopping, milling, ball milling, grinding, trimming, mixing, or a combination thereof of the plant biomass.

[0509] Any material containing suitable polysaccharides can be plant biomass. Since various oligosaccharides are used in the food, cosmetic, and nutraceutical industries, there is no particular limitation on the polysaccharides applicable to this method. Plant biomass suitable for generating the oligosaccharide profiles of the present disclosure can include, for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan), xyloglucan, and mixed-linkage glucan, and / or mannan (such as glucomannan, galactomannan, or galactoglucomannan). However, any plant biomass on which it can act appropriately is envisioned.

[0510] Thus, the plant biomass can be grains, husks, pods, seed coats, and / or other seed materials; seaweeds; corn stover, straw, bagasse, miscanthus, sorghum bagasse, switchgrass, bamboo, and / or other monocotyledonous tissues; water hyacinth, leaf tissue, roots, and / or other plant materials; and / or any combination of suitable plant biomass. In some embodiments, the plant biomass can include sugarcane, corn stover, corn cobs, wheat bran, wheat straw, hardwood, or softwood. In certain embodiments, the plant biomass can include corn cobs.

[0511] Step (b) is a washing cycle (or incubation cycle) pretreatment of the plant biomass that can be carried out after the physical pretreatment step. The purpose of step (b) can be to dissolve and remove monosaccharides and / or disaccharides from the biomass. For example, the monosaccharides and / or disaccharides can include, but are not limited to, free sucrose, maltose, lactose, glucose, fructose, or galactose. The removal of free disaccharides such as sucrose can be meaningful because disaccharides cannot then be easily removed from the oligosaccharide fraction, for example, using filtration methods but usually resulting in an equivalent loss of other disaccharides.

[0512] The washing cycle of step (i) can be carried out within a temperature range, for example, 5 to 150 °C, 10 to 100 °C, or 15 to 50 °C. In some cases, the washing cycle can be carried out at room temperature, for example, about 15 to 25 °C or about 20 to 22 °C. Higher temperatures can cause the monosaccharides and / or disaccharides to dissolve more quickly; however, too high a temperature may be more difficult to achieve in an effective and cost-effective manner and may damage the biomass compounds or dissolve compounds that are not desired to be dissolved in this step.

[0513] The washing cycle of step (i) can occur within a certain time range. For example, a large amount of biomass can be exposed to this step for a longer time, which can be adjusted accordingly. For example, the time can be from 0.5 minutes to 72 hours, from 1 minute to 12 hours, from 5 minutes to 24 hours, or from 10 minutes to 3 hours. In certain embodiments, this step can be carried out batchwise or continuously.

[0514] In some embodiments, the washing cycle of step (i) can include washing the plant biomass in water (i.e., water supplied at a neutral pH of about 7). On the other hand, the washing cycle of step (i) can include heating the plant biomass in water (i.e., water supplied at a neutral pH of about 7). During the washing cycle, the supplied neutral water can become slightly acidic as monosaccharides and / or disaccharides dissolve.

[0515] In certain embodiments, the washing cycle of step (i) can include heating the plant biomass in an alkaline solution having a pH of 7.1 to 14, 7.5 to 12, or 8 to 11. The solution can comprise any one or be suitably composed of the following: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, ammonia water. In various embodiments, the base can be sodium hydroxide. Combinations of the listed bases are also contemplated.

[0516] In various cases, the washing cycle of step (i) can include heating the plant biomass in an acidic solution having a pH of 1 to 6.9, 2 to 6.5, or 4 to 6. The solution can comprise any organic acid or inorganic acid or be suitably composed thereof, such as an acid selected from the following: sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, and oxalic acid. In certain cases, the acid can be sulfuric acid. Combinations of the listed acids are also contemplated.

[0517] In certain cases, some or all of the monosaccharides and / or disaccharides in the plant biomass are dissolved and removed during step (b). Other contaminants, such as other soluble sugars and minerals, can also be removed during this step.

[0518] Step (b) can be repeated to remove monosaccharides and / or disaccharides that were not removed from the plant biomass during the washing cycle. In various cases, step (b) can be performed at least twice, at least three times, at least four times, or at least five times. Step (c) is a thermochemical pretreatment of the plant biomass that can be carried out after the physical washing pretreatment step. As used herein, "thermochemical" generally refers to heating the plant biomass in a chemical substance to a temperature above room temperature (room temperature is, for example, about 15 to 25 °C or about 20 to 22 °C), such as heating in water, an acid, or a base solution. The purpose of the thermochemical step can be to help speed up the overall process because it can chemically modify the chemical composition of the plant biomass. For example, it can break the hydrogen bonds between sugars, making it easier for the enzymes in subsequent steps to break down the sugars.

[0519] The heating can be in a temperature range such as 50 to 150 °C, 60 to 130 °C, 65 to 120 °C, or 70 to 110 °C. Higher temperatures can allow for faster chemical and / or physical modification. However, too high a temperature may be more difficult to achieve in an effective and cost-effective manner.

[0520] The heating can occur within a certain time range. In particular, a large amount of biomass can be exposed to heating for a longer period of time, which can be adjusted accordingly. For example, the heating of the plant biomass can be for 5 minutes to 72 hours, 15 minutes to 24 hours, 30 minutes to 12 hours, or 1 hour to 4 hours.

[0521] In some embodiments, the thermochemical pretreatment can include heating the plant biomass in water (i.e., at a neutral pH of about 7).

[0522] In certain embodiments, the thermochemical treatment can include heating the plant biomass in an alkaline solution having a pH of 7.1 to 14, 9 to 13, or 10 to 13. The solution can contain any one of the following bases or be suitably composed thereof: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and ammonia water. In various examples, the base can be sodium hydroxide. Combinations of the listed bases are also contemplated.

[0523] In some cases, the thermochemical treatment can include heating the plant biomass in an acidic solution having a pH of 1 to 6.9, 2 to 6.5, or 4 to 6. The solution can contain any organic or inorganic acid or be suitably composed thereof, such as an acid selected from the following: sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, and oxalic acid. In certain cases, the acid can be sulfuric acid. Combinations of the listed acids are also contemplated.

[0524] For step (c), multiple different consecutive thermochemical treatment steps are also contemplated. For example, step (c) can be performed at least twice, at least three times, at least four times, or at least five times.

[0525] In some cases, the washing of step (b) can be carried out in water, while the pretreatment of step (c) can be carried out in alkali.

[0526] After the pretreatment step, step (d) can include an enzymatic reaction to form one or more oligosaccharides from plant biomass. The polysaccharides present in the plant biomass can be partially cleaved by enzymes into oligosaccharides (e.g., useful oligosaccharides), leaving partially cleaved or uncleaved polysaccharides, which can include cellulose, xylan (e.g., glucuronoxylan, arabinoxylan, or glucuronoarabinoxylan), mannan (e.g., glucomannan, galactomannan, or galactoglucomannan), mixed-linkage glucan, xyloglucan, chitin, chitosan, or lignocellulose.

[0527] The reaction can be carried out in solution and / or suspension. The reaction can be carried out in a suitable reaction vessel. In some cases, the reaction can be carried out at a temperature or temperature profile suitable for a particular combination of enzyme and plant biomass, the reaction can be allowed to proceed for a certain period of time until the product reaches the desired concentration or until some other requirement is met, and the product is separated or otherwise collected. The period can be from about 1 minute to about 6 days, from about 0.5 days to about 5 days, or from about 16 hours to about 96 hours. Alternatively, the reaction can be allowed to proceed until no further catalysis occurs.

[0528] To ensure optimal contact between the enzyme and the plant biomass, the reaction mixture can be stirred continuously or intermittently. Stirring can take the form of rhythmic movement of the entire reaction vessel, a blower or other stirring device, bubble injection, or any other stirring method.

[0529] The enzymatic reaction can be microbial fermentation. The temperature and reaction time can be suitable for the growth of the microorganism used. The microbial organism can be genetically altered to produce an enzyme suitable for producing the oligosaccharides of the present disclosure. The microorganism can be, for example, a bacterium (e.g., Escherichia coli), or a fungus, such as Saccharomyces cerevisiae or Trichoderma reesei.

[0530] Also embodied in the present disclosure is an expression vector suitable for modifying a subject microorganism such that it produces the enzyme or enzyme mixture of the present disclosure. If desired, the expression vector can be a plasmid or any other nucleic acid capable of inducing enzyme production, and the expression vector can contain one or more of the following regulatory sequences to control the expression of the foreign enzyme: regulatory sequences of heat shock genes, regulatory sequences of virulence genes, and regulatory sequences of sporulation genes.

[0531] The enzymatic reaction can be carried out at a temperature or temperature profile suitable for the enzyme and substrate used. For example, the enzymatic reaction can be carried out at a constant temperature of about 10 °C to about 100 °C, about 20 °C to about 70 °C, or about 30 °C to about 40 °C. If the enzymatic reaction takes the form of microbial fermentation, the temperature can be suitable for this. For example, the enzymatic reaction can include the growth of Escherichia coli and / or the temperature can be constant and about 37 °C.

[0532] The pH of the solution or suspension can affect the activity of the enzyme. Control of the pH can ensure that the enzymatic reaction proceeds at a suitable rate. The enzymatic reactions of the present disclosure can be carried out within a pH range of about 2 to about 10, about 3 to about 8, or about 4 to about 6.

[0533] The enzymatic reaction can be allowed to continue until 5 - 75%, 5 - 70%, 5 - 65%, 5 - 55%, more, or 10 - 50% of the undigested polysaccharide-containing plant biomass remains. This can be monitored or checked by reducing end assays (e.g., anthrone assay) and / or chromatography (e.g., thin layer chromatography and high performance anion exchange chromatography).

[0534] A variety of enzymes can be suitable for the enzymatic reaction of the present method. For example, "lytic polysaccharide monooxygenase" and "LPMO", where the "lytic polysaccharide monooxygenase" and "LPMO" refer to a class of enzymes that can use a copper-containing moiety and an oxygen source (such as dioxygen molecule, peroxide, or any other oxygen source); and a suitable reducing agent to oxidatively cleave polysaccharides. Thus, when using LPMO, the enzymatic reaction can be carried out under aerobic conditions. Suitable reducing agents are not particularly limited, but examples include ascorbic acid, gallic acid, cysteine, NADH, NADPH, pyrogallol, dithiothreitol, cyanoborohydride, borohydride, photosynthetic pigments, lignin, ligninol, and combinations of cellobiose and cellobiose dehydrogenase. A variety of photosynthetic pigments can be used, such as thylakoids and purified fractions or chlorophyllin, and light can be provided. The LPMO can be selected from the following families: AA9, AA10, AA11, AA13, AA14, and AA15. The LPMO can be PaLPMO9E (SEQ ID NO:1), which is an AA9 LPMO originally isolated from the ascomycete fungus (Podospora anserina). The LPMO can be an AA9 LPMO from Trichoderma reesei (SEQ ID NO:23).

[0535] Aerobic conditions can include the addition of oxygen, which can be provided by aerating the substrate mixture with an oxygen-containing gas such as air. Aeration can be carried out by introducing oxygen-containing bubbles into the aqueous substrate mixture through various systems such as air injectors, aeration frits, membrane systems or internal circulation air-lift reactors. The concentration of molecular oxygen in the enzymatic reaction can be from about 4 mg / L to about 14 mg / L.

[0536] Another enzyme that can be used in the method is "cellulase", which has hydrolytic activity towards cellulose, for example, endo-1,4-β-glucanase, cellobiohydrolase and / or β-glucosidase activity. The enzyme is capable of cleaving glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. In this process, cellulose produces products including glucose and cellooligosaccharides. β-Glucanases include enzymes selected from the GH5, GH7 and GH12 enzymes, such as those derived from Aspergillus niger (SEQ ID NO: 12, 13 and 14) and Trichoderma reesei (SEQ ID NO: 24 and 25).

[0537] Another enzyme is "cellobiohydrolase", which has hydrolytic activity towards cellulose and mainly produces cellobiose as a product. Cellobiose is a disaccharide and is a type of cellooligosaccharide. The enzyme is capable of cleaving glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. Cellobiohydrolases can be from the GH6 and GH7 families, such as the Cel6A or Cel7A enzymes from Trichoderma reesei (SEQ ID NO: 10 and 11).

[0538] Another enzyme is "β-glucosidase", which has hydrolytic activity towards cellulose and mainly produces glucose as a product. Such enzymes are capable of cleaving glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. β-Glucosidases can include GH3 β-glucosidases, such as those derived from Trichoderma reesei (SEQ ID NO: 22).

[0539] Another class of enzymes is lichenase, which can be selected from the GH5, GH7, GH8, GH9, GH12, GH16, GH17 or GH26 families. In some cases, the lichenase can be a GH16 enzyme. The GH16 enzyme can be derived from Bacillus subtilis (SEQ ID NO: 2). This enzyme is capable of acting on, for example, mixed-linkage glucans (which are glucans that are a mixture of β-1,3 and β-1,4 linkages), and can cleave them at the β-1,4 glycosidic bond. In cases where the lichenase acts on mixed-linkage glucans, the resulting β-glucans can mainly fall within the size range of about 3 to about 7 residues, and thus they can be used in the food, cosmetic and nutritional industries. Mixed-linkage glucans are abundant in members of the grass and horsetail families, and thus, grassy biomass such as straw has a high level of mixed-linkage glucans and can be effectively acted upon by lichenase.

[0540] Another enzyme is xylanase, which acts on, for example, plant biomass containing a xylan backbone. The xylanase can be, for example, a glucuronoxylanase, an arabinoxylanase or a glucuronarabinoxylanase. This enzyme can be active against a variety of polymers having a xylan backbone, such as glucuronoxylan, arabinoxylan and glucuronarabinoxylan. These polymers are abundant in various plant biomass, for example, both hardwoods and softwoods can contain suitable polysaccharides, where hardwoods typically contain glucuronoxylan and softwoods typically contain glucuronoarabinoxylan. In some embodiments, the xylanase can include GH5 xylanases derived from Clostridium thermocellum (SEQ ID NO: 3) and Saprolegnia ferax (SEQ ID NO: 4), and GH30 xylanases derived from Erwinia chrysanthemi (SEQ ID NO: 5), Bacillus subtilis (SEQ ID NO: 6), Bacteroides ovatus (SEQ ID NO: 7) and Trichoderma reesei (SEQ ID NO: 15).

[0541] Other enzymes useful in the present disclosure can include xyloglucanase and xyloglucan endo-glucanases (XEGs), which are produced by a variety of organisms, including plant pathogenic microorganisms. Xyloglucanase and xyloglucan endo-glucanases are capable of acting on xyloglucan, which is a hemicellulose β-1,4 glucan chain abundant in the primary cell walls of higher plants, which is modified with xylose and some of the xylose residues are further modified with other residues such as galactose. When a suitable xyloglucanase or XEG acts on xyloglucan, the product contains xyloglucan oligosaccharides of a length of the backbone that can be used in the food, cosmetic and nutritional industries. The xyloglucanase can include a GH5 xyloglucanase derived from Bacteroides ovatus (SEQ ID NO: 8) and a GH74 xyloglucanase derived from Trichoderma reesei.

[0542] Since any given natural plant biomass may contain a mixture of different polysaccharides, sometimes a mixture of different enzymes may be beneficial. Such a mixture can contain one or more of any other enzyme. For example, such a mixture can contain LPMO and endoglucanase, xylanase and lichenase, cellobiohydrolase and mannanase, or endoglucanase and cellobiohydrolase, where the enzyme partners are present in a molar ratio of 1:100 to 100:1.

[0543] In some cases, one or more enzymes can be a mixture of different enzymes, such as a crude enzyme preparation or a semi-crude enzyme preparation. As used herein, the term "crude enzyme preparation" generally refers to a soluble preparation extracted from a microbial fermentate that undergoes minimal processing after extraction. For example, typically the preparation can only undergo filtration to remove insoluble components. As used herein, the term "semi-crude enzyme preparation" generally refers to a soluble preparation extracted from a microbial fermentate that undergoes some processing after extraction, such as the preparation can undergo filtration to remove insoluble components, increase enzyme concentration, and / or nanofiltration to remove small molecular weight compounds.

[0544] In some cases, the crude enzyme preparation or semi-crude enzyme preparation can be from bacteria or fungi. For example, the preparation can be from fungi, such as filamentous cellulolytic fungi, such as from species of Trichoderma or Aspergillus. The enzyme can be a crude enzyme preparation or a semi-crude enzyme preparation from a strain of Trichoderma reesei.

[0545] In step (e), the oligosaccharides can be separated from the enzymatic reaction mixture in a variety of ways. It can be separated based on solubility such that only a composition of soluble sugars is extracted for further processing, and / or by chromatographic separation to produce a composition with a narrower band of oligosaccharide chain lengths. Separation can be, for example, based on precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration. In the case of separation based on solubility, the sugar profile present in the separated composition can depend on the initial enzymatic reaction, as the solubility of different sugars decreases at different rates with length.

[0546] Also contemplated within the scope of the present disclosure is the further processing of all or part of the resulting oligosaccharides to produce further products, which are then incorporated into food, cosmetics, or nutraceuticals. The further processing can include any chemical, physical, or enzymatic steps, such as reduction, such as reductive amination carried out in a suitable place; oxidation, caramelization, modification with Schiff bases or by the Maillard reaction or by any combination of these steps, and can provide different products with improved properties for the desired purpose. For example, caramelization characteristics, calorific value, flavor, and color can be modified. The oligosaccharides can also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration.

[0547] The composition formed in step (e) can contain various oligosaccharides and can contain different amounts depending on the desired properties. In various cases, the composition can contain at least 20% dry weight or at least 30% dry weight of cellooligosaccharides with a degree of polymerization of 2 to 6, the composition can contain at least 20% dry weight or at least 30% dry weight of xylooligosaccharides with a degree of polymerization of 2 to 12, the composition can contain at least 20% dry weight or at least 30% dry weight of mixed-linkage glucan oligosaccharides with a degree of polymerization of 2 to 5, the composition can contain at least 20% dry weight or at least 30% dry weight of mannan oligosaccharides with a degree of polymerization of 2 to 12, the composition can contain at least 20% dry weight or at least 30% dry weight of xyloglucan oligosaccharides with a degree of polymerization of 4 to 12, and / or the composition can contain at least 20% dry weight or at least 30% dry weight of chitooligosaccharides with a degree of polymerization of 2 to 12. In some embodiments, the composition can contain up to 100% dry weight of the above oligosaccharides and the polysaccharides described herein, so the above embodiments in which the oligosaccharides are present at at least 20% dry weight do not include all six types of oligosaccharides.

[0548] In some cases, the composition can contain sugars present at at least 50%, 60%, 70%, 80%, 90%, 95%, 99% or 99.5% dry weight. The composition can consist essentially of sugars. For example, the composition can have other substances at less than 0.5% dry weight or less than 0.3% dry weight, such as 0.1% dry weight.

[0549] In various cases, the composition can contain at least two oligosaccharides. The amount of each oligosaccharide can vary according to the desired properties of the resulting food, cosmetic or nutritional product. Two oligosaccharides can be present in a ratio of 1:9 to 9:1 or 1:2 to 2:1. Further, the composition can contain three oligosaccharides, four oligosaccharides, five oligosaccharides or six oligosaccharides.

[0550] In some embodiments, the composition can contain cellooligosaccharides, such as a combination of cellooligosaccharides and xylooligosaccharides. In certain embodiments, the composition can contain a combination of cellooligosaccharides and mannan oligosaccharides.

[0551] The produced composition can be used in the applications of oligosaccharides, sugars, bulking sweeteners, low-intensity sweeteners or other related food ingredients for conventional use. For example, as a sweetener, bulking agent, added dietary fiber or humectant. Of particular note is the use of reduced sucrose in food. The composition can be incorporated into cakes, bread or other baked goods; put into chocolates or other confections, such as toffee, fudge, meringue, jam, jelly or caramel; or incorporated into beverages, for example to provide good flavor or color characteristics or to increase the dietary fiber content. In some cases, the composition can be incorporated into animal feed, for example as a separated component or by directly using the enzymatic reaction mixture as a raw material.

[0552] The compositions or ingredients as described herein can be used to modify one or more properties of the final product. Such properties include, but are not limited to, sweetness, texture, mouthfeel, adhesiveness, glossiness, smoothness, moistness, viscosity, color, hygroscopicity, flavor, fluffiness, water retention, caramelization, surface texture, crystallization, structural properties, calorie reduction, glycemic index reduction, glycemic load reduction, fiber increase, sugar reduction, and solubility. These can be possible improvements over the currently used different types of sugars, sugar substitutes, and / or other such compounds.

[0553] In the cosmetic industry, the ingredient can improve texture and moisturization, act as an ultraviolet light absorbing molecule, maintain the structure of gels or creams, and / or act as a fluffiness agent. Additionally, the ingredient can be used in nutritional compositions because the dietary fiber they provide has been shown to promote digestive health, regulate gut flora well, and provide other benefits to health. In such cases, the ingredients herein can also be used as ingredients in probiotic beverages or other prebiotic or probiotic formulations.

[0554] The detailed description will be further supplemented with reference to the following numbered embodiments. 1) A method for producing a food, cosmetic, or nutritional ingredient, the ingredient comprising one or more oligosaccharides, wherein the method comprises the following steps: a) physical pretreatment of plant biomass comprising monosaccharides and / or disaccharides; b) a washing cycle, which comprises the steps of: (i) washing the plant biomass to dissolve at least a portion of the monosaccharides and / or disaccharides, and (ii) removing at least a portion of the monosaccharides and / or disaccharides; c) thermochemical pretreatment of the plant biomass; d) formation of one or more oligosaccharides by an enzymatic reaction, the enzymatic reaction comprising the step of contacting one or more polysaccharide-cleaving enzymes with the plant biomass in solution or suspension; e) separating the one or more oligosaccharides from the enzymatic reaction mixture and using the one or more oligosaccharides to form an ingredient. 2) The method according to numbered embodiment 1, wherein the physical pretreatment step comprises cutting, chopping, grinding, ball milling, polishing, trimming, or mixing the plant biomass. 3) The method according to numbered embodiment 1 or 2, wherein the washing cycle of step (i) is carried out in water, acid, or base. 4) The method according to any of the preceding numbered embodiments, wherein the washing cycle of step (i) is carried out at a temperature of 5 to 150 °C, 10 to 100 °C, or 15 to 50 °C. 5) The method according to any of the preceding numbered embodiments, wherein the washing cycle of step (i) is carried out for a time ranging from 0.5 minutes to 72 hours, 1 minute to 12 hours, 5 minutes to 24 hours, or 10 minutes to 3 hours. 6) The method according to any of the preceding numbered embodiments, wherein the thermochemical pretreatment comprises heating the plant biomass in a solution of water, acid, or base. 7) The method according to numbered embodiment 6, wherein the heating of the plant biomass is carried out at a temperature of 50 to 150 °C, 60 to 130 °C, 65 to 120 °C, or 70 to 110 °C. 8) The method according to numbered embodiment 6 or 7, wherein the heating of the plant biomass is carried out for 5 minutes to 72 hours, 15 minutes to 24 hours, 30 minutes to 12 hours, or 1 hour to 4 hours. 9) The method according to any of numbered embodiments 6 to 8, wherein the pH of the solution is 7.1 to 14, 7.5 to 12, or 8 to 11. 10) The method according to numbered embodiment 9, wherein the solution comprises sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, ammonia water, ammonium sulfate, or ammonium hydroxide. 11) The method according to any of numbered embodiments 6 to 8, wherein the pH of the solution is 1 to 6.9, 2 to 6.5, or 4 to 6. 12) The method according to numbered embodiment 11, wherein the solution comprises sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, or oxalic acid. 13) The method according to any of numbered embodiments 1 to 12, wherein the plant biomass is sugarcane, corn stover, corncob, wheat bran, wheat straw, hardwood, or softwood.14) The method according to any one of numbered embodiments 1 to 13, wherein the plant biomass comprises cellulose, chitin, chitosan, xylan, xyloglucan, mixed-linkage glucan, mannan or lignocellulose. 15) The method according to any one of numbered embodiments 1 to 14, wherein the one or more polysaccharide-cleaving enzymes are one of the following: cellulase, xylanase, xyloglucanase, endoglucanase, cellobiohydrolase, mannanase, lichenase or lytic polysaccharide monooxygenase (LPMO), such as selected from AA9, AA10, AA11, AA13, AA14 and AA15. 16) The method according to any one of numbered embodiments 1 to 15, wherein the one or more polysaccharide-cleaving enzymes are prepared from Trichoderma reesei fungi. 17) The method according to any one of numbered embodiments 1 to 16, wherein the one or more oligosaccharides comprise β-glucan, cellooligosaccharide, MLG oligosaccharide, mannan oligosaccharide or xylooligosaccharide. 18) The method according to any one of numbered embodiments 1 to 17, wherein the polysaccharide-cleaving enzyme is operably linked to a catalytic module or a non-catalytic module, for example wherein the polysaccharide-cleaving enzyme is operably linked to a non-catalytic module and the non-catalytic module is a carbohydrate-binding module. 19) The method according to any one of numbered embodiments 1 to 18, wherein after separating the one or more oligosaccharides, the one or more oligosaccharides are subjected to chemical, physical or enzymatic treatment, such as reduction, oxidation, caramelization or Maillard reaction.

[0555] The detailed description will be further supplemented with reference to the following numbered embodiments. 1) A method for producing a composition for human consumption, the method comprising: (a) physically treating plant biomass; (b) subjecting the physically treated plant biomass to a incubation cycle, the incubation cycle comprising: (i) incubating the physically treated plant biomass in an incubation solution having a pH of 6.6 to 7.4 to dissolve monosaccharides and / or disaccharides from the physically treated plant biomass; and (ii) removing a portion of the dissolved monosaccharides and / or disaccharides from the incubation solution; (c) thermochemically treating the incubated plant biomass under one of the following conditions: (i) an acidic solution having a pH of 2 to 6.5, or (ii) a basic solution having a pH of 7.5 to 12; (d) contacting one or more polysaccharide-cleaving enzymes with the thermochemically treated plant biomass in a solution or suspension to form one or more oligosaccharides; and (e) enriching the solution or suspension to increase the concentration of one or more oligosaccharides to form the composition. 2) The method according to numbered embodiment 1, further comprising removing at least a portion of the monosaccharides and / or disaccharides from the incubation solution in step (b)(ii). 3) The method according to any one of the preceding numbered embodiments, wherein the thermochemically treated plant biomass does not contain or substantially does not contain monosaccharides. 4) The method according to any of the preceding numbered embodiments, further comprising purifying one or more oligosaccharides from the solution or suspension. 5) The method according to any of the preceding numbered embodiments, further comprising repeating step (b). 6) The method according to numbered embodiment 5, wherein step (b) is carried out two, three, four, or five times.

[0556] 7) The method according to any of the preceding numbered embodiments, further comprising repeating step (c).

[0557] 8) The method according to numbered embodiment 7, wherein step (c) is carried out two, three, four or five times. 9) The method according to any of the preceding numbered embodiments, further comprising discarding a portion of the dissolved monosaccharides and / or disaccharides removed in step (b). 10) The method according to any of the preceding numbered embodiments, wherein a portion of the dissolved monosaccharides and / or disaccharides removed in step (b) is not combined with a portion of one or more oligosaccharides of step (e) to form a composition. 11) The method according to any of the preceding numbered embodiments, wherein the composition is substantially free of monosaccharides. 12) The method according to any of the preceding numbered embodiments, wherein the composition is substantially free of disaccharides. 13) The method according to any of the preceding numbered embodiments, wherein the one or more oligosaccharides comprise at least one of the following: i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2 to 12; iii) arabinoxylooligosaccharides with a DP of 3 to 15; iv) mannanooligosaccharides with a DP of 2 to 12; v) mixed-linkage glucan oligosaccharides with a DP of 2 to 5; vi) xyloglucan oligosaccharides with a DP of 4 to 12; or vii) chitooligosaccharides with a DP of 2 to 12. 14) The method according to numbered embodiment 13, wherein the composition comprises at least two of the oligosaccharides listed in (i) to (vii). 15) The method according to numbered embodiment 14, wherein the composition comprises at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other. 16) The method according to any of the preceding numbered embodiments, wherein the monosaccharides and / or disaccharides comprise at least one of sucrose, glucose, maltose, lactose, glucose, fructose or galactose. 17) The method according to any of the preceding numbered embodiments, wherein the physical treatment of step (a) comprises at least one of cutting, chopping, grinding, ball milling, milling, trimming or mixing the plant biomass. 18) The method according to any of the preceding numbered embodiments, wherein the incubation of step (b) is carried out in an incubation solution containing water. 19) The method according to any of the preceding numbered embodiments, wherein the incubation of step (b) is carried out at a temperature of 15°C to 95°C. 20) The method according to any of the preceding numbered embodiments, wherein the incubation of step (b) is carried out for 15 minutes to 1 hour. 21) The method according to any of the preceding numbered embodiments, wherein the thermochemical treatment of step (c) comprises heating the physically treated plant biomass in an acidic solution or a basic solution. 22) The method according to numbered embodiment 21, wherein the heating is carried out at a temperature of 50°C to 150°C. 23) The method according to numbered embodiment 21 or 22, wherein the heating is carried out for 30 minutes to 4 hours. 24) The method according to any of the preceding numbered embodiments, wherein the incubated plant biomass is subjected to thermochemical treatment in a basic solution with a pH of 8 to 11.25) The method according to embodiment 24, wherein the alkaline solution comprises at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, ammonia water, ammonium sulfate, or ammonium hydroxide. 26) The method according to any of the preceding numbered embodiments, wherein the thermochemically treated plant biomass is carried out in an acidic solution having a pH of 4 to 6. 27) The method according to embodiment 26, wherein the acidic solution comprises at least one of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, or oxalic acid. 28) The method according to any of the preceding numbered embodiments, wherein the plant biomass comprises at least one of sugarcane, corn stover, corn cob, wheat bran, wheat straw, hardwood, or softwood. 29) The method according to any of the preceding numbered embodiments, wherein the plant biomass comprises at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed-link glucan, mannan, or lignocellulose. 30) The method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide-cleaving enzymes comprise at least one of cellulase, xylanase, xyloglucanase, endoglucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase (LPMO). 31) The method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide-cleaving enzymes comprise at least one of AA9, AA10, AA11, AA13, AA14, or AA15. 32) The method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide-cleaving enzymes are prepared from Trichoderma reesei fungi. 33) The method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide-cleaving enzymes are operably linked to a catalytic module. 34) The method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide-cleaving enzymes are operably linked to a non-catalytic module. 35) The method according to embodiment 34, wherein the non-catalytic module is a carbohydrate-binding module. 36) A method for producing a component for human consumption, the method comprising: (a) pretreating plant biomass, wherein the pretreatment comprises: (i) physically treating the plant biomass; (ii) incubating the plant biomass in an incubation solution having a pH of 6.6 to 7.4 to dissolve a portion of the monosaccharides and / or disaccharides and removing a portion of the dissolved monosaccharides and / or disaccharides; and (iii) thermochemically treating the plant biomass in one of: (i) an acidic solution having a pH of 2 to 6.5 or (ii) an alkaline solution having a pH of 7.5 to 12; (b) contacting one or more polysaccharide-cleaving enzymes with the pretreated plant biomass in a solution or suspension to form one or more oligosaccharides; (c) separating a portion of the one or more oligosaccharides to form the component.

[0558] Example

[0559] The following illustrative embodiments represent implementations of the compositions and methods described herein and are not meant to be limiting in any way.

[0560] Example 1 - Exemplary Process

[0561] The following steps can be carried out to produce the ingredients provided herein:

[0562] 1. Physical pretreatment of plant biomass: 100 g of ground corn cobs were mixed with water to a solid concentration of 10% (w / w) and mixed for 60 minutes at room temperature. Mixing was stopped at the end of 60 minutes and the liquid was filtered off while retaining the solid.

[0563] 2. Resuspend the solids in water at a solid concentration of 10% (w / w). Start mixing, heat to 95 °C, and mix for 60 minutes at 95 °C.

[0564] 3. At the end of the 60-minute heating, 6 g of sodium hydroxide (0.2 - 3% of the weight of the corn cobs) was added and stirring was continued. Heat to 95 °C and mix for 60 minutes to break down the hemicellulose present in the corn cobs. At the end of 60 minutes, heating was stopped and the mixture was cooled to 50 °C.

[0565] 4. Hydrolysis: Cellulolytic enzymes (e.g., from Trichoderma reesei) were added and incubated at 50 °C at pH 5.5 (adjusted with 1 mol / L sulfuric acid and / or 1 mol / L sodium hydroxide) for 72 hours.

[0566] 5. Separation of biomass: At the end of hydrolysis, the liquid was separated from the product by a solid-liquid separator.

[0567] 6. Separation of enzymes after enzymatic hydrolysis: The liquid fraction in the slurry contains enzymes, oligosaccharides, water, and salts that need to be separated. A 10 kDa hollow fiber membrane was used to separate the enzyme protein and other macromolecules.

[0568] 7. Removal of salts using an ion exchange column at ≤ 45 °C.

[0569] a. Cation column: Strong acid cation exchange resin, cross-linked polystyrene matrix, sulfonate functional group, Na+ counterion.

[0570] b. Anion column: Macroporous, weak base anion exchange resin, cross-linked polystyrene matrix, dimethyl tertiary amine functional group, OH- counterion.

[0571] 8. Oligosaccharide concentration: The desired oligosaccharides were selectively concentrated at room temperature by nanofiltration.

[0572] 9. Concentration: The liquid was concentrated to 40 - 75% at 60 - 80 °C.

[0573] 10. Spray drying: Spray drying, where the inlet temperature is 130 - 160 °C and the outlet temperature is 65 - 85 °C.

[0574] Example 2 - Removal of soluble sugars through a washing cycle (i.e., incubation cycle)

[0575] The following steps were carried out to remove soluble sugars from plant biomass:

[0576] 1. Water was added to 100 mg of four types of plant biomass (i.e., sugarcane, wheat, corncob, and willow) to a concentration of 10% (w / v), and the suspension was incubated at 45 °C for 30 minutes. Then the suspension was centrifuged and the supernatant was removed.

[0577] 2. Step 1 was repeated 5 times.

[0578] 3. 2.5 μl of each supernatant fraction was analyzed by thin layer chromatography (TLC).

[0579] Figure 1 The thin layer chromatography showed the presence of soluble sugars washed out from the four types of plant biomass in five consecutive washing cycles or incubation cycles (1, 2, 3, 4, and 5). Unwanted monosaccharides and disaccharides (such as glucose, sucrose, and maltose) were marked with arrows. The results of TLC showed that for all plant biomass, the supernatant removed after the first washing cycle had abundant monosaccharides and disaccharides present in it. Thus, the washing cycle successfully removed monosaccharides and disaccharides from the plant biomass. The supernatants from subsequent washing cycles had significantly fewer monosaccharides and disaccharides present in them (if any), indicating that the plant biomass had minimal monosaccharides and disaccharides remaining in it after the washing cycle as provided herein.

[0580] Example 3 - Absence of washed soluble sugars in enzymatic hydrolysis

[0581] The following steps were carried out to show the absence of soluble sugars in enzymatic hydrolysis:

[0582] 1. The washed corncob and willow plant biomass from Example 2 were each incubated in 1% (w / v) NaOH at 99 °C for 30 minutes and then cooled. 100 mg of unwashed corncob and willow biomass were also each incubated in 1% (w / v) NaOH at 99 °C for 30 minutes and then cooled.

[0583] 2. 150 μl of each resulting suspension was each mixed with 150 μl of 1 M ammonium acetate (pH 5.5) and 150 μl of an enzyme composition containing β - xylanase and cellobiase. Then the suspension was incubated at 50 °C for 16 hours to allow the enzymes to react with the plant biomass.

[0584] 3. Take 2.5 μl of each supernatant and analyze it by TLC.

[0585] Figure 2 Thin layer chromatography of shows the enzymatic digestion products of four types of biomass that have been washed (+) or unwashed (-) in Example 2. The results show that after enzymatic digestion, the final products include glucose, sucrose, and maltose for unwashed corncob and willow plant biomass. However, the final products after enzymatic digestion do not include glucose, sucrose, and maltose for washed corncob and willow plant biomass.

[0586] Example 4 - Adding polymers to oligosaccharide solutions can cause them to dry into a hard glaze

[0587] The following steps were performed to show that adding polymers to oligosaccharide solutions can cause them to dry into a hard glaze:

[0588] 1. Pipette 100 μl of 10 - 320 mM (10, 20, 40, 80, 160, and 320 mM) cellobiose ± 1% w / v birchwood xylan onto a glass plate.

[0589] 2. Dry the samples at 37 °C.

[0590] 3. Scratch the samples with a knife to test whether the components sink into the solid glaze. Using cellobiose alone, no firm glaze was formed, and the dried powder easily cracked when pressure was applied with the knife. In contrast, compositions with 1% w / v xylan and 80 mM or less of cellobiose were dried to form a solid, off - white, translucent surface that was strong enough to leave a mark when scratched with a knife but did not crack. When the cellobiose concentration was 160 mM or higher (5.5% w / v, or 550% w / w compared to xylan), the morphology of the glaze reverted to the state without the presence of xylan. That is, no firm glaze was formed, and the dried powder easily cracked when pressure was applied with the knife (see Figure 3 ).

[0591] Example 5 - Verification of a composition containing two oligosaccharides and a polysaccharide in food

[0592] The following steps were performed to demonstrate a composition containing two oligosaccharides and a polysaccharide in food:

[0593] 1. Dissolve 4 g of birchwood xylan in 75 ml of water and boil.

[0594] 2. Add 12 g of cellobiose and 24 g of xylo - oligosaccharides (predominant degree of polymerization (DP) of 2 - 6) in 3 g increments and dissolve while boiling.

[0595] 3. Reduce the mixture to 50 ml by heating to form a concentrated solution, which has the consistency and appearance of cloudy honey but a lower sweetness.

[0596] 4. Mix 10 mL of the mixture with 12 g of oats to make a pancake bar / cereal bar mixture. Additionally, mix 10 mL of the mixture with 6 g of fruit and 6 g of nuts to make a cereal bar mixture.

[0597] 5. Bake the samples at 100 °C for 10 minutes, then let them cool and dry overnight.

[0598] As Figure 4 shown, in small figure A (pancake bar / cereal bar) and small figure B (fruit bar and nut bar), the produced pancake and fruit and nut bars have an ideal texture and consistency consistent with pancake bars / cereal bars and fruit and nut bars made from known syrups commonly used in baking.

[0599] For the pancake bar / cereal bar, the thick solution from step 3 helps bind the mixture in step 4 together. The combined effect of the ingredients and the oats' properties produces a granular surface texture consistent with pancake bars / cereal bars produced using conventional sugar. This ingredient produces a firm, chewy, moist, and sticky texture, resulting in a mouthfeel that matches what is desired for these types of foods, but this mouthfeel does not exist in oats alone. The product has a slightly sweet taste and no bitter or off-flavors, which may be characteristic of high-intensity sweeteners.

[0600] For the fruit bar and nut bar, the thick solution from step 3 helps bind the mixture in step 4 together. It also adds a smooth, shiny pulp surface, which is a core part of the aesthetic quality of such foods, and this pulp surface would not be produced if the ingredients consisted only of oligosaccharides (i.e., oligosaccharides without polysaccharides). This ingredient produces a firm, chewy, moist, and sticky texture, resulting in a mouthfeel that matches what is desired for these types of foods, but this mouthfeel does not exist in either nuts or fruit alone. The product has a slightly sweet taste and no bitter or off-flavors, which may be characteristic of high-intensity sweeteners.

[0601] Example 6 - Process for Preparing Ingredients

[0602] The following steps can be performed to prepare the ingredients provided herein:

[0603] 1. At 90 °C, heat 15% w / v of corn cobs (ground through a 1 mm aperture filter) in 1% w / v NaOH for 1 hour to dissolve a portion of the polymer components of the biomass.

[0604] 2. Adjust the pH to 5.5 with sulfuric acid.

[0605] 3. Extract the volume of the reaction liquid components that only contain 15% of the total reaction volume ("soluble polymer"); retain the remaining 85% volume of the reaction, including all insoluble biomass fractions ("remaining biomass").

[0606] 4. Add a cellulase mixture (e.g., an enzyme mixture from Trichoderma reesei, including cellulase, xylanase, arabinofuranosidase, LPMO, etc.) to the remaining biomass to 0.5% w / v and incubate at 50 °C for 24 hours.

[0607] 5. Separate the soluble oligomeric reaction products from the undigested insoluble polymers by filtration.

[0608] 6. Purify the oligomeric reaction products by sequentially using microfiltration, ultrafiltration, and ion exchange chromatography (e.g., cross-flow filtration on a ceramic membrane; the filtration can be carried out at a maximum feed pressure of 3 bar, using an Inside Céram candle filter with a 0.45 μm cut-off provided by TAMI Industries (TiO2, x L1178mm HD 6mm, 8 channels per membrane)).

[0609] 7. Purify the soluble polymer by using ultrafiltration (e.g., a 10 kDa spiral wound membrane (Snyder ST-2B-6338, PES, feed spacer thickness 31 mm) operating on an Alfa Laval ultrafiltration unit).

[0610] 8. Recombine the solutions formed in steps 6 and 7 and further purify and concentrate by using nanofiltration to form the composition.

[0611] Example 7 - Viscosity measurement of different solutions

[0612] Perform the following steps to measure the viscosity of different solutions:

[0613] 1. Three sugar solutions containing cellobiose (Cell 2 ), xylooligosaccharides (XOS) mainly with DP-6, and polymeric beechwood xylan (BWX) are made by boiling the sugars in water. The final concentrations are:

[0614] a. Sample 1: 0.33 g / ml Cell2, 0.66 g / ml XOS, 0.13 g / ml BWX;

[0615] b. Sample 2: 0.17 g / ml Cell2, 0.33 g / ml XOS, 0.07 g / ml BWX;

[0616] c. Sample 3: 0.54 g / ml Cell 2 , 0.52 g / ml XOS, 0.07 g / ml BWX.

[0617] 2. The samples were tested using a Brookfield HDB VE rotational viscometer using standard test procedures. 400 mL of the sample was taken in a tall beaker to ensure that no container effects occurred. The instrument was operated according to the manufacturer's instructions, with the following ranges: rotational viscometry using spindle code 61, spindle speed 100 rpm, at 22 °C.

[0618] Sample number Viscosity (cps) 1 393 2 13 3 26

[0619] The consistency of Sample 1 was similar to thick honey that needed to be mixed to dilute into an aqueous solution. In contrast, Samples 2 and 3 were more fluid and could be easily mixed into an aqueous solution. The results showed that the effect of the polysaccharide concentration on the viscosity of the composition was greater than the total concentration of oligosaccharides and polysaccharides. Samples 2 and 3 had the same polysaccharide concentration, but the total oligosaccharide and polysaccharide concentration of Sample 3 was twice that of Sample 2. The viscosity of Sample 3 was twice the viscosity of Sample 2, conforming to the linear relationship between the total concentration and viscosity. However, as the polysaccharide concentration increased, the viscosity value increased exponentially. The polysaccharide concentration of Sample 1 was twice that of Sample 3, and their total oligosaccharide and polysaccharide concentrations were the same, but the viscosity of Sample 1 was fifteen (15) times greater than that of Sample 3.

[0620] Example 8 - Preparation of a water-soluble liquid product / component

[0621] The following steps were performed to produce a water-soluble liquid product / component (Sample 4):

[0622] 1. 100 g of ground corncobs was heated in 1 L of deionized water containing 2.5 g of sodium chlorite at 80 °C for 1.5 hours with continuous stirring. The residual volume was reconstituted to 900 mL by adding 200 mL of deionized water containing an additional 5 g of sodium chlorite, and stirring and heating were continued at 80 °C for 1 hour.

[0623] 2. The solution was filtered under vacuum through a 2 mm pore size ceramic filter funnel until the filtrate was clear.

[0624] 3. The retained solids were incubated in 1 L of 0.5 M sodium hydroxide with 0.1% (w / v) sodium borohydride at 50 °C with shaking at 115 rpm for 17 hours.

[0625] 4. Then the pH was adjusted to 7 with concentrated sulfuric acid and dialyzed against tap water in a 12,000 Dalton cut-off dialysis tube for 24 hours.

[0626] 5. The contents of the dialysis tube were transferred to a 2-L beaker and the insoluble fraction was allowed to sediment by gravity.

[0627] 6. Decant the supernatant twice and evaporate and concentrate it to a volume of 120 mL at 80 °C.

[0628] 7. After adding 3 volumes of ethanol, precipitate the water-soluble polymer by centrifugation. Discard the resulting supernatant and air-dry the precipitate to a constant weight at room temperature.

[0629] 8. Add the oligosaccharide to the final 10% w / w cellobiose, 75% w / w xylo-oligosaccharide, 15% of the extracted water-soluble polymer and mix to homogeneity in a Waring Xtreme blender at the lowest power setting. Recover 94 g of solid from the blender.

[0630] 9. Unexpectedly, all 94 g of the solid dissolved in 60 mL of water at 50 °C with gentle constant stirring (about 100 rpm), indicating a solubility greater than 150 g / 100 g.

[0631] The samples generated (e.g., at steps 8 and 9 above) are designated as Sample 4.

[0632] Example 9 - Physicochemical properties of the water-soluble liquid product of Example 8

[0633] Flow characteristics: The flow characteristics of the water-soluble liquid product (Sample 4) according to the present disclosure described in Example 8 are detailed in Table 1 together with comparative compositions of water, 20% w / v glucose, 40% w / v glucose, 60% w / v glucose, 80% w / v glucose and ≥99% glycerol (supplied by Fisher G / 0650 / 17). Glucose solutions were prepared by weighing 6 g, 12 g, 18 g and 24 g of D-glucose respectively and making up to 30 mL with 90 °C water. The flow characteristics were measured by timing the flow rate of 5 mL and, where appropriate, 20 mL of liquid from a vertically placed syringe (BDP lastipak 300613) filled with 20 mL of the test liquid under gravity at room temperature.

[0634] Table 1: Flow characteristics

[0635] Sample Flow time for 5 mL (seconds) Flow time for 20 mL (seconds) Water 2 13 20% w / v glucose 2.3 13.5 40% w / v glucose 2.5 15 60% w / v glucose 3 20 80% w / v glucose 5 38 ≥99% glycerol 256 Not measured Sample 4 237 Not measured

[0636] The increased time required for the sample to flow out of the bottom of the syringe (i.e., lower flow rate) is related to an increase in the viscosity of the sample. The lower the flow rate, the more syrupy / viscous the liquid sample. The measured flow rate of Sample 4 is similar to that of glycerol and lower than all the glucose solutions tested. This property of Sample 4 makes it more suitable as a binder in foods (such as cereal bars) than glucose solutions and water and provides sweetness to the product.

[0637] Color: The color of Sample 4 corresponds to No. 30 in the Standard Reference Method (SRM), which is a recommended method by the American Society of Brewing Chemists (ASBC Methods of Analysis, Beer 10. Spectrophotometric Color Method Approved 1958, rev. 2015. American Society of Brewing Chemists, St. Paul, MN, U.S.A). Briefly, the absorbance of the sample was measured in a cell with a path length of 1 cm at a wavelength of 430 nm. The resulting absorbance value was multiplied by 12.7 to yield the color value. Due to the turbidity of Sample 4, the absorbance could not be measured and was evaluated by optical comparison with SRM No. 30 (dark red / brown).

[0638] Anion exchange chromatography: Sample 4 was analyzed by high-performance anion exchange chromatography (HPAEC) using a Thermo Fisher Scientific DIONEX ICS-6000 system equipped with a CarboPac PA200 analytical column (3 x 250 mm) and a CarboPac PA200G guard column (3 x 50 mm), as well as a Dionex ED electrochemical detector. Data was acquired using Chromeleon 7 software.

[0639] Eluents A (ultrapure water), B (250 mM NaOH), and C (250 mM NaOH + 1 M sodium acetate) were used to prepare the mobile phase with the gradient characteristics shown in Table 2.

[0640] Table 2: Gradient characteristics

[0641]

[0642] The sample for analysis was prepared by diluting Sample 4 100-fold and passing it through a 0.45 μm syringe filter, with an injection volume of 10 μl for the analyte.

[0643] HPAEC analysis (see Figure 5)It was confirmed that Sample 4 is a mixture of monosaccharides, disaccharides, and other oligosaccharides composed of glucose and xylose. This is in contrast to syrups commonly used in the food industry (such as corn syrup and high fructose corn syrup, which mainly contain monosaccharides of glucose and fructose). Therefore, compared with corn syrup and / or high fructose corn syrup, the product in Sample 4 is expected to have fewer calories, a lower glycemic index, and contain fiber when used in food.

[0644] Example 10 - Cold-Pressed Fruit and Grain Bar

[0645] The cold-pressed fruit and grain bar is prepared as follows:

[0646] 1. Heat 120 g of Sample 4 from Example 8 together with 30 g of coconut oil, and add one-quarter (1 / 4) teaspoon of cinnamon.

[0647] 2. After foaming, remove the mixture from the heat source, and add 40 g of oats, 40 g of dried dates, 10 g of puffed rice, and 10 g of seeds.

[0648] 3. Mix the ingredients well until all ingredients are coated, then transfer the mixture to a freezer bag and place it in the freezer. Roll the contents of the bag to a thickness of 7 - 10 mm, and freeze overnight at 4°C, then cut into rectangles.

[0649] The resulting product (as Figure 6 shown) is a chewy, sticky grain bar that solidifies loosely and contains oats and puffed rice as well as a perceivable sweetness delivered by Sample 4 and chopped dates.

[0650] Example 11 - Producing Ingredients in a Large-Scale Manufacturing Process

[0651] The following steps can be used to produce the ingredient in a large-scale manufacturing process:

[0652] 1. Physical pretreatment of plant biomass: In a suspension, mix 100 kg of ground corn cobs with water at a solid concentration of 15% (w / w). Start mixing and heat to 95°C, and mix at 95°C for 60 minutes. Add 6 kg of sodium hydroxide (0.2 - 3% based on the weight of the corn cobs) and continue stirring. Heat to 95°C and mix for 60 minutes to release the hemicellulose present in the corn cobs. At the end of 60 minutes, cool to 50°C and adjust the pH to 5.5 with sulfuric acid.

[0653] 2. Removal of partial soluble polysaccharides: Remove the partial soluble phase, equivalent to 5 - 30% of the total xylan. Neutralize with sulfuric acid and concentrate and purify by ultrafiltration. Remove any precipitated polymers.

[0654] 3. Hydrolysis: Add a cellulolytic enzyme (e.g., from Trichoderma reesei) to the ground corn cob mixture and incubate at about 50°C for 12 - 72 hours.

[0655] 4. Separation of biomass: At the end of hydrolysis, the liquid is separated from the product by a solid-liquid separator (e.g., a filter press or a decanter centrifuge).

[0656] 5. Separation of enzymes after enzymatic hydrolysis: The liquid fraction from the slurry contains enzymes, oligosaccharides, water, and salts that need to be separated. A 3 kDa or 10 kDa hollow fiber membrane is used to separate the enzyme protein and other macromolecules.

[0657] 6. Removal of salts using an ion exchange column at ≤ 45 °C.

[0658] a. Cation column: Strong acid cation exchange resin, cross-linked polystyrene matrix, sulfonate functional group, and Na + counterion.

[0659] b. Anion column: Macroporous, weak base anion exchange resin, cross-linked polystyrene matrix, dimethyl tertiary amine functional group, and OH - counterion.

[0660] 7. Oligosaccharide concentration: The desired oligosaccharides are selectively concentrated by nanofiltration at room temperature.

[0661] 8. Concentration: Optionally, the liquid is concentrated to 40 - 75% at 60 - 80 °C.

[0662] 9. Reconstitution: The purified soluble polymer is combined with the oligomers produced by the enzyme in a dry weight ratio of 5:95 - 20:80.

[0663] 10. Spray drying: The resulting solution is spray dried with an inlet temperature of 130 - 160 °C and an outlet temperature of 65 - 85 °C.

[0664] Example 12 - Use of the liquid composition in the manufacture of extruded cereal bars

[0665] The following steps can be performed to use the liquid composition provided herein to manufacture extruded cereal bars:

[0666] 1. A 120 kg solution containing 10.5 kg xylan, 57 kg xylooligosaccharides, and 7.5 kg cellobiose is heated with 130 kg of coconut oil and transferred to a high-speed mixer. 200 kg of oatmeal and 25 kg of a chopped date / raisin mixture are added and mixed well. This is pulsed through a dough mixer and set at 20 psi through a dough feed system.

[0667] 2. The mixture is conveyed through a conveyor belt and a ramshorn and baked at 180 °C for 20 minutes. Then, the product is transferred to a cookie cutting line by an oven mover and then enters variable form filling and sealing packaging when cooled to < 5 °C.

[0668] 3. The resulting product is a soft, sticky cereal bar that is loose and filled with oats. The sweetness is delivered by a liquid solution containing 10.5 kg of xylan, 57 kg of xylo-oligosaccharides, 7.5 kg of cellobiose, and chopped dates. The liquid solution containing 10.5 kg of xylan, 57 kg of xylo-oligosaccharides, and 7.5 kg of cellobiose, and coconut oil both act as binders, holding the other ingredients in the bar together and giving the bar structure.

[0669] Example 13 - Use of Liquid Ingredients in the Manufacture of Extruded Breakfast Cereals

[0670] The following steps can be carried out to manufacture extruded breakfast cereals using the liquid ingredients provided herein:

[0671] 1. Mix cereal flour (about 85 - 75% w / v) with a solution (about 15 - 25% v / v) containing 22.5 g of xylan, 30 g of cellobiose, and 97.5 g of xylo-oligosaccharides per 100 g of water, and any additives for fortification (such as preservatives, vitamins, and minerals) to form a dough. This is extruded using a twin-screw extruder that cooks the product using a combination of heat and humidification and / or steam and mechanical shear, forming the shape of the product by pushing it through a nozzle. The product is then puffed to a light texture and golden brown color and cooled.

[0672] 2. A light, crispy, shaped breakfast cereal product is obtained. The liquid solution containing 22.5 g of xylan, 30 g of cellobiose, and 97.5 g of xylo-oligosaccharides imparts sweetness to the product and helps form the structure of the dough before extrusion.

[0673] Example 14 - Use of Liquid Ingredients in the Manufacture of Tomato Ketchup

[0674] The following steps can be carried out to manufacture tomato ketchup using the liquid ingredients provided herein:

[0675] 1. Mix 4 onions and 250 g of celery in a food processor until chopped. They are fried gently in 5 tablespoons of vegetable oil for 5 minutes. Add 4 cloves of garlic and cook for another 5 minutes. Add 1 teaspoon of ground coriander, 1 short cinnamon stick, 1 teaspoon of all spice, 1 / 2 teaspoon of black pepper, and 2 teaspoons of celery salt and cook for one more minute.

[0676] 2. Add 2 kg of cooked chopped tomatoes, 3 tablespoons of tomato ketchup, 1 / 2 teaspoon of chili sauce, 200 mL of white wine vinegar, and 285 mL of a solution containing 22.5 g of xylan, 30 g of cellobiose, and 97.5 g of xylo-oligosaccharides per 100 g of water to the mixture. Bring the mixture to a boil again and cook uncovered for 1 hour until the tomatoes are soft. Discard the cinnamon stick, blend the sauce mixture until smooth, and then sieve it.

[0677] 3. The resulting product is a smooth and rich tomato sauce. A liquid solution containing 22.5 g of xylan, 30 g of cellobiose, and 97.5 g of xylooligosaccharides sweetens the product and adds body to the sauce, helping to thicken and puff up the sauce.

[0678] Example 15 - HPAEC Chromatogram of Sugars in Water after Corncob Washing

[0679] The corncobs were incubated in water at room temperature at a concentration of 100 g / L ("washing" in Table 3), and then the water was poured out. The water was added back to the original total volume and heated to 90 °C for 60 minutes ("wetting" in Table 3), and then heated in dilute NaOH at 90 °C for 60 minutes ("pretreatment" in Table 3).

[0680] HPAEC was performed on the washed, wetted, and pretreated samples, and the peaks of the sugars were identified (see Figure 7 , e.g., the chromatogram from "washing"). As shown in Table 3, at the start of the process, approximately 2% of the corncobs were glucose that could be washed out, and more glucose might be washable. It was also noted that the pH dropped to 4.5 after the washing step.

[0681] Table 3

[0682] Step Glucose (g / l) Washing 2.43g / l Wetting 1.43g / l Pretreatment 0.02g / l

[0683] Example 16 - Quantification of Sugars and Organic Acids

[0684] To quantify the effect of the pre - washing step on the process, three batches of corncobs were processed separately according to the procedure outlined in Figure 10A . The samples were analyzed for sugars by HPLC ( Figure 10B ), and for organic acids by HPLC ( Figure 10C ). The differences in the sugar and organic acid compositions separated from different samples (samples A - E and from 5 minutes (1 / 12 h) to 4 h) indicated the effect of washing. If more than 150 mL was taken out of the 600 mL for each wash, the effect of pre - washing might be greater. Thus, these data indicated the direction that washing could take, rather than the limit of washing.

[0685] As shown, glucose, fructose, and sucrose all decreased with pre - washing. Fructose and glucose were mostly decomposed during the NaOH treatment, but sucrose was resistant to the NaOH treatment. Since sucrose generally cannot be removed from other disaccharides by filtration, sucrose can be effectively removed by washing. Table 4 shows the comparison of sugars in sample Ds ("no washing" and "two washes"), and Table 5 shows the comparison of sugars in the 4 - hour samples ("no washing" and "two washes").

[0686] Table 4

[0687]

[0688] Table 5

[0689]

[0690] Before the start of the "alkali cooking" step (e.g., the thermochemical step), pre-washing results in a reduction of approximately 50% of glucose and fructose and approximately 80% of sucrose. At the end of hydrolysis, pre-washing results in a minor difference in small sugars. The concentration of xylose sugars appears to increase (as evidenced by the negative change), while the concentrations of glucose and cellobiose decrease.

[0691] The large amount of organic acids detected are not wash products, but rather pretreatment products. However, the unwashed biomass appears to have a higher total organic acid loading compared to the twice-washed material.

[0692] For the steps that cause the pretreatment stage, the acid concentration of the washed biomass also appears to be lower. Table 6 shows the comparison of sample Ds, and Table 7 shows the comparison of the four-hour samples.

[0693] Table 6

[0694]

[0695]

[0696] Table 7

[0697]

[0698] The effect of washing on the organic acid content is evident both at the end of the wetting stage (sample D) and at the end of the hydrolysis reaction (4-hour sample), as shown by the "change" (calculated by subtracting "twice-washed" from "no wash") (see, for example, Tables 6 and 7). At the end of the wetting stage (sample D), all other acids were detected except for lactic acid and formic acid (n / d), and they were at a lower concentration for the material washed twice.

[0699] At the end of hydrolysis, the acetate content is higher (acetate is released during hydrolysis). The other acids of the washed biomass are still lower than those of the unwashed (except for lactic acid, which is present at a low concentration).

[0700] The visual observation of the samples shows in Figure 10D After two washes, the corncobs release fewer colored compounds and the juice is lighter. Without being bound by any particular theory, the colored compounds may be phenols and organic acids released during washing.

[0701] Example 17 - Comparison of Cold-Pressed Cereal Bars

[0702] Prepare cold-pressed cereal bars according to the previous formulation (see Example 10).

[0703] Compare the use of soluble polysaccharides and insoluble polysaccharides in the cereal bars. The soluble and insoluble polysaccharides are as follows:

[0704] Soluble polysaccharide: 60 mL of water containing 94 g of dry ingredients, the composition of the dry ingredients being 10% dry w / w cellobiose, 75% xylooligosaccharides and 15% extracted water-soluble polymer (sample 4 as described in Example 8 above).

[0705] Insoluble polysaccharide: 60 mL of water containing 94 g of dry ingredients, the composition of the dry ingredients being 10% dry w / w cellobiose, 75% xylooligosaccharides and 15% microcrystalline cellulose.

[0706] Refer to as Figure 11A shown, although the cereal bars made with insoluble polysaccharides look like solid bars when placed on the table, however, due to their soft texture and the components not being well combined together, they start to fall apart when picked up from the table. In contrast, the cereal bars made with soluble polysaccharides can be easily handled and maintain their shape.

[0707] Use a TA-XTPlusC texture analyzer (Stable Micro systems, UK) and measure the hardness and stickiness of the cereal bars using "ExponentC" software. Place a sample with dimensions of 9.6 cm x 3.8 cm x 1 cm (L x W x H) in the center under the probe. An aluminum cylindrical probe with a diameter of 6 mm is used for the penetration test in the "return to start" mode and a 30 kg load cell. Once the probe triggers on the surface, it penetrates the sample at a speed of 2 mm / s for a distance of 2 mm. At this point (2 mm depth), record the force value and use the force value as a measure of the "hardness" of the sample. Then the probe withdraws from the sample, and at this time, record the maximum withdrawal force or "stickiness". The speed before the test is 1 mm / s and the speed after the test is 10 mm / s.

[0708] The results show that the hardest bars are those made with soluble ingredients, while for the insoluble ingredients, lower values are obtained ( Figure 11B ). Similarly, the bars containing soluble polysaccharides have higher stickiness, while the bars containing insoluble polysaccharides have lower stickiness. These results confirm the visual and tactile observations.

[0709] Table 8: Results

[0710] Sample Hardness (g) Viscosity (g) Cereal bar made of soluble polysaccharide 169.19 -20.33 Cereal bar made of insoluble polysaccharide 47.43 -11.24

[0711] Table 9: Texture analyzer settings

[0712] Mode Measuring compression force Options: Restore to start Speed before test: 1 mm / s Test speed 2 mm / s Speed after test 10.0 mm / s Distance: 2 mm Trigger type: Automatic - 20 g Tare mode: Automatic Data acquisition rate: 400 pps

[0713] The hardness of the cereal bars was further measured using a TA-XTPlusC texture analyzer (Stable Micro systems, UK) with "ExponentC" software. A sample with dimensions of 9.6 cm x 3.8 cm x 1 cm (L x W x H) was placed in the center under the knife. A knife edge, "Return to Start" mode, and a 30 kg load cell were used in the cutting test. Once the knife triggered on the surface, it penetrated the sample by 5 mm at a test speed of 2 mm / s. The maximum force measured during the cutting test w...

Claims

1. A method for producing a composition, the method comprising: a) applying a physical pretreatment to biomass to reduce the average size of the biomass; b) applying a wash cycle pretreatment to the physically pretreated biomass, wherein the wash cycle exposes the physically pretreated biomass to water and dissolves one or more monosaccharides, one or more disaccharides, and one or more organic acids from the physically pretreated biomass, wherein the wash cycle pretreatment comprises: (i) incubating the physically pretreated biomass in an aqueous solution to dissolve one or more organic acids and monosaccharides and / or disaccharides from the physically pretreated biomass; and (ii) removing a portion of the dissolved monosaccharides, disaccharides, and organic acids from the aqueous solution, wherein 50% or more of the dissolved monosaccharides and disaccharides are removed from the aqueous solution; c) applying a strong pretreatment to the wash cycle-pretreated biomass from step (b) to increase the digestibility of the biomass, the strong pretreatment comprising a thermochemical treatment performed at a temperature of at least 75 °C; d) contacting one or more polysaccharide-cleaving enzymes with the strongly pretreated biomass from step c) in a solution or suspension to form one or more oligosaccharides; and e) enriching the solution or suspension to increase the concentration of the one or more oligosaccharides to form the composition; wherein the wash cycle pretreatment is performed at a temperature lower than the temperature of the strong pretreatment.

2. The method according to claim 1, wherein the composition comprises less than 50% dry weight of disaccharides.

3. The method according to claim 1, wherein the one or more organic acids comprise at least one of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid, or acetic acid.

4. The method according to claim 1, wherein the strong pretreatment is a thermochemical treatment comprising incubating the wash cycle-pretreated biomass in an alkaline solution having a pH of at least 7.

5.

5. The method according to any one of claims 1 to 4, wherein the strongly pretreated biomass after step c) comprises less than 10% w / w monosaccharides.

6. A method for forming a composition, the method comprising: (a) separating one or more soluble polysaccharides from biomass, the one or more soluble polysaccharides comprising xylan, (b) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides, wherein the one or more enzymes comprise at least one of cellulase, xylanase, xyloglucanase, endoglucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase, wherein the one or more oligosaccharides comprise at least one of the following: i) cellooligosaccharides having a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides having a DP of 2 to 12; iii) arabinoxylooligosaccharides having a DP of 3 to 15; iv) manooligosaccharides having a DP of 2 to 12; v) mixed-link glucan oligosaccharides having a DP of 2 to 5; vi) xyloglucan oligosaccharides having a DP of 4 to 12; or vii) chitooligosaccharides having a DP of 2 to 12; (c) separating the one or more oligosaccharides; and (d) Combining a portion of the one or more soluble polysaccharides from step (a) with a portion of the one or more oligosaccharides from step (c) to form the composition, wherein the composition comprises at least 20% dry weight of the one or more oligosaccharides and at least 2% dry weight of the one or more soluble polysaccharides.

7. The method according to claim 6, further comprising combining the composition with a liquid to form a liquid composition.

8. An edible composition comprising: Soluble polysaccharides, the soluble polysaccharides comprising xylan; and Oligosaccharides, the oligosaccharides comprising: i) cellooligosaccharides with a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2 to 12; iii) arabinoxylooligosaccharides with a DP of 3 to 15; wherein the concentration of cellooligosaccharides with a polymer of 2 in the cellooligosaccharide mixture is at least 30% w / w, and wherein the ratio of the amounts of soluble polysaccharides and oligosaccharides in the composition is from 1:100 to 1:

1.

9. A method for producing a composition, the method comprising: a) Applying a physical pretreatment to the biomass to reduce the average size of the biomass; b) Applying a washing cycle pretreatment to the physically pretreated biomass, wherein the washing cycle exposes the physically pretreated biomass to water and dissolves one or more monosaccharides, one or more disaccharides, and one or more organic acids from the physically pretreated biomass, wherein the washing cycle pretreatment comprises: (i) Incubating the physically pretreated biomass in an aqueous solution to dissolve one or more organic acids and monosaccharides and / or disaccharides from the physically pretreated biomass; and (ii) Removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution, wherein 50% or more of the dissolved monosaccharides and / or disaccharides are removed from the aqueous solution; c) Applying a strong pretreatment to the washing cycle pretreated biomass from step (b) to increase the digestibility of the biomass, the strong pretreatment comprising a thermochemical treatment performed at a temperature of at least 75°C; d) Separating one or more soluble polysaccharides from the biomass; e) Contacting one or more polysaccharide-cleaving enzymes with the strongly pretreated biomass from step (c) in solution or suspension to form one or more oligosaccharides; and f) Enriching the solution or suspension to increase the concentration of the one or more oligosaccharides to form the composition; wherein the washing cycle pretreatment is performed at a temperature lower than the temperature of the strong pretreatment.

10. The method according to claim 9, the method further comprising step g) combining a portion of the one or more soluble polysaccharides from step (d) with one or more oligosaccharides from step (f) to form another composition.

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