Full-calorie, slowly digestible carbohydrate composition

Abstract

The present invention relates to a digestible carbohydrate composition comprising at least 65% (w / w) glucose-based saccharides on a dry basis, the glucose-based saccharides having reducing ends and D-glucose monomers linked by alternating α1-6 glycosidic and α1-3 glycosidic bonds, with maltose units at the reducing ends, and 0.1-30% (w / w) fructose equivalents on a dry basis, the glucose-based saccharides having an average degree of polymerization greater than 12.

Description

Technical Field

[0001] [Background Art] Carbohydrates have functions specific to the human diet. Carbohydrates provide technical functions such as texturizers or bulking agents. In some cases, carbohydrates can be regarded as sweeteners. Carbohydrates also provide energy and regulate blood glucose levels.

[0002] Intake of full-calorie carbohydrates such as glucose syrup, maltodextrin, sugar, and starch hydrolyzates typically results in a high blood glucose peak.

[0003] Current α-glucan components available on the market such as glucose syrup, isomaltooligosaccharide, and starch hydrolyzates are also not very slowly digestible in that their maximum concentration (Cmax) and the area under the rising curve (iAUC) are not sufficiently reduced compared to those of glucose. Furthermore, unpurified α-glucan components such as commercially available scromolt have a high content of monosaccharides, especially fructose, which contributes to added sugars. Furthermore, raw starches such as wheat starch, which are known to have reduced Cmax and iAUC compared to glucose, are not sufficiently soluble and, especially for liquid applications, these properties are lost by heat treatment.

[0004] Other α-glucan components with higher molecular weights such as resistant dextrin, polydextrose, dextran, and reuteran may not be full-calorie because they are difficult to digest.

[0005] It is clear that there is a need to provide to the market an improved, full-calorie and slowly digestible carbohydrate composition that does not have the above-mentioned drawbacks.

[0006] [Summary of the Invention] The inventors of the present application have surprisingly found a composition that does not have the drawbacks of prior art carbohydrate compositions. It is defined as containing 5 to 10% or less sugar (monosaccharides and disaccharides), with little contribution from added sugars. The contribution of fructose is little or none. Further, it is full calorie (3.5 to 4 kcal / g).

[0007] The composition is also suitable for a ready-to-drink (RTD) or ready-to-use (RTU) liquid matrix.

[0008] The composition is also suitable for ready-to-mix or powder applications.

[0009] In a first aspect, the present invention relates to a digestible carbohydrate composition, the digestible carbohydrate composition comprising a. glucose-based saccharides of at least 65% (w / w) on a dry weight basis, having a reducing end and D-glucose monomers linked by alternating α1-6 glycosidic bonds and α1-3 glycosidic bonds, with a receptor molecule present at the reducing end, the glucose-based saccharides, b. fructose equivalents of 0.1 to 30% (w / w) on a dry weight basis, wherein the glucose-based saccharides have an average degree of polymerization of more than 12, and the receptor molecule is preferably a maltose unit.

[0010] In a second aspect, the present invention relates to a food product or beverage comprising the digestible carbohydrate composition.

[0011] In a third aspect, the present invention relates to a method for reducing postprandial glucose in a subject, comprising administering an effective amount of the food product or beverage or the digestible carbohydrate composition to a subject in need thereof.

Brief Description of the Drawings

[0012]

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Mode for Carrying Out the Invention

[0013] Digestible Carbohydrate Composition The present invention relates to a digestible carbohydrate composition, which comprises glucose-based saccharides of at least 65% (w / w) on a dry weight basis, having a reducing end and D-glucose monomers linked alternately by α1-6 glycosidic bonds and α1-3 glycosidic bonds, with a receptor molecule present at the reducing end, glucose-based saccharides, and fructose equivalents of less than 30% (w / w) on a dry weight basis. The glucose-based saccharides have an average degree of polymerization of more than 12, and the receptor molecule is preferably a maltose unit.

[0014] In particular, the present invention relates to a digestible carbohydrate composition, which a. Glucose-based saccharides of at least 65% (w / w) on a dry weight basis, having a reducing end and D-glucose monomers linked alternately by α1-6 glycosidic bonds and α1-3 glycosidic bonds, with a receptor molecule present at the reducing end, glucose-based saccharides, and b. Fructose equivalents of 0.1 to 30% (w / w) on a dry weight basis, and the glucose-based saccharides have an average degree of polymerization of more than 12, and the receptor molecule is preferably a maltose unit. The receptor molecule is preferably a carbohydrate or a carbohydrate derivative.

[0015] The receptor molecule can be selected from sugars or sugar alcohols having a free hydroxyl group at one or more of carbon position numbers 2, 3, and 6 that can accept a glucose unit from sucrose.

[0016]

[0017] ​The carbohydrate receptor is preferably a saccharide selected from the group consisting of maltose units, isomaltose units, maltitol units, (iso)maltotriose units, and methyl-α-D-glucan units. Other preferred receptor molecules are glucose units, gentiobiose units, raffinose units, melibiose units, isomaltitol units, isomaltooligosaccharide units, teanderose, kojibiose units, glucosyl trehalose units, cellobiose units, maltotetraose units, nigerose units, lactose units, panose units, or mixtures thereof.

[0018] Preferably, the receptor molecule is a maltose unit.

[0019] The average degree of polymerization is determined by either GPC-RI (gel permeation chromatography with refractive index detection) or GPC-MALLS (gel permeation chromatography with multi-angle light scattering), or HPAEC-PAD (high performance anion exchange chromatography using pulsed amperometric detection).

[0020] Preferably, the average degree of polymerization (or weight average degree of polymerization) is determined by HPAEC-PAD.

[0021] The average degree of polymerization can be 12 to 90, or 12 to 70, or 12 to 50, or 12 to 35, or 12 to 30, or 12 to 20, or 12 to 18 (HPAEC-PAD).

[0022] The average degree of polymerization can be greater than 13, 14, 15, or 16 (HPAEC-PAD). In some embodiments, the average degree of polymerization can be greater than 17 (HPAEC-PAD).

[0023] The average degree of polymerization can be less than 90, 70, 50, 35, 20, 19, or 18 (HPAEC-PAD).

[0024] The composition typically has little involvement with added sugars in the form of monosaccharides and disaccharides. Preferably, the composition contains less than 10% (w / w), 9% (w / w), 8% (w / w), 7% (w / w), 6% (w / w), or 5% (w / w) of added sugars in the form of monosaccharides and disaccharides.

[0025] In some embodiments, the composition has 65 - 99% (w / w) of glucose - based saccharides on a dry weight basis.

[0026] In some embodiments, the composition has 75 - 95% (w / w) of glucose - based saccharides on a dry weight basis.

[0027] In some embodiments, the composition has 85 - 90% (w / w) of glucose - based saccharides on a dry weight basis.

[0028] In some embodiments, the composition has at least 70% (w / w), preferably at least 75% (w / w) of glucose - based saccharides on a dry weight basis.

[0029] In some embodiments, the composition has at least 80% (w / w), preferably at least 85% (w / w) of glucose - based saccharides on a dry weight basis.

[0030] The composition typically provides only a small amount of fructose equivalents on a dry weight basis.

[0031] In some embodiments, the composition contains less than 25% (w / w) of fructose equivalents on a dry weight basis.

[0032] In some embodiments, the composition contains less than 20% (w / w) of fructose equivalents on a dry weight basis.

[0033] In some embodiments, the composition contains less than 10% (w / w) of fructose equivalents on a dry weight basis. Preferably, the composition contains less than 9% (w / w), less than 8% (w / w), less than 7% (w / w), less than 6% (w / w), less than 5% (w / w), less than 3% (w / w), less than 2% (w / w), less than 1% (w / w), or less than 0.5% (w / w) of fructose equivalents on a dry weight basis.

[0034] In some embodiments, the composition contains 0.1 - 25% (w / w), or 0.1 - 20% (w / w), or 0.1 - 15% (w / w), or 0.1 - 10% (w / w), or 0.1 - 9% (w / w), or 0.1 - 8% (w / w), or 0.1 - 7% (w / w), or 0.1 - 6% (w / w), or 0.1 - 5% (w / w), or 0.1 - 3% (w / w), or 0.1 - 2% (w / w), or 0.1 - 1% (w / w), or 0.1 - 0.5% (w / w) of fructose equivalents on a dry weight basis.

[0035] In some embodiments, the fructose equivalents are leucrose, fructose, and / or sucrose. In some embodiments, the fructose equivalents are leucrose. In some embodiments, the fructose equivalents are fructose. In some embodiments, the fructose equivalents are sucrose. In some embodiments, the fructose equivalents are leucrose and fructose. In some embodiments, the fructose equivalents are leucrose and sucrose. In some embodiments, the fructose equivalents are fructose and sucrose. In some embodiments, the fructose equivalents are leucrose, fructose, and sucrose.

[0036] In one embodiment, the average molecular weight of the composition is greater than 2 kDa.

[0037] In one embodiment, the average molecular weight of the composition is 2 - 14.58 kDa.

[0038] The glucose-based saccharide of the composition further contains an α1-4 bond at one end.

[0039] Typical compositions of the present invention include a glucose-based saccharide having alternating α1-6 bonds and α1-3 bonds, 1.5 to 2.5 grams of fructose equivalents per 33 grams of total carbohydrates, and 30.5 to 31.5 grams of glucose-based carbohydrates per 33 grams of total carbohydrates, and the glucose-based saccharide has an average degree of polymerization of 12 to 13.

[0040] Another typical composition of the present invention includes a glucose-based saccharide having alternating α1-6 bonds and α1-3 bonds, 2.5 to 3.5 grams of fructose equivalents per 33 grams of total carbohydrates, and 29.5 to 30.5 grams of glucose-based carbohydrates per 33 grams of total carbohydrates, and the glucose-based saccharide has an average degree of polymerization of 17 to 18.

[0041] Typically, the composition is full-calorie. In some embodiments, the composition provides at least 3 kcal / gram, or 3.5 kcal / gram, or at most 4 kcal / gram, or 3.5 to 4 kcal / gram.

[0042] The composition induces a lower blood glucose response in a subject compared to maltodextrin or glucose syrup. Typically, during a 3-hour period immediately after ingestion of the composition, the blood glucose response in the subject is lower. The blood glucose response can be measured, for example, by the incremental area under the curve (iAUC) as described herein.

[0043] The composition has the further advantage of being digested slowly. Postprandial hydrogen production can be used as an indirect measure of digestibility. When indigestible carbohydrates reach the colon, they are fermented by colonic bacteria. This fermentation produces gases such as hydrogen and methane, which can be measured in the subject's breath.

[0044] In one embodiment, the composition does not result in more than 20 ppm of hydrogen in the breath during a 240-minute period immediately following ingestion of 33 grams of the composition by a subject. In one embodiment, 33 grams of the composition is digested more slowly than 33 grams of maltodextrin syrup or glucose syrup.

[0045] The gastrointestinal tolerance of the composition is very high. In one embodiment, the composition does not result in one or more of diarrhea, abdominal cramps, vomiting, audible bowel sounds, or flatulence in a subject during a 180-minute period immediately following ingestion of, for example, 33 g of the composition.

[0046] In some embodiments, the digestible carbohydrate composition according to the invention is for use in reducing the blood glucose response in a subject, preferably a human subject.

[0047] In one embodiment, the composition is for use in reducing the blood glucose response in a subject, the composition comprising a glucose-based saccharide having alternating α1-6 linkages and α1-3 linkages, 2.5 to 3.5 grams of fructose equivalents per 33 grams of total carbohydrate, and 29.5 to 30.5 grams of glucose-based carbohydrates per 33 grams of total carbohydrate, and having an average DP of 17 to 18.

[0048] In one embodiment, the composition is for use in reducing the blood glucose response of a subject during a 60-minute, 120-minute, or 180-minute period immediately following ingestion of the composition, as compared to the blood glucose response to glucose syrup in the subject.

[0049] The blood glucose response can be reduced by up to 45% in the subject during a 60-minute period immediately following ingestion. The blood glucose response can be reduced by up to 38% in the subject during a 120-minute period immediately following ingestion. The blood glucose response can be reduced by up to 33% in the subject during a 180-minute period immediately following ingestion.

[0050] Preferably, the subject is a human subject.

[0051] Food product or beverage The present invention also relates to a food product or beverage comprising the digestible carbohydrate composition described herein. The digestible carbohydrate composition can be in the form of a powder, such as a ready-to-mix powder for beverages. In other embodiments, the digestible carbohydrate composition can be in the form of a liquid, such as a syrup.

[0052] Preferably, the beverage is a ready-to-drink (RTD) or heat-treated beverage.

[0053] In some embodiments, the food product or beverage is a nutritional supplement.

[0054] In some embodiments, the food product or beverage is a nutritional formulation.

[0055] The nutritional formulation can be in any oral nutritional form, such as, for example, a health drink, a commercial beverage, optionally a juice, a milkshake, a yogurt drink, a smoothie or a soft drink including a soy-based beverage, a nutritional bar, etc., or can be dispersed in any type of food, such as a baked product, a cereal bar, a dairy product bar, a snack food, a soup, a breakfast cereal, a muesli, a candy, a tablet, a cookie, a biscuit, a cracker (such as a rice cracker), and a dairy product.

[0056] The nutritional formulation can be a nutritional bar.

[0057] The nutritional formulation can further comprise a source of fat, protein, and other carbohydrates.

[0058] The nutritional supplement or nutritional preparation can be, for example, in the form of tablets, capsules, lozenges, or liquids. The nutritional supplement or nutritional preparation may further contain a protective hydrophilic colloid (such as gum, protein, modified starch, etc.), binder, film-forming agent, encapsulating agent / material, partition / shell material, matrix compound, coating, emulsifier, surfactant, solubilizing agent (such as oil, fat, wax, lecithin, etc.), adsorbent, carrier, filler, co-compound, dispersant, wetting agent, processing aid (solvent), fluidizing agent, flavoring agent, bulking agent, gelling agent, and gel-forming agent.

[0059] In some embodiments, the nutritional preparation or nutritional supplement is for use as a drug.

[0060] In some embodiments, the nutritional preparation or nutritional supplement is for managing blood glucose control or reducing blood glucose response in a human subject, such as a healthy human subject.

[0061] In some embodiments, the nutritional preparation or nutritional supplement is for an infant, child, or adolescent human subject.

[0062] In some embodiments, the nutritional preparation or nutritional supplement is for a human subject with diabetes and / or prediabetes.

[0063] In some embodiments, the nutritional preparation or nutritional supplement is suitable for a human subject under emergency treatment.

[0064] In some embodiments, the nutritional preparation or nutritional supplement is suitable for a weight loss product for a human subject.

[0065] In some embodiments, the food product is a pet food product.

[0066] Method for reducing postprandial glucose The present invention also relates to a method for reducing postprandial glucose in a subject, comprising administering to the subject in need thereof an effective amount of a digestible carbohydrate composition or a food product or beverage described herein.

[0067] In some embodiments, the subject is a human subject.

[0068] In some embodiments, the subject is a companion animal subject such as a dog or a cat.

[0069] Definitions All percentages described herein are by dry weight based on the total dry weight of the composition, unless otherwise indicated. As used herein, "about," "approximately," and "substantially" refer to numbers within a numerical range, e.g., within -10% to +10% of the reference number, preferably within -5% to +5% of the reference number, more preferably within -1% to +1% of the reference number, and most preferably within -0.1% to +0.1% of the reference number. All numerical ranges herein are to be understood to include all integers or fractions within the range. Further, these numerical ranges are to be construed as supporting claims directed to any number or sub-set of numbers within this range. For example, the disclosure of 1 to 10 is to be construed as supporting ranges such as 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, and the like.

[0070] As used in this disclosure and the appended claims, the singular forms "a," "an," and "the" include plural referents unless otherwise indicated. Thus, for example, reference to "a component" or "the component" includes two or more components.

[0071] The terms "comprise", "comprises", and "comprising" are to be construed as non-exclusive and capable of including other elements. Similarly, the terms "include", "including", and "or" are all to be construed as including all, provided such construction is not clearly precluded by the context. However, the compositions disclosed herein may not include elements not specifically disclosed herein. Thus, the disclosure of embodiments using the term "comprising" includes the disclosure of embodiments "consisting essentially of" the specified components and embodiments "consisting of" the specified components.

[0072] Digestible carbohydrate composition: A carbohydrate composition that is digested in the small intestine and does not reach the colon does not result in hydrogen production in the breath upon ingestion by a subject, preferably a human subject.

[0073] Carbohydrate composition: All carbohydrates including digestible carbohydrates and sugars.

[0074] Glucose-based saccharides: Carbohydrates of various sizes and molecular weights (MW) containing glucose monomers as constituent units linked by α-glycosidic bonds.

[0075] Maltose unit: A disaccharide composed of two glucose units linked by an α1-4 glycosidic bond.

[0076] Reducing end: The reducing end of a carbohydrate is a monosaccharide having a free anomeric carbon not involved in a glycosidic bond and can thus be converted to an open-chain form.

[0077] Fructose equivalents: Any carbohydrate containing monosaccharides and disaccharides, which is composed of at least one fructose monomeric constituent unit and is subjected to fructose metabolism in the small intestine upon ingestion by a subject, preferably a human subject. Examples are leucrose, sucrose, and fructose.

[0078] Average degree of polymerization (DP): The average number of monosaccharide constituent units per chain of glucose-based saccharides in each carbohydrate composition.

[0079] Maltose-alternan-oligosaccharide (MAOS): An example of a glucose-based saccharide having maltose at the reducing end.

[0080] Monosaccharides: Any class of monosaccharides that can function as constituent units of larger carbohydrate structures. Monosaccharides cannot be hydrolyzed to obtain more simple sugars. Examples of monosaccharides include glucose and fructose. Disaccharides: Any sugar containing two monosaccharides as constituent units, such as sucrose, leucrose, and maltose.

[0081] Added sugars: All of the above monosaccharides and disaccharides present in a composition Blood glucose response or postprandial glucose: The glucose concentration measured in blood or interstitial tissue after ingestion of a carbohydrate by a subject, preferably a human subject.

[0082] The terms "food", "food product", and "food composition" mean a product or composition intended for ingestion by an individual such as a human and providing at least one nutrient to such an individual. As used herein, these terms encompass foods in any form, including both liquids (e.g., beverages) and solids. The compositions of the present disclosure, including many of the embodiments described herein, can contain, consist of, or consist essentially of the elements disclosed herein, as well as any additional or optional raw materials, components, or elements described herein or not described herein that are useful in a diet.

[0083] "Beverage" is a substantially homogeneous liquid that is at least 85% by weight water, in some embodiments at least 90% by weight water or at least 95% by weight water. A "ready-to-drink" beverage is in liquid form that can be consumed without further addition of liquid and is preferably sterile. Reconstitution and dilution can each include the addition of water and / or milk to a powder or concentrate, and in some embodiments, the method includes a reconstitution or dilution step.

[0084] Those skilled in the art will understand that all features of the invention disclosed herein can be freely combined. In particular, the features described for the products of the invention may be combined with the methods of the invention, and vice versa. Furthermore, features described for different embodiments of the invention may be combined. If well-known equivalents exist for a particular feature, such equivalents can be incorporated as specifically recited herein.

[0085] Further advantages and features of the invention are apparent from the figures and non-limiting examples.

Examples

[0086] Example 1 Production of a digestible carbohydrate composition Prior art methods are known, for example, those of International Publication No. 0047727 (A2) and International Publication No. 2009095278 (A2), and include steps P1 - P4 of Figure 1a. In step P1, the bioconversion of sucrose and maltose is carried out in a batch reactor at T = 37°C for about 20 hours. The sucrose:maltose ratio is selected to be 7:1 (w / w or mol / mol). Step P1 is carried out in the batch reactor for bioconversion shown in Figure 2a.

[0087] P2 is a step of ultrafiltration to remove alternan polymer (alternan polysaccharide) and alternan sucrase enzyme (AlSu). This step is carried out in the ultrafiltration device shown in Figure 2a.

[0088] In P3, fructose is removed by nanofiltration. This step is carried out in the nanofiltration device shown in Fig. 2a. Here, water is added and the mixture of water and fructose is removed.

[0089] In the final step P4, the product from P3 is concentrated by evaporation. This step is carried out in the evaporator of Fig. 2a, and maltose - alternan - oligosaccharide (MAOS) is obtained.

[0090] The method of the present invention includes, in a specific embodiment, three steps S1 to S3 shown in Fig. 1b.

[0091] Compared with the prior - art method of Fig. 1a, the bioconversion in step S1 includes the continuous supply of sucrose and the continuous removal of fructose. Semi - continuous supply and removal are possible. The mass ratio of sucrose (total amount added) to maltose is 19:1 (19 kg sucrose / 1 kg maltose), and the duration is about 72 hours. Since fructose has already been removed in step S1, the prior - art step P3 can be omitted. By removing fructose, a higher sucrose - to - maltose ratio can be used, and a higher degree of polymerization of maltose - alternan - oligosaccharide can be achieved.

[0092] In Process S1, bioconversion is carried out in the reactor of Figure 2b, where maltose and alternansucrase enzyme (AlSu) are present in water. As described above, the bioconversion is carried out with a continuous supply of sucrose, continuous removal of fructose, a duration of about 72 hours (variable), T = 37 °C, and a sucrose:maltose ratio of 19:1 (w / w or mol / mol). Sucrose is added as a feed (dissolved in water) and the contents of the reactor are stirred. In the bioconversion within the reactor, alternan oligosaccharides containing the receptor molecule maltose (also called maltose alternan oligosaccharides (MAOS)) are formed as the main product, and alternan polymer, fructose, and leucrose are formed as by-products. The contents are continuously circulated from the reactor through a membrane cell (diafiltration cell), and water, fructose, and leucrose are removed by membrane filtration. In this example, the membrane filtration is nanofiltration carried out as constant volume diafiltration. The leucrose content is reduced to about 30% to less than 10% compared to the prior art. The removed water is replaced by the water feed stream.

[0093] The reactor and the membrane cell form a combination of a combined bioconversion and nanofiltration device, also called a reactor system.

[0094] The method step S2 of this embodiment corresponds to the step P2 of the prior art. Here, the alternan polysaccharide (alternan polymer) and alternansucrase enzyme (AlSu) as by-products are removed by ultrafiltration. This step is beneficial if more alternan polymer is formed as required or to lead to the desired DPw of the remaining alternan species.

[0095] The method step S3 of this embodiment corresponds to the step P4 of the prior art. In this step, the product is concentrated by evaporation.

[0096] The composition of the reaction solution used during production is shown in Table 1. The 2.1 L solution amounts to approximately 5.6 L in total with the water of the system (dead volume).

[0097] The method is carried out for the first hour without depletion in order to minimize the risk of decreasing maltose passing through the membrane. Thereafter, fructose is constantly depleted by the nanofiltration membrane Filmtec NF270-2540 (DOW). Once the chain elongation is complete, the nanofiltration module is replaced with a TRISEP2540-UE50-QXF ultrafiltration module (Microdyn Nadir). This separated the maltose-alternan-oligosaccharide (MAOS) fraction from the longer aged chains and the enzyme. The membranes used in this method and the method parameters used are summarized in Table 2. The filtrate was finally concentrated to a dry matter content of >72%.

[0098]

Table 1

[0099]

Table 2

[0100] Figure 3 shows the increase in chain length over the course of the method. The values were recorded by GPC-RI measurement. The relationship for calculating DPw from the Mw values in Figure 3 is as follows. DPw = Mw / (162 Da). Thus, it can be seen that a DPw of approximately 15.4 is reached at the end of the method (2500 / 162).

[0101] A total of 19 kg of sucrose per kg of maltose was used to reach an average chain length DPw of approximately 15.

[0102] Comparative example: Alternan oligosaccharides were prepared according to the method shown in Figure 1a. A ratio of 21:1 (kg sucrose: kg maltose) was used and a DPw of 9.9 was obtained. GPC-RI was used for the measurement of DPw, but the above-described method-protocol was not exactly used. Nevertheless, it is shown that, compared to the results in Figure 3, alternan with a higher DPw is obtained by the present invention.

[0103] The DPw values of two samples obtained by the method of the present invention analyzed by HPAEC-PAD method are summarized in Table 4.

[0104] Example 2 Structural analysis of the digestible carbohydrate composition prepared according to Example 1 Monosaccharides (glucose, fructose, leucrose, sucrose, maltose) were quantified using a Dionex ICS-3000 DC device equipped with an HPLC carbohydrate column (CarboPac PA1 column, 4×250 mm, no guard at 30 °C), an inert styrene divinylbenzene polymer (Dionex Corporation, 2010), and a gold triple potential pulsed amperometric detector (PAD). Eluent A (300 mM aqueous NaOH solution), eluent B (MiliQ water), and eluent C (500 mM CH3COONa in 150 mM NaOH) were used as the mobile phase in gradient mode for a total run time of 35 minutes.

[0105] The monosaccharides present in the test product on a dry weight basis are summarized in Table 3 below.

[0106]

Table 3

[0107] The DPw value was measured by HPAEC-PAD after reducing and hydrolyzing glucose-based saccharides. A 2 milliliter solution containing 6 g / mL of the digestible carbohydrate composition was treated with 0.2 mL of a NaBH4 solution (40 mg / mL) in 0.5 M ammonia at 40 °C for 30 minutes. Then, the reduced sample was hydrolyzed with 0.5 mL of 2 M trifluoroacetic acid and heated at 121 °C for 1 hour to release monomers. The released monomers were quantified by injecting the sample solution into a Thermo Scientific(™) Dionex(™) ICS-6000 ion chromatography system equipped with CarboPac(™) MA1 and supplying eluent (water and 1000 mM NaOH) at 0.4 mL / min. The DP value was calculated by the following formula.

[0108]

Number

[0109] The DP values of the samples are summarized in Table 4 below.

[0110]

Table 4

[0111] The molecular weights of DCC-1, DCC-2 P1, and DCC-3 P2 were also quantitatively analyzed using thin layer chromatography with a silica TLC plate, a chloroform:acetic acid:water:ethanol mixture (30:35:25:20) as the mobile phase, and a diphenylamine-aniline reagent as the visualizing agent (Figure 4).

[0112] The glycosidic bond profile of glucose-based oligosaccharides was measured by GC-MS using partially methylated alditol acetates. Briefly, the sample was dissolved in anhydrous DMSO, deprotonated by the addition of n-butyllithium (Sigma 230707), and methylated with methyl iodide (Sigma 289566). Subsequently, the methylated sample was hydrolyzed with 2N TFA (60 minutes at 121 °C). The hydrolyzed sample was evaporated under nitrogen stream and redissolved in 1M ammonium hydroxide, and the aldehyde groups were reduced with a DMSO solution containing sodium borate (20 mg / ml). Glacial acetic acid was added dropwise to stop the reaction, and acetylation was performed by adding 1-methylimidazole and acetic anhydride. Partially methylated alditol acetates in acetone were quantified by GCM (7890A-5975C MSD, Agilent Technologies, Inc., Santa Clara, CA, USA) using a Supelco 24111-U SP-2380 capillary column (injector volume 0.5 μL, injector temperature 250 °C, detector temperature 250 °C, carrier gas, helium: 30 mL / min, split ratio 40:1, temperature program, 3 minutes at 100 °C, 4 °C / min to 270 °C for 20 minutes). Electron impact spectra were acquired at 69.9 eV over a mass range of 50 - 550 Da.

[0113]

Table 5

[0114] The higher value of the 1,6-glycosidic bond is explained by the raffinose content in the digestible carbohydrate composition that contributes to the amount of 1,5,6-tri-O-acetyl-1-deuterio-2,3,4-tri-O-methyl-D-glucitol observed. In addition, raffinose, and monomeric glucose, contribute to the amount of terminal Glc in the digestible carbohydrate composition.

[0115] Example 3 Production of Digestible Carbohydrate Composition In this example, the parameters of the method described in Example 1 for DCC-3 were changed as shown in the following table. Also, the alternansucrase was divided into four equal parts and placed in the reactor, and the first part was present before supplying sucrose to the reactor.

[0116]

Table 6

[0117] Enzyme activity means the total units of the alternansucrase enzyme used in the method.

[0118] The reaction time of the method of the present invention (not including further method steps such as removal of alternan polysaccharide and alternansucrase enzyme by further membrane filtration, or concentration of the retentate obtained in further membrane filtration) can be shortened by increasing the amount of sucrose, increasing the sucrose supply rate, increasing the enzyme activity, and increasing the temperature.

[0119] The glycosidic bond profile of the glucose-based oligosaccharide was measured according to Example 2. The results are shown in Table 6 below.

[0120]

Table 7

[0121] Example 4 Results of a preliminary crossover study In a pre-randomized controlled crossover trial, a digestible carbohydrate composition (DCC-1) having a degree of polymerization of 7 was tested in 16 healthy volunteers. 25 g of DCC-1 dissolved in 300 mL of water was ingested, and the postprandial glucose response was measured over 2 hours (Figures 6 and 7). As a control, 25 g of glucose syrup was ingested.

[0122] In this test, the intake of DCC-1 resulted in a lower blood glucose response compared to glucose syrup, but there were no significant differences in tmax, iCmax, and iAUC. Furthermore, no severe gastrointestinal discomfort was reported by the test participants. These results suggest that although most of the ingested α-glucan was digested, the increase in the degree of polymerization (DP7) was slight compared to glucose syrup, and thus no significant advantage for the blood glucose response was obtained.

[0123] Example 5 Clinical Trial - Method Sixteen participants were made to ingest different test products containing different types of carbohydrates with different structures and compositions, and a monocentric, controlled, randomized, double-blind, crossover trial was conducted. The participation of subjects in this research project was voluntary, and they could terminate their participation at any time without the need to justify withdrawing consent or ending participation. If a participant withdrew, the coded data collected up to that point was used and anonymized after analysis.

[0124] The purpose of this study was to test the effects of different α-glucans on postprandial glucose response (PPGR) and hydrogen (H2) production and to provide insights into their digestibility.

[0125] The first objective was to determine whether the tested α-glucans induced a lower glucose response compared to maltodextrin.

[0126] The second objective was to indirectly determine whether the tested α-glucans were completely absorbed in the small intestine and induced gastrointestinal discomfort.

[0127] The glucose response was evaluated for 3 hours after the intake of the test product. The area under the rising curve for 3 hours (3h-iAUC) was used as the primary evaluation item to address the first objective.

[0128] As secondary evaluation items, the following parameters were analyzed: Other parameters derived from PPGR (i.e., cross-sectional value, iCmax, Cmax, tmax, partial iAUC, AUC); Digestibility by measuring exhaled H2 during the 4 hours after ingestion of the test product. From these H2 curves, parameters of 4h-iAUC, partial iAUC, partial AUC, Cmax, and cross-sectional value were derived. All these values were derived from H2 data corrected for CO2. Similar parameters were derived from H2 data not corrected for CO2 and CH4 data (regardless of CO2 correction); Gastrointestinal tolerance at 3 hours after product ingestion by visual analog scale for each target symptom: 1) diarrhea, 2) abdominal cramps, 3) vomiting, 4) audible bowel sounds, 5) flatulence or gas.

[0129] Hyperglycemia was measured using a flash glucose monitoring (FGM, Free Style Libre (registered trademark), Abbott) device for minimally invasive real-time monitoring of blood glucose levels in interstitial fluid. FGM was developed and validated for use in adults with type 1 or type 2 diabetes [1 - 4] and measures interstitial glucose every 15 minutes for 14 days.

[0130] The breath hydrogen test is widely used for the detection of carbohydrate malabsorption. Its principle is mainly based on the detection of hydrogen in exhaled breath due to bacterial fermentation of carbohydrates in the colon. To obtain rapid and reliable indirect information on carbohydrate absorption, a breath analyzer (Lactotest 202, M.E.C Belgium) that measures H2 was used in this test.

[0131] The test products were α-glucans with various degrees of polymerization (DP) and glucose binding, and thus had different degrees of digestibility. The digestible carbohydrate composition 2 (DCC-2) had a DP of 12.4, and the digestible carbohydrate composition 3 (DCC-3) had a DP of 17.3. As a reference, a fully digestible and full-calorie maltodextrin (glucose syrup) was administered. 33 g of total carbohydrates were included in the test products in either powder (glucose syrup) or syrup (digestible carbohydrate composition) form. The day before the visit for the test, the test products were dissolved in 300 mL of water and stored at 4 °C overnight. On the morning before ingestion, the beverage was warmed to room temperature and then provided to the participants.

[0132] The subject population consisted of sufficiently healthy men and women (based on medical history) aged 18 to 45 years with a BMI of 20 to 29.9 kg / m2. Subjects were considered ineligible to participate if they presented one or more of the following exclusion criteria: Women who are pregnant or lactating; Concomitant medications, colonoscopy, enema, or other bowel cleansing procedures that may affect the test procedure and evaluation, such as antibiotics, antacids, or other drugs that affect transit time, within 4 weeks before the test; Serious medical / surgical events within the past 3 months that may interfere with the test procedure and evaluation; History of abnormal intestinal transit and chronic constipation with spontaneous bowel movements less than 3 times per week on average, or chronic or recurrent diarrhea with spontaneous bowel movements more than 3 times per day; Known food allergies and intolerances to the test products; Medically known skin allergies to adhesives and plasters; Alcohol intake exceeding 2 cups per day. One cup is 0.4 dl for hard alcohol, 1 dl for red or white wine, or 3 dl for beer; Smokers; Volunteers who cannot be expected to comply with the protocol; Subjects having a hierarchical relationship with members of the test team.

[0133] Notifications were made within Nestle entities and contact was established with potential candidates. For volunteers who wished to participate and were eligible, the research team provided a sufficient oral briefing (V0) on the purpose, methods, and potential risks / disadvantages of the trial. Candidates were invited to a medical screening visit for confirmation of eligibility and signing of the informed consent.

[0134] At least 24 hours before the first test product intake, one FGM device was attached to the non-dominant arm of each volunteer. Subsequently, participants were asked to fast for at least one day for each beverage and participate in different test visits so that they could consume a low-fiber diet the day before each test visit. After all measurements were completed, the sensor was removed on the last day of the trial.

[0135] On the day before each test visit, the subjects were required to refrain from consuming alcohol. Drugs such as aspirin or dietary supplements containing vitamin C, accelerators, laxatives, and antibiotics that could affect FGM measurements were not to be taken four weeks before the trial and throughout the entire trial period. Fermentable foods such as complex carbohydrates had to be avoided on the day before the breath test. The last meal on the day before the test had to be fiber-free and not an excessive amount that would cause over-fullness. Participants were also asked to avoid chewing gum from 8 pm on the day before each test visit.

[0136] The general research scheme is described in Figure 5. The subjects arrived at the metabolism department at 8 pm in a fasting state since 8 am the previous day. To minimize interference with the breath test, the subjects had to avoid chewing gum, avoid using perfume, and were required to brush their teeth thoroughly at home before each visit to the metabolism department.

[0137] Readings from the FGM device were taken before and after ingestion of the product. The average of the two measurements was considered the baseline value. Readings were also taken at 30, 60, 90, 120, 150, 180, 210, and 240 minutes. Breath samples for measuring hydrogen production were taken before product ingestion and then at 30, 60, 90, 120, 150, 180, 210, and 240 minutes immediately after the glucose reading. Subjects were asked to complete a questionnaire regarding gastrointestinal symptoms 180 minutes after product ingestion and after the corresponding glucose reading and breath test were completed. After these procedures, a fiber-free breakfast consisting of white bread, honey, and coffee or tea or water was provided. Any other food items, including chewing gum, were prohibited during the test period, and only water was permitted.

[0138] The minimum sample size of N = 10 subjects who completed the test was required by ISO-26642 for determining and classifying the glycemic index (GI) of food products [5]. For α-glucans, study 17.08.BIO showed that N = 16 is required to detect a 35% reduction compared to the maltodextrin control (SD = 50%) of the PPGR 3h-iAUC at α = 5% (two-sided) and a power of 80%. This pilot study used smaller adult-sized portions (i.e., 25 g), so the calculated sample size is conservative (i.e., variability in healthy subjects is generally reduced with larger adult-sized portions). This sample size is also suitable for H2-related endpoints, as shown by the fact that N = 16 enables distinguishing a difference of 0.35 log ppm at α = 5% and a power of 80% [6]. As a conclusion, the sample size for this study was set at N = 16.

[0139] For the primary evaluation item (i.e., 3h-iAUC derived from PPGR), the test product was compared with the maltodextrin control using a paired t-test (in accordance with the logic of ISO-26642). To control the false discovery rate (FDR) at α = 5%, the Benjamini-Hochberg procedure was applied [7]. Sensitivity analysis was performed using a mixed model to account for potential systematic positions or carry-over effects [8]. For all other pairwise comparisons and all other endpoints, the analysis was the same, but no correction for multiplicity was made.

[0140] Example 6 Results of the clinical trial Sixteen healthy volunteers (6 females and 10 males) with an average age of 31.4 ± 5.9 years, an average BMI of 23.0 ± 1.6 kg / m2, and an average fasting blood glucose level of 4.8 ± 0.5 mmol / L were recruited.

[0141] Figures 8 and 9 show the glucose response and incremental glucose response at 4 hours after a meal, respectively, for all test products. Compared with glucose syrup, both glucose-based saccharides resulted in significantly lower iCmax, but no significant difference was observed in tmax. The time required for the blood glucose level to return to the baseline value was not significantly longer compared to the reference. The 1h-iAUC was significantly lower for both test products, but only DCC-3 had significantly lower 2h- and 3h-iAUC compared to glucose syrup.

[0142] When comparing the glucose curve shape of α-glucan with the reference, it was observed that both α-glucans had a lower glucose peak and a slower return to baseline. The decrease in blood glucose level after the peak was gentler than that of the reference, and the blood glucose level remained slightly higher for a longer period, suggesting that α-glucan can be digested and utilized slightly more slowly than maltodextrin. Furthermore, the ingestion of α-glucan resulted in a lower hypoglycemia after returning to the baseline compared to the control.

[0143] Figure 10 shows the mean relative 1h-, 2h- and 3h-iAUC of the test products compared to glucose syrup. DCC-2 results in a reduction of 27% (p<0.05), 16% and 13% in 1h-, 2h- and 3h-iAUC respectively, compared to the reference. DCC-3, which has a higher degree of polymerization than DCC-2, results in a greater reduction in blood glucose, with 45% (p<0.05) for 1h-iAUC, 38% (p<0.05) for 2h-iAUC and 33% (p<0.05) for 3h-iAUC, compared to the reference.

[0144] DCC-2 and DCC-3 contain 2.1 and 2.9 g of fructose equivalents respectively, and thus, unlike glucose syrup, are not pure glucose-based. The dark grey bars in Figure 10 are the corrected data of the relative iAUC predicted if the product were a pure glucose-based. After correction, the reduction in iAUC for DCC-2 remains significant only for 1h-iAUC (24%), while the reduction in 1h-, 2h- and 3h-iAUC for DCC-3 is significant (42%, 34% and 28% respectively). Even with the corrected predicted values, the reduction in blood glucose level is significant, and it can be concluded that the main factor for PPGR reduction is not fructose.

[0145] Postprandial breath H2 was used to indirectly measure the product digestibility. When indigestible carbohydrates reach the colon, they are fermented by colonic bacteria. This fermentation produces gases such as H2 and CH4, which can be measured in the breath of the subjects. After ingestion of the test products, no significant differences were observed among them, and there were no significant differences compared to the completely digestible control product (Figures 11 and 12). The general range of hydrogen in fasting breath is 7 ± 3 ppm, and the maximum value at 240 minutes is found within this range. Only DCC-2 has a higher value than other products, but it remains below the threshold of 20 ppm for poor carbohydrate absorption, decreases during the test, and reaches a level similar to other products. At 180 minutes after breakfast ingestion, a slight increase in hydrogen production was observed, and the lactose test suggests the detection of a further small amount of H2, which may result from the presence of a very small amount of complex carbohydrates or slightly incomplete absorption of simple carbohydrates (e.g., fructose). Therefore, all α-glucans seem to be highly digestible and do not induce carbohydrate fermentation like fiber.

[0146] The gastrointestinal tolerance to the product was evaluated using a visual analog scale for five different symptoms: abdominal cramps, bowel sounds, diarrhea, flatulence, and vomiting (Table 6). Generally, few people reported discomfort, and no severe events were reported. In fact, bowel sounds were the symptom with the highest score. The scores from those who reported discomfort were mostly low. The average z-score remained low and was the highest for the reference product, glucose syrup. Therefore, in healthy subjects, the gastrointestinal tolerance to α-glucan is good, suggesting that most of it is digestible.

[0147]

Table 8

[0148] As expected, all of the α-glucans tested resulted in a lower postprandial blood glucose response compared to the fully digestible maltodextrin control. α-Glucan structures with alternating α1-3 / 6 linkages result in a significant reduction in glucose response compared to glucose syrup when the molecular weight is higher than 1.6 kDa (DP>10). Indeed, in healthy volunteers, a glucose-based saccharide with a DP of 17.3 results in a 38% (p<0.01) reduction in glucose response (2h-iAUC).

[0149] Unlike fibers that cause colonic fermentation due to incomplete digestion, the results of this clinical trial suggest that α-glucans are mostly digestible, as none of the products tested produced breath hydrogen as a result of colonic fermentation. Furthermore, the assessment of gastrointestinal tolerance was not different from that reported for the reference product, indicating a very low level of discomfort.

[0150] References 1. Distiller,L.A.,I.Cranston,and R.Mazze,First Clinical Experience with Retrospective Flash Glucose Monitoring(FGM)Analysis in South Africa:Characterizing Glycemic Control with Ambulatory Glucose Profile.J Diabetes Sci Technol,2016.10(6):p.1294-1302。 2. Bonora,B.,et al.,Head-to-head comparison between flash and continuous glucose monitoring systems in outpatients with type 1 diabetes.J Endocrinol Invest,2016.39(12):p.1391-1399。 3. Schierenbeck, F., A. Franco-Cereceda, and J. Liska, Accuracy of 2 Different Continuous Glucose Monitoring Systems in Patients Undergoing Cardiac Surgery. J Diabetes Sci Technol, 2017. 11(1): p. 108-116。 4. Akintola, A. A., et al., Accuracy of Continuous Glucose Monitoring Measurements in Normo-Glycemic Individuals. PLoS One, 2015. 10(10): p. e0139973。 5. Internal Standard Organization, Food Products - Determination of the Glycaemic Index (GI) and Recommendations for Food Classification. ISO 26642. 2010。 6. Grysman, A., T. Carlson, and T. M. Wolever, Effects of sucromalt on postprandial responses in human subjects. Eur J Clin Nutr, 2008. 62(12): p. 1364-71。 7. Benjamini, Y. and Y. Hochberg, Controlling the False Discovery Rate: a Practical and Powerful Approach to Multiple Testing. J Roy Stat Soc, 1995. 57: p. 289-300。 8. Senn, S., Cross-over Trials in Clinical Research. 2002, Chichester: New York: J. Wiley。

Claims

**Claim 1** a. A glucose-based saccharide that is at least 65% (w / w) on a dry weight basis, having a reducing end and D-glucose monomers linked alternately by α1-6 glycosidic bonds and α1-3 glycosidic bonds, and a receptor molecule is present at the reducing end, a glucose-based saccharide, and b. A fructose equivalent that is 0.1 to 30% (w / w) on a dry weight basis and is at least one selected from the group consisting of leucrose, fructose, and sucrose, a fructose equivalent, comprising The glucose-based saccharide has an average degree of polymerization of 12 to 90 when measured by HPAEC-PAD, and the receptor molecule is selected from a sugar or a sugar alcohol having a free hydroxyl group at one or more of carbon position numbers 2, 3, and 6 that can accept a glucose unit from sucrose, a digestible carbohydrate composition. **Claim 2** The digestible carbohydrate composition according to claim 1, wherein the average degree of polymerization is 17 to 90 when measured by HPAEC-PAD. **Claim 3** The digestible carbohydrate composition according to claim 1 or 2, wherein the composition has at least 80% (w / w) of glucose-based saccharides on a dry weight basis. **Claim 4** The digestible carbohydrate composition according to any one of claims 1 to 3, wherein the composition has an α1-6 branch point on glucose that is trimerized by alternately α1-6 glycosidic bonds and α1-3 glycosidic bonds. **Claim 5** The digestible carbohydrate composition according to any one of claims 1 to 4, wherein the composition contains 0.1 to 10% (w / w) of fructose equivalent on a dry weight basis. **Claim 6** The digestible carbohydrate composition according to any one of claims 1 to 5, wherein the receptor molecule is a maltose unit. **Claim 7** The digestible carbohydrate composition according to any one of claims 1 to 6, for use in reducing blood glucose response in a subject. **Claim 8** A food product comprising the digestible carbohydrate composition according to any one of claims 1 to 6. **Claim 9** The food product according to claim 8, wherein the food product is a beverage. **Claim 10** The food product according to claim 8 or 9, wherein the food product is a dietary supplement. **Claim 11** The food product according to any one of claims 8 to 10, wherein the food product is a nutritional preparation. **Claim 12** The food product according to claim 11, wherein the nutritional preparation is for a) a human subject with diabetes and / or prediabetes, or b) a human subject who is an infant, child, or adolescent.

13. The food product according to claim 11 or 12, wherein the nutritional preparation is for a human subject under emergency treatment.

14. The food product according to any one of claims 8 to 11, wherein the food product is a pet food product.

15. A digestible carbohydrate composition according to any one of claims 1 to 6 for reducing postprandial glucose in a subject.

16. The digestible carbohydrate composition according to claim 15, wherein the subject is a human subject.

17. The digestible carbohydrate composition according to claim 15, wherein the subject is a companion animal subject.

18. A food product according to any one of claims 8 to 14 for reducing postprandial glucose in a subject.

19. The food product according to claim 18, wherein the subject is a human subject.

20. The food product according to claim 18, wherein the subject is a companion animal subject.

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