Methods and related compositions for processing biomass to produce oligosaccharides

Abstract

Provided herein is a method for forming a composition for human consumption. The method may include separating one or more soluble polysaccharides from biomass, producing one or more oligosaccharides from biomass, and combining one or more separated soluble polysaccharides with the produced oligosaccharides to form a composition. Also provided is a method for pretreating biomass. The method may include applying physical pretreatment to biomass, applying gentle pretreatment to physically pretreated biomass, and applying strong pretreatment to gently pretreated biomass. Also provided is a composition for human consumption.

Description

[0001] Cross-references

[0002] This application claims the benefit of UK patent application number 1911762.1 filed on August 16, 2019, UK patent application number 1911764.7 filed on August 16, 2019, and UK patent application number 2002315.6 filed on February 19, 2020, the entire contents of which are incorporated herein by reference. Background Art

[0003] Sugary foods and drinks are an important part of culture and lifestyle habits around the world, but the sugar they contain has been linked to obesity, diabetes, poor dental health and destructive behaviors. For these reasons, consumer preferences have shifted away from sugary foods, and governments are increasingly implementing regulations to encourage reduced sugar consumption.

[0004] 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 by non-natural resources, and are often accompanied by a slightly bitter or other unpleasant taste in their sweetness, which cannot attract consumers. In addition, although many sweeteners can simulate the sweetness of sugar in food and beverages, few sweeteners can simulate the broad role that sugar plays in food, such as increasing volume, adjusting texture, providing structure, acting as a preservative, and adjusting color and flavor through caramelization and Maillard reaction. In addition, many bulk sweeteners that can simulate these physical properties of sugar all have gastrointestinal tolerance problems, which limits their use to levels far below those required for replacing sugar in a standard Western diet.

[0005] Dietary fiber is an important part of a positive diet and helps maintain a healthy digestive system and a well-regulated intestinal flora. This fiber includes 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 production process. Summary of the invention

[0006] Other aspects and advantages of the present disclosure will become apparent to those skilled in the art through the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be appreciated, the present disclosure can have other different embodiments, and its several details can be modified in various obvious aspects, all without departing from the present disclosure. Therefore, the drawings and descriptions should be considered to be illustrative in nature, rather than restrictive.

[0007] In some embodiments, methods of producing ingredients for human consumption are described herein. A method of producing an ingredient for human consumption may include: (a) isolating one or more soluble polysaccharides from biomass; (b) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides; (c) isolating 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 ingredient.

[0008] In some cases, the method may include purifying the isolated one or more soluble polysaccharides.

[0009] In some cases, the method may include purifying the isolated one or more oligosaccharides.

[0010] In some cases, the methods may include treating the biomass to solubilize one or more soluble polysaccharides.

[0011] In some cases, the method may include purifying the isolated one or more soluble polysaccharides.

[0012] In some cases, the treating comprises a thermochemical treatment.

[0013] In some cases, the thermochemical treatment includes at least one of a hot water treatment or a hot alkaline treatment.

[0014] In some cases, the hot alkaline treatment uses an alkali at a pH of 10 to 14.

[0015] In some cases, the hot alkaline treatment uses at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, or aqueous ammonia.

[0016] In some cases, the treatment may be performed at a temperature of 30°C to 180°C.

[0017] In some cases, the treatment may be performed for 10 minutes to 24 hours.

[0018] In some cases, the one or more soluble polysaccharides and / or the one or more oligosaccharides are dried prior to step (d).

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

[0020] In some cases, the ingredient is soluble in water.

[0021] In some cases, the solubility of the ingredient in water may be at least 80 g of the ingredient per 100 g of water at 50°C.

[0022] In some cases, the method includes combining the component with a liquid to form a liquid component.

[0023] In some cases, the viscosity of the liquid component may be similar to the viscosity of corn syrup.

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

[0025] In some cases, the liquid component has fewer calories per gram than corn syrup or high fructose corn syrup.

[0026] In some cases, the liquid component has a lower glycemic index than corn syrup or high fructose corn syrup.

[0027] In some cases, the liquid comprises water.

[0028] In some cases, the liquid component comprises at least 20% dry weight of at least one oligosaccharide and at least 2% dry weight of at least one polysaccharide.

[0029] In some cases, the viscosity of the liquid component 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.

[0030] In some cases, the liquid component comprises at least 2% xylan by dry weight.

[0031] In some cases, the liquid component comprises at least 2% mannan by dry weight.

[0032] In some cases, the liquid component comprises at least 2% by dry weight of the cellulose derivative.

[0033] In some cases, the liquid component has a polysaccharide concentration of 0.1% to 50% w / v.

[0034] In some cases, the liquid component comprises polysaccharides and oligosaccharides in an amount ratio of 1:100 to 1:1.

[0035] In some cases, the one or more soluble polysaccharides comprise at least one of mannan, xylan, mixed bond glucan, lignocellulose, hemicellulose, a cellulose derivative, chitosan, or xyloglucan.

[0036] In some cases, the cellulose derivative comprises at least one of cellulose acetate, hydroxyethyl cellulose, or hydroxymethyl cellulose.

[0037] In some cases, the biomass comprises at least one of sugar cane biomass, corn biomass, wheat biomass, hardwood biomass, or softwood biomass.

[0038] In some cases, 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) xylo-oligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; vi) xyloglucan-oligosaccharides having a DP of 4 to 12; or vii) chito-oligosaccharides having a DP of 2 to 12.

[0039] In some cases, the ingredient comprises at least two of the oligosaccharides listed in (i) to (vii).

[0040] In some cases, the composition comprises at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other.

[0041] In some embodiments, compositions for human consumption are described herein. The compositions for human consumption may include: soluble polysaccharides; and oligosaccharides, which may include at least one of: (i) cellooligosaccharides having a degree of polymerization (DP) of 2 to 6; (ii) xylo-oligosaccharides having a DP of 2 to 12; (iii) manno-oligosaccharides having a DP of 2 to 12; (iv) arabinoxylo-oligosaccharides having a DP of 3 to 15; (v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; or (vi) chito-oligosaccharides having a DP of 2 to 12, wherein the composition contains less than 5% insoluble polysaccharides by dry weight.

[0042] In some cases, the composition may be substantially free of insoluble polysaccharides.

[0043] In some cases, the composition may be soluble in water.

[0044] 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.

[0045] In some cases, the composition further comprises a liquid, thereby forming a liquid component.

[0046] In some cases, the liquid may be water.

[0047] In some cases, the liquid component comprises at least 20% dry weight of at least one oligosaccharide and at least 2% dry weight of at least one polysaccharide.

[0048] In some cases, the viscosity of the liquid component 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.

[0049] In some cases, the liquid component comprises at least 2% xylan by dry weight.

[0050] In some cases, the liquid component comprises at least 2% mannan by dry weight.

[0051] In some cases, the liquid component comprises at least 2% by dry weight of the cellulose derivative.

[0052] In some cases, the liquid component has a polysaccharide concentration of 0.1% to 50% w / v.

[0053] In some cases, the liquid component comprises polysaccharides and oligosaccharides in an amount ratio of 1:100 to 1:1.

[0054] In some cases, the one or more soluble polysaccharides comprise at least one of mannan, xylan, mixed bond glucan, lignocellulose, hemicellulose, a cellulose derivative, chitosan, or xyloglucan.

[0055] In some cases, the cellulose derivative comprises at least one of cellulose acetate, hydroxyethyl cellulose, or hydroxymethyl cellulose.

[0056] In some cases, the biomass comprises at least one of sugar cane biomass, corn biomass, wheat biomass, hardwood biomass, or softwood biomass.

[0057] In some cases, 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) xylo-oligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; vi) xyloglucan-oligosaccharides having a DP of 4 to 12; or vii) chito-oligosaccharides having a DP of 2 to 12.

[0058] In some cases, the composition comprises at least two of the oligosaccharides listed in (i) to (vii).

[0059] In some cases, the composition comprises at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other.

[0060] In some embodiments, methods of producing ingredients for human consumption are described herein. The methods of producing ingredients for human consumption may include: (a) applying physical pretreatment to biomass to reduce the average size of the biomass; (b) applying 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.

[0061] In some cases, the mild pretreatment can be an incubation cycle.

[0062] In some cases, the strong pretreatment may be a thermochemical treatment, which may include incubating the mildly pretreated biomass in one of an acidic solution or an alkaline solution.

[0063] In some cases, the method may further comprise removing at least 25% or 50% of the dissolved monosaccharides and / or disaccharides from the incubation solution in step (b)(ii).

[0064] In some cases, the heavily pretreated biomass composition after step (c) comprises less than 10% w / w monosaccharides.

[0065] In some cases, the method may further comprise purifying the one or more oligosaccharides from the solution or suspension.

[0066] In some cases, the heavily pretreated biomass composition after step (c) comprises less than 20% w / w monosaccharides.

[0067] In some cases, the method may further include repeating step (b).

[0068] In some cases, step (b) may be performed two, three, four, or five times.

[0069] In some cases, the method may further include repeating step (c).

[0070] In some cases, step (c) may be performed two, three, four, or five times.

[0071] In some cases, the method may further comprise concentrating a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

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

[0073] In some cases, a portion of the dissolved monosaccharides and / or disaccharides removed in step (b) may not be combined with a portion of the one or more oligosaccharides of step (e) to form the component.

[0074] In some cases, the ingredient comprises less than 15% monosaccharides by dry weight.

[0075] In some cases, the component comprises less than 50% disaccharide by dry weight.

[0076] In some cases, the composition may be substantially free of monosaccharides.

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

[0078] In some cases, 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) xylo-oligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; vi) xyloglucan-oligosaccharides having a DP of 4 to 12; or vii) chito-oligosaccharides having a DP of 2 to 12.

[0079] In some cases, the ingredient comprises at least two of the oligosaccharides listed in (i) to (vii).

[0080] In some cases, the composition comprises at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other.

[0081] In some cases, the component comprises at least one of sucrose, maltose, lactose, glucose, fructose, or galactose at less than 50% total dry weight w / w of the total dry weight w / w of all oligosaccharides of i-vii.

[0082] In some cases, the monosaccharide and / or disaccharide comprises at least one of sucrose, maltose, lactose, glucose, fructose, or galactose.

[0083] In some cases, step (b) dissolves one or more organic acids in addition to monosaccharides and / or disaccharides.

[0084] In some cases, 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.

[0085] In some cases, the total weight of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid and acetic acid may 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.

[0086] In some cases, the physical pretreatment of step (a) comprises at least one of chipping, chopping, milling, ball milling, grinding, trimming, or mixing the biomass.

[0087] In some cases, the mild pretreatment of step (b) is performed in an aqueous solution comprising water.

[0088] In some cases, the mild pretreatment of step (b) is carried out at a temperature of 5°C to 150°C.

[0089] In some cases, the mild pretreatment of step (b) may be performed for 15 minutes to 1 hour.

[0090] In some cases, the strong pretreatment of step (c) comprises heating the mildly pretreated biomass in an acidic solution or an alkaline solution.

[0091] In some cases, the heating may be performed at a temperature of 50°C to 150°C.

[0092] In some cases, the heating can be performed for 30 minutes to 4 hours.

[0093] In some cases, step (c) comprises treating the mildly pretreated biomass in an alkaline solution having a pH of 8 to 11.

[0094] In some cases, the alkaline solution comprises at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, aqueous ammonia, ammonium sulfate, or ammonium hydroxide.

[0095] In some cases, step (c) comprises treating the mildly pretreated biomass in an acidic solution having a pH of 4 to 6.

[0096] In some cases, 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.

[0097] In some cases, the biomass comprises at least one of sugar cane, corn stover, corn cobs, wheat bran, wheat straw, hardwood, or softwood.

[0098] In some cases, the biomass comprises at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed-linkage glucan, mannan, or lignocellulose.

[0099] In some cases, the one or more polysaccharide-cleaving enzymes comprise at least one of a cellulase, a xylanase, a xyloglucanase, an endoglucanase, a cellobiohydrolase, a mannanase, a lichenase, or a lytic polysaccharide monooxygenase (LPMO).

[0100] In some cases, the one or more polysaccharide-cleaving enzymes comprise at least one of AA9, AA10, AA11, AA13, AA14, or AA15.

[0101] In some cases, the one or more polysaccharide-cleaving enzymes may be produced by a filamentous fungus, such as Trichoderma reesei.

[0102] In some cases, the one or more polysaccharide-cleaving enzymes can be operably linked to a catalytic module.

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

[0104] In some cases, the non-catalytic module can be a carbohydrate binding module.

[0105] In some cases, a water-soluble composition for human consumption may include at least one of the following oligosaccharides: i) cellooligosaccharides having a degree of polymerization (DP) of 2 to 6; ii) xylo-oligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; vi) xyloglucan-oligosaccharides having a DP of 4 to 12; or vii) chito-oligosaccharides having 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 monosaccharides or disaccharides is less than 10% of the total dry weight of the oligosaccharides having a DP of 2 to 12.

[0106] In some cases, the composition may include at least two of the oligosaccharides listed in (i) to (vii). In some cases, the composition may also include 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 may include at least two of the organic acids. In some cases, the composition may include at least two of the oligosaccharides listed in (i) to (vii) in a ratio of 1:9 to 1:1 relative to each other.

[0107] In some cases, the composition may include at least one monosaccharide or disaccharide selected from the group consisting of glucose, fructose, or sucrose; and at least one organic acid selected from the group consisting of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid, or acetic acid, wherein the total weight of the organic acids is greater than 10% of the total weight of the monosaccharides or disaccharides.

[0108] In some cases, a method of producing an ingredient 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, wherein the mild pretreatment includes: (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 the one or more oligosaccharides; and (g) combining a portion of the one or more soluble polysaccharides from step (d) with a portion of the one or more oligosaccharides from step (f) to form the ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0109] The novel features of the present invention are particularly set forth in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by referring to the following detailed description and accompanying drawings which set forth illustrative embodiments in which the principles of the present invention are utilized, wherein:

[0110] Figure 1 Depicted are the results of thin layer chromatography (TLC) analysis showing the mono- and disaccharide content of the indicated biomass after 1-5 washing cycles (incubation pretreatment).

[0111] Figure 2 Depicted are the results of TLC analysis comparing the levels of mono- and disaccharides in the enzymatic hydrolysate mixture when washed (+) and not (-) (incubation pretreatment).

[0112] Figure 3 Depicted are the results of preparing glazes using compositions of the present disclosure and comparative compositions.

[0113] Figure 4 Depicted are baked products utilizing the liquid ingredients of the present disclosure.

[0114] Figure 5 High performance anion exchange chromatography (HPAEC) analysis of sample 4 is depicted.

[0115] Figure 6 Depicted are cereal bar products produced using the liquid ingredients of the present disclosure.

[0116] Figure 7 Depicted is the HPAEC chromatogram of sugars from washed corncobs.

[0117] Figure 8 is a simplified flow chart describing a method of processing biomass according to some embodiments of the present disclosure.

[0118] Fig. 9 is a simplified flow diagram depicting another method of processing biomass according to some embodiments of the present disclosure.

[0119] Fig. 10A is a diagram depicting a method of processing biomass.

[0120] Fig. 10B The measurement of sugars in multiple samples is described.

[0121] Fig. 10C Depicted are the measurements of organic acids in multiple samples.

[0122] Fig. 10D Describes the basis Fig. 10A Visual observation of samples treated by the method.

[0123] Fig.11A A comparison between cereal bars made with soluble polysaccharides and cereal bars made with insoluble polysaccharides is described.

[0124] Fig. 11B Describes the use of penetration testing Fig.11A The hardness and stickiness of the cereal bar.

[0125] Fig. 11C Depicts the use of a cutting test Fig.11A The hardness of the cereal bar. DETAILED DESCRIPTION

[0126] introduction

[0127] Provided herein is a method for generating one or more compositions. The method may include separating one or more soluble polysaccharides from biomass or raw materials (e.g., 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 oligosaccharides of one or more enrichments or separations to form the composition.

[0128] The texture of composition can be continuous and smooth. In addition, this composition can have some characteristics, so that this composition can be used as sweetener and / or sugar substitute. The property of this composition can include sweet taste, smooth texture, desirable mouthfeel, bonding ability, sizing ability or the ability, wettability, viscosity, bulking ability and / or caramelization ability of forming juice. Compared with corn syrup or high fructose corn syrup, this composition can also have lower calorie, reduced glycemic index, reduced glycemic load, increased fiber and / or reduced sugar.

[0129] Also provided herein is a method for producing a composition for incorporating into food, nutrition and / or cosmetics, wherein the method may include carrying out one or more pretreatment steps on the biomass. For example, the method may include a first pretreatment step, a second pretreatment step, a third pretreatment step or an additional pretreatment step carried out on the biomass. The pretreatment step may be carried out in a specified order.

[0130] The biomass for producing one or more oligosaccharides can be plant biomass. The example of plant biomass includes but is not limited to: sugar cane, corn stalks, corn cobs, wheat bran, wheat straw, hardwood or softwood. In some cases, the biomass can include cellulose, chitin, chitosan, xylan, xyloglucan, mixed bond glucan, mannan or lignocellulose.

[0131] Biomass can be digested into one or more oligosaccharides. Biomass digestion can be performed enzymatically. Enzymatic digestion can be performed after one or more pre-treatment steps. One or more pre-treatment steps can be performed to reduce the size of biomass and / or increase the surface area of ​​biomass that can be used for digestion. One or more pre-treatment steps can include one or more washing steps, dissolving steps or pre-digestion treatments. In some cases, one or more pre-treatment steps can be performed to reduce the monosaccharides and / or disaccharides present in the biomass. In some cases, one or more pre-treatment steps can be performed to reclaim the soluble polysaccharide fraction from the biomass.

[0132] 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 the biomass during one or more pre-treatment steps. Therefore, after one or more pre-treatment steps are completed, monosaccharides and / or disaccharides may not be produced or substantially not be produced. In other words, the pre-treated biomass may not contain or substantially not contain monosaccharides and / or disaccharides. This can improve the efficiency of the method for producing the composition of food, nutrition and / or cosmetics as provided herein. For example, due to having removed a portion of monosaccharides and / or disaccharides from the biomass during one or more pre-treatment steps, less component purification (e.g., to remove monosaccharides and / or disaccharides) may be required. Specifically, less filtering steps or less stringent filtering steps may be required during purification to produce the composition as disclosed herein.

[0133] The first pretreatment step (pretreatment step 1) can include physically treating the biomass (e.g., cutting the biomass). The second pretreatment step (pretreatment step 2 or mild pretreatment) can include subjecting the physically treated biomass to an incubation cycle or a wash cycle. The incubation cycle can include incubating the physically treated biomass (from pretreatment 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. Pretreatment step 2 can be a mild pretreatment step. For example, the conditions (e.g., solution, temperature, time, etc.) of pretreatment step 2 can be milder than the conditions of pretreatment step 3 described in further detail below.

[0134] 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 biomass (for example, by enzyme). Pretreatment step 3 can also improve the entry of enzymes to biomass. In various cases, pretreatment step 3 can occur in an alkaline solution (for example, 1% w / v NaOH solution) at a temperature higher than room temperature (for example, about 90°C to about 110°C). In addition, pretreatment step 3 can be carried out by keeping about 30 minutes to 1 hour at a desired temperature. For example, the effective temperature--90°C in the present embodiment--keeps 1 hour (this 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 step after the pretreatment step as described in further detail below, the solution may not be discarded. In some cases, pretreatment step 3 may include thermochemical treatment. That is, pretreatment step 3 may be performed in an acidic or alkaline solution and / or pretreatment step 3 may be performed at a temperature higher than room temperature.

[0135] In various cases, after one or more pretreatment steps, the method for producing an ingredient for human consumption may 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 may also include enriching the solution or suspension to increase the concentration of one or more oligosaccharides to form the ingredient (e.g., an ingredient for human consumption).

[0136] Compared with some other methods, the pretreatment step can improve the efficiency of the method. Because less downstream processing may be required, the efficiency can be improved. In some cases, downstream processing can include a process for removing monosaccharides and / or disaccharides from pretreated biomass. In some embodiments, it may be difficult to remove disaccharides separately, such as from an intermediate solution or fraction produced by an enzyme digestion method. Therefore, it may be more effective to remove disaccharides during the pretreatment step.

[0137] The step of downstream processing can include ion exchange chromatography, ultrafiltration, microfiltration, nanofiltration, etc. The part of the nanofiltration step can be to remove excessive monosaccharides from the oligosaccharide mixture. This nanofiltration can be performed multiple times to reach the required monosaccharide level. When the monosaccharide in the pretreated biomass is less (for example, when the monosaccharide has been removed by washing or incubation), the quantity of the nanofiltration step can be reduced. In addition, ultrafiltration can not distinguish between required disaccharides (for example cellobiose) and unwanted disaccharides usually. Therefore, removing unwanted disaccharides during pretreatment and producing required disaccharides during the enzyme treatment step can also reduce the number of steps involved or required during downstream processing.

[0138] 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 as examples only. Without departing from the present invention, those skilled in the art may make many changes, modifications and substitutions. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed.

[0139] 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 a plurality of samples, including mixtures thereof.

[0140] As used herein, the term "about" can mean within 1 or more than 1 standard deviation. Alternatively, "about" can mean a range of up to 10%, up to 5%, or up to 1% of a given value. For example, about can mean up to ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of a given value.

[0141] As used herein, "food" and "foodstuff" generally refer to any edible item that can be consumed by humans or any other animals. It can be food, feed, drink, or an ingredient for producing any of the above.

[0142] As used herein, "nutraceuticals" generally refer to any composition introduced into a human or other animal by ingestion, injection, absorption or any other method in order to provide nutrition to the human or other animal. Such nutraceuticals may be used in the form of beverages with added dietary fiber, prebiotic additives, pills or other capsules, or any other suitable use.

[0143] As used herein, "cosmetic" generally refers to any composition intended for use on humans or other animals to increase their aesthetic appeal or prevent future loss of aesthetic appeal, as well as any other composition generally referred to as cosmetics. Aesthetic appeal is not limited to visual aesthetics, but also applies to structural or any other aspects of attractiveness. Cosmetics can be mascara, foundation, lip gloss, eye shadow, eyeliner, primer, lipstick, blush, nail polish, bronzer, or any other makeup; shampoo, conditioner, styling mousse, styling gel, hair spray, hair dye, hair wax, or any other hair product; moisturizer, exfoliant, sunscreen, cleanser, toothpaste, cream, lotion, ointment, or any other composition that effectively modifies teeth, skin, hair, or other parts of the body in some aesthetic way. In addition, cosmetics can be compositions used as components of facial masks, brushes, curling irons, other styling devices, other solid structures, or any other suitable compositions.

[0144] As used herein, "ingredient" generally refers to any composition suitable for incorporation into a food, cosmetic or nutraceutical, which may include those used directly as a product itself. It may be a dry ingredient or a liquid ingredient unless it is specifically referred to as "dry" or "liquid". This includes compositions that may 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, prior to any further purification, optimization, drying, dissolution or any other such steps, as well as final compositions obtained from the methods.

[0145] As used herein, "polysaccharide" generally refers to any sugar polymer greater than about 20 residues in length. Polysaccharides can be highly branched, slightly branched or unbranched. Polysaccharides can include any combination of glycosidic bonds of any type, 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), such as any combination of the above monomers modified with acetyl or other groups. Polysaccharides can be cellulose or hemicellulose polymers, and the hemicellulose polymers envisioned include xylan, glucuronic acid xylan, arabinoxylan, glucomannan and xyloglucan. In some embodiments, cellulose polymers can be cellulose.

[0146] 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 may contain one or more of the following polysaccharides associated together: cellulose, xylan, mannan, and mixed bond glucan.

[0147] As used herein, "highly branched," "lightly branched," and "unbranched" generally refer to the number of side chains per backbone in a saccharide. Highly branched saccharides have an average of 4 to 10 side chains per 10 backbone residues, lightly branched saccharides have an average of 1 to 3 side chains per 10 backbone residues, and unbranched saccharides have only one backbone and no side chains. The average is calculated by dividing the number of side chains in the saccharide by the number of backbone residues.

[0148] As used herein, "sugar" generally refers to any polysaccharide and / or oligosaccharide, such as monosaccharides and / or disaccharides.

[0149] As used herein, "oligosaccharides" generally refer to sugar polymers having a chain length of less than or equal to about 20 sugar residues. Oligosaccharides may be highly branched, lightly branched, or unbranched; and may 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 derivatives thereof). Suitable derivatives include the above monomers containing acetyl or other groups.

[0150] As used herein, "monosaccharides" and "disaccharides" generally refer to sugar compounds consisting of one or two residues, respectively. Monosaccharides are compounds such as glucose, glucosamine, xylose, galactose, fucose, fructose, glucuronic acid, arabinose, galacturonic acid; or their diastereomers or other derivatives. Suitable derivatives include acetyl or other groups. Disaccharides are compounds consisting of two monosaccharides linked by any glycosidic bond.

[0151] As used herein, "cello-oligosaccharide" generally refers to an oligosaccharide composed of one or more glucose residues linked by β-1,4-glycosidic bonds, and may be chemically related by oxidation, reduction, esterification, epimerization, or another chemical modification.

[0152] As used herein, "xylooligosaccharides" generally refer to oligosaccharides consisting primarily of xylose residues, typically 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 may be chemically associated therewith by oxidation, reduction, esterification, epimerization, further glycosylation or another chemical modification.

[0153] As used herein, "arabinoxylo-oligosaccharide" generally refers to an oligosaccharide composed of xylose residues, typically linked by β-(1→4)-linkages substituted with arabinose side chains, typically linked by (1→2)-linkages or (1→3)-linkages, and may be chemically related thereto by oxidation, reduction, esterification, epimerization, further glycosylation or another chemical modification.

[0154] 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 may be chemically associated therewith by oxidation, reduction, esterification, epimerization, or another chemical modification.

[0155] 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 which may be chemically related by oxidation, reduction, esterification, epimerization or another chemical modification;

[0156] As used herein, "chitooligosaccharide" generally refers to an oligosaccharide composed of one or more glucosamine and / or N-acetyl-glucosamine residues, and may be chemically related thereto by oxidation, reduction, esterification, epimerization or another chemical modification.

[0157] As used herein, "cellulose" generally refers to a polysaccharide consisting of glucose residues linked by β-1,4-glycosidic bonds and its derivatives. As used herein, "xylan" generally refers to a polysaccharide consisting of a skeleton 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 bond glucan" generally refers to a polysaccharide consisting 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 consisting 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 consisting of glucosamine and / or N-acetyl-glucosamine residues. The polysaccharides of cellulose, xylan, mixed bond glucan, mannan, chitin or chitosan may include chemical variants that have been modified by oxidation, reduction, esterification, epimerization or another chemical modification.

[0158] As used herein, "soluble", "solubility" and grammatical variations thereof generally refer to solubility in an aqueous solution (eg, water). As used herein, "dissolve" generally refers to the incorporation of a solid into an aqueous solution or liquid to form a solution.

[0159] 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 may 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%, based on the weight of the composition. The suspension may include a suitable solvent, which may be water.

[0160] As used herein, "viscosity" generally refers to a quantity that represents the magnitude of internal friction in a fluid, measured by the force per unit area resisting uniform flow. Viscosity can be measured by a variety of methods, but unless otherwise indicated, the values ​​given herein refer to the values ​​obtained by using a Brookfield HDB VE rotational viscometer using standard test methods, operating in accordance with the manufacturer's instructions for the range, and collecting a 400 mL sample in a tall beaker to ensure that container effects do not occur.

[0161] As used herein, "dissolved" generally refers to the incorporation of a solid into a liquid to form a solution.

[0162] I. Pretreatment

[0163] Physical pretreatment

[0164] Biomass can be subjected to mechanical or physical pretreatment to digest biomass into one or more oligosaccharides. Mechanical and / or physical pretreatment can be the first pretreatment step in the biomass digestion process. Alternatively, a mechanical or physical pretreatment step can be performed after another pretreatment step. For example, a mechanical or physical pretreatment step can be performed after another pretreatment step (such as a washing pretreatment step).

[0165] The biomass may be subjected to mechanical or physical pretreatment to reduce the size of the biomass. Examples of mechanical or physical pretreatment steps include, but are not limited to, cutting, chopping, grinding, ball milling, grinding, trimming, mixing and / or steam explosion of the biomass. More than one physical pretreatment may be performed on the biomass.

[0166] Dissolution Steps

[0167] The biomass may be subjected to a dissolution step. The dissolution step may be a mild pretreatment step or a part of a mild pretreatment step, which dissolves the polysaccharide fraction or monosaccharide and / or disaccharide fraction from the biomass. The dissolution pretreatment step may be a washing step, an incubation step, a thermochemical step or a chemical treatment step. The dissolution pretreatment may be performed before a physical or mechanical pretreatment step. The dissolution pretreatment may be performed after a physical or mechanical pretreatment step.

[0168] In some embodiments, a dissolution step may be performed to remove a portion of soluble polysaccharides. Soluble polysaccharides may be added to one or more oligosaccharides, for example, after the soluble polysaccharide is purified. The soluble polysaccharide fraction may be used to produce food types with desired taste, texture, quality, viscosity and odor. The presence of soluble polysaccharides may also contribute to the production of a better quality product that does not produce a precipitate (or granular) in the food.

[0169] The dissolving step may be a chemical or thermochemical treatment of the biomass. The chemical or thermochemical treatment may include one or more aqueous solutions. The aqueous solution may contain one or more salts, acids, bases or ions. The aqueous solution may include one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, ammonia, dilute sulfuric acid, dilute acetic acid, dilute hydrochloric acid or dilute phosphoric acid.

[0170] The aqueous solution may be an alkaline solution having a pH of 10 to 14. The aqueous solution may be an alkaline solution having 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 may be an acid solution having a pH of 2 to 6. The aqueous solution may be an acid solution having 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.

[0171] The dissolving step may be performed at a temperature of 30°C to 180°C. The dissolving step may be performed at a temperature of at least 30°C. The dissolving step may be performed at a temperature of up to 180°C. The dissolving step may be performed at a temperature of 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 dissolving step may be performed at a temperature of at least 30°C, 60°C, 90°C, 120°C, 150°C, or 180°C.

[0172] The duration of the dissolution step can be varied depending on the biomass used, the desired soluble polysaccharide component and / or the complexity of the desired soluble polysaccharide. The dissolution step can be performed from about 10 minutes to about 24 hours. The dissolution step can be performed for at least 10 minutes. The dissolution step can be performed for up to 60 minutes. The dissolution step can be performed for 10 minutes to 30 minutes, 10 minutes to 60 minutes, or 30 minutes to 60 minutes. The dissolution step can be performed for at least 10 minutes, 30 minutes, or 60 minutes. The dissolution step can be performed for 1 hour to 24 hours. The dissolution step can be performed for at least 1 hour. The dissolution step can be performed for up to 24 hours. The dissolving step can be performed 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 dissolving step can be performed for at least 1 hour, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours.

[0173] 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 may be removed and / or purified. In some cases, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80%, or 100% of the soluble polysaccharides may be removed and / or purified.

[0174] Mild pretreatment

[0175] Biomass can be subjected to a mild pretreatment step. The mild pretreatment step can dissolve polysaccharide fractions or monosaccharide and / or disaccharide fractions from the biomass. In some cases, the mild pretreatment step can include at least a portion or all of the dissolving step. In other words, the dissolving step can be a part or 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 once or multiple times. The mild pretreatment can be performed 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.

[0176] The mild pretreatment step may be an incubation step or a washing step. The biomass (physically treated or untreated) may be incubated in an aqueous solution. The aqueous solution may be water, or the aqueous solution may contain salts, acids, bases, ions, alcohols and / or other chemicals. The pH of the aqueous solution may be between 6.2 and 8.5. The pH of the aqueous solution may be at least 6.2. The pH of the aqueous solution may be at most 8.5. The pH of the aqueous solution may be between 6.2 and 6.5, 6.2 and 7, 6.2 and 7.2, 6.2 and 7.5, 6.2 and 7.7, 6.2 and 8, 6.2 and 8.2, 6.2 and 8.5, 6.5 and 7, 6.5 and 7.2, 6.5 and 7.5, 6.5 and 7.7, 6.5 and 8, 6.5 and 8.2, 6.5 and 8.5, 7 and 7.2, 7 and 7.5 The pH of the aqueous solution may be 6.2, 6.5, 7, 7.2, 7.5, 7.7, 8, 8.2, or 8.5.

[0177] The pH of the aqueous solution may be from 9 to 12. The pH of the aqueous solution may be at least 9. The pH of the aqueous solution may be up to 12. The pH of the aqueous solution may be from 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 may be 9, 9.5, 10, 10.5, 11, 11.5, or 12.

[0178] The incubation step may be performed for 15 minutes to 60 minutes. The incubation step may be performed for at least 15 minutes. The incubation step may be performed for up to 60 minutes. The incubation step may be performed 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 may be performed for at least 15 minutes, 30 minutes, 45 minutes or 60 minutes. The incubation step may be performed for 1 hour to 24 hours. The incubation step may be performed for at least 1 hour. The incubation step may be performed for up to 24 hours. The incubation step may be performed 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 may be performed for at least 1 hour, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours.

[0179] After the mild pretreatment step, dissolved polysaccharides, monosaccharides and / or disaccharides may be removed from the aqueous solution. In some cases, after the mild pretreatment step, monosaccharides and / or disaccharides may be removed from the aqueous solution.

[0180] 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 may be removed and / or purified. In some cases, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80%, or 100% of the dissolved polysaccharides can be removed and / or purified following the mild pretreatment step.

[0181] After the mild pretreatment step, monosaccharides and / or disaccharides may be removed from the solution. A portion of the dissolved monosaccharides and / or disaccharides may be removed from the aqueous solution after the incubation step. In some cases, at least 5% of the dissolved monosaccharides and / or disaccharides may be removed. In some cases, up to 100% of the dissolved monosaccharides and / or disaccharides may 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 may be removed. In some cases, after the gentle pretreatment step, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80% or 100% of the dissolved monosaccharides and / or disaccharides may be removed and / or purified. After the incubation step, the monosaccharide and / or disaccharide portions may be discarded. In some cases, the portion of dissolved monosaccharides and / or disaccharides removed in this step may not be combined with a portion of one or more oligosaccharides produced in the biomass treatment.

[0182] After the mild pretreatment step, the aqueous solution may be removed from the biomass. After the mild pretreatment step, a portion of the aqueous solution may be removed from the biomass. After the mild pretreatment step, 5% to 100% of the aqueous solution may be removed from the biomass. After the mild pretreatment step, at least 5% of the aqueous solution may be removed from the biomass. After the mild pretreatment step, up to 98% of the aqueous solution may be removed from the biomass. After the 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 %. The present invention relates to a biomass pretreatment method comprising the steps of: (a) preparing biomass from a biomass comprising: a ...

[0183] Strong preprocessing

[0184] Biomass can be subjected to a strong pretreatment step. The strong pretreatment step can dissolve the polysaccharide fraction or the monosaccharide fraction and / or the disaccharide fraction from the biomass. The strong pretreatment step can be performed to make the biomass more easily digested by enzymes. Strong pretreatment can help to destroy the hydrogen bonds in the biomass. The strong pretreatment step can be a washing step, a thermochemical step or a chemical treatment step. Strong pretreatment can be performed before a physical or mechanical pretreatment step. Strong pretreatment steps can be performed after a physical or mechanical pretreatment step. Strong pretreatment steps can be performed before a mild pretreatment step. Strong pretreatment steps can be performed after a mild pretreatment step. Strong pretreatment steps can be performed once or multiple times. Strong pretreatment can be performed 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.

[0185] The strong pretreatment step can be a thermochemical treatment. Chemical or thermochemical treatment can include one or more aqueous solutions. The aqueous solution can include one or more salts, acids, bases or ions. The aqueous solution can be an alkaline solution that includes one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide or ammonia. The aqueous solution can be an acidic solution that includes 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 discarded.

[0186] Thermochemical pretreatment may be performed at a pH of about 2 to 6.5. Thermochemical pretreatment may be performed at a pH of at least 2. Thermochemical pretreatment may be performed at a pH of up to 6.5. Thermochemical pretreatment may be performed at the following pHs: 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. Thermochemical pretreatment may be performed at a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, or 6.5.

[0187] Thermochemical pretreatment may be performed at a pH of 7.5 to 12. Thermochemical pretreatment may be performed at a pH of at least 7.5. Thermochemical pretreatment may be performed at a pH of up to 12. Thermochemical pretreatment may be performed at a pH of 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, Thermochemical pretreatment may be performed at a pH of about 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12. Thermochemical pretreatment may be performed at a pH of about 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12.

[0188] The thermochemical pretreatment may be performed at a temperature of 50°C to 150°C. The thermochemical pretreatment may be performed at a temperature of at least 50°C. The thermochemical pretreatment may be performed at a temperature of up to 150°C. The thermochemical pretreatment may be performed 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 9 Thermochemical pretreatment may be performed at a temperature of at least 50°C, 75°C, 80°C, 90°C, 100°C, 120°C, 130°C or 150°C.

[0189] The thermochemical treatment may be performed for 0.5 hours to 4 hours. The thermochemical treatment may be performed for at least 0.5 hours. The thermochemical treatment may be performed for up to 4 hours. The thermochemical treatment may be performed 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 Thermochemical treatment may be performed for at least 0.5 hour, 0.75 hour, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours or 4 hours. Thermochemical treatment may be performed for at least 0.5 hour, 0.75 hour, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours or 4 hours.

[0190] Thermochemically treated biomass may contain less than 1% w / w to 30% w / w monosaccharides. Thermochemically treated biomass may contain 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, %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 monosaccharides. The thermochemically treated biomass may comprise 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.

[0191] Thermochemically treated biomass may contain 5% w / w to 50% w / w disaccharides. Thermochemically treated biomass may contain 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. / 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, 2 Thermochemically treated biomass may comprise disaccharides 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. Thermochemically treated biomass may comprise disaccharides 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.

[0192] II. Enzyme Treatment and Downstream Processing

[0193] The method of the present disclosure may also include contacting one or more polysaccharide cutting enzymes with the biomass of thermochemical treatment in a solution or suspension to form one or more oligosaccharides. In addition, these methods may include enrichment solution or suspension to increase the concentration of one or more oligosaccharides to form the composition. One or more oligosaccharides may be purified from a solution or suspension provided herein.

[0194] One or more steps of the method for forming or making the composition can be an enzymatic reaction, wherein one or more enzymes are placed in a suitable reaction vessel together with one or more raw materials or biomass (e.g., plant biomass) and a suitable solvent, and the one or more raw materials or biomass may be soluble or insoluble in water. As used herein, unless otherwise indicated, the term "plant biomass" may be replaced with the term "raw material" or "biomass" (e.g., biomass not derived from plants).

[0195] A variety of enzymes are suitable for enzymatic reactions. Any enzyme that produces oligosaccharides when acting on a raw material containing polysaccharides is suitable. For example, the enzymatic reaction can include cellulase, endoglucanase, cellobiohydrolase, cleavage polysaccharide monooxygenase (LPMO), lichenase, xyloglucan endoglucanase (XEG), mannanase, chitinase, xylanase and / or one or more suitable enzymes.

[0196] In various cases, the enzymatic reaction may comprise a cellulolytic preparation from a species such as Trichoderma reesei, which may be purified and / or pretreated and / or may be 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 may be provided to the enzymatic reaction as purified enzymes, semi-purified mixtures derived from some natural source or laboratory-grown cultures, in the form of microbial strains engineered to produce enzymes, or in any other suitable form. Fusion of these enzymes with other enzymes or with non-enzyme modules such as carbohydrate-binding modules (CBMs) is also contemplated. For example, an LPMO fused to a CBM, a xylanase fused to a CBM, or a xylanase fused to an LPMO may be used.

[0197] Aerobic conditions can be used for one or more enzymatic reactions. Aerobic conditions can include adding oxygen, which can be provided by aerating the substrate mixture with an oxygen-containing gas (such as air). Oxygen-containing bubbles can be introduced into the aqueous substrate mixture for aeration by various systems (such as air ejectors, aeration frits, membrane systems, or internal circulation airlift reactors). In some cases, the molecular oxygen concentration in the enzymatic reaction can be from about 4 mg / L to about 14 mg / L.

[0198] Another exemplary enzyme is a lichenase, which can be selected from 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 bond glucans (which are glucans containing a mixture of β-1,3 bonds and β-1,4 bonds), and can cut them at β-1,4 glycosidic bonds. In the case where the lichenase acts on mixed bond glucans, the β-glucans produced can mainly fall within the size range of 3 to about 7 residues, so they are particularly suitable for food, cosmetics and nutrition products industries. Mixed bond glucans are abundant in members of the grass and horsetail families, and therefore, grass raw materials (such as straw) generally have high levels of mixed bond glucans and can be effectively acted upon by lichenases. Lichenases can include GH5 lichenases derived from Bacillus subtilis.

[0199] Another kind of alternative enzyme is xylanase, and this xylanase can act on the raw material that for example comprises xylan backbone.Xylanase can be for example glucuronoxylanase, arabinoxylanase or glucuronyl arabinoxylanase.This enzyme can act on the various polymers with xylan backbone, such as glucuronoxylan, arabinoxylan and glucuronyl arabinoxylan.These polymers are usually very abundant in the raw material of various plant sources, for example, hardwood and softwood can both comprise suitable polysaccharide, and wherein hardwood usually comprises glucuronoxylan, and softwood usually comprises arabinoglucuronoxylan. Xylanases can include GH5 xylanases from Ruminiclostridium thermocellum and Gonapodya prolifera, and GH30 xylanases from Dickea chrysanthemi, Bacillus subtilis, Bacteroides ovatus, and Trichoderma reesei.

[0200] Another kind of alternative enzyme is mannanase, and this mannanase can act on the raw material that for example comprises mannan backbone.Mannanase can be, for example, mannanase, glucomannanase, galactomannanase or galactoglucomannanase.This enzyme can be active to various polymers with mannan backbone, for example mannan, glucomannan, galactomannan or galactoglucomannan (galactoglucomannan).These polymers are usually very abundant in the raw materials of various plant sources, for example, hardwood and softwood can comprise suitable polysaccharide.Suitable mannanase can comprise the GH5 mannanase that is derived from Trichoderma reesei and Aspergillus niger (Aspergillus niger), and the GH26 mannanase that is derived from Aspergillus niger.

[0201] Other enzymes can include xyloglucanase and xyloglucan endoglucanase (XEG), which are produced by many organisms (including plant pathogenic microorganisms). Xyloglucanase and XEG can act 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 appropriate xyloglucanase or XEG act on xyloglucan, the product can include xyloglucan oligosaccharides with a useful length backbone in the food, cosmetics and nutrition industry. Suitable xyloglucanase can include GH5 xyloglucanase derived from Bacteroides ovatus and GH74 xyloglucanase derived from Trichoderma reesei.

[0202] The enzymatic reaction can be carried out in a 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 regime 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 a desired concentration or until some other requirements are met.

[0203] To ensure optimal contact between the enzyme and the starting materials, the reaction mixture may be stirred continuously or intermittently. Agitation may take the form of (i) rhythmic movement of the entire reaction vessel, (ii) a blower or other stirring device, (iii) bubble sparging, or (iv) any other suitable stirring method.

[0204] The enzymatic reaction can be a microbial fermentation. The temperature and reaction time can be suitable for the growth of the microorganism used. The microorganism can be genetically modified to produce an enzyme suitable for producing an oligosaccharide composition. The microorganism can be a bacterium, such as Escherichia coli, or a fungus, such as Saccharomyces cerevisiae or Trichoderma reesei.

[0205] In some embodiments, an expression vector suitable for modifying the subject microorganism can be used so 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: a regulatory sequence of a heat shock gene, a regulatory sequence of a toxic gene, a regulatory sequence of a spore formation gene, or any other suitable regulatory sequence.

[0206] The enzymatic reaction can be carried out at a temperature or temperature regime 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 a microbial fermentation, the temperature may be suitable for this. For example, the enzymatic reaction may include the growth of E. coli, and / or the temperature may be substantially constant and about 37°C.

[0207] The pH of a solution or suspension can affect the activity of an enzyme. Control of pH can help ensure that the enzymatic reaction proceeds at an appropriate rate. The enzymatic reaction can be carried out in a pH range of 2 to 10, 3 to 8, or 4 to 6.

[0208] 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 of time can be 1 minute to 6 days, 0.5 day to 5 days, or 16 hours to 96 hours. Alternatively, the reaction can be allowed to proceed until no further catalysis occurs.

[0209] One or more raw materials added to the enzymatic reaction can comprise polysaccharide.Such polysaccharide can be produced by the independent reaction carried out in reaction vessel simultaneously or substantially simultaneously.The polysaccharide existing in the enzymatic reaction can be partially cut into useful oligosaccharides, leave part cutting or uncut polysaccharide by enzyme, it can include but not limited to Mierocrystalline cellulose, xylan (such as glucuronic acid xylan, arabinoxylan or glucuronic acid arabinoxylan), mannan (such as glucomannan, galactomannan or galactoglucomannan), mixed bond glucan, xyloglucan chitin, chitosan or lignocellulose.

[0210] 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 undigested polysaccharide-containing feedstock remains. This can be monitored or checked by a reducing end assay (such as an anthrone assay) and / or chromatography (such as thin layer chromatography and / or high performance anion exchange chromatography).

[0211] Any material comprising suitable polysaccharide can form a part for raw material. Because food, cosmetics and nutrition products industry use various oligosaccharides conventionally, the polysaccharide that is suitable for participating in enzymatic reaction is not particularly limited. The raw material that is applicable to produce oligosaccharide spectrum can include for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronic acid xylan, arabinoxylan and glucuronic acid arabinoxylan) and / or mannan (such as glucomannan, galactomannan or galactoglucomannan), however, it is envisioned that any raw material that can work suitably. Raw material can include sugar cane, corn stalk, corn cob, wheat bran, wheat straw, hardwood, softwood or any other suitable biomass or plant biomass.

[0212] The raw materials containing such polysaccharides are also not particularly limited, as most plant materials are rich in such polymers. Thus, the raw materials may include plant biomass, such as grains, husks, bean pods, seed coats and / or other seed materials; seaweed; corn stalks, rice straw, bagasse, miscanthus, sorghum residue, switchgrass, bamboo and / or other monocot tissues; water hyacinth, leaf tissue, roots and / or other plant matter; hardwood, hardwood chips, hardwood pulp, softwood, softwood chips, softwood pulp, paper, pulp, cardboard and / or other wood raw materials; crab shells, squid biomass, shrimp shells and / or other marine biomass, and / or any combination of suitable raw materials. The raw materials may include wheat straw or wood. Since any given natural raw material may contain a mixture of different polysaccharides, a mixture of different enzymes will sometimes be beneficial. Such a mixture may contain one or more of any suitable enzymes as discussed herein. For example, such a mixture can include LPMO and endoglucanase, xylanase and lichenase, cellobiohydrolase and mannanase or endoglucanase and cellobiohydrolase. In some embodiments, the enzyme partner can be present in a molar ratio of, for example, 1:100 to 100:1. In addition, since many suitable raw materials are difficult to degrade, it is envisioned that the raw materials are pre-treated.

[0213] After the enzymatic reaction proceeds 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 enzyme used, the process can be performed in a variety of ways. Since the reaction mixture usually comprises a mixture of soluble oligosaccharides and insoluble polysaccharides, the reaction mixture can be filtered to remove insoluble matter and prepare the resulting soluble oligosaccharides for further processing.

[0214] Oligosaccharides can also be separated from polysaccharides in a variety of ways. Oligosaccharides can be separated based on solubility, thereby only extracting the composition of soluble sugars for further processing, and / or by chromatographic separation to produce a composition with narrower oligosaccharide chain length bands. For example, 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 the sugar present in the isolated composition will depend on the original enzymatic reaction usually, because different polysaccharides usually reduce solubility with length at different rates.

[0215] It is also envisioned that all or part of the oligosaccharides produced may be further processed to produce further products before incorporation into food, cosmetics or nutritional products. The further processing may include any chemical, physical or enzymatic steps, such as reduction, such as reductive amination when appropriate; oxidation, caramelization, modification with Schiff bases or by Maillard reaction or by any combination of these steps, and different products that achieve or improve performance for the desired purpose may be provided. For example, caramelization characteristics, calorific value, flavor and color may be modified. The oligosaccharides may also be purified, for example by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0216] It is also contemplated that all or part of the polysaccharide fraction produced may be further processed prior to incorporation into food, cosmetics or nutritional products to produce products with improved properties. Such further processing may include any chemical, physical or enzymatic steps, such as alkylation or acid treatment. The polysaccharide may also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0217] In some cases, after modification and / or purification of the oligosaccharide and polysaccharide fractions, all or part of the fractions can then be reconstituted at a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, e.g., 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 component and the modifications and purifications that have been applied to the fractions. It may not be necessary for all of the oligosaccharides and polysaccharides isolated from the enzymatic reaction to be reconstituted.

[0218] The fractions may be reconstituted in a variety of ways, for example by mixing a solution comprising all or part of the oligosaccharide fraction with a solution and / or suspension comprising all or part of the polysaccharide fraction, which may be further dried, lyophilized or otherwise concentrated. The fractions may also be reconstituted by mixing a dry form comprising all or part of the oligosaccharide fraction produced by drying, lyophilizing or concentrating in some other manner with a dry form comprising all or part of the polysaccharide fraction produced by drying, lyophilizing or concentrating in some other manner.

[0219] The oligosaccharide component of the final composition can include one or more oligosaccharides of any type. For example, the oligosaccharide component can include cellooligosaccharides, xylo-oligosaccharides, mixed-linkage glucan oligosaccharides, manno-oligosaccharides, xyloglucan oligosaccharides, chito-oligosaccharides, arabinoxylo-oligosaccharides, or derivatives of any of the above oligosaccharides.

[0220] Any such dry or liquid composition may be considered as an ingredient suitable for incorporation into a food, cosmetic or nutraceutical at any stage of the process. This includes compositions that may be considered as intermediates during the process, such as compositions formed after reconstitution of the oligosaccharide and polysaccharide fractions prior to any further purification, optimization, drying, dissolution or any other such step, as well as final compositions obtained from the process.

[0221] As described herein, the dry composition can be formed by drying and / or freeze-drying. The dry composition can be dissolved in a solution of various liquids including water, syrups, pastes, solvents, alcohols, etc. to form a liquid composition ingredient suitable for incorporation into food, cosmetics, or nutritional products. The liquid composition is particularly useful for foods that require a silky texture, such as candy, chocolate, and yogurt.

[0222] In some embodiments, following modification and / or purification of the oligosaccharide fraction and the polysaccharide fraction, all or part of the fractions can then be reconstituted at a ratio of 1:100 to 1:1 polysaccharide:oligosaccharide, e.g., 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 component and the modifications and purification that have been applied to the fractions.

[0223] After obtaining the composition of the oligosaccharide product suitable for the application under consideration, further processing and / or separation can be carried out. Food, cosmetics or nutritional products derived from the composition can provide a wide range of potential uses. The composition as described herein can be used in applications where oligosaccharides, sugars, bulking sweeteners, low-intensity sweeteners or other related food ingredients are routinely used.

[0224] The polysaccharide cleaving enzyme can be one of the following: a cellulase, a xylanase, a xyloglucanase, an endoglucanase, a cellobiohydrolase, a mannanase, a lichenase or a lytic polysaccharide monooxygenase (LPMO), for example, selected from AA9, AA10, AA11, AA13, AA14 and AA15. The polysaccharide cleaving enzyme can be prepared by Trichoderma reesei fungus, and / or the enzymatic reaction is carried out until 5-75%, 5-65% or 5-50% undigested polysaccharide-containing feedstock remains.

[0225] The polysaccharide cleaving enzyme can be operatively linked to a catalytic or non-catalytic module, for example wherein the polysaccharide cleaving enzyme can be operatively linked to a non-catalytic module and the non-catalytic module is a carbohydrate binding module.

[0226] In various embodiments, after separation of the one or more oligosaccharides and the one or more polysaccharides, the one or more oligosaccharides and the 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.

[0227] In some embodiments, the composition may comprise three or more oligosaccharides of different molecular weights, wherein the method may comprise forming the three or more oligosaccharides by an enzymatic reaction, wherein the enzymatic reaction comprises the steps of contacting one or more polysaccharide cleaving enzymes with one or more starting materials in a solution or suspension.

[0228] III. Food, cosmetic or nutritional supplement ingredients

[0229] Composition

[0230] The polysaccharide component of the composition can include one or more polysaccharides of any type. For example, the polysaccharide can include cellulose, lignocellulose, xylan, mixed bond glucan, mannan, xyloglucan, chitin, chitosan, or any derivative of the above polysaccharides.

[0231] The composition or ingredient may comprise a variety of oligosaccharides. The composition may include different amounts of oligosaccharides, for example, depending on the desired properties of the composition. In some cases, the composition may comprise at least 20% by dry weight, for example at least 30% by dry weight, of cellooligosaccharides having a degree of polymerization of 2 to 6, and / or the composition may comprise at least 20% by dry weight, for example at least 30% by dry weight, of xylooligosaccharides having a degree of polymerization of 2 to 12, and / or the composition may comprise at least 20% by dry weight, for example at least 30% by dry weight, of mixed bond glucan oligosaccharides having a degree of polymerization of 2 to 5, and / or the composition may comprise at least 20% by dry weight, for example at least 30% by dry weight, of At least 30% by dry weight of manno-oligosaccharides with a degree of polymerization of 2 to 12, and / or the composition may comprise at least 20% by dry weight, e.g. at least 30% by dry weight, of xyloglucan-oligosaccharides with a degree of polymerization of 4 to 12, and / or the composition may comprise at least 20% by dry weight, e.g. at least 30% by dry weight, of chito-oligosaccharides with a degree of polymerization of 2 to 12, and / or the composition may comprise at least 20% by dry weight, e.g. at least 30% by dry weight, of arabinoxylo-oligosaccharides with a degree of polymerization of 3 to 15. In certain embodiments, it is understood that the composition may comprise up to 100% by dry weight of the above oligosaccharides, and thus the above embodiments (wherein the oligosaccharides are present at least 20% by dry weight) do not comprise all seven types of oligosaccharides.

[0232] In various embodiments, the composition or ingredient may comprise 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 may comprise 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 may comprise 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 may comprise 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 may comprise 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.

[0233] In various embodiments, the composition or ingredient may comprise about 20% to about 90% w / w xylooligosaccharides having a degree of polymerization of 2 to 5. In certain embodiments, the composition or ingredient may comprise 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 xylooligosaccharides having a degree of polymerization of 2 to 5. For example, the composition may comprise at least 30% w / w xylooligosaccharides 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 xylooligosaccharides having a degree of polymerization of 2 to 5.

[0234] In certain embodiments, the composition or ingredient may comprise from about 0.1% to about 15% w / w arabinoxylo-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 arabinoxylo-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 arabinoxylo-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 arabinoxylo-oligosaccharides having a degree of polymerization of 3 to 15.

[0235] In some embodiments, provided herein is the use of an oligosaccharide mixture in forming a food, cosmetic or nutraceutical, wherein the oligosaccharide mixture comprises two oligosaccharides selected from the following list:

[0236] i) cellooligosaccharides having a degree of polymerization of 2 to 6;

[0237] ii) xylo-oligosaccharides having a degree of polymerization of 2 to 12;

[0238] iii) mixed-linkage dextran oligosaccharides having a degree of polymerization of 2 to 5;

[0239] iv) manno-oligosaccharides having a degree of polymerization of 2 to 12;

[0240] v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 10;

[0241] vi) chitosan oligosaccharides having a degree of polymerization of 2 to 12; and / or

[0242] vii) Arabinoxylo-oligosaccharides having a degree of polymerisation 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.

[0243] In certain cases, the arabinoxylo-oligosaccharide may comprise at least 0.1% arabinose residues.The arabinoxylo-oligosaccharide may comprise at least 0.1%, 0.2%, 0.5%, 1%, 5% or 10% arabinose residues.

[0244] The amount of each oligosaccharide may vary depending on the desired properties of the resulting food, cosmetic or nutraceutical. For example, the two oligosaccharides may 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.

[0245] Oligosaccharide mixture can also include the third oligosaccharide.The oligosaccharide mixture can include the third oligosaccharide and the fourth oligosaccharide.The oligosaccharide mixture can include the third oligosaccharide, the fourth oligosaccharide and the fifth oligosaccharide.The oligosaccharide mixture can also include the third oligosaccharide, the fourth oligosaccharide, the fifth oligosaccharide and the sixth oligosaccharide.The oligosaccharide mixture can also include the third oligosaccharide, the fourth oligosaccharide, the fifth oligosaccharide, the sixth oligosaccharide and the seventh oligosaccharide.These oligosaccharides can be selected from the list identical with at least two oligosaccharides provided above.

[0246] The oligosaccharide mixture of at least two oligosaccharides can include cellooligosaccharides, for example, a combination of cellooligosaccharides and xylooligosaccharides. Optional compositions can include a combination of cellooligosaccharides and manno-oligosaccharides. In some embodiments, the oligosaccharide mixture can include cellooligosaccharides, xylooligosaccharides and arabinoxylo-oligosaccharides combined with each other.

[0247] The oligosaccharide mixture of at least two oligosaccharides may further comprise a polysaccharide, for example a cellulosic polysaccharide, such as cellulose; or a polysaccharide derivative, for example a cellulose derivative, such as carboxymethyl cellulose; or a polysaccharide aggregate, for example a portion of lignocellulosic biomass. In some cases, the ratio in the combination may be 1:100 to 1:1 polysaccharide / polysaccharide derivative / polysaccharide aggregate: oligosaccharide, for example 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4 or 1:60 to 1:5. In this way, the ratio between the first oligosaccharide, the second oligosaccharide and the polysaccharide may be 2:2:1 to 30:30:1, for example about 3:3:1.

[0248] Combination of oligosaccharides

[0249] Composition can comprise the mixture of one or more oligosaccharides.The mixture of oligosaccharides can comprise two forms or types of oligosaccharides, for example, cello-oligosaccharides and xylo-oligosaccharides.The oligosaccharide mixture can comprise three forms of oligosaccharides, for example, cello-oligosaccharides, manno-oligosaccharides and xylo-oligosaccharides.The oligosaccharide mixture can comprise four forms of oligosaccharides, for example, cello-oligosaccharides, manno-oligosaccharides, mixed bond glucan oligosaccharides and chito-oligosaccharides.

[0250] The oligosaccharide mixture may include two forms of oligosaccharides, for example, a first oligosaccharide and a second oligosaccharide. The oligosaccharide mixture may include about 5% w / w of the first oligosaccharide and about 95% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 10% w / w of the first oligosaccharide and about 90% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 15% w / w of the first oligosaccharide and about 85% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 20% w / w of the first oligosaccharide and about 80% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 25% w / w of the first oligosaccharide and about 75% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 30% w / w of the first oligosaccharide and about 70% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 35% w / w of the first oligosaccharide and about 65% w / w of the second oligosaccharide. The oligosaccharide mixture may include about 40% w / w of the first oligosaccharide and about 50% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 45% w / w of the first oligosaccharide and 55% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 50% w / w of the first oligosaccharide and 50% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 55% w / w of the first oligosaccharide and 45% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 60% w / w of the first oligosaccharide and 30% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 65% w / w of the first oligosaccharide and 35% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 70% w / w of the first oligosaccharide and 30% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 75% w / w of the first oligosaccharide and 25% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 80% w / w of the first oligosaccharide and 20% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 85% w / w of the first oligosaccharide and 15% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 90% w / w of the first oligosaccharide and 10% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 95% w / w of the first oligosaccharide and 5% w / w of the second oligosaccharide. In some cases, the first oligosaccharide may be a cello-oligosaccharide, and the second oligosaccharide may be a xylo-oligosaccharide. In some cases, the first oligosaccharide may be a cello-oligosaccharide, and the second oligosaccharide may be a manno-oligosaccharide. In some embodiments, the first oligosaccharide may be a xylo-oligosaccharide, and the second oligosaccharide may be a manno-oligosaccharide. Other combinations of the first oligosaccharide and the second oligosaccharide are also within the scope of the present disclosure.

[0251] The oligosaccharide mixture may include three forms of oligosaccharides, such as the first oligosaccharide, the second oligosaccharide and the third oligosaccharide. The oligosaccharide mixture may include 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 include 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 include 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 include 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 include 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 manno-oligosaccharide, the second oligosaccharide may be xylo-oligosaccharide, and the third oligosaccharide may be cello-oligosaccharide. In some examples, the first oligosaccharide can be a xyloglucan-oligosaccharide, the second oligosaccharide can be a xyloglucan-oligosaccharide, and the third oligosaccharide can be a cellooligosaccharide.Other combinations of the first oligosaccharide, the second oligosaccharide, and the third oligosaccharide are also within the scope of the present disclosure.

[0252] Oligosaccharide mixture can comprise two or more kinds of oligosaccharides, the first oligosaccharide and the second oligosaccharide that is different from the first oligosaccharide.For example, the first oligosaccharide can be xylo-oligosaccharide or cello-oligosaccharide or manno-oligosaccharide or other oligosaccharides provided herein, and the second oligosaccharide can be xylo-oligosaccharide or cello-oligosaccharide or manno-oligosaccharide or other oligosaccharides that are not used as the first oligosaccharide.In other words, the first oligosaccharide can be different from the second oligosaccharide (for example, the first oligosaccharide can be an oligosaccharide different from the second oligosaccharide).In the mixture, the ratio of the first oligosaccharide and the second oligosaccharide can be about 1:1,1:2,1:3,1:4,1:5,1:6,1:7,1:8 or 1:9.

[0253] The ratio of the first oligosaccharide to the second oligosaccharide in the mixture can be about 2:1, 2:3, 2:5, 2:7 or 2:9. Oligosaccharides can be cellooligosaccharides, manno-oligosaccharides, xyloglucan-oligosaccharides, mixed bond oligosaccharides, chitosan oligosaccharides, arabinoxylo-oligosaccharides or other oligosaccharides provided herein, wherein the first oligosaccharide is selected to be an oligosaccharide different from the second oligosaccharide. In other words, the first oligosaccharide can be an oligosaccharide different from the second oligosaccharide.

[0254] The ratio of the first oligosaccharide to the second oligosaccharide in the mixture can be about 3: 1, 3: 2, 3: 4, 3: 5, 3: 7 or 3: 8. The oligosaccharide can be a cellooligosaccharide, a manno-oligosaccharide, a xylo-oligosaccharide, a xyloglucan-oligosaccharide, a mixed-linkage oligosaccharide, a chito-oligosaccharide, an arabinoxylo-oligosaccharide or other oligosaccharide provided herein, wherein the first oligosaccharide is selected to be an oligosaccharide different from the second oligosaccharide.

[0255] In the oligosaccharide mixture comprising two or more oligosaccharides, the ratio of the first oligosaccharide to the second oligosaccharide can be 1:9 to 9:1, 1:4 to 4:1, 1:3 to 3:1, or 2:3 to 3:2. The oligosaccharide can be a cellooligosaccharide, a manno-oligosaccharide, a xylo-oligosaccharide, a xyloglucan-oligosaccharide, a mixed bond oligosaccharide, a chitosan oligosaccharide, an arabinoxylo-oligosaccharide or other oligosaccharide provided herein, wherein the first oligosaccharide is selected to be an oligosaccharide different from the second oligosaccharide.

[0256] In some cases, a 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 cellobiose, xylobiose, mannobiose (e.g., Man-β-1,4-Man), Glc-β-1,4-Man, Man-β-1,4-Glc, laminarin, gentiobiose, sophorose, maltose, lactose, or sucrose. In some cases, the composition or ingredient may 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 some cases, the composition or ingredient may 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 xylotetrose, cellotetraose, monoarabinosylated xylotriose, monoglucuronosylated xylotriose, diarabinosylated xylobiose, diglucuronosylated xylobiose, maltotetraose, mannotetraose (e.g., 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-Gl c-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 some cases, The composition or ingredient may 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 xylopentose, cellopentaose, monoarabinosylated xylotetrose, monoglucuronosylated xylotetrose, diarabinosylated xylotriose, diglucuronosylated xylotriose, maltopentaose, mannopentaose (e.g., Man-β-1,4-Man-β-1,4-Man-β-1,4-Man-β-1,4-Man), a mixed linkage glucan-derived pentasaccharide, or a mannan-derived pentasaccharide.

[0257] The composition or ingredient may include 1% to 50%, 5% to 40%, 10% to 30%, or 15% to 25% w / w cellobiose. The composition or ingredient may include 2.5% to 90%, 5% to 80%, 10% to 70%, or 20% to 60% w / w xylobiose. The composition or ingredient may include 2.5% to 75%, 5% to 50%, 10% to 40%, or 20% to 30% w / w xylotriose.

[0258] Oligosaccharide compositions with different degrees of polymerization

[0259] 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.

[0260] The concentration of xylooligosaccharides with a degree of polymerization of 2 in the xylooligosaccharide mixture may be from 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.

[0261] The concentration of xylo-oligosaccharides with a degree of polymerization of 3 in the xylo-oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0262] The concentration of xylo-oligosaccharides with a degree of polymerization of 4 in the xylo-oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0263] The concentration of xylo-oligosaccharides with a degree of polymerization of 5 in the xylo-oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0264] The concentration of xylo-oligosaccharides with a degree of polymerization of 6 in the xylo-oligosaccharide mixture may be about 5% to about 25% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0265] The concentration of xylo-oligosaccharides with a degree of polymerization of 7 in the xylo-oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 7 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0266] The concentration of xylo-oligosaccharides with a degree of polymerization of 8 in the xylo-oligosaccharide mixture may be from about 1% to about 15% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 8 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0267] The concentration of xylooligosaccharides with a degree of polymerization of 9 in the xylooligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of xylooligosaccharides with a degree of polymerization of 9 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

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

[0269] The concentration of xylo-oligosaccharides with a degree of polymerization of 11 in the xylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 11 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0270] The concentration of xylo-oligosaccharides with a degree of polymerization of 12 in the xylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of xylo-oligosaccharides with a degree of polymerization of 12 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0271] The concentration of the cellooligosaccharide with a degree of polymerization of 2 in the cellooligosaccharide mixture may be from about 2% to about 80% w / w. The concentration of the cellooligosaccharide 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 the cellooligosaccharide 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.

[0272] The concentration of cellooligosaccharides with a degree of polymerization of 3 in the cellooligosaccharide mixture may be about 2% to about 20% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0273] The concentration of cellooligosaccharides with a degree of polymerization of 4 in the cellooligosaccharide mixture may be about 5% to about 20% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0274] The concentration of cellooligosaccharides with a degree of polymerization of 5 in the cellooligosaccharide mixture may be about 5% to about 20% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0275] The concentration of cellooligosaccharides with a degree of polymerization of 6 in the cellooligosaccharide mixture may be about 5% to about 25% w / w. The concentration of cellooligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0276] The concentration of manno-oligosaccharides with a degree of polymerization of 2 in the manno-oligosaccharide mixture may be from about 2% to about 30% w / w. The concentration of manno-oligosaccharides 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.

[0277] The concentration of manno-oligosaccharides with a degree of polymerization of 3 in the manno-oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0278] The concentration of manno-oligosaccharides with a degree of polymerization of 4 in the manno-oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0279] The concentration of manno-oligosaccharides with a degree of polymerization of 5 in the manno-oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0280] The concentration of manno-oligosaccharides with a degree of polymerization of 6 in the manno-oligosaccharide mixture may be about 5% to about 25% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0281] The concentration of manno-oligosaccharides with a degree of polymerization of 7 in the manno-oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 7 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0282] The concentration of manno-oligosaccharides having a degree of polymerization of 8 in the manno-oligosaccharide mixture may be from about 1% to about 15% w / w. The concentration of manno-oligosaccharides having a degree of polymerization of 8 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0283] The concentration of manno-oligosaccharides with a degree of polymerization of 9 in the manno-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 9 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0284] The concentration of manno-oligosaccharides having a degree of polymerization of 10 in the manno-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of manno-oligosaccharides having a degree of polymerization of 10 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0285] The concentration of manno-oligosaccharides with a degree of polymerization of 11 in the manno-oligosaccharide mixture may be about 2% to about 15% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 11 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0286] The concentration of manno-oligosaccharides with a degree of polymerization of 12 in the manno-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of manno-oligosaccharides with a degree of polymerization of 12 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0287] The concentration of xyloglucan oligosaccharides with a degree of polymerization of 4 in the xyloglucan oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of xyloglucan oligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0288] The concentration of xyloglucan oligosaccharides with a degree of polymerization of 5 in the xyloglucan oligosaccharide mixture may be from about 5% to about 20% w / w. The concentration of xyloglucan oligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0289] The concentration of xyloglucan oligosaccharides with a degree of polymerization of 6 in the xyloglucan oligosaccharide mixture may be from about 5% to about 25% w / w. The concentration of xyloglucan oligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0290] The concentration of xyloglucan oligosaccharides with a degree of polymerization of 7 in the xyloglucan oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of xyloglucan oligosaccharides with a degree of polymerization of 7 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0291] The concentration of xyloglucan oligosaccharides having a degree of polymerization of 8 in the xyloglucan oligosaccharide mixture may be from about 1% to about 15% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization of 8 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0292] The concentration of xyloglucan oligosaccharides with a degree of polymerization of 9 in the xyloglucan oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of xyloglucan oligosaccharides with a degree of polymerization of 9 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

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

[0294] The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 2 in the mixed bond glucan oligosaccharide mixture may be from about 2% to about 30% w / w. The concentration of mixed bond glucan oligosaccharides 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.

[0295] The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 3 in the mixed bond glucan oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0296] The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 4 in the mixed bond glucan oligosaccharide mixture may be from about 5% to about 20% w / w. The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0297] The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 5 in the mixed bond glucan oligosaccharide mixture may be from about 5% to about 20% w / w. The concentration of mixed bond glucan oligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0298] The concentration of chitosan oligosaccharides with a degree of polymerization of 2 in the chitosan oligosaccharide mixture may be from about 2% to about 30% w / w. The concentration of chitosan oligosaccharides 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.

[0299] The concentration of chitosan oligosaccharides with a degree of polymerization of 3 in the chitosan oligosaccharide mixture may be about 2% to about 20% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0300] The concentration of chitosan oligosaccharides with a degree of polymerization of 4 in the chitosan oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0301] The concentration of chitosan oligosaccharides with a degree of polymerization of 5 in the chitosan oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0302] The concentration of chitosan oligosaccharides with a degree of polymerization of 6 in the chitosan oligosaccharide mixture may be about 5% to about 25% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0303] The concentration of chitosan oligosaccharide with a degree of polymerization of 7 in the chitosan oligosaccharide mixture may be about 2% to about 20% w / w. The concentration of chitosan oligosaccharide with a degree of polymerization of 7 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0304] The concentration of chitosan oligosaccharides with a degree of polymerization of 8 in the chitosan oligosaccharide mixture may be from about 1% to about 15% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 8 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0305] The concentration of chitosan oligosaccharides with a degree of polymerization of 9 in the chitosan oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 9 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

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

[0307] The concentration of chitosan oligosaccharides with a degree of polymerization of 11 in the chitosan oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 11 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0308] The concentration of chitosan oligosaccharides with a degree of polymerization of 12 in the chitosan oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of chitosan oligosaccharides with a degree of polymerization of 12 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0309] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 3 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 3 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0310] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 4 in the arabinoxylo-oligosaccharide mixture may be about 5% to about 20% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 4 may be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0311] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 5 in the arabinoxylo-oligosaccharide mixture may be from about 5% to about 20% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 5 may be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0312] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 6 in the arabinoxylo-oligosaccharide mixture may be from about 5% to about 25% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 6 may be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0313] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 7 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 20% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 7 may be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0314] The concentration of arabinoxylo-oligosaccharides having a degree of polymerization of 8 in the arabinoxylo-oligosaccharide mixture may be from about 1% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides having a degree of polymerization of 8 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0315] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 9 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 9 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0316] The concentration of arabinoxylo-oligosaccharides having a degree of polymerization of 10 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides having a degree of polymerization of 10 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0317] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 11 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 11 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0318] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 12 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 12 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0319] The concentration of arabinoxylo-oligosaccharides having a degree of polymerization of 13 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides having a degree of polymerization of 13 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0320] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 14 in the arabinoxylo-oligosaccharide mixture may be from about 2% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 14 may be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

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

[0322] The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 3 to 12 in the arabinoxylo-oligosaccharide mixture may be from about 0.1% to about 15% w / w. The concentration of arabinoxylo-oligosaccharides with a degree of polymerization of 3 to 12 may 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.

[0323] Compositions having combinations of monosaccharides, polysaccharides and / or oligosaccharides

[0324] In some embodiments, the composition or ingredient (eg, an ingredient for human consumption) is soluble in water. The solubility of the ingredient in water may be at least 80 g of the ingredient per 100 g of water at 50°C.

[0325] Ingredients can be combined with liquids to form liquid ingredients. In some cases, the viscosity of the liquid ingredients can be comparable or similar to the viscosity of corn syrup. In some other cases, the viscosity of the liquid ingredients can be comparable or similar to the viscosity of high fructose corn syrup. For example, the viscosity of the liquid ingredients can be 5cps to 100,000cps, 8,000cps to 100,000cps, 10,000cps to 50,000cps or 15,000cps to 25,000cps. In addition, the calories per gram of the liquid ingredients are lower than corn syrup or high fructose corn syrup. The glycemic index of the liquid ingredients is lower than corn syrup or high fructose corn syrup.

[0326] In some embodiments, the liquid may include water or any other suitable liquid. The liquid component may include at least 5%, 10%, 20%, 30%, 40% or 50% dry weight of at least one oligosaccharide. In addition, the liquid component may contain at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% dry weight of at least one polysaccharide. For example, the liquid component may 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.

[0327] The liquid component comprises at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% xylan by dry weight. The liquid component may comprise at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% mannan by dry weight. The liquid component may comprise at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% cellulose derivative by dry weight.

[0328] In various cases, the polysaccharide concentration of the liquid composition 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 composition can be 0.1% to 50% w / v. The liquid composition 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 composition can contain a ratio of the amount of polysaccharides to oligosaccharides of 1:100 to 1:1.

[0329] The one or more soluble polysaccharides may comprise one or more of mannan, xylan, mixed bond glucan, lignocellulose, hemicellulose, cellulose derivatives, chitosan, xyloglucan or any other suitable soluble polysaccharide. The cellulose derivative may comprise at least one of cellulose acetate, hydroxyethyl cellulose, hydroxymethyl cellulose or any other suitable cellulose derivative.

[0330] The biomass may comprise at least one of sugar cane biomass, corn biomass, wheat biomass, hardwood biomass, softwood biomass, or any other suitable biomass.

[0331] In some cases, the composition for human consumption may include soluble polysaccharides and oligosaccharides, the oligosaccharides comprising at least one of: (i) cellooligosaccharides having a degree of polymerization (DP) of 2 to 6; (ii) xylo-oligosaccharides having a DP of 2 to 12; (iii) manno-oligosaccharides having a DP of 2 to 12; (iv) arabinoxylo-oligosaccharides having a DP of 3 to 15; (v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; or (vi) chito-oligosaccharides having a DP of 2 to 12. The composition may include less than 5% dry weight of soluble polysaccharides. In some cases, the composition may include less than 1%, 2%, 5%, 7.5%, 10% or 20% dry weight of soluble polysaccharides. In some embodiments, the composition may be free of or substantially free of insoluble polysaccharides.

[0332] Composition can comprise the combination of polysaccharide and oligosaccharide.In some embodiments, composition can comprise the combination of oligosaccharide and soluble polysaccharide.Polysaccharide (or soluble polysaccharide) source in such composition can comprise cellulose, such as biomass, for example the undigested component of partially digested biomass, for example the undigested biomass from the reaction identical with the reaction producing oligosaccharide.Polysaccharide in undigested biomass can comprise lignin, polyphenol, cellulose, lignocellulose or any other suitable polysaccharide described herein.Polysaccharide (for example, soluble polysaccharide) can be added to oligosaccharide mixture to improve the gastrointestinal tolerance of oligosaccharide mixture.Oligosaccharide consumption may cause gastrointestinal discomfort, including diarrhea, discomfort and abdominal distention.Compared with commercially available sugar composition or sugar composition mainly comprising monosaccharide and / or disaccharide, composition as described herein can have improved gastrointestinal tolerance, for example less or without discomfort, abdominal distention, diarrhea or gastrointestinal discomfort. For example, a subject who ingests one or more compositions provided herein can have improved gastrointestinal tolerability, e.g., less or no discomfort, bloating, diarrhea, or gastrointestinal discomfort, compared to if or when the subject ingests a commercially available sugar composition or a sugar composition comprising primarily monosaccharides and / or disaccharides.

[0333] The concentration of undigested biomass in the composition may be 1% to 50% w / w. The concentration of undigested biomass in the composition may 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.

[0334] The concentration of the soluble polysaccharide in the composition may be 1% to 50% w / w. The concentration of the soluble polysaccharide in the composition may 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 may 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 may 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 may be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

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

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

[0337] In some embodiments, the composition may comprise at least 5% w / w cellooligosaccharides and at least 5% w / w of a second oligosaccharide (e.g., at least 5% w / w xylo-oligosaccharides, manno-oligosaccharides, mixed-linkage glucan-oligosaccharides, xyloglucan-oligosaccharides, chito-oligosaccharides, arabinoxylo-oligosaccharides, or any other suitable oligosaccharide).

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

[0339] The concentration of chitosan oligosaccharide in the composition may be 1% to 50% w / w. The concentration of chitosan oligosaccharide in the composition may 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 50%, 5% 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.

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

[0341] The concentration of the mixed linkage glucan oligosaccharide in the composition may be 1% to 50% w / w. The concentration of the mixed linkage glucan oligosaccharide in the composition may 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 may 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 may 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 may be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

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

[0343] The composition may include one or more polysaccharides (e.g., one or more soluble polysaccharides) and one or more oligosaccharides. The composition may include a polysaccharide and one type of oligosaccharide. The composition may include a polysaccharide or multiple polysaccharides and two forms of oligosaccharides. The composition may include a polysaccharide or multiple polysaccharides and three forms of oligosaccharides. The composition may include a polysaccharide or multiple polysaccharides and four forms of oligosaccharides. The composition may include a polysaccharide or multiple polysaccharides and five forms of oligosaccharides. The oligosaccharide may be a xylo-oligosaccharide, a cellooligosaccharide, a manno-oligosaccharide, a mixed bond glucan oligosaccharide, a xyloglucan oligosaccharide, a chitosan oligosaccharide, an arabinoxylo-oligosaccharide, or any other suitable oligosaccharide described herein.

[0344] Composition can comprise about 1% to 50%w / w polysaccharide (such as the type of soluble polysaccharide with undigested biomass or extraction), and about 5% to about 95%w / w oligosaccharide.The composition of 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.Oligosaccharide in such mixture can exist greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%w / w.Oligosaccharide can be a mixture of one or more oligosaccharides.For example, as described elsewhere herein, composition can comprise 5%w / w undigested biomass and 50%w / w oligosaccharide mixture. In another instance, as described elsewhere herein, the composition may comprise 2.5% w / w of a soluble polysaccharide and 50% w / w of an oligosaccharide mixture.

[0345] Composition can comprise about 5%w / w polysaccharide (such as in undigested biomass type), and about 5% to about 95%w / w oligosaccharide.Oligosaccharide in such mixture can be greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%w / w existence.Oligosaccharide can be a mixture of one or more oligosaccharides.For example, as described elsewhere herein, composition can comprise 5%w / w undigested biomass and 50%w / w oligosaccharide mixture.In another case, as described elsewhere herein, composition can comprise 5%w / w soluble polysaccharide and 50%w / w oligosaccharide mixture.

[0346] Composition can comprise about 7%w / w polysaccharide (such as in undigested biomass type), and about 5% to about 93%w / w oligosaccharide. 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 in such mixture. Oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, composition can comprise 7%w / w undigested biomass and 50%w / w oligosaccharide mixture. In another case, as described elsewhere herein, composition can comprise 7%w / w soluble polysaccharide and 50%w / w oligosaccharide mixture.

[0347] Composition can comprise about 10%w / w polysaccharide (such as in undigested biomass type), and about 5% to about 90%w / w oligosaccharide. Oligosaccharide can 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 mixture. Oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, composition can comprise 10%w / w undigested biomass and 50%w / w oligosaccharide mixture. In another case, as described elsewhere herein, composition can comprise 10%w / w polysaccharide and 50%w / w oligosaccharide mixture.

[0348] Composition can comprise about 12%w / w polysaccharide (such as in undigested biomass type), and about 5% to about 95%w / w oligosaccharide. Oligosaccharide can 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 mixture. Oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, composition can comprise 12%w / w undigested biomass and 50%w / w oligosaccharide mixture. In another case, as described elsewhere herein, composition can comprise 12%w / w soluble polysaccharide and 50%w / w oligosaccharide mixture.

[0349] Composition can comprise about 15%w / w polysaccharide (such as in undigested biomass type), and about 5% to about 85%w / w oligosaccharide. Oligosaccharide can 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 mixture. Oligosaccharide can be a mixture of one or more oligosaccharides. For example, as described elsewhere herein, composition can comprise 15%w / w undigested biomass and 50%w / w oligosaccharide mixture. In another case, as described elsewhere herein, composition can comprise 15%w / w soluble polysaccharide and 50%w / w oligosaccharide mixture.

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

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

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

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

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

[0355] In some embodiments, the composition or ingredient may include less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w monosaccharide. For example, the composition may include less than 20% w / w monosaccharide. The composition may include 10% to 40%, 15% to 30%, 18% to 25% or about 20% w / w monosaccharide. In some embodiments, the composition or ingredient may include less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w glucose. For example, the composition may include less than 10% w / w glucose. The composition may include 10% to 40%, 15% to 30%, 18% to 25% or about 20% w / w glucose. In some embodiments, the composition or ingredient may include less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w xylose. For example, the composition may comprise less than 10% w / w xylose.The composition may comprise 10% to 40%, 15% to 30%, 18% to 25%, or about 20% w / w xylose.

[0356] In certain cases, the ratio of glucose residues to xylose residues (eg, glucose:xylose) within a composition or ingredient can be 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.

[0357] In certain embodiments, the composition may include less than 30%, 40%, 50%, 60%, 65%, 70%, 75% or 80% w / w disaccharides. For example, the composition may include less than 70% w / w disaccharides. The composition may include 10% to 95%, 15% to 90%, 20% to 80%, 30% to 70% or 40% to 60% w / w disaccharides. The composition may include 5% to 95%, 10% to 92.5%, 15% to 90%, 20% to 70%, 30% to 60% or 40% to 50% disaccharides. In various embodiments, the composition may include at least 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15% or 20% w / w trisaccharides. For example, the composition may include at least 5% w / w trisaccharides. In various embodiments, the composition may comprise at least 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15% or 20% w / w trisaccharides. For example, the composition may comprise at least 5% w / w trisaccharides. The composition may comprise 1% to 75%, 2.5% to 60%, 5% to 50%, 10% to 40% or 20% to 30% trisaccharides. 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 tetrasaccharides. For example, the composition may comprise at least 1% w / w tetrasaccharides. 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 pentasaccharides. For example, the composition may comprise at least 0.1% w / w pentasaccharide.

[0358] Use of the composition as an ingredient

[0359] In some embodiments, the composition is an ingredient (e.g., in a food). In certain embodiments, the ingredient comprises sugars present at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5% by dry weight. The ingredient may consist essentially of sugars. For example, the ingredient may have less than 0.5%, 0.3%, or 0.1% other substances by dry weight.

[0360] As described elsewhere herein, the composition may comprise a mixture of oligosaccharides. The composition may comprise at least two oligosaccharides. For example, the composition may comprise three oligosaccharides. The composition may comprise four oligosaccharides. The composition may comprise five oligosaccharides. The composition may comprise six oligosaccharides. The composition may comprise seven oligosaccharides.

[0361] In some embodiments, the ingredient comprises cellooligosaccharides, such as a combination of cellooligosaccharides and xylo-oligosaccharides.Alternative ingredients may comprise a combination of cellooligosaccharides and manno-oligosaccharides.

[0362] The ingredients may be used to prepare the final product. The ingredients may also be treated in some physical or chemical manner before or during incorporation into the food, cosmetic or nutraceutical. The ingredients may be incorporated directly into the product, or the ingredients may be incorporated into, for example, a dough, cake mix, chocolate mix or other food precursor; into a cosmetic base composition; or into a nutraceutical and, for example, cooked or otherwise treated in a manner that may cause chemical modification, textural change, color change or other modification.

[0363] Food, cosmetics or nutritional products can be produced by ingredients as described herein. For example, in the food industry, the sugar preparations produced by the current method can be used as sweeteners, leavening agents, dietary fiber or humectants added. The ingredients can be used as sugar substitutes. The ingredients can be incorporated into cakes, breads or other baked goods, or incorporated into chocolate or other candies, such as toffee, soft candy, meringue, jam, jelly or caramel; or incorporated into beverages, for example, to provide good taste or color properties or increase dietary fiber content. In some cases, the ingredients can be incorporated into animal feed as raw materials, for example, as separated ingredients or by directly utilizing enzyme reaction mixtures.

[0364] In the cosmetic industry, sugars can be used as ingredients because they can improve texture and moisture retention, act as UV absorbing molecules, maintain gel or cream structure and / or act as leavening agents. The compositions described herein can be incorporated into nutraceutical compositions because the dietary fiber they provide can promote digestive health, good regulation of intestinal flora, and other health benefits. In this case, they can also be used as ingredients in probiotic beverages or other prebiotic or probiotic preparations.

[0365] The compositions or ingredients 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, adhesion, glazing, smoothness, wettability, viscosity, color, hygroscopicity, flavor, bulkiness, water retention, caramelization, surface texture, crystallization, structural properties, and solubility.

[0366] In some cases, the compositions and / or ingredients described herein can provide comparable or better properties for the final product compared to the same properties provided by a sugar mixture mainly comprising monosaccharides and / or disaccharides. The control composition can be a sugar commonly used in consumables, such as a monosaccharide composition, such as glucose, fructose, etc.; a disaccharide composition, such as sucrose or an artificial sugar composition. 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 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.

[0367] In some cases, the compositions described herein can be used as sweetener compositions. Sweetener compositions can be used alone or as an ingredient in a final product. Compared with the same amount of control compositions, the compositions described herein can provide about the same level of sweetness or a higher sweetness, wherein the control compositions mainly comprise monosaccharides and / or disaccharides. Compositions described herein can be used to replace control compositions as sweeteners in final products. In some cases, the sweetness of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than the same amount of control compositions.

[0368] The compositions described herein can provide comparable flavor profiles or better flavor profiles compared to the same amount of control compositions, wherein the control compositions mainly comprise monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control compositions as flavor enhancers in the final product. In some cases, the flavor of the compositions can exceed 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% compared to the same amount of control compositions.

[0369] The compositions described herein can provide a comparable texture profile or a better texture profile compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a texture enhancer in the final product.

[0370] The compositions described herein can provide comparable or better bonding profiles compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as an adhesion enhancer in a final product.

[0371] The compositions described herein can provide comparable or better light-up spectra compared to the same amount of a control composition, wherein the control composition comprises mainly monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition as a light-up enhancer in the final product.

[0372] The compositions described herein can provide comparable or better moisture than the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein can be used to replace the control composition to provide moisture in the final product.

[0373] The compositions described herein can provide a comparable color spectrum or a better color spectrum compared to the same amount of a control composition, 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.

[0374] The compositions described herein can provide comparable dissolution profiles or better dissolution profiles compared to the same amount of control compositions, wherein the control compositions primarily comprise monosaccharides and / or disaccharides. The compositions described herein can be used to replace control compositions as dissolution enhancers 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 the same amount of control compositions.

[0375] The compositions described herein can provide a comparable mouthfeel or a better mouthfeel compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0376] The compositions described herein can provide comparable viscosity or better viscosity compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0377] The compositions described herein can provide comparable or better hygroscopicity than the same amount of a control composition, wherein the control composition comprises primarily 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 the same amount of the control composition.

[0378] The compositions described herein can provide comparable or better water retention than the same amount of a control composition, wherein the control composition comprises primarily 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 the same amount of the control composition.

[0379] The compositions described herein can provide a lower calorie composition compared to the same amount of a control composition, wherein the control composition primarily comprises monosaccharides and / or disaccharides. In some cases, the calorie count of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% lower than the same amount of the control composition.

[0380] The compositions described herein can provide a lower glycemic index compared to the same amount of a control composition, wherein the control composition comprises primarily 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 the same amount of the control composition.

[0381] The compositions described herein can provide comparable or better leavening than the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0382] The compositions described herein can provide comparable caramelization or better caramelization compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0383] The compositions described herein can provide comparable surface texture or better surface texture compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0384] The compositions described herein can provide comparable crystallization or better crystallization compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0385] The compositions described herein can provide comparable textural properties compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0386] The compositions described herein can provide less aftertaste compared to the same amount of a control composition, wherein the control composition comprises primarily monosaccharides and / or disaccharides.

[0387] Different oligosaccharide compositions may have improved dissolution profiles, hygroscopicity profiles and taste profiles compared to the oligosaccharide used alone.

[0388] Compositions as described herein or composition can be used to increase the fiber content of final products such as food or nutritional products.Compared with the control composition of the same amount, 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 any negative or substantially negative effects on any other properties, such as taste, sweetness, mouthfeel, texture, adhesiveness or any other properties 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 the control composition of the same amount.

[0389] The ingredients can be used to modify the properties of a final product, such as a food or a nutraceutical or a cosmetic. To modify the properties of the final product, the final product may additionally comprise a polysaccharide, such as a cellulosic polysaccharide, such as cellulose; or a polysaccharide derivative, such as a cellulose derivative, such as carboxymethyl cellulose; or a polysaccharide aggregate, such as a portion of a lignocellulosic biomass. In some cases, the final product may comprise greater than 0% to 40% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight, for example, greater than 1% to 30% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight, for example greater than 5% to 25% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight, for example greater than 10% to 20% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight.

[0390] The concentration of the composition comprising the mixture of polysaccharides and oligosaccharides in the final product may be anywhere between 0.1% and 40% w / w. The concentration of the composition comprising the mixture of polysaccharides and oligosaccharides in the final product may 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 ... 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 The concentration of the composition comprising a mixture of polysaccharides and oligosaccharides in the final product may be about 0.1%, about 0.5%, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, 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 comprising a mixture of polysaccharides and oligosaccharides in the final product may 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 comprising a mixture of polysaccharides and oligosaccharides in the final product may be at least 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30% or 35% w / w. The concentration of the composition comprising a mixture of polysaccharides and oligosaccharides in the final product may be at most 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35% or 40% w / w.

[0391] In some cases, oligosaccharide mixtures (e.g., cellooligosaccharides and xylooligosaccharides) can form at least 20%, 30%, 40%, 50%, 60% or 70% w / w edible compositions or ingredients. For example, a combination of 50% w / w cellooligosaccharides and xylooligosaccharides can form an edible composition or ingredient.

[0392] In some cases, the ingredients may include maize cob extract (MCE). MCE may be an oligosaccharide mixture mainly consisting of arabinoxy-oligosaccharides, xylo-oligosaccharides, cello-oligosaccharides and cellulose. In some cases, the oligosaccharides may be indigestible or substantially indigestible. Arabinoxy-oligosaccharides may be oligomers connected by β-(1→4)-bonds of a xylose backbone substituted by arabinose side chains. Arabinoxy-oligosaccharides may be indigestible. Arabinoxy-oligosaccharides may 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 arabinoxy-oligosaccharides may be 3 to 15.

[0393] In various cases, xylooligosaccharides can be oligomers of xylose backbones connected by β-(1→4)-bonds. Xylooligosaccharides can be indigestible. Xylooligosaccharides can be produced by hydrolyzing arabinoxylan. In addition, the DP of xylooligosaccharides can be 2 to 8.

[0394] In some cases, cellooligosaccharides can be oligomers whose glucose backbones are 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)-linked glucose units). In addition, the DP of cellooligosaccharides can be 2 to 4, with most having a DP of 2.

[0395] IV. Exemplary Embodiments

[0396] Exemplary methods for extracting soluble polysaccharides for subsequent conjugation with produced oligosaccharides

[0397] Figure 8 is a simplified flow chart illustrating an embodiment of a method of extracting soluble polysaccharides for subsequent combination with produced oligosaccharides to form an ingredient.

[0398] In the illustrated method, extracting or removing at least a portion of soluble polysaccharide 140 before enzyme treatment 150 can ensure to retain or preserve at least a portion of soluble polysaccharide to combine 144 with the oligosaccharide produced, to form composition 105. In some other methods (not shown), soluble polysaccharide can be digested by one or more enzymes, because soluble polysaccharide is not extracted before enzyme treatment. The composition 105 of the illustrated method can be a sweetener or a sugar substitute, which can keep substantially soluble or completely soluble. Therefore, composition 105 can be delivered as a syrupy product (for example, viscous liquid). In some cases, composition 105 can be following substitute or partial substitute: corn syrup, high fructose corn syrup, maple syrup, honey, honey, golden syrup, molasses, glucose syrup, fructose syrup, agave syrup, date syrup, brown rice syrup, coconut syrup, corn syrup or other suitable liquid sweeteners in syrup food.

[0399] As depicted, biomass 107 (e.g., corn cobs or any other suitable biomass) can be subjected to physical pretreatment 110 (e.g., by cutting or any other suitable method of physically pretreating biomass). The physically treated biomass 112 can then undergo or be subjected to thermochemical pretreatment 130 (e.g., 15% w / v cut corn cobs can be heated in 1% w / v NaOH for 1 hour). In some embodiments, thermochemical pretreatment 130 can be followed by a neutralization step (not shown) and then extraction of soluble compounds or materials 140 from the physically pretreated biomass 112. Extraction of soluble compounds or materials 140 can include removal of a 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, 15% of the liquid portion (including soluble compounds or materials 146) can then be extracted or removed from the thermochemically pretreated biomass 112. The extracted portion including soluble compounds or materials 146 (eg, soluble polysaccharides) may then undergo one or more purification steps 142b (eg, ultrafiltration) to enrich the soluble polysaccharides 147 .

[0400] In addition, the liquid portion that is not extracted in step 140 may include soluble compounds or materials 146 and insoluble compounds or materials 148. For example, in the case where 15% of the liquid portion is extracted as described above, the remaining portion of the physically pretreated biomass 112 (including 85% of the liquid portion) may include soluble polysaccharides and insoluble polysaccharides. Then, the solution including soluble compounds or materials 146 and insoluble compounds or materials 148 may undergo or be subjected to enzyme treatment 150, as disclosed herein. For example, one or more polysaccharide cutting enzymes may be added to a solution including soluble compounds or materials 146 and insoluble compounds or materials 148 to 0.5% w / v and incubated at 50° C. for 24 hours. Then, the enzyme-treated biomass 151 may be processed (e.g., filtered) 152 to remove at least a portion of the undigested biomass. In some cases, the removed undigested biomass may be disposed of or discarded. The digested biomass 141 can then be subjected to 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 component 105.

[0401] Exemplary methods of pretreating biomass to remove monosaccharides and / or disaccharides

[0402] Fig. 9is a simplified flow diagram illustrating an embodiment of a method for pre-treating biomass to remove mono- and / or disaccharides prior to enzymatic treatment.

[0403] Fig. 9 Embodiments of the invention may include aspects similar to Figure 8 For example, Fig. 9 Embodiments include those which may be similar to Figure 8 The physical pretreatment step 110 of the physical pretreatment 210 of the embodiment shown can have similar features. Therefore, similar features are represented by similar reference numerals, wherein a leading numeral is added to increase each reference numeral by 100. For example, the physical pretreatment in Figure 8 In the figure, it is represented as "110"; similar physical pretreatment is Fig. 9 Therefore, the above disclosures on similar identification features may not be repeated below. Fig. 9 Specific features of the methods and associated components or steps shown may not be shown or identified by reference numerals in the drawings, or may not be specifically discussed in the subsequent written description. However, the features may clearly be the same or substantially the same as features depicted in other embodiments and / or described with respect to the embodiments. Therefore, the relevant descriptions of these features also apply to Fig. 9 The characteristics of the method and related components or steps. Figure 8 Any suitable combination of the features described in the method shown and its variants can be used for Fig. 9 The disclosed mode is also applicable to other embodiments depicted in the subsequent drawings and / or described below.

[0404] As shown, the gentle pretreatment 220 (e.g., washing or incubation cycles provided herein) of the physically pretreated biomass 212 can include removing 224 soluble compounds 246. In some cases, the soluble compounds 246 can include monosaccharides and / or disaccharides. Then, the removed soluble monosaccharides and / or disaccharides can be discarded and / or abandoned. Therefore, a gentle pretreatment 220 can be performed or executed to remove soluble monosaccharides and / or disaccharides from the biomass 207.

[0405] Biomass 207 may be subjected to physical pretreatment 210 (e.g., cutting), and then the physically pretreated biomass 212 may be subjected to or subjected to mild pretreatment 220, such as washing or incubation (e.g., 30 minutes in water at 25°C). Soluble sugars 246 (e.g., soluble monosaccharides and / or disaccharides) may then be removed from the solution comprising mildly pretreated biomass 226. Mildly pretreated biomass 226 may then be subjected to or subjected to strong pretreatment 230. In some cases, strong pretreatment 230 may be a thermochemical pretreatment. For example, mildly pretreated biomass 226 may be treated in 1% w / v NaOH at 100°C for 60 minutes. In some embodiments, strong pretreatment 230 may be followed by a neutralization step (not shown) before enzyme treatment 250. Strongly pretreated biomass 232 may then be treated with enzyme 250, as discussed herein. In addition, enzyme-treated biomass 251 may then be subjected to downstream processing 260 to produce component 205.

[0406] Other exemplary embodiments of extracting soluble polysaccharides for combination with oligosaccharides

[0407] In some cases, the present disclosure relates to new methods for processing plant biomass materials to produce food, cosmetic, or nutraceutical ingredients.

[0408] Sugary foods and drinks are an important part of culture and lifestyle habits around the world, but the sugar they contain has been linked to obesity, diabetes, poor dental health and destructive behavior in people. For these reasons, consumer preferences have shifted away from sugary foods, and governments are increasingly implementing regulations to encourage less sugar consumption.

[0409] As a result, the industry has been searching for decades for suitable low-calorie sweeteners to replace sugar in food and beverages. Unfortunately, many sugar substitutes are produced from non-natural sources and often have a slightly bitter or other unpleasant taste along with their sweetness, both of which are unappealing to consumers. In addition, while many sweeteners can mimic the sweetness of sugar in food and beverages, few can mimic the broad range of effects that sugar has in food and beverages, such as adding bulk, adjusting texture, providing structure, acting as a preservative, and adjusting color and flavor through caramelization and Maillard reactions.

[0410] Dietary fiber is an important part of a positive diet and helps maintain a healthy digestive system and a well-regulated intestinal flora. This fiber consists of sugars of varying 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.

[0411] Biomass is a good source of sugars that can be used to replace sugar and add fiber to foods. However, there remains a need to optimize methods for obtaining these sugars from biomass and processing them into compositions that can be used as food, cosmetic or nutraceutical ingredients.

[0412] First, sugars generally need to be produced by controlled decomposition. Different amounts of different sized sugars in an ingredient can affect its nutritional value, as well as other properties such as hygroscopicity, which in turn affects properties such as the texture of a product made using the ingredient. It is also desirable for ingredients to include polysaccharides, as polysaccharides can improve gastrointestinal tolerability. However, due to the speed at which some particularly desirable polysaccharides decompose during enzymatic reactions in previously known methods, these desirable polysaccharides are difficult to isolate and then incorporate into ingredients.

[0413] In addition, foods (such as candy, chocolate and yogurt) that require smooth textures generally require that the ingredients are soluble to achieve smooth textures. Ingredients comprising insoluble polymer materials can produce granular textures. However, due to the insolubility of some polysaccharides, it may be difficult to prepare compositions comprising fully soluble polysaccharides, particularly in one-pot processes prepared by monolithic biomass. Typically, soluble polysaccharides decompose faster than insoluble polysaccharides, so after exposing plant biomass to enzymes, as previously known methods, soluble polysaccharides are fully decomposed and only insoluble polysaccharides are left.

[0414] Surprisingly, methods have been identified herein that can allow the isolation of polysaccharides and their incorporation into soluble food, cosmetic or nutraceutical ingredients containing oligosaccharides, thereby maintaining the benefits of increasing the gastrointestinal tolerability of the ingredient and allowing its use in foods with a smooth texture. Polysaccharides can be isolated from the same plant biomass as other desired sugars by providing an efficient and streamlined production process.

[0415] Therefore, 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:

[0416] a) providing a plant biomass comprising one or more soluble polysaccharides and one or more insoluble polysaccharides;

[0417] b) treating the plant biomass to solubilize the one or more soluble polysaccharides;

[0418] c) removing a portion of the dissolved one or more soluble polysaccharides;

[0419] d) reacting the remaining plant biomass with one or more enzymes to form one or more oligosaccharides; e) removing the one or more oligosaccharides; and

[0420] f) combining a portion of the solubilized one or more soluble polysaccharides from step (c) with the one or more oligosaccharides from step (e) to form a component.

[0421] Therefore, there is also provided a food, cosmetic or nutraceutical ingredient obtainable by the method of the present disclosure.

[0422] In another aspect of the present disclosure, there is provided a food, cosmetic or nutritional liquid ingredient comprising at least one oligosaccharide selected from the group consisting of:

[0423] i) cellooligosaccharides having a degree of polymerization of 2 to 6;

[0424] ii) xylo-oligosaccharides having a degree of polymerization of 2 to 12;

[0425] iii) manno-oligosaccharides having a degree of polymerization of 2 to 12;

[0426] iv) mixed-linkage dextran oligosaccharides having a degree of polymerization of 2 to 5;

[0427] v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 12; and

[0428] vi) chitosan oligosaccharides having a degree of polymerization of 2 to 12;

[0429] and at least one polysaccharide selected from the group consisting of:

[0430] i) xylan;

[0431] ii) mannan;

[0432] iii) cellulose derivatives;

[0433] iv) mixed bond dextran;

[0434] v) xyloglucan; and

[0435] vi) chitosan;

[0436] wherein the liquid component comprises at least 20% by dry weight of at least one oligosaccharide and at least 2% by dry weight of at least one polysaccharide, and wherein the viscosity of the liquid component is from 5 to 100,000 cps.

[0437] Preparing food, cosmetic or nutraceutical ingredients in the manner provided herein can allow efficient use of biomass by incorporating oligomeric and polymeric materials from the same biomass source to prepare soluble ingredients. In addition, these methods can allow purification, derivatization or other modifications, and control of the ratio of oligomeric and polymeric, which can improve the functional properties, nutritional properties and tolerance of the ingredients.

[0438] Any material comprising suitable polysaccharides can be plant biomass. Because food, cosmetics and nutrition products industry use multiple oligosaccharides, there is no particular restriction on the polysaccharide applicable to the method. Plant biomass applicable to the oligosaccharide spectrum of the present disclosure can include, for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronic acid xylan, arabinoxylan and glucuronic acid arabinoxylan), xyloglucan, mixed bond 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 that plant biomass comprises can include any of: mannan, mixed bond glucan, lignocellulose, hemicellulose, some cellulose derivatives (such as, cellulose acetate, hydroxyethyl cellulose and hydroxymethyl cellulose), and chitosan. In some embodiments, plant biomass comprises hemicellulose. In certain embodiments, hemicellulose comprises xylan and / or mannan.

[0439] Thus, plant biomass can be grains, husks, pods, seed coats, and / or other seed materials; seaweed; corn stalks, straw, bagasse, miscanthus, sorghum residue, switchgrass, bamboo, and / or other monocotyledonous tissues; water hyacinth, leaf tissue, roots, and / or other plant matter; and / or any combination of suitable plant biomass. In some cases, plant biomass includes or is suitably composed of: sugarcane biomass (such as bagasse), corn biomass (such as corn cobs or corn stalks), wheat biomass (such as wheat straw or wheat bran), hardwood, or softwood. In some cases, plant biomass includes corn cobs, bagasse, wheat straw, or straw.

[0440] In various cases, in step (b), "treatment" is a thermochemical treatment of plant biomass. As used herein, "thermochemistry" generally refers to heating plant biomass in a chemical substance to above room temperature (room temperature can be about 20°C to 22°C), such as heating in a solution comprising water, an alkali, or an ionic solvent. Thermochemical steps can physically and chemically modify the chemical composition of plant biomass. For example, free hydroxide ions from water or alkali can break hydrogen bonds between sugars, thereby dissolving certain types of sugars (e.g., hemicellulose), and can better make it easier for enzymes in subsequent steps to decompose sugars. These broken hydrogen bonds may be located between monomers of the same sugar chain, which contributes to the tertiary structure of the chain. Broken hydrogen bonds may also be located between monomers of different sugar chains, which contributes to the quaternary structure of more than one chain. Subsequently, the treatment can produce one or more polysaccharides soluble (i.e., polysaccharides that are particularly susceptible to breaking hydroxide ions, especially, for example, hemicellulose) to dissolve into the chemical substance used. One or more insoluble polysaccharides (such as cellulose) are insoluble in the chemical substance.

[0441] The heating of treatment step (b) may be within a temperature range, suitably 30° C. to 180° C., 50° C. to 150° C. or 70° C. to 120° C. Higher temperatures may facilitate faster dissolution of soluble polysaccharides, however, excessively high temperatures may be more difficult to achieve in an efficient and cost-effective manner and may chemically modify biomass components, including sugars (e.g., in an undesirable manner).

[0442] The heating can be carried out over a range of time scales, and in particular large amounts of biomass can be exposed to heating for longer periods of time, which can be adjusted accordingly. For example, the heating of plant biomass can be from 1 minute to 72 hours, from 10 minutes to 24 hours, from 20 minutes to 12 hours, or from 25 minutes to 8 hours.

[0443] In some cases, thermochemical treatment may include heating the plant biomass in water, ie, at a neutral pH of about pH 7.

[0444] In some cases, the thermochemical treatment may include heating the plant biomass in an alkaline solution having a pH of 8 to 14, 9 to 14, or 10 to 14. The solution may comprise, or suitably consist of, any of the following bases: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and aqueous ammonia. In various cases, the base may be sodium hydroxide. Combinations of the listed bases are also contemplated.

[0445] A plurality of different sequential thermochemical treatment steps are also contemplated. For example, there may be two sequential thermochemical treatments, there may be three sequential thermochemical treatments, or there may be four or more sequential thermochemical treatments. In some cases, the biomass may be thermochemically treated in a neutral aqueous solution and then thermochemically treated in an alkaline aqueous solution.

[0446] After the treatment step, step (c) may include removing a portion of the dissolved one or more soluble polysaccharides. The purpose of this step may be to separate and remove soluble polysaccharides from the plant biomass so that they are not decomposed and thus lost in subsequent enzymatic reactions. This may enable the use of these polysaccharides when forming ingredients in subsequent steps. Depending on the amount required in the final ingredient, all of the dissolved polysaccharides or a portion of the dissolved polysaccharides may be removed. The soluble polysaccharides may be removed using simple steps, such as filtering the chemical substance in which the soluble polysaccharide is dissolved.

[0447] Step d) comprises reacting the remaining plant biomass (which may be in the form of a solution and / or 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 (e.g., useful oligosaccharides) by one or more enzymes, possibly leaving partially cleaved or uncleaved polysaccharides, which may include cellulose, xylan (e.g., glucuronoxylan, arabinoxylan or glucuronylarabinoxylan), mannan (e.g., glucomannan, galactomannan or galactoglucomannan), mixed bond glucan, xyloglucan, chitin, chitosan or lignocellulose.

[0448] The enzyme reaction can be carried out in solution and / or suspension in a suitable reaction vessel. The enzyme reaction can be carried out at a temperature or temperature regime 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 a desired concentration, or until some other requirement has been met.

[0449] To ensure optimal contact between the enzyme and the plant biomass, the reaction mixture may be agitated continuously or intermittently. Agitation may take the form of rhythmic movement of the entire reaction vessel, a blower or other agitation device, bubble sparging, or any other agitation method.

[0450] The enzymatic reaction can be a microbial fermentation. The temperature and reaction time can be adapted to the growth of the microorganism used. The microbial organism can be genetically modified 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).

[0451] Further embodied in the present disclosure is an expression vector suitable for modifying a subject microorganism so that the microorganism produces an 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: a regulatory sequence of a heat shock gene, a regulatory sequence of a toxic gene, and a regulatory sequence of a spore formation gene.

[0452] The enzymatic reaction can be carried out at a temperature or temperature regime 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 a microbial fermentation, the temperature may be suitable for this, for example, the enzymatic reaction may include the growth of E. coli and / or the temperature may be constant and about 37°C.

[0453] The pH value of a solution or suspension can affect the activity of the enzyme. Controlling the pH can ensure that the enzymatic reaction is carried out at a suitable rate. The enzymatic reaction of the present disclosure can be carried out at a pH in the range of about 2 to about 10, about 3 to about 8, or about 4 to about 6.

[0454] The enzymatic reaction can be allowed to continue for a period of time, then quenched, and the product can be separated or otherwise collected. The time period can be from about 1 minute to about 6 days, from about 0.5 day to about 5 days, or from about 16 hours to about 96 hours. Alternatively, the reaction can be allowed to proceed until catalysis no longer occurs.

[0455] 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 anthrone assays) and / or chromatography (e.g., thin layer chromatography and high performance anion exchange chromatography). The reaction can be carried out until all polysaccharides are converted to oligosaccharides.

[0456] There are many enzymes suitable for the enzymatic reaction of the present method. For example, "cleavage polysaccharide monooxygenase" and "LPMO", which refers to a class of enzymes that can use a copper-containing part and use an oxygen source (such as dioxygen molecules, peroxides or any other oxygen source); and a suitable reducing agent to oxidatively cut polysaccharides. Therefore, 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, lignin alcohol, and a combination of cellobiose and cellobiose dehydrogenase. A variety of photosynthetic pigments can be used. In some embodiments, thylakoids and purified fractions or chlorophyllins 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, the LPMO may be PaLPMO9E (SEQ ID NO: 1), which is an AA9 LPMO originally isolated from an ascomycete fungus (Podospora anserina), or the LPMO may be AA9 LPMO from Trichoderma reesei (SEQ ID NO: 23).

[0457] Aerobic conditions may include the addition of oxygen, which may be provided by aerating the substrate mixture with an oxygen-containing gas such as air. Aeration may be performed by introducing oxygen-containing bubbles into the aqueous substrate mixture via various systems such as air injectors, aeration frits, membrane systems, or internal circulation airlift reactors. The concentration of molecular oxygen in the enzymatic reaction may be from about 4 mg / L to about 14 mg / L.

[0458] Another enzyme that can be used in the method is "cellulase", which has hydrolytic activity on cellulose, for example, endo-1,4-β-glucanase, cellobiohydrolase and / or β-glucosidase activity. The enzyme is capable of cutting glycosidic bonds in one or more forms of cellulose (including cellulose found in plant biomass). In the process, cellulose produces products including glucose and cellooligosaccharides. In some cases, the β-glucanase may include an enzyme selected from 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).

[0459] Another enzyme is "cellobiohydrolase", which has hydrolytic activity on cellulose and produces primarily cellobiose as a product. Cellobiose is a disaccharide and a cellooligosaccharide. Such enzymes are capable of cleaving glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. In various cases, the cellobiohydrolase can be from the GH6 and GH7 families, Cel6A or Cel7A enzymes derived from Trichoderma reesei (SEQ ID NOs: 10 and 11, respectively).

[0460] Another enzyme is a "β-glucosidase," which has hydrolytic activity on cellulose and produces primarily 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. In some embodiments, the β-glucosidase may include a GH3 β-glucosidase, such as an enzyme derived from Trichoderma reesei (SEQ ID NO: 22).

[0461] Another class of enzymes is lichenase, which can be selected from 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). The enzyme can act on, for example, mixed-linkage glucans (which are glucans containing a mixture of β-1,3 bonds and β-1,4 bonds), and can cut them at β-1,4 glycosidic bonds. In the case where the lichenase acts on mixed-linkage glucans, the β-glucans produced can mainly fall within the size range of about 3 to about 7 residues, so they are particularly used in the food, cosmetics and nutrition industries. Mixed-linkage glucans are abundant in members of the grass and horsetail families, so grass biomass (such as straw) has a high level of mixed-linkage glucans and can be effectively acted on by lichenase.

[0462] Another class of enzymes is xylanase, which acts on plant biomass such as comprising a xylan backbone. Xylanase can be, for example, glucuronoxylanase, arabinoxylanase or glucuronyl arabinoxylanase. Enzyme can be active to a variety of polymers with a xylan backbone, such as glucuronoxylan, arabinoxylan and glucuronyl arabinoxylan. These polymers are abundant in various plant biomass, and for example hardwood and softwood can both comprise suitable polysaccharides, wherein hardwood generally comprises glucuronoxylan and softwood generally comprises arabinoglucuronoxylan. In some cases, xylanase can include the GH5 xylanase derived from Clostridium thermocellum (SEQ ID NO:3) and Polysaccharide (SEQ ID NO:4), and the GH30 xylanase 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).

[0463] Other enzymes useful in the present disclosure may include xyloglucanase and xyloglucan endoglucanase (XEG), which are produced by a variety of organisms (including plant pathogenic microorganisms). Xyloglucanase and xyloglucan endoglucanase can act on xyloglucan, which is a hemicellulose β-1,4 glucan chain abundant in the primary cell wall of higher plants, which is modified by xylose, and some xylose residues are further modified by other residues (such as galactose). When suitable xyloglucanase or XEG act on xyloglucan, the product includes xyloglucan oligosaccharides with a length backbone that can be used in the food, cosmetics and nutrition industry. In some cases, xyloglucanase may include GH5 xyloglucanase (SEQ ID NO:8) derived from Bacteroides ovatus and GH74 xyloglucanase derived from Trichoderma reesei.

[0464] Since any given natural plant biomass may comprise a mixture of different polysaccharides, a mixture of different enzymes may be useful sometimes. This mixture may comprise one or more other enzymes. For example, this mixture may comprise LPMO and endoglucanase, xylanase and lichenase, cellobiohydrolase and mannanase or endoglucanase and cellobiohydrolase, wherein the enzyme partner exists in a molar ratio of, for example, 1:100 to 100:1.

[0465] In some cases, one or more enzymes can be a mixture of different enzymes, for example 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 fermentation product, which is subjected to minimum processing after extraction, and for example, conventional preparations can only be subjected to filtration to remove insoluble components. As used herein, the term "semi-crude enzyme preparation" generally refers to a soluble preparation extracted from a microbial fermentation product, which is subjected to some processing after extraction, and for example, preparations can be subjected to filtration to remove insoluble components, increase enzyme concentration, and / or nanofiltration to remove small molecular weight compounds.

[0466] In some cases, crude enzyme preparation or semi-crude enzyme preparation can be from bacteria or fungi. In some embodiments, crude enzyme preparation or semi-crude enzyme preparation can be from fungi, such as filamentous cellulose decomposing fungi, such as from Trichoderma or Aspergillus species. In certain embodiments, enzyme can be crude enzyme preparation or semi-crude enzyme preparation from Reesei mold strain.

[0467] In step (e), one or more oligosaccharides formed in step (d) are removed, which can be done in a variety of ways. It can be separated based on solubility so that only the composition of soluble sugars is extracted for further processing, and / or separated by chromatography to produce a composition with a narrower band of oligosaccharide chain length. Separation can be based on precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration. In the case of separation based on solubility, the sugar spectrum present in the separated composition can depend on the initial enzymatic reaction, because the solubility of different sugars decreases at different rates with length.

[0468] It is also contemplated within the scope of the present disclosure that all or part of the removed one or more oligosaccharides are further processed to produce further products, which are then combined with one or more dissolved polysaccharides to form a component. This further processing may include any chemical, physical or enzymatic step, such as reduction, such as reductive amination performed where appropriate; oxidation, caramelization, modification with Schiff bases or by Maillard reaction or by any combination of these steps, and may provide different products with improved properties for the desired purpose. For example, caramelization characteristics, calorific value, flavor and color may be modified. The oligosaccharides may also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0469] It is also contemplated within the scope of the present disclosure that the one or more soluble polysaccharides that are fully or partially dissolved are further processed to produce a product with improved properties and then combined with one or more removed oligosaccharides to form a component. The further processing may include any chemical, physical or enzymatic steps, such as alkylation or acid treatment. The polysaccharides may also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0470] In some cases, after modification and / or purification of the oligosaccharides and polysaccharides, all or part of the oligosaccharides and polysaccharides are combined, such as in step (f), the ratio of polysaccharide:oligosaccharide can be 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 can depend on the desired properties of the final component and the modification and purification that has been applied to the sugars. In some embodiments, it is not necessary to reconstitute all of the removed oligosaccharides and polysaccharides.

[0471] In step (f), the combination can be carried out in a variety of ways, for example, by mixing a solution comprising all or part of one or more soluble polysaccharides and a solution and / or suspension comprising 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 mixing a dry form comprising all or part of the one or more removed oligosaccharides produced by spray drying, freeze-drying or concentrating in some other way (after removing them in step (e)), with a dry form comprising all or part of the one or more removed polysaccharides produced by condensation in some other way (after removing them in step (c)). Alternatively, one of (i) one or more soluble polysaccharides or (ii) the removed oligosaccharides can be in dry form, while the other is in solution when they are combined.

[0472] When the ingredients are in dry form, the method may further comprise the step (g) of mixing and dissolving the ingredients in a liquid to form a liquid ingredient. In various cases, the liquid may be an aqueous solution, such as water.

[0473] The second aspect of the present disclosure, the ingredients formed in step (f) and the liquid ingredients formed in step (g) may contain various oligosaccharides and contain different amounts depending on the desired properties. In some cases, the ingredients and liquid ingredients 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 ingredients may contain at least 20% dry weight or at least 30% dry weight of xylo-oligosaccharides with a degree of polymerization of 2 to 12, the ingredients may contain at least 20% dry weight or at least 30% dry weight of mixed bond glucan oligosaccharides with a degree of polymerization of 2 to 5, the ingredients may contain at least 20% dry weight or at least 30% dry weight of manno-oligosaccharides with a degree of polymerization of 2 to 12, the ingredients 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 ingredients 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 comprise up to 100% by dry weight of the above oligosaccharides and the polysaccharides described herein, thus the above embodiments wherein the oligosaccharides are present at least 20% by dry weight do not comprise five or six types of oligosaccharides.

[0474] The ingredients and liquid ingredients may contain at least 50%, 60%, 70%, 80%, 90%, 95%, 99% or 99.5% sugar present by dry weight. The ingredients and liquid ingredients may consist essentially of sugar. For example, the ingredients may have less than 0.5% dry weight or less than 0.3% dry weight, such as 0.1% dry weight of other substances.

[0475] 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 required to be in syrup, its viscosity may 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 using a standard test procedure (where a 400 mL sample is taken in a tall beaker to ensure that no container effect occurs). The instrument was operated according to the manufacturer's instructions for the range (rotational viscometer using spindle code 61, spindle speed 100 rpm, at 22°C).

[0476] In some cases, the flow rate of the liquid component may 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 through per unit time. Unless otherwise noted, the flow rate values ​​specified herein are measured by the following steps: the timing of the flow rate of 5 mL of the liquid component from a vertical standing syringe (BD Plastipak 300613) filled with 20 mL of the test liquid is measured under gravity at room temperature.

[0477] In various cases, the oligosaccharide concentration of the liquid component may 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.

[0478] In some cases, the liquid component may have a polysaccharide concentration of 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.

[0479] In some cases, the total concentration of oligosaccharides and polysaccharides of 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.

[0480] In another aspect, composition and liquid component can comprise at least two kinds of oligosaccharides.The amount of every kind of oligosaccharide can change according to the desired property of gained food, cosmetics or nutrition product.Two kinds of oligosaccharides can exist with the ratio of 1:9 to 9:1 or 1:2 to 2:1.In addition, composition and liquid component can comprise three kinds of oligosaccharides, they can comprise four kinds of oligosaccharides, they can comprise five kinds of oligosaccharides, or they can comprise six kinds of oligosaccharides.

[0481] The ingredients and liquid ingredients may comprise cellooligosaccharides, such as a combination of cellooligosaccharides and xylo-oligosaccharides. Alternatively, the ingredients and liquid ingredients may comprise a combination of cellooligosaccharides and manno-oligosaccharides.

[0482] One or more soluble polysaccharides in the composition can be particularly soluble in water or alkali. For example, the soluble polysaccharide used in the present disclosure can include hemicellulose, such as xylan, mannan, mixed bond glucan and some cellulose derivatives (such as cellulose acetate, hydroxyethyl cellulose and hydroxymethyl cellulose) and chitosan. In some embodiments, one or more soluble polysaccharides can include hemicellulose. In certain embodiments, hemicellulose can include xylan and / or mannan.

[0483] In some cases, the ingredients and liquid ingredients may contain at least 2% dry weight or at least 3% dry weight of xylan. In various cases, the ingredients and liquid ingredients may contain at least 2% dry weight or at least 3% dry weight of mannan. In some cases, the ingredients and liquid ingredients may contain at least 2% dry weight or at least 3% dry weight of cellulose derivatives. In some cases, the ingredients and liquid ingredients may contain at least 2% dry weight or at least 3% dry weight of mixed bond glucans. In various cases, the ingredients and liquid ingredients may contain at least 2% dry weight or at least 3% dry weight of xyloglucans. In some cases, the ingredients and liquid ingredients may contain at least 2% dry weight or at least 3% dry weight of chitosan.

[0484] In some cases, the ingredients and liquid ingredients may contain 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.

[0485] In some cases, the ratio of polysaccharide:oligosaccharide of the ingredient and the liquid ingredient 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.

[0486] The produced ingredients and liquid ingredients can be used for applications of conventionally used oligosaccharides, sugars, bulking sweeteners, low-intensity sweeteners or other related food ingredients. For example, as a sweetener, bulking agent, added dietary fiber or humectant. Of particular note is the use of reduced sucrose in foods. It can be incorporated into cakes, breads or other baked goods; chocolate or other candies such as toffees, soft candies, meringues, jams, jellies or caramels; or incorporated into beverages, for example to provide a good taste or color profile or to increase the dietary fiber content. Alternatively, the ingredient can be incorporated into animal feed, for example as an isolated ingredient or by directly utilizing the enzymatic reaction mixture as a raw material.

[0487] Compositions or ingredients as described herein can be used to change one or more properties of the final product. Such properties include, but are not limited to, sweetness, texture, mouthfeel, adhesiveness, glossiness, smoothness, wettability, viscosity, color, hygroscopicity, flavor, fluffiness, water retention, caramelization, surface texture, crystallization, structural properties, reduced calories, reduced glycemic index, reduced glycemic load, increased fiber, reduced sugar content, and solubility. These can be possible improvements to the current use of different types of sugars, sugar substitutes, and / or other such compounds.

[0488] In the cosmetics industry, the ingredients can improve texture and moisture retention, act as UV absorbing molecules, maintain gel or cream structure, and / or act as bulking agents. In addition, the ingredients and liquid ingredients can be used in nutritional compositions because they have been shown to provide dietary fiber that promotes digestive health, good regulation of intestinal flora, and other health benefits. In this case, the ingredients provided herein can also be used as ingredients in probiotic beverages or other prebiotic or probiotic preparations.

[0489] The detailed description is further supplemented with reference to the following numbered embodiments. 1) A method for producing a food, cosmetic or nutritional ingredient, the food, cosmetic or nutritional ingredient comprising one or more oligosaccharides and one or more soluble polysaccharides, the method comprising the following steps: (a) providing plant biomass comprising one or more soluble polysaccharides and one or more insoluble polysaccharides; (b) treating the plant biomass to dissolve the 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 the 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) A method according to numbered embodiment 1, wherein the treatment in step (b) is a thermochemical treatment. 3) A method according to numbered embodiment 2, wherein the thermochemical treatment is a hot water treatment or a hot alkali treatment. 4) A method according to numbered embodiment 3, wherein the alkali treatment uses an alkali with a pH of 10 to 14. 5) The method according to numbered embodiment 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 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 numbered embodiment 9, wherein the method further comprises the step of: (g) mixing and dissolving the ingredients in a liquid to form a liquid ingredient, wherein the viscosity of the liquid ingredient is 5 to 100,000 cps. 11) Method 10 according to numbered embodiment, wherein the concentration of oligosaccharides and polysaccharides in the liquid component is 1 to 200% w / v. 12) Method according to any of the preceding numbered embodiments, wherein the one or more soluble polysaccharides comprise at least one selected from the group consisting of mannans, mixed bond glucans, lignocellulose, hemicellulose, certain cellulose derivatives (e.g., cellulose acetate, hydroxyethyl cellulose, and hydroxymethyl cellulose), and chitosan. 13) Method according to numbered embodiment 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 of 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) cello-oligosaccharides having a degree of polymerization of 2 to 6; ii) xylo-oligosaccharides having a degree of polymerization of 2 to 12; iii) manno-oligosaccharides having a degree of polymerization of 2 to 12; iv) mixed-bond 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) chito-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 bond glucan; v) xyloglucan; and vi) chitosan; wherein the liquid component 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 component 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 component according to numbered embodiment 16, wherein the liquid component comprises at least two of the oligosaccharides listed in (i) to (vi). 18) The liquid component according to numbered embodiments 16 or 17, wherein the liquid component comprises at least 20% dry weight of cellooligosaccharides having a degree of polymerization of 2 to 6. 19) The liquid component according to any one of numbered embodiments 16 to 18, wherein the liquid component comprises at least 20% dry weight of xylo-oligosaccharides having a degree of polymerization of 2 to 12. 20) A liquid ingredient according to any one of numbered embodiments 16 to 19, wherein the liquid ingredient comprises at least 20% dry weight of manno-oligosaccharides having a degree of polymerization of 2 to 12. 21) A 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) A 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) A 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) A liquid ingredient according to any one of numbered embodiments 16 to 23, wherein the liquid ingredient has a polysaccharide concentration of 0.1 to 50% (w / v). 25) A liquid ingredient according to any one of numbered embodiments 16 to 24, wherein the liquid ingredient has an oligosaccharide concentration of 1 to 200% (w / v). 26) A liquid ingredient according to any one of numbered embodiments 16 to 25, wherein the liquid ingredient has a polysaccharide and oligosaccharide concentration of 1-200% (w / v).27) A liquid ingredient according to any one of numbered embodiments 16 to 26, wherein the liquid ingredient comprises polysaccharides and oligosaccharides in a ratio of 1:100 to 1:1. 28) A liquid ingredient according to any one of numbered embodiments 16 to 27, wherein the liquid ingredient comprises two oligosaccharides in a ratio of 1:9 to 9:1. 29) Use of the liquid ingredient according to any one of numbered embodiments 16 to 28 in food, cosmetics or nutritional products.

[0490] Other Exemplary Embodiments of Pretreating Biomass to Remove Monosaccharides and / or Disaccharides

[0491] In some cases, the present disclosure relates to novel methods for physically and thermochemically processing plant biomass materials to produce food, cosmetic, or nutraceutical ingredients.

[0492] Sugary foods and drinks are an important part of culture and lifestyle habits around the world, but the sugar they contain has been linked to obesity, diabetes, poor dental health and destructive behaviors. For these reasons, consumer preferences have shifted away from sugary foods, and governments are increasingly implementing regulations to encourage less sugar consumption.

[0493] As a result, the industry has been searching for decades for suitable low-calorie sweeteners to replace sugar in food and beverages. Unfortunately, many sugar substitutes are produced from non-natural sources and often have a slightly bitter or other unpleasant taste along with their sweetness, both of which are unappealing to consumers. In addition, while many sweeteners can mimic the sweetness of sugar in food and beverages, few can mimic the broad range of effects that sugar has in food, such as adding volume, adjusting texture, providing structure, acting as a preservative, and adjusting color and flavor through caramelization and Maillard reactions.

[0494] Dietary fiber is an important part of a positive diet and helps maintain a healthy digestive system and a well-regulated intestinal flora. This fiber consists of sugars of varying 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.

[0495] Biomass is a good source of sugar, can be used for replacing sugar and adding fiber to food. Compositions made of raw materials have been provided. However, it is still necessary to optimize the method of obtaining these sugars from biomass and processing these sugars into compositions that can be used as food, cosmetics or nutrition ingredients on a large scale and commercial scale. Enzyme decomposition of a large amount of plant biomass can require a considerable time. In addition, sugar usually needs to be produced by controlled decomposition. For this reason, enzyme decomposition is needed, because the product size can be accurately controlled, ensuring that little monosaccharide is not produced and / or produced.

[0496] The method that this paper provides can be economical and effectively produce food, cosmetics or nutrition ingredients from plant biomass starting materials, which is faster than the method previously used, produces purer final products, and can be used for large-scale and commercial scale. The method can be carried out by following: perform pre-washing step to remove endogenous monosaccharide and / or disaccharide from biomass, and adopt thermochemical pretreatment step to ensure that the controlled decomposition and release of the sugar that manufacturing composition can need are realized. In a word, these steps can ensure that in the enzyme front processing process, do not produce and / or produce little monosaccharide. This can improve efficiency to the greatest extent and limit the amount of purification after required reaction.

[0497] Thus, in another aspect of the present disclosure, there is provided a method for producing a food, cosmetic or nutraceutical ingredient comprising one or more oligosaccharides, wherein the method comprises the following steps:

[0498] a) physical pretreatment of plant biomass containing monosaccharides and / or disaccharides;

[0499] b) a wash cycle (also referred to herein as an incubation cycle) comprising the steps of: (i) washing (e.g., incubating) the plant biomass to solubilize at least a portion of the monosaccharides and / or disaccharides, and (ii) removing at least a portion of the monosaccharides and / or disaccharides;

[0500] c) thermochemical pretreatment of plant biomass;

[0501] d) forming one or more oligosaccharides by an enzymatic reaction comprising the step of contacting one or more polysaccharide-cleaving enzymes with the plant biomass in solution or suspension;

[0502] e) isolating (also referred to herein as enriching or separating) one or more oligosaccharides from the enzymatic reaction mixture and using the one or more oligosaccharides to form an ingredient.

[0503] Preparation of food, cosmetic or nutraceutical 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 control of the ratio of oligomeric and polymeric materials, which can improve the functional properties, nutritional properties and tolerability of the ingredients.

[0504] Steps (a), (b) and (c) of the methods of the present disclosure are all steps of "pretreatment" performed on plant biomass starting material. As used herein, "pretreatment" generally refers to steps performed on plant biomass before contacting the polysaccharide cleaving enzyme with the plant biomass.

[0505] Step (a) is a physical pretreatment of plant biomass, the purpose of which can be to physically decompose the plant biomass in preparation for subsequent steps. The physical step can help to accelerate the entire method because it can increase the available surface area of ​​the plant biomass, making the chemical substances used in the subsequent steps, for example, active to more plant biomass at the same time. The physical pretreatment step can include cutting, chopping, grinding, ball milling, grinding, dressing, mixing or a combination thereof of the plant biomass.

[0506] Any material comprising suitable polysaccharides can be plant biomass. Because food, cosmetics and nutrition industry use multiple oligosaccharides, there is no particular restriction on the polysaccharide applicable to the method. Plant biomass applicable to the oligosaccharide spectrum producing the present disclosure can include for example cellulose, lignocellulose, chitin, chitosan, xylan (such as, glucuronic acid xylan, arabinoxylan and glucuronic acid arabinoxylan), xyloglucan and mixed bond glucan, and / or mannan (such as, glucomannan, galactomannan, or galactoglucomannan). However, it is envisioned that any plant biomass can be suitably acted on thereon.

[0507] Thus, plant biomass can be grains, husks, bean pods, seed coats, and / or other seed materials; seaweed; corn stalks, rice straw, bagasse, miscanthus, sorghum residue, switchgrass, bamboo, and / or other monocotyledonous tissues; water hyacinth, leaf tissue, roots, and / or other plant matter; and / or any combination of suitable plant biomass. In some embodiments, plant biomass can include sugar cane, corn stalks, corn cobs, wheat bran, wheat straw, hardwood, or softwood. In certain embodiments, plant biomass can include corn cobs.

[0508] Step (b) is the 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 biomass. For example, 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 may be meaningful because disaccharides can not be easily removed from the oligosaccharide fraction subsequently, for example, filtering methods can be used but usually cause the equivalent loss of other disaccharides.

[0509] The washing cycle of step (i) can be carried out at 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 monosaccharides and / or disaccharides to dissolve more quickly, however, temperatures that are too high may be more difficult to achieve in an efficient and cost-effective manner and may damage biomass compounds or dissolve compounds that are not desired to be dissolved in this step.

[0510] The washing cycle of step (i) can occur within a certain time range, for example, a large amount of biomass can be exposed to the longer time of this step, which can be adjusted accordingly. For example, the time can be 0.5 minutes to 72 hours, 1 minute to 12 hours, 5 minutes to 24 hours or 10 minutes to 3 hours. In certain embodiments, this step can be carried out in batches or continuously.

[0511] In some embodiments, the wash cycle of step (i) may include washing the plant biomass in water at room temperature, i.e., water supplied at a neutral pH of about pH 7. In another aspect, the wash cycle of step (i) may include heating the plant biomass in water, i.e., water supplied at a neutral pH of about pH 7. During the wash cycle, the supplied neutral water may become slightly acidic as the monosaccharides and / or disaccharides dissolve.

[0512] In certain embodiments, the washing cycle of step (i) may comprise 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 may comprise or suitably consist of any one selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, aqueous ammonia. In various embodiments, the base may be sodium hydroxide. Combinations of the listed bases are also contemplated.

[0513] In various cases, the washing cycle of step (i) may comprise 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 may comprise or suitably consist of any organic or inorganic acid, such as one selected from the group consisting of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid and oxalic acid. In some cases, the acid may be sulfuric acid. Combinations of the listed acids are also contemplated.

[0514] In some 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, may also be removed during this step.

[0515] Step (b) can be repeated to remove monosaccharides and / or disaccharides that are 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 performed after the physical washing pretreatment step. As used herein, "thermochemistry" generally refers to heating the plant biomass in a chemical substance to a temperature higher than room temperature (room temperature is, for example, about 15 to 25 ° C or about 20 to 22 ° C), such as heating in water, acid or alkaline solutions. The purpose of the thermochemical step can be to help speed up the entire method because it can chemically modify the chemical composition of the plant biomass, for example, it can break the hydrogen bonds between sugars, and the enzyme in the subsequent steps can more easily decompose sugars.

[0516] Heating can be in a temperature range of, for example, 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 modifications, however, temperatures that are too high can be more difficult to achieve in an efficient and cost-effective manner.

[0517] Heating can occur over a range of time, particularly large amounts of biomass can be exposed to heating over longer periods of time, which can be adjusted accordingly. For example, the heating of plant biomass can be from 5 minutes to 72 hours, from 15 minutes to 24 hours, from 30 minutes to 12 hours, or from 1 hour to 4 hours.

[0518] In some embodiments, thermochemical pretreatment may include heating the plant biomass in water (ie, at a neutral pH of about pH 7).

[0519] In certain embodiments, the thermochemical treatment may 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 may comprise 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 aqueous ammonia. In various embodiments, the base may be sodium hydroxide. Combinations of the listed bases are also contemplated.

[0520] In some cases, the thermochemical treatment may 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 may contain or suitably consist of any organic or inorganic acid, such as one selected from sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, and oxalic acid. In some cases, the acid may be sulfuric acid. Combinations of the listed acids are also contemplated.

[0521] For step (c), a plurality of different sequential thermochemical treatment steps are also envisaged. For example, step (c) may be performed at least twice, at least three times, at least four times or at least five times.

[0522] 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.

[0523] After the pretreatment step, step (d) may include an enzymatic reaction to form one or more oligosaccharides from the plant biomass. The polysaccharides present in the plant biomass may be partially cleaved into oligosaccharides (e.g., useful oligosaccharides) by an enzyme, leaving partially cleaved or uncleaved polysaccharides, which may include cellulose, xylan (e.g., glucuronoxylan, arabinoxylan or glucuronylarabinoxylan), mannan (e.g., glucomannan, galactomannan or galactoglucomannan), mixed bond glucan, xyloglucan, chitin, chitosan or lignocellulose.

[0524] Reaction can be carried out in solution and / or suspension.Reaction can be carried out in suitable reaction vessel.In some cases, reaction can be carried out under the temperature or temperature scheme that is suitable for the specific combination of enzyme and plant biomass, can allow described reaction to carry out certain time, until product reaches desired concentration or until satisfying some other requirements, and separate product or otherwise collect product.This time period can be about 1 minute to about 6 days, about 0.5 day to about 5 days or about 16 hours to about 96 hours.Alternately, can allow described reaction to carry out until catalysis no longer occurs.

[0525] To ensure optimal contact between the enzyme and the plant biomass, the reaction mixture may be stirred continuously or intermittently. Agitation may take the form of rhythmic movement of the entire reaction vessel, a blower or other agitation device, bubble sparging, or any other agitation method.

[0526] The enzymatic reaction can be a microbial fermentation. The temperature and reaction time can be adapted to the growth of the microorganism used. The microbial organism can be genetically modified 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.

[0527] Also embodied in the present disclosure are expression vectors suitable for modifying the subject microorganisms so that they produce the enzymes or enzyme mixtures 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 toxic genes, and regulatory sequences of spore formation genes.

[0528] The enzymatic reaction can be carried out at a temperature or temperature regime 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 a microbial fermentation, the temperature can be suitable for this, for example the enzymatic reaction can include the growth of E. coli and / or the temperature can be constant and about 37°C.

[0529] The pH of a solution or suspension can affect the activity of the enzyme. Control of pH can ensure that the enzymatic reaction is carried out at a suitable rate. The enzymatic reaction of the present disclosure can be carried out in a range of pH from about 2 to about 10, from about 3 to about 8, or from about 4 to about 6.

[0530] The enzymatic reaction can be allowed to continue until 5-75%, 5-70%, 5-65%, 5-55%, more or 10-50% of undigested polysaccharide-containing plant biomass remains. This can be monitored or checked by reducing end assays (e.g., anthrone assays) and / or chromatography (e.g., thin layer chromatography and high performance anion exchange chromatography).

[0531] A variety of enzymes may be suitable for the enzymatic reaction of the present method. For example, "cleavage polysaccharide monooxygenase" and "LPMO", which "cleavage polysaccharide monooxygenase" and "LPMO" refer to a class of enzymes that can use a copper-containing part and use an oxygen source (such as dioxygen molecules, peroxides or any other oxygen source); and a suitable reducing agent to oxidatively cut polysaccharides. Therefore, 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, lignin alcohol, and a combination of cellobiose and cellobiose dehydrogenase. A variety of photosynthetic pigments can be used, such as thylakoids and purified fractions or chlorophyllins, and light can be provided. LPMO can be selected from the following families: AA9, AA10, AA11, AA13, AA14 and AA15. The LPMO may be PaLPMO9E (SEQ ID NO: 1), which is an AA9 LPMO originally isolated from an ascomycete fungus (Podospora anserina). The LPMO may be an AA9 LPMO from Trichoderma reesei (SEQ ID NO: 23).

[0532] Aerobic conditions may include the addition of oxygen, which may be provided by aerating the substrate mixture with an oxygen-containing gas such as air. Aeration may be performed by introducing oxygen-containing bubbles into the aqueous substrate mixture via various systems such as air injectors, aeration frits, membrane systems, or internal circulation airlift reactors. The concentration of molecular oxygen in the enzymatic reaction may be from about 4 mg / L to about 14 mg / L.

[0533] Another enzyme that can be used in this method is "cellulase", which has hydrolytic activity on cellulose, for example, endo-1,4-β-glucanase, cellobiohydrolase and / or β-glucosidase activity. The enzyme is capable of cutting 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-glucanase includes an enzyme selected from GH5, GH7 and GH12 enzymes, such as derived from Aspergillus niger (SEQ ID NO: 12, 13 and 14) and Trichoderma reesei (SEQ ID NO: 24 and 25).

[0534] Another enzyme is "cellobiohydrolase", which has hydrolytic activity on cellulose and produces primarily cellobiose as a product. Cellobiose is a disaccharide and a cellooligosaccharide. The enzyme is capable of cleaving glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. Cellobiohydrolase can come from the GH6 and GH7 families, Cel6A or Cel7A enzymes derived from Trichoderma reesei (SEQ ID NOs: 10 and 11).

[0535] Another enzyme is a "β-glucosidase," which has hydrolytic activity on cellulose and produces primarily 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. The β-glucosidase may include a GH3 β-glucosidase, such as derived from Trichoderma reesei (SEQ ID NO: 22).

[0536] Another class of enzymes is lichenase, which can be selected from GH5, GH7, GH8, GH9, GH12, GH16, GH17 or GH26 families. In some cases, lichenase can be GH16 enzyme. GH16 enzyme can be derived from Bacillus subtilis (Bacillus subtilis) (SEQ ID NO:2). The enzyme can act on, for example, mixed bond glucan (which is a glucan containing a mixture of β-1,3 bonds and β-1,4 bonds), and can cut them at β-1,4 glycosidic bonds. In the case where lichenase acts on mixed bond glucan, the β-glucan produced can mainly fall within the size range of about 3 to about 7 residues, so they can be used in food, cosmetics and nutrition products industries. Mixed bond glucan is abundant in members of grass and horsetail families, and therefore, grass biomass (such as straw) has a high level of mixed bond glucan, and can be effectively acted on by lichenase.

[0537] Another enzyme is a xylanase, which acts on plant biomass that for example comprises a xylan backbone. Xylanase can be for example a glucuronoxylanase, an arabinoxylanase or a glucuronyl arabinoxylanase. The enzyme can be active to a variety of polymers with a xylan backbone, such as glucuronoxylan, arabinoxylan and glucuronyl arabinoxylan. These polymers are abundant in various plant biomass, and for example hardwood and softwood can comprise suitable polysaccharides, wherein hardwood comprises glucuronoxylan conventionally, and softwood comprises glucuronyl arabinoxylan conventionally. In some embodiments, the xylanases may include GH5 xylanases derived from Clostridium thermocellum (SEQ ID NO:3) and Hydroxytricha acuminatum (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).

[0538] Other enzymes useful in the present disclosure may include xyloglucanase and xyloglucan endoglucanase (XEGs), which are produced by a variety of organisms (including plant pathogenic microorganisms). Xyloglucanase and xyloglucan endoglucanase can act on xyloglucan, which is a hemicellulose β-1,4 glucan chain abundant in the primary cell wall of higher plants, which is modified by xylose, and some xylose residues are further modified by other residues (such as galactose). When a suitable xyloglucanase or XEG acts on xyloglucan, the product includes xyloglucan oligosaccharides of the length main chain that can be used in the food, cosmetics and nutrition industry. Xyloglucanase may include GH5 xyloglucanase (SEQ ID NO:8) derived from Bacteroides ovatus and GH74 xyloglucanase derived from Trichoderma reesei.

[0539] Because any given natural plant biomass may comprise a mixture of different polysaccharides, a mixture of different enzymes may be useful sometimes. This mixture may comprise one or more other enzymes. For example, this mixture may comprise LPMO and endoglucanase, xylanase and lichenase, cellobiohydrolase and mannanase, or endoglucanase and cellobiohydrolase, wherein the enzyme partner exists in a mol ratio of 1:100 to 100:1.

[0540] In some cases, one or more enzymes can be a mixture of different enzymes, for example 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 fermentation product, which is subjected to minimum processing after extraction. For example, conventional preparations can only be subjected to filtration to remove insoluble components. As used herein, the term "semi-crude enzyme preparation" generally refers to a soluble preparation extracted from a microbial fermentation product, which is subjected to some processing after extraction, for example preparations can be subjected to filtration to remove insoluble components, increases enzyme concentration, and / or nanofiltration to remove small molecular weight compounds.

[0541] In some cases, crude enzyme preparations or semi-crude enzyme preparations can be from bacteria or fungi. For example, the preparation can be from a fungus, such as a filamentous cellulose decomposing fungus, such as from a Trichoderma or Aspergillus species. The enzyme can be a crude enzyme preparation or semi-crude enzyme preparation from a Trichoderma reesei strain.

[0542] In step (e), oligosaccharides can be isolated from the enzymatic reaction mixture in a variety of ways. It can be separated based on solubility so that only the composition of soluble sugars is extracted for further processing, and / or separated by chromatography to produce a composition with a narrower band of oligosaccharide chain length. Separation can be based on precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration, for example. In the case of separation based on solubility, the sugar spectrum present in the separated composition can depend on the initial enzymatic reaction, because the solubility of different sugars decreases at different rates with length.

[0543] It is also contemplated within the scope of the present disclosure that all or part of the oligosaccharides produced are further processed to produce further products which are then incorporated into foods, cosmetics or nutritional products. The further processing may include any chemical, physical or enzymatic steps, such as reduction, such as reductive amination where appropriate; oxidation, caramelization, modification with Schiff bases or by Maillard reaction or by any combination of these steps, and may provide different products with improved properties for the desired purpose. For example, the caramelization characteristics, calorific value, flavor and color may be modified. The oligosaccharides may also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0544] The composition formed in step (e) may contain various oligosaccharides and in different amounts depending on the desired properties. In various cases, the composition may contain at least 20% dry weight or at least 30% dry weight of cellooligosaccharides having a degree of polymerization of 2 to 6, the composition may contain at least 20% dry weight or at least 30% dry weight of xylo-oligosaccharides having a degree of polymerization of 2 to 12, the composition may contain at least 20% dry weight or at least 30% dry weight of mixed bond glucan-oligosaccharides having a degree of polymerization of 2 to 5, the composition may contain at least 20% dry weight or at least 30% dry weight of manno-oligosaccharides having a degree of polymerization of 2 to 12, the composition may contain at least 20% dry weight or at least 30% dry weight of xyloglucan-oligosaccharides having a degree of polymerization of 4 to 12, and / or the composition may contain at least 20% dry weight or at least 30% dry weight of chitosan-oligosaccharides having a degree of polymerization of 2 to 12. In some embodiments, an ingredient may comprise up to 100% by dry weight of the above oligosaccharides and the polysaccharides described herein, thus the above embodiments wherein oligosaccharides are present at at least 20% by dry weight do not comprise all six types of oligosaccharides.

[0545] In some cases, the ingredient may contain at least 50%, 60%, 70%, 80%, 90%, 95%, 99% or 99.5% sugar present by dry weight. The ingredient may consist essentially of sugar. For example, the ingredient may have less than 0.5% dry weight or less than 0.3% dry weight, such as 0.1% dry weight of other substances.

[0546] In various cases, the composition can include at least two oligosaccharides. The amount of each oligosaccharide can vary according to the desired properties of the resulting food, cosmetics or nutrition products. The two oligosaccharides can be present in a ratio of 1:9 to 9:1 or 1:2 to 2:1. Further, the composition can include three oligosaccharides, four oligosaccharides, five oligosaccharides or six oligosaccharides.

[0547] In some embodiments, the ingredients may include cellooligosaccharides, such as a combination of cellooligosaccharides and xylo-oligosaccharides. In certain embodiments, the ingredients may include a combination of cellooligosaccharides and manno-oligosaccharides.

[0548] The produced ingredients can be used for applications of conventionally used oligosaccharides, sugars, bulking sweeteners, low-intensity sweeteners or other related food ingredients. For example, as a sweetener, bulking agent, added dietary fiber or humectant. Of particular note is the use of reduced sucrose in food. The ingredients can be incorporated into cakes, breads or other baked goods; put into chocolate or other candies such as toffee, soft candy, meringue, jam, jelly or caramel; or incorporated into beverages, for example to provide a good taste or color characteristic or to increase the dietary fiber content. In some cases, the ingredient can be incorporated into animal feed, for example, as an isolated ingredient or by directly utilizing the enzymatic reaction mixture as a raw material.

[0549] Compositions or ingredients as described herein can be used to change one or more characteristics of the final product. The characteristics include, but are not limited to, sweetness, texture, mouthfeel, adhesion, glossiness, smoothness, wettability, viscosity, color, hygroscopicity, flavor, fluffiness, water retention, caramelization, surface texture, crystallization, structural properties, reduced calories, reduced glycemic index, reduced glycemic load, increased fiber, reduced sugar content, and solubility. These can be possible improvements to the current use of different types of sugars, sugar substitutes, and / or other such compounds.

[0550] In the cosmetics industry, the ingredients can improve texture and moisture retention, act as UV absorbing molecules, maintain gel or cream structure, and / or act as bulking agents. In addition, the ingredients can be used in nutraceutical compositions because the dietary fiber they provide has been shown to promote digestive health, good regulation of intestinal flora, and other health benefits. In this case, the ingredients herein can also be used as ingredients in probiotic beverages or other prebiotic or probiotic preparations.

[0551] 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 following steps: (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) 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 a solution or suspension; e) separating one or more oligosaccharides from the enzymatic reaction mixture and using the one or more oligosaccharides to form an ingredient. 2) A method according to numbered embodiment 1, wherein the physical pretreatment step comprises cutting, chopping, grinding, ball milling, grinding, trimming or mixing the plant biomass. 3) A method according to numbered embodiments 1 or 2, wherein the washing cycle of step (i) is carried out in water, acid or base. 4) A 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) A method according to any of the preceding numbered embodiments, wherein the washing cycle of step (i) is carried out for a time range of 0.5 minutes to 72 hours, 1 minute to 12 hours, 5 minutes to 24 hours, or 10 minutes to 3 hours. 6) A method according to any of the preceding numbered embodiments, wherein the thermochemical pretreatment comprises heating the plant biomass in a solution of water, an acid, or an alkali. 7) A 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) A method according to numbered embodiments 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 one 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, aqueous ammonia, ammonium sulfate, or ammonium hydroxide. 11) The method according to any one 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 one of numbered embodiments 1 to 12, wherein the plant biomass is sugar cane, corn stover, corn cob, wheat bran, wheat straw, hardwood, or softwood.14) A method according to any one of numbered embodiments 1 to 13, wherein the plant biomass comprises cellulose, chitin, chitosan, xylan, xyloglucan, mixed bond glucan, mannan or lignocellulose. 15) A 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) A method according to any one of numbered embodiments 1 to 15, wherein the one or more polysaccharide cleaving enzymes are prepared by Trichoderma reesei fungus. 17) A method according to any one of numbered embodiments 1 to 16, wherein the one or more oligosaccharides comprise β-glucan, cellooligosaccharide, MLG oligosaccharide, manno-oligosaccharide or xylo-oligosaccharide. 18) A 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) A 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.

[0552] The detailed description will be further supplemented with reference to the following numbered embodiments. 1) A method for producing an ingredient for human consumption, the method comprising: (a) physically treating plant biomass; (b) subjecting the physically treated plant biomass to an 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) an alkaline 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 the one or more oligosaccharides to form the ingredient. 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 of the numbered preceding embodiments, wherein the thermochemically treated plant biomass contains no or substantially no monosaccharides. 4) The method according to any of the numbered preceding embodiments, further comprising purifying one or more oligosaccharides from the solution or suspension. 5) The method according to any of the numbered preceding embodiments, further comprising repeating step (b). 6) The method according to numbered embodiment 5, wherein step (b) is performed two, three, four or five times. 7) The method according to any of the numbered preceding embodiments, further comprising repeating step (c). 8) The method according to numbered embodiment 7, wherein step (c) is performed two, three, four or five times. 9) The method according to any of the numbered preceding 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 numbered preceding embodiments, wherein a portion of the dissolved monosaccharides and / or disaccharides removed in step (b) is not combined with a portion of the one or more oligosaccharides of step (e) to form a component. 11) A method according to any of the preceding numbered embodiments, wherein the component is substantially free of monosaccharides. 12) A method according to any of the preceding numbered embodiments, wherein the component is substantially free of disaccharides. 13) A method according to any of the preceding numbered embodiments, 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) xylo-oligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed-linkage glucan-oligosaccharides having a DP of 2 to 5; vi) xyloglucan-oligosaccharides having a DP of 4 to 12; or vii) chito-oligosaccharides having a DP of 2 to 12.14) A method according to numbered embodiment 13, wherein the component comprises at least two of the oligosaccharides listed in (i) to (vii). 15) A method according to numbered embodiment 14, wherein the component comprises at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other. 16) A method according to any of the preceding numbered embodiments, wherein the monosaccharide and / or disaccharide comprises at least one of sucrose, glucose, maltose, lactose, glucose, fructose or galactose. 17) A 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, grinding, trimming or mixing the plant biomass. 18) A method according to any of the preceding numbered embodiments, wherein the incubation of step (b) is carried out in an incubation solution comprising water. 19) A 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) A 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) A 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 an alkaline solution. 22) A method according to numbered embodiment 21, wherein the heating is performed at a temperature of 50°C to 150°C. 23) A method according to numbered embodiment 21 or 22, wherein the heating is performed for 30 minutes to 4 hours. 24) A method according to any of the preceding numbered embodiments, wherein the incubated plant biomass is thermochemically treated in an alkaline solution having a pH of 8 to 11. 25) A method according to numbered embodiment 24, wherein the alkaline solution comprises at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, aqueous ammonia, ammonium sulfate, or ammonium hydroxide. 26) A method according to any of the preceding numbered embodiments, wherein the incubated plant biomass is thermochemically treated in an acidic solution having a pH of 4 to 6. 27) A method according to numbered 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) A method according to any of the preceding numbered embodiments, wherein the plant biomass comprises at least one of sugar cane, corn stover, corn cobs, wheat bran, wheat straw, hardwood, or softwood. 29) A method according to any of the preceding numbered embodiments, wherein the plant biomass comprises at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed-linkage glucan, mannan, or lignocellulose. 30) A method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide-cleaving enzymes comprise at least one of a cellulase, a xylanase, a xyloglucanase, an endoglucanase, a cellobiohydrolase, a mannanase, a lichenase, or a lytic polysaccharide monooxygenase (LPMO).31) A 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) A method according to any of the preceding numbered embodiments, wherein the one or more polysaccharide cleaving enzymes are prepared by Trichoderma reesei fungus. 33) A 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) A 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) A method according to numbered embodiment 34, wherein the non-catalytic module is a carbohydrate binding module. 36) A method for producing an ingredient for human consumption, the method comprising: (a) pre-treating plant biomass, wherein the pre-treating 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 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 the following: (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 pre-treated plant biomass in a solution or suspension to form one or more oligosaccharides; and (c) separating a portion of the one or more oligosaccharides to form the ingredient.

[0553] Example

[0554] The following illustrative examples represent embodiments of the compositions and methods described herein and are not meant to be limiting in any way.

[0555] Example 1 - Exemplary Process

[0556] The following steps can be performed to produce the compositions provided herein:

[0557] 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 at room temperature for 60 minutes. At the end of 60 minutes, mixing was stopped and the liquid was filtered off while retaining the solids.

[0558] 2. Resuspend the solids in water at a solids concentration of 10% (w / w). Start mixing, heat to 95°C, and mix at 95°C for 60 minutes.

[0559] 3. At the end of 60 minutes of heating, add 6 g of sodium hydroxide (0.2-3% of the weight of the corn cob) and continue stirring. Heat to 95°C and mix for 60 minutes to break down the hemicellulose present in the corn cob. At the end of 60 minutes, stop heating and cool to 50°C.

[0560] 4. Hydrolysis: Add cellulolytic enzymes (eg, from Trichoderma reesei) and incubate at 50°C at pH 5.5 (adjusted with 1 mol / L sulfuric acid and / or 1 mol / L sodium hydroxide) for 72 hours.

[0561] 5. Separation of biomass: At the end of hydrolysis, the liquid is separated from the product by a solid-liquid separator.

[0562] 6. Enzyme separation after enzymatic hydrolysis: The liquid fraction in the slurry includes the enzymes, oligosaccharides, water and salts that need to be separated. Use 10 kDa hollow fiber membranes to separate enzyme proteins and other macromolecules.

[0563] 7. Use ion exchange columns to remove salt at ≤45°C.

[0564] a. Cation column: Strong acidic cation exchange resin, cross-linked polystyrene matrix, sulfonate functional group, Na + Counter ion.

[0565] b. Anion column: macroporous, weakly basic anion exchange resin, cross-linked polystyrene matrix, dimethyl tertiary amine functional group, OH - Counter ion.

[0566] 8. Oligosaccharide concentration: Selectively concentrate the desired oligosaccharides by nanofiltration at room temperature.

[0567] 9. Concentration: Concentrate the liquid to 40-75% at 60-80°C.

[0568] 10. Spray drying: Spray drying, where the inlet temperature is 130-160°C and the outlet temperature is 65-85°C.

[0569] Example 2 - Removal of soluble sugars by washing cycles (ie, incubation cycles)

[0570] The following steps are performed to remove soluble sugars from plant biomass:

[0571] 1. Water was added to 100 mg of four types of plant biomass (ie, sugarcane, wheat, corncob, and willow) to a concentration of 10% (w / v) and incubated at 45°C for 30 minutes, and then the suspension was centrifuged and the supernatant was removed.

[0572] 2. Repeat step 1 5 times.

[0573] 3. Analyze 2.5 μl of each supernatant fraction by thin layer chromatography (TLC).

[0574] Figure 1The thin layer chromatography of shows the presence of soluble sugars washed out from four types of plant biomass in five continuous washing cycles or incubation cycles (1, 2, 3, 4 and 5). Unwanted monosaccharides and disaccharides (such as glucose, sucrose and maltose) are indicated by arrows. The result of TLC shows that for all plant biomass, the supernatant removed after the first washing cycle has abundant monosaccharides and disaccharides present therein. Therefore, the washing cycle successfully removes monosaccharides and disaccharides from plant biomass. The supernatant from subsequent washing cycles has significantly less monosaccharides and disaccharides present therein (if any), which shows that plant biomass has minimal monosaccharides and disaccharides remaining therein after the washing cycles as provided herein.

[0575] Example 3 - No washed soluble sugars present in enzymatic hydrolysis

[0576] Perform the following steps to demonstrate the absence of soluble sugars from the enzymatic hydrolysis:

[0577] 1. Washed corn cobs 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 corn cobs and willow biomass were also each incubated in 1% (w / v) NaOH at 99°C for 30 minutes and then cooled.

[0578] 2. 150 μl of the resulting suspensions were each mixed with 150 μl of 1 M ammonium acetate (pH 5.5) and 150 μl of an enzyme composition comprising beta xylanase and cellobiose degrading enzymes. The suspensions were then incubated at 50° C. for 16 hours to allow the enzymes to react with the plant biomass.

[0579] 3. Take 2.5 μl of each supernatant and analyze by TLC.

[0580] Figure 2 The thin layer chromatography of the results showed the enzymatic digestion products of the four types of biomass that were washed (+) or unwashed (-) in Example 2. The results showed that after enzymatic digestion, the final products included glucose, sucrose, and maltose for the unwashed corn cob and willow plant biomass. However, the final products after enzymatic digestion did not include glucose, sucrose, and maltose for the washed corn cob and willow plant biomass.

[0581] Example 4 - Adding polymer to oligosaccharide solution can dry it into hard glaze

[0582] The following steps were performed to show that adding polymers to oligosaccharide solutions can cause them to dry into hard slurries:

[0583] 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.

[0584] 2. The samples were dried at 37°C.

[0585] 3. The sample was scored with a knife to test whether the ingredients sank into a solid slurry. With cellobiose alone, no firm slurry was formed, and the dried powder easily broke when pressure was applied with a knife. In contrast, with the addition of 1% w / v xylan and 80 mM or less of cellobiose, the composition was dried to form a solid, off-white, translucent surface that was strong enough to leave a mark 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 slurry returned to a state in which no xylan was present. That is, no firm slurry was formed, and the dried powder easily broke when pressure was applied with a knife (see Figure 3 ).

[0586] Example 5 - Demonstration of a composition comprising two oligosaccharides and a polysaccharide in a food product

[0587] The following steps were performed to demonstrate the inclusion of a combination of both oligosaccharides and polysaccharides in a food product:

[0588] 1. Dissolve 4g birchwood xylan in 75ml water and boil.

[0589] 2. Add 12 g cellobiose and 24 g xylo-oligosaccharide (mainly degree of polymerization (DP) 2-6) in 3 g increments and dissolve and boil.

[0590] 3. Reduce the mixture to 50 ml by heating to form a concentrated solution which has the consistency and appearance of cloudy honey but is less sweet.

[0591] 4. 10 mL of the mixture was mixed with 12 g oats to make a flapjack bar / cereal bar mix and another 10 mL of the mixture was mixed with 6 g fruit and 6 g nuts to make a cereal bar mix.

[0592] 5. Bake the sample at 100°C for 10 minutes and then leave to cool and dry overnight.

[0593] like Figure 4 As shown, Panel A (flapjack bars / cereal bars) and Panel B (fruit and nut bars), the produced flapjacks and fruit and nut bars have an ideal texture and consistency consistent with flapjack bars / cereal bars and fruit and nut bars made with known syrups commonly used in baking.

[0594] For the flapjack / cereal bars, the thickened solution from step 3 helps bind the mixture together in step 4. The combination of the ingredients and the properties of the oats produces a grainy surface texture consistent with flapjack / cereal bars produced using conventional sugar. The ingredients produce a firm, chewy, moist, sticky texture that produces a mouthfeel consistent with the desired mouthfeel of these types of foods, but not present in oats alone. The product has a slightly sweet taste without bitterness or off-flavors, which can be characteristic of high-intensity sweeteners.

[0595] For fruit and nut bars, the concentrated solution from step 3 helps bind the mixture in step 4 together. It also adds a smooth, shiny syrupy top, which is a core part of the aesthetic quality of such foods and would not be produced if the ingredients contained only oligosaccharides (i.e., oligosaccharides without polysaccharides). The ingredient creates a firm, chewy, moist, and sticky texture that produces a mouthfeel consistent with the desired mouthfeel of these types of foods, but which is not present in nuts or fruits alone. The product is slightly sweet, without bitterness or off-flavors, which can be characteristic of high-intensity sweeteners.

[0596] Example 6 - Process for preparing ingredients

[0597] The following steps can be performed to prepare the ingredients provided herein:

[0598] 1. 15% w / v corn cobs (ground through a 1 mm pore size filter) were heated in 1% w / v NaOH at 90°C for 1 hour to dissolve a portion of the polymer component of the biomass.

[0599] 2. Adjust pH to 5.5 with sulfuric acid.

[0600] 3. Extract a volume containing only the liquid components of the reaction ("soluble polymer") that accounted for 15% of the total reaction volume; retain the remaining 85% volume of the reaction, including any insoluble biomass fractions ("remaining biomass").

[0601] 4. To the remaining biomass, add a cellulolytic enzyme cocktail (e.g., an enzyme cocktail from Trichoderma reesei, including cellulases, lignocellulases, and arabinofuranosidases, LPMO, etc.) to 0.5% w / v and incubate at 50°C for 24 hours.

[0602] 5. Separate the soluble oligomeric reaction products from the undigested insoluble polymers by filtration.

[0603] 6. By sequential use of microfiltration, ultrafiltration and ion exchange chromatography (e.g., cross-flow filtration on ceramic membranes; filtration can be performed at a feed pressure of up to 3 bar using Inside Céram candle filters (TiO2, x L 1178mm HD 6mm, 8 channels per membrane), to perform) purification of oligomeric reaction products.

[0604] 7. Purify the soluble polymer by using ultrafiltration (e.g., 10 kDa spiral wound membrane (Snyder ST-2B-6338, PES, feed gap thickness 31 mm) run on an Alfa Laval ultrafiltration unit).

[0605] 8. The solutions formed in steps 6 and 7 were recombined and further purified and concentrated by using nanofiltration to form fractions.

[0606] Example 7 - Viscosity measurements of different solutions

[0607] Perform the following steps to measure the viscosity of different solutions:

[0608] 1. Three sugar solutions containing cellobiose (Cell2), xylo-oligosaccharides (XOS) mainly DP-6 and polymerized beechwood xylan (BWX) were prepared by boiling sugars in water. The final concentrations were:

[0609] a. Sample 1: 0.33 g / ml Cell2, 0.66 g / ml XOS, 0.13 g / ml BWX;

[0610] b. Sample 2: 0.17 g / ml Cell2, 0.33 g / ml XOS, 0.07 g / ml BWX;

[0611] c. Sample 3: 0.54 g / ml Cell2, 0.52 g / ml XOS, 0.07 g / ml BWX.

[0612] 2. Test the samples using a Brookfield HDB VE rotational viscometer using standard test procedures. Take 400 mL of sample in a tall beaker to ensure that no container effects occur. The instrument is operated according to the manufacturer's instructions and the range is as follows: Rotational viscometry using spindle code 61, spindle speed 100 rpm, at 22°C.

[0613] Sample No. Viscosity(cps) 1 393 2 13 3 26

[0614] 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 polysaccharide concentration had a greater effect on the viscosity of the composition than the total concentration of oligosaccharides and polysaccharides. Samples 2 and 3 had the same polysaccharide concentration, but sample 3 had twice the total oligosaccharide and polysaccharide concentrations of sample 2. The viscosity of sample 3 was twice the viscosity of sample 2, which conformed to the linear relationship between total concentration and viscosity. However, as the polysaccharide concentration increased, the viscosity values ​​increased exponentially. Sample 1 had twice the polysaccharide concentration of sample 3, which had the same total oligosaccharide and polysaccharide concentrations, but the viscosity of sample 1 was fifteen (15) times greater than that of sample 3.

[0615] Example 8 - Preparation of water-soluble liquid products / ingredients

[0616] The following steps were performed to produce a water-soluble liquid product / ingredient (Sample 4):

[0617] 1. Heat 100 g of ground corn cobs in 1 L of deionized water containing 2.5 g of sodium chlorite at 80°C for 1.5 hours with constant stirring. Make up the residual volume to 900 mL by adding 200 mL of deionized water containing another 5 g of sodium chlorite and heat at 80°C for 1 hour with constant stirring.

[0618] 2. Filter the solution through a 2 mm pore size ceramic filter funnel under vacuum until the filtrate is clear.

[0619] 3. The retained solids were incubated in 1 L of 0.5 M sodium hydroxide 0.1% (w / v) sodium borohydride at 50°C with shaking at 115 rpm for 17 hours.

[0620] 4. The pH was then adjusted to 7 with concentrated sulfuric acid and dialyzed against tap water in 12,000 Dalton cut-off dialysis tubing for 24 hours.

[0621] 5. Transfer the contents of the dialysis tubing to a 2-L beaker and allow the insoluble fraction to settle by gravity.

[0622] 6. Pour off the supernatant twice and evaporate to a volume of 120 mL at 80°C.

[0623] 7. After adding 3 volumes of ethanol, the water-soluble polymer was precipitated by centrifugation. The resulting supernatant was discarded and the precipitate was air-dried at room temperature to constant weight.

[0624] 8. Add the oligosaccharides to the final 10% w / w cellobiose, 75% w / w xylo-oligosaccharides, 15% extracted water soluble polymers and mix in a Waring Xtreme mixer at the lowest power setting until homogeneous. 94 g of solids were recovered from the mixer.

[0625] 9. Unexpectedly, all 94 g of 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.

[0626] The sample generated (eg, at steps 8 and 9 above) is referred to as sample 4.

[0627] Example 9 - Physicochemical properties of the water-soluble liquid product of Example 8

[0628] Flow properties: The flow properties of the water-soluble liquid product according to the present disclosure described in Example 8 (Sample 4) are detailed in Table 1 along 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 30 mL with 90°C water. Flow properties 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.

[0629] Table 1: Flow characteristics

[0630] sample 5mL flow time (seconds) 20mL flow time (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 Undetermined Sample 4 237 Undetermined

[0631] The increase in the time required for the sample to flow out of the bottom of the syringe (i.e., with a lower flow rate) is related to the increase in sample viscosity. The lower the flow rate, the more syrupy / viscous and viscous the liquid sample is. The measured flow rate of sample 4 is similar to glycerol and lower than all the glucose solutions tested. This property of sample 4 makes it more suitable as a binder in food (such as cereal bars) than glucose solutions and water, and provides sweetness to the product.

[0632] Color: The color of sample 4 corresponds to No. 30 in the Standard Reference Method (SRM), which is the recommended method of 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, USA). In short, the absorbance of the sample was measured at a wavelength of 430 nm in a cell with a path length of 1 cm. The resulting absorbance value was multiplied by 12.7 to generate a 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).

[0633] Anion Exchange Chromatography: Sample 4 was analyzed by high-performance anion exchange chromatography (HPAEC) using a Thermo Fisher Scientific DIONEXICS-6000 system equipped with a CarboPac PA200 analytical column (3 x 250 mm) and a CarboPac PA200G guard column (3 x 50 mm) and a Dionex ED electrochemical detector. Data were acquired using Chromeleon 7 software.

[0634] Eluents A (ultrapure water), B (250 mM NaOH) and C (250 mM NaOH + 1 M sodium acetate) were used to prepare mobile phases with the gradient characteristics shown in Table 2.

[0635] Table 2: Gradient characteristics

[0636] Time (min) Flow (ml / min) ％A ％B ％C 0 0.5 75 25 0 3 0.5 75 25 0 6 0.5 50 50 0 15 0.5 50 42.5 7.5 20 0.5 0 0 100 23 0.5 75 25 0 26 0.5 75 25 0

[0637] Samples for analysis were prepared by diluting sample 4 100-fold and passing through a 0.45 μm syringe filter, with an injection volume of 10 μl of analyte.

[0638] HPAEC analysis (see Figure 5) confirmed that Sample 4 is a mixture of monosaccharides, disaccharides and other oligosaccharides consisting of glucose and xylose. This is in contrast to syrups commonly used in the food industry (e.g., 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.

[0639] Example 10 - Cold Pressed Fruit Cereal Bar

[0640] Cold pressed fruit cereal bars are prepared as follows:

[0641] 1. Heat 120 g of Sample 4 from Example 8 with 30 g of coconut oil and add one quarter (1 / 4) teaspoon of cinnamon.

[0642] 2. Once bubbling, remove the mixture from the heat and add 40g oats, 40g dried dates, 10g crispy rice and 10g seeds.

[0643] 3. Mix the ingredients well until all are coated, then transfer the mixture to a freezer bag and place in the freezer. Roll the contents of the bag to a thickness of 7-10mm and freeze at 4°C overnight, then cut into rectangles.

[0644] The resulting product (such as Figure 6 ) is a chewy, sticky cereal bar that is loosely solidified and contains oats and crispy rice with a perceived sweetness delivered by Sample 4 and chopped dates.

[0645] Example 11 - Production of ingredients in a large-scale manufacturing process

[0646] The following steps can be used to produce this ingredient in a large-scale manufacturing process:

[0647] 1. Physical pretreatment of plant biomass: In suspension, 100 kg of ground corncobs were mixed with water at a solid concentration of 15% (w / w). Mixing was started and heated to 95°C and mixed at 95°C for 60 minutes. 6 kg of sodium hydroxide (0.2-3% by weight of corncobs) was added and stirring was continued. Heat to 95°C and mix for 60 minutes to release the hemicellulose present in the corncobs. At the end of 60 minutes, cool to 50°C and adjust the pH to 5.5 with sulfuric acid.

[0648] 2. Removal of some soluble polysaccharides: Remove part of the soluble phase, equivalent to 5-30% of the total xylan. Neutralize with sulfuric acid and concentrate and purify by ultrafiltration. Remove any precipitated polymers.

[0649] 3. Hydrolysis: Add cellulolytic enzymes (eg, from Trichoderma reesei) to the ground corncob mixture and incubate at about 50°C for 12-72 hours.

[0650] 4. Separation of biomass: At the end of the hydrolysis, the liquid is separated from the product by a solid-liquid separator (eg, filter press or decanter centrifuge).

[0651] 5. Enzyme separation after enzymatic hydrolysis: The liquid fraction from the slurry contains the enzymes, oligosaccharides, water and salts that need to be separated. Use 3kDa or 10kDa hollow fiber membranes to separate enzyme proteins and other macromolecules.

[0652] 6. Remove salt using an ion exchange column at ≤45°C.

[0653] a. Cation column: Strong acid cation exchange resin, cross-linked polystyrene matrix, sulfonate functional group and Na + Counter ion.

[0654] b. Anion column: macroporous, weakly basic anion exchange resin, cross-linked polystyrene matrix, dimethyl tertiary amine functional groups and OH - Counter ion.

[0655] 7. Oligosaccharide concentration: Selectively concentrate the desired oligosaccharides by nanofiltration at room temperature.

[0656] 8. Concentration: You can choose to concentrate the liquid to 40-75% at 60-80℃.

[0657] 9. Reconstitution: Combine the purified soluble polymer with the enzyme-produced oligomers at a dry weight ratio of 5:95-20:80.

[0658] 10. Spray drying: The obtained solution is spray dried, wherein the inlet temperature is 130-160°C and the outlet temperature is 65-85°C.

[0659] Example 12 - Use of liquid ingredients in the manufacture of extruded cereal bars

[0660] The following steps can be performed to make extruded cereal bars using the liquid ingredients provided herein:

[0661] 1. 120 kg solution containing 10.5 kg xylan, 57 kg xylooligosaccharides and 7.5 kg cellobiose was heated with 130 kg coconut oil and transferred to a high speed mixer. 200 kg oatmeal and 25 kg chopped date / raisin mixture were added and mixed thoroughly. This was pulsed through a dough machine and set at 20 psi through the dough feed system.

[0662] 2. The mixture is transferred via a conveyor belt and ramshorn and baked at 180°C for 20 minutes. The product is then transferred to a biscuit cutting line via an oven mover and then into a flexible form fill and seal package when cooled to <5°C.

[0663] 3. The resulting product is a soft, sticky cereal bar that is loose and full of oats. The sweetness is delivered by a liquid solution containing 10.5 kg xylan, 57 kg xylo-oligosaccharides, 7.5 kg cellobiose and chopped dates. The liquid solution containing 10.5 kg xylan, 57 kg xylo-oligosaccharides, 7.5 kg cellobiose and coconut oil all act as a binder, holding the other ingredients in the bar together and giving the bar structure.

[0664] Example 13 - Use of a liquid ingredient in the manufacture of extruded breakfast cereal

[0665] The following steps can be performed to make extruded breakfast cereals using the liquid ingredients provided herein:

[0666] 1. Cereal flour (about 85-75% w / v) is mixed with a solution (about 15-25% v / v) containing 22.5g xylan, 30g cellobiose and 97.5g xylo-oligosaccharide per 100g 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 uses a combination of heat and humidification and / or steam and mechanical shear to cook the product, forming the shape of the product by pushing the product through a nozzle. The product is then puffed to a light texture and golden brown and cooled.

[0667] 2. A light, crispy, shaped breakfast cereal product is obtained. The liquid solution containing 22.5 g xylan, 30 g cellobiose and 97.5 g xylo-oligosaccharide gives the product a sweet taste and helps to form the structure of the dough before extrusion.

[0668] Example 14 - Use of liquid ingredients in the manufacture of ketchup

[0669] The following steps can be performed to make tomato sauce using the liquid ingredients provided herein:

[0670] 1. Blend 4 onions and 250g celery in a food processor until finely chopped. Fry them in 5 tablespoons of vegetable oil over low heat 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, half (1 / 2) teaspoon of ground black pepper and 2 teaspoons of celery salt and cook for another minute.

[0671] 2. Add 2 kg of cooked chopped tomatoes, 3 tablespoons of tomato paste, half (1 / 2) teaspoon of hot pepper 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. Return the mixture to a boil, uncovered, for 1 hour, until the tomatoes are soft. Discard the cinnamon stick and blend the sauce mixture until smooth, then sieve.

[0672] 3. The resulting product was a smooth, rich tomato sauce. The liquid solution containing 22.5 g xylan, 30 g cellobiose and 97.5 g xylo-oligosaccharides sweetened the product and added body to the sauce, helping to thicken and bulk it.

[0673] Example 15 - HPAEC Chromatography of Sugars in Water after Corn Cob Washing

[0674] Corncobs were incubated in room temperature water at a concentration of 100 g / L ("Washing" in Table 3), and the water was then poured off. Water was added to the original total volume and heated to 90°C for 60 minutes ("Wetting" in Table 3), and then heated at 90°C in dilute NaOH for 60 minutes ("Pretreatment" in Table 3).

[0675] HPAEC was performed on washed, wetted, and pretreated samples, and the sugar peaks were identified (see Figure 7 , such as the chromatogram from "Washing"). As shown in Table 3, at the beginning of the process, about 2% of the corncob was glucose that could be washed out, and more glucose could be washed out. It was also noted that the pH after the washing step dropped to 4.5.

[0676] Table 3

[0677] step Glucose (g / l) washing 2.43g / l Wetting 1.43g / l Preprocessing 0.02g / l

[0678] Example 16 - Quantification of Sugars and Organic Acids

[0679] In order to quantify the effect of the pre-washing step on the process, Fig. 10A Three batches of corn cobs were processed according to the procedure outlined in . The samples were analyzed by HPLC for sugars ( Fig. 10B ), and HPLC analysis of organic acids ( Fig. 10C ). The differences in the sugar and organic acid compositions isolated from the different samples (samples AE and 5 minutes (1 / 12 hour) to 4 hours) indicate the effect of washing. If more than 150 mL was taken out of the 600 mL per wash, the effect of pre-washing may be greater. Therefore, these data indicate the direction that washing can take, rather than the limits of washing.

[0680] As shown in the figure, glucose, fructose and sucrose all reduce along with pre-washing.Fructose and glucose are mostly decomposed during NaOH treatment, but sucrose is resistant to NaOH treatment.Because sucrose can not be removed from other disaccharides by filtration usually, sucrose can be effectively removed by washing.Table 4 shows the comparison of sugar in sample Ds ("no washing" and "two washings"), and Table 5 shows the comparison of sugar in 4 hour samples ("no washing" and "two washings").

[0681] Table 4

[0682]

[0683] Table 5

[0684]

[0685] Before the "alkali cooking" step (e.g., thermochemical step) begins, pre-washing results in a reduction of about 50% of glucose and fructose and about 80% of sucrose. At the end of the hydrolysis, pre-washing results in a slight difference in small sugars. The concentration of xylose sugars appears to increase (evidenced by negative changes), while the concentrations of glucose and cellobiose decrease.

[0686] The large amounts of organic acids detected were not products of washing but rather pretreatment. However, the unwashed biomass appeared to have a higher total loading of organic acids compared to the twice washed material.

[0687] For the steps leading up to the pretreatment stage, the acid concentration also appears to be lower for the washed biomass.Table 6 shows a comparison of the samples Ds and Table 7 shows a comparison of the four hour samples.

[0688] Table 6

[0689]

[0690] Table 7

[0691]

[0692] The effect of washing on the organic acid content was evident both at the end of the wetting phase (Sample D) and at the end of the hydrolysis reaction (4 hour sample) as shown by "Change" (calculated by "No Wash" minus "Two Washes") (see, e.g., Tables 6 and 7). At the end of the wetting phase (Sample D), all other acids except lactic acid and formic acid (n / d) were detected and were at lower concentrations for the twice washed material.

[0693] At the end of the hydrolysis, the acetate content was higher (hydrolysis releases acetate). The other acids of the washed biomass were still lower than those of the unwashed (except for lactic acid, which was present in low concentrations).

[0694] Visual observation of the samples showed Fig. 10D After two washes, the corncobs released less colored compounds and the juice was lighter. Without being bound by any particular theory, the colored compounds may be phenols and organic acids released during washing.

[0695] Example 17 - Comparison of Cold Pressed Cereal Bars

[0696] Cold pressed cereal bars were prepared according to the previous recipe (see Example 10).

[0697] Soluble and insoluble polysaccharides were used for comparison in cereal bars. The soluble and insoluble polysaccharides were as follows:

[0698] Soluble polysaccharides: 60 mL water containing 94 g dry ingredients consisting of 10% dry w / w cellobiose, 75% xylooligosaccharides and 15% extracted water-soluble polymers (as sample 4 described in Example 8 above).

[0699] Insoluble polysaccharides: 60 mL water, containing 94 g dry ingredients consisting of 10% dry w / w cellobiose, 75% xylo-oligosaccharides and 15% microcrystalline cellulose.

[0700] References Fig.11A As shown in Figure 1, although cereal bars made with insoluble polysaccharides look like solid bars when placed on a table, they begin to fall apart when picked up from the table into the hand due to their soft texture and the ingredients not being well bound together. In contrast, cereal bars made with soluble polysaccharides can be easily handled and retain their shape.

[0701] The hardness and stickiness of cereal bars were measured using a TA-XTPlusC texture analyzer (Stable Micro systems, UK) using the "ExponentC" software. A sample of 9.6cm x 3.8cm x 1cm (L x W x H) was placed centrally below the probe. A 6mm diameter aluminum cylindrical probe was used for penetration testing in the "recovery to start" mode and a 30kg load cell. Once the probe is triggered on the surface, it penetrates the sample a distance of 2mm at a speed of 2mm / s. At this point (2mm depth), the force value is recorded and used as a measure of the "hardness" of the sample. The probe then withdraws from the sample, and the maximum withdrawal force or "stickiness" is recorded at this time. The speed before the test is 1mm / s, and the speed after the test is 10mm / s.

[0702] The results show that the hardest bars are those made with soluble components, while for insoluble components, lower values ​​( Fig. 11B ). Likewise, the sticks containing soluble polysaccharides had higher stickiness, while the sticks containing insoluble polysaccharides had lower stickiness. These results confirmed the visual and tactile observations.

[0703] Table 8: Results

[0704] sample Hardness(g) Viscosity(g) Cereal bars made from soluble polysaccharides 169.19 -20.33 Cereal bars made from insoluble polysaccharides 47.43 -11.24

[0705] Table 9: Texture Analyzer Settings

[0706] model Measuring compression force Options: Restore to start Speed ​​before test: 1mm / s Test speed 2mm / s Speed ​​after test 10.0mm / s distance: 2mm Trigger Type: Automatic - 20g Tare mode: automatic Data acquisition rate: 400pps

[0707] The hardness of the cereal bars was further measured using a TA-XTPlusC texture analyzer (Stable Micro systems, UK) using the "ExponentC" software. A sample measuring 9.6cm x 3.8cm x1cm (L x W x H) was placed in the center below the knife. A knife edge, "recovery to start" mode and a 30kg load cell were used in the cutting test. Once the knife was triggered on the surface, it penetrated the sample 5mm at a test speed of 2mm / s. The maximum force measured during the cutting test was recorded as the hardness of the sample. The speed before the test was 1.5mm / s and the speed after the test was 10mm / s.

[0708] The results obtained using the cutting method confirmed the results obtained using the penetration method described above. The results showed that the hardest rods were those made with soluble polysaccharides, while lower values ​​were obtained for rods containing insoluble polysaccharides, which were softer ( Fig. 11C ).

[0709] Table 10: Results

[0710] sample Hardness(kg) Cereal bars made from soluble polysaccharides 0.73 Cereal bars made from insoluble polysaccharides 0.34

[0711] Table 11: Texture Analyzer Settings

[0712] model Measuring compression force Options: Back to start Speed ​​before test: 1.5mm / s Test speed 2.5mm / s Speed ​​after test 10.0mm / s distance: 5mm Trigger Type: Automatic - 25g Tare mode: automatic Data acquisition rate: 400pps

[0713] Example 18 - Viscosity of Sugar Compositions Containing Insoluble Polysaccharides and Sugar Compositions Containing Soluble Polysaccharides

[0714] Prepare the following samples:

[0715] Sample 1: 60 mL water containing 94 g dry ingredients consisting of 10% dry w / w cellobiose, 75% xylo-oligosaccharides and 15% extracted water-soluble polymers.

[0716] Sample 2: 60 mL water containing 94 g dry ingredients consisting of 10% dry w / w cellobiose, 75% xylo-oligosaccharides and 15% microcrystalline cellulose.

[0717] "Sample 4" of Example 8, "soluble polysaccharide" of Example 17 and "Sample 1" of Example 18 are substantially the same and / or interchangeable. In addition, "Sample 2" of Example 18 and "insoluble polysaccharide" of Example 17 are substantially the same and / or interchangeable.

[0718] The samples were analyzed for flow properties as described in Example 9. The data demonstrate that the soluble polysaccharide modulates the viscosity of the oligosaccharide composition (Table 12). This allows for fine tuning of the viscosity properties of the solution in a manner not normally achievable using oligosaccharides alone.

[0719] Table 12

[0720]

[0721] Sequence Listing

[0722] LPMO

[0723] AA9 LPMO from Podospora anserina (SEQ ID NO: 1).

[0724] GenBank ID CAP67740

[0725] 1 mkgllsvaal slavsevsah yifqqlstgs tkhgvfqyir qntnynspvt dlssndlrcn

[0726] 61 eggasgantq tvtvragdsf tfhldtpvyh qgpvsvylsk apgsassydg sgtwfkikdw

[0727] 121 gptfpggqwt lagsytaqlp scitdgeyll riqslgihnp ypagtpqfyi scaqikvtgg

[0728] 181 gsvnpsgvai pgafkatdpg ytaniysnfn sytvpgpsvf scgsngggss pvepqpqptt

[0729] 241 tlvtstrapv atqpagcava kwgqcggngw tgcttcaags tcntqnayyh qcv

[0730] Lichenase

[0731] GH16 lichenase from Bacillus subtilis subsp. subtilis strain 168 (SEQ ID NO: 2).

[0732] GenBank ID CAA86922.1

[0733] 1 mpylkrvllll lvtglfmslf avtatasaqt ggsffdpfng ynsgfwqkad gysngnmfnc

[0734] 61 twannvsmt slgemrlalt spaynkfdcg enrsvqtygy glyevrmkpa kntgivssff

[0735] 121 tytgptdgtp wdeidieflg kdttkvqfny ytngagnhek ivdlgfdaan ayhtyafdwq

[0736] 181 pnsikwyvdg qlkhtatnqi pttpgkimmn lwngtgvdew lgsyngvnpl yahydwvryt

[0737] 241 kk

[0738] Xylanase

[0739] GH5 arabinoxylanase from Clostridium thermocellum (SEQ ID NO: 3).

[0740] GenBank ID ABN53395.1

[0741] 1 mgasiktsik irtvafvsii aialsilsfi pnrayaspqr grprlnaart tfvgdngqpl

[0742] 61 rgpytstewt aaapydqiar vkelgfnavh lyaecfdpry papgskapgy avneidkive

[0743] 121 rtrelglylv itignganng nhnaqwardf wkfyapryak ethvlyeihn epvawgppys

[0744] 181 sstanppgav dmeidvyrii rtyapetpvl lfsyavfggk ggaaealkdi rafnkavfgn

[0745] 241 enavwtneav afhgyagwqe ttiaveellk agypcfmtey aggawgsgmg gldveltyel

[0746] 301 erlgvswltf qyipptgvsd dvtkpeyfsa lvensglswt pdygnwpaar gvygngglar

[0747] 361 etatwinnfl tgttrieaed fdwggngvsy ydtdsvnvgg qyrpdegvdi ektsdtgggy

[0748] 421 nvgwisegew leytirvrnp gyynlslrva gisgsrvqvs fgnqdktgvw elpatggfqt

[0749] 481 wttatrqvfl gaglqklrin alsggfnlnw ielspistgt ipdgtykfln rangktlqev

[0750] 541 tgnnsiitad ykgiteqhwk iqhigggqyr issagrgwnw nwwmgfgtvg wwgtgsstcf

[0751] 601 iisptgdgyy rivlvgdgtn lqissgdpsk iegkafhgga nqqwailpvs apafptglsa

[0752] 661 vldssgntan ltwnaapgan synvkrstks ggpyttiatn itstnytdtg vatgtkyyyv

[0753] 721 vsavsngvet lnsaeailqy pkltgtvigt qg...

Claims

1. A method of producing an ingredient for human consumption, the method comprising: (a) applying a physical pretreatment to the biomass to reduce the average size of the biomass; (b) applying a wash cycle pretreatment to the physically pretreated biomass obtained from step (a), wherein the wash cycle pretreatment exposes the physically pretreated biomass to water at a temperature of 15° C. to 50° C. to dissolve and remove monosaccharides and / or disaccharides and one or more organic acids in 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 and / or disaccharides from the aqueous solution, wherein 50% or more of the monosaccharides and / or disaccharides are removed from the aqueous solution; (c) applying a strong pretreatment to the wash cycle pretreated biomass obtained from step (b) to improve digestibility of the biomass, wherein the strong pretreatment comprises a thermochemical treatment at a temperature of at least 75° C. and optionally a pH of at least 7.5; (d) contacting one or more polysaccharide-cleaving enzymes with the heavily pretreated biomass obtained from step (c) in 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 fraction.

2. 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 at a pH of at least 7.

5.

3. The process according to any one of claims 1 to 2, wherein the heavily pretreated biomass after step (c) comprises less than 10% w / w monosaccharides.

4. The method according to any one of claims 1 to 2, further comprising purifying the one or more oligosaccharides from the solution or suspension.

5. The method according to claim 1 or 2, wherein the heavily pretreated biomass after step (c) comprises less than 20% w / w monosaccharides.

6. The method of claim 1, further comprising repeating step (b).

7. The method of claim 6, wherein step (b) is performed two, three, four or five times.

8. The method of claim 1, further comprising repeating step (c).

9. The method of claim 8, wherein step (c) is performed two, three, four or five times.

10. The method of claim 1, further comprising concentrating a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

11. The method of claim 1, further comprising discarding a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

12. The method of claim 1, wherein a portion of the dissolved monosaccharides and / or disaccharides removed in step (b) is not combined with a portion of the one or more oligosaccharides of step (e) to form a component.

13. The method of claim 1, wherein the component comprises less than 15% monosaccharides by dry weight.

14. The method of claim 1, wherein the composition comprises less than 50% disaccharides by dry weight.

15. The method of claim 1, wherein the ingredient is free of monosaccharides.

16. The method of claim 1, wherein the composition is free of disaccharides.

17. The method of claim 1, wherein the one or more oligosaccharides comprise at least one of: i) cellooligosaccharides having a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed-linkage dextran 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.

18. The method of claim 17, wherein the ingredient comprises at least two of the oligosaccharides listed in i) to vii).

19. The method according to claim 18, wherein the composition comprises two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other.

20. The method according to any one of claims 17 to 19, wherein the component comprises at least one of sucrose, maltose, lactose, glucose, fructose or galactose accounting for less than 50% total dry weight w / w of the total dry weight w / w of all oligosaccharides of i) to vii).

21. The method of claim 1, wherein the monosaccharide and / or disaccharide comprises at least one of sucrose, maltose, lactose, glucose, fructose or galactose.

22. The method of claim 21, wherein the one or more organic acids comprises at least one of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid, or acetic acid.

23. The process of claim 22, wherein the total weight of oxalic acid, tartaric acid, succinic acid, formic acid, citric acid, malic acid, lactic acid and acetic acid is greater than 10% of the total weight of sucrose, maltose, lactose, glucose, fructose and galactose in the portion of the monosaccharides and / or disaccharides removed in step (b).

24. The method of claim 1, wherein the physical pretreatment of step (a) comprises at least one of chipping, chopping, grinding, or mixing the biomass.

25. The method of claim 1, wherein the wash cycle pretreatment of step (b) is performed for 15 minutes to 1 hour.

26. The method of claim 1, wherein the strong pretreatment of step (c) comprises heating the wash cycle pretreated biomass in an alkaline solution.

27. The method of claim 26, wherein the heating is performed for 30 minutes to 4 hours.

28. The method of claim 1, wherein step (c) comprises treating the wash cycle pretreated biomass in an alkaline solution having a pH of 8 to 11.

29. The method of claim 28, wherein the alkaline solution comprises at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, aqueous ammonia, ammonium sulfate, or ammonium hydroxide.

30. The method of claim 1, wherein the biomass comprises at least one of sugar cane, corn stover, corn cobs, wheat bran, wheat straw, hardwood, or softwood.

31. The method of claim 1, wherein the biomass comprises at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed bond glucan, mannan, or lignocellulose.

32. The method of claim 1, wherein the one or more polysaccharide-cleaving enzymes comprise at least one of a cellulase, a xylanase, a xyloglucanase, a mannanase, a lichenase, or a lytic polysaccharide monooxygenase (LPMO).

33. The method of claim 1, wherein the one or more polysaccharide cleaving enzymes comprise at least one of AA9, AA10, AA11, AA13, AA14, or AA15.

34. The method of claim 1, wherein the one or more polysaccharide-cleaving enzymes are produced by a filamentous fungus.

35. The method of claim 1, wherein the one or more polysaccharide cleaving enzymes are operably linked to a catalytic module.

36. The method of claim 1, wherein the one or more polysaccharide cleaving enzymes are operably linked to a non-catalytic module.

37. The method of claim 36, wherein the non-catalytic module is a carbohydrate binding module.

38. A method of producing an ingredient for human consumption, the method comprising: (a) applying a physical pretreatment to the biomass to reduce the average size of the biomass; (b) applying a wash cycle pretreatment to the physically pretreated biomass obtained from step (a), wherein the wash cycle pretreatment exposes the biomass to water at a temperature of 15° C. to 50° C. to dissolve and remove monosaccharides and / or disaccharides and one or more organic acids in the 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 and / or disaccharides from the aqueous solution, wherein 50% or more of the monosaccharides and / or disaccharides are removed from the aqueous solution; (c) applying an intensive pretreatment to the wash cycle pretreated biomass obtained from step (b) to dissolve polysaccharides and increase digestibility of the plant biomass, wherein the intensive pretreatment comprises a thermochemical treatment at a temperature of at least 75° C. and optionally a pH of at least 7.5; (d) separating one or more dissolved polysaccharides from the biomass; (e) contacting the remaining biomass obtained from step (d) with one or more enzymes to form one or more oligosaccharides; (f) separating the one or more oligosaccharides; and (g) combining a portion of the one or more soluble polysaccharides from step (d) with a portion of the one or more oligosaccharides from step (f) to form the component.

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