Nutritional composition for promoting absorption of amino acids and minerals and application thereof

Abstract

The invention discloses a nutritional composition for promoting absorption of amino acid and mineral substances and application, and belongs to the technical field of food. The nutritional composition provided by the invention comprises alpha-lactalbumin and breast milk oligosaccharide (especially 6 '-SL and / or 3-FL), and can help to promote absorption of amino acids and mineral substances by a body, especially help to promote transmembrane transport capability of intestinal cells of the body to amino acids and mineral substances, thereby improving absorption efficiency.

Description

TECHNICAL FIELD

[0001] The present application relates to a nutritional composition for promoting the absorption of amino acids and minerals and uses thereof, belonging to the field of food technology. BACKGROUND

[0002] Amino acids are essential building blocks for the growth and development of infants. They are not only the basic units of proteins, but also participate in almost all physiological processes. During the rapid growth stage of infants, the body has a high demand for amino acids, as these small molecules are the key materials for building muscles, bones, organs, and the nervous system. For example, lysine is crucial for the growth of bones and muscles, and tryptophan is the precursor for the synthesis of the neurotransmitter serotonin, which affects mood and sleep regulation. In addition, certain amino acids such as arginine and glutamine play important roles in the immune system, helping infants establish a strong immune barrier. However, the digestive system of infants is not fully developed, and the efficiency of amino acid absorption is relatively low, which can lead to nutritional deficiencies and affect their growth and development and health. Calcium and zinc are extremely important nutrients for the growth and development of infants, and they play a key role in bone health, immune function, growth rate, and cognitive development. However, the nutritional needs of infants are different from adults, and their digestive and absorptive capacity is not fully mature, so special attention should be paid to their calcium and zinc nutrition. Studies have shown that the absorption efficiency of calcium and zinc in breast milk is significantly higher than that in cow's milk formula. The absorption rate of calcium in breast milk can reach 76%, while the absorption rate of cow's milk formula is usually between 40% - 60%; the absorption efficiency of zinc in breast milk is about 80%, while the absorption efficiency of zinc from cow's milk formula is about 30%.

[0003] Currently, there have been some studies on the absorption of amino acids. For example, reference 1 provides an infant formula for improving the bioavailability of stable amino acids and its application. The composition contains α-lactalbumin, linoleic acid, and prebiotics, which can improve the digestion and absorption rate of acid-stable amino acids, and is beneficial to the healthy growth of infants. Reference 2 discloses a plant lactobacillus that can improve the absorption of amino acids, with the accession number CGMCC NO. 25190. Reference 3 discloses an amino acid absorption promoter, which contains at least one or more matobianic acids and salts of matobianic acids.

[0004] For how to promote the absorption of minerals, reference 4 discloses a synergistic composition for improving mineral bioavailability, including lutein, casein phosphopeptide or 1,3 dioleic acid-2-palmitic acid glycerol triester, preferably in combination with one or more non-digestible oligosaccharides, which can be used for preventing and / or treating mineral deficiency and related diseases, preferably anemia, the minerals include iron and / or calcium. Reference 5 discloses a plant lactobacillus plantarum with a preservation number of CGMCC No. 32404, which can promote mineral absorption and transport.

[0005] At present, in order to ensure the nutrition supply of infants and young children, and solve the problem that the absorption of nutrients in infant formula powder is lower than that in breast milk, the addition amount of nutritional ingredients is mainly increased. However, the excessive addition of some nutrients, such as protein, may cause infant overweight. Therefore, it is a direction worth paying attention to to optimize the nutritional formula, develop efficient nutritional supplements, and improve the utilization efficiency of these key nutrients. In the face of the differences between formula milk powder and breast milk, researchers are constantly developing new functional raw materials in order to make formula milk powder closer to breast milk in terms of nutritional composition and function.

[0006] Although previous studies have shown that various optional additives in infant formula milk powder have different promoting effects on nutrient absorption, the sufficiency of these studies still needs to be further improved. At present, the academic circle has rich research on the nutritional absorption function of single component, but there is still a lack of clear understanding of whether the combination of multiple components can produce synergistic effect, especially in promoting the simultaneous digestion and absorption of multiple nutrients such as amino acids and minerals.

[0007] Therefore, the present application aims to develop a nutritional composition through scientific combination of multiple components, in order to achieve more effective promotion of the simultaneous digestion and absorption of multiple essential nutrients by the body, thereby making up for the shortcomings of existing research and providing a scientific basis for the optimization of infant formula milk powder.

[0008] Reference:

[0009] Reference 1: CN118901799A;

[0010] Reference 2: CN116286462A;

[0011] Reference 3: JP2024122087A;

[0012] Reference 4: CN119836289A;

[0013] Reference 5: CN120424829A. SUMMARY

[0014] Problem to be solved by the invention

[0015] Although previous studies have shown that various optional additives in infant formula can promote the absorption of nutrients in different ways, the adequacy of these studies still needs to be further improved. At present, the academic community has conducted extensive research on the nutritional absorption function of single ingredients, but there is still a lack of clear understanding of whether the combination of multiple ingredients can produce a synergistic effect, especially in promoting the simultaneous digestion and absorption of multiple nutrients such as amino acids and minerals. Alpha-lactalbumin, 3-FL and 6'-SL are important nutritional ingredients in breast milk, which play an important role in the intestinal health and immune system development of infants, but specific research on their relationship with amino acid and mineral absorption has not been reported.

[0016] The present application aims to achieve more effective promotion of the co-digestion and absorption of multiple essential nutrients in the body by scientific combination of alpha-lactalbumin, 6'-SL and 3-FL, thereby making up for the inadequacy of existing research and providing a new approach for the optimization of infant formula.

[0017] Solution to the problem

[0018] [1]. The present application first provides a nutritional composition for promoting the absorption of amino acids and minerals, which comprises the necessary active ingredients as shown in (a) and (b) below:

[0019] (a) alpha-lactalbumin;

[0020] (b) one or more human milk oligosaccharides;

[0021] wherein, in the nutritional composition, the mass ratio of the necessary active ingredients as shown in (a) to the necessary active ingredients as shown in (b) is (10-20):(1-25).

[0022] [2]. The nutritional composition according to [1], wherein the human milk oligosaccharides include 3-fucosyllactose and / or 6'-sialyllactose;

[0023] provided that, if the human milk oligosaccharides include 3-fucosyllactose and 6'-sialyllactose, the mass ratio of the alpha-lactalbumin, the 6'-sialyllactose and the 3-fucosyllactose is (10-20):(1-5):(5-20).

[0024] [3]. The nutritional composition according to [1] or [2], wherein the nutritional composition further comprises whey protein other than alpha-lactalbumin.

[0025] [4]. Use of the nutritional composition according to any one of [1]-[3] in the preparation of food that helps to promote the absorption of amino acids and minerals.

[0026] [5]. The use according to [4], wherein the amino acid comprises a free amino acid.

[0027] [6]. The use according to [4] or [5], wherein the mineral comprises calcium and / or zinc.

[0028] [7]. The use according to any one of [4] to [6], wherein the promoting absorption of amino acid and mineral comprises promoting expression of amino acid transporter protein, calcium transporter protein and / or zinc transporter protein.

[0029] [8]. The use according to any one of [4] to [7], wherein the food product is selected from the group consisting of infant food, children food, adolescent food, pregnant and lactating women food, adult food and elderly food.

[0030] [9]. The use according to any one of [4] to [8], wherein the food product comprises any one or more of the following ingredients: plant product ingredient, animal milk product ingredient, animal meat product ingredient, functional additive ingredient and any acceptable adjuvant.

[0031]

[10] . The use according to any one of [4] to [9], characterized in that the content of the a-lactalbumin is 0.5% to 4.5% and the content of the human milk oligosaccharide is 0.01% to 2.5% in the food product, based on the total dry matter content of the food product.

[0032] Effects of the invention

[0033] The present application provides a composition containing a-lactalbumin and human milk oligosaccharide (especially 6'-SL and / or 3-FL), and surprisingly finds that the above-mentioned composition can help to promote the absorption of amino acid and mineral by the body, especially can help to promote the transmembrane transport capacity of amino acid and mineral by the intestinal cells of the body, thereby improving the absorption efficiency.

[0034] Specifically, the composition can improve the transport ratio of free total amino acid, mineral (calcium, zinc), promote the expression of amino acid transporter protein (SLC6A14), calcium transporter protein (TRPV6) and zinc transporter protein (ZnTl). And when the mass of a-lactalbumin and human milk oligosaccharide is within a certain range, there is a synergistic effect, preferably when the mass of a-lactalbumin and 6'-SL and / or 3-FL is within a certain range, there is a better synergistic effect between the two or three. DETAILED DESCRIPTION

[0035] Various exemplary embodiments, features, and aspects of the present invention will be described in detail below. The term "exemplary" as used herein means "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as superior to or better than other embodiments.

[0036] Furthermore, to better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In other instances, methods, means, apparatus, and steps well known to those skilled in the art have not been described in detail in order to highlight the spirit of the present invention.

[0037] Unless otherwise stated, all units used in this specification are international standard units, and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors that are unavoidable in industrial production.

[0038] In this invention, "comprising," "having," "including," or "containing" can mean included or open-ended, and does not exclude additional, uncited elements or method steps. At the same time, "comprising," "having," "including," or "containing" can also mean closed-ended, excluding additional, uncited elements or method steps.

[0039] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0040] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "implementation," etc., refer to specific elements (e.g., features, structures, properties, and / or characteristics) related to that embodiment, which are included in at least one of the embodiments described herein and may or may not be present in other embodiments. Furthermore, it should be understood that these elements may be combined in any suitable manner in various embodiments.

[0041] In this specification, "optional" and "optionally" mean that the events or circumstances described below may or may not occur, and the description includes both cases where the events or circumstances occur and cases where the events or circumstances do not occur.

[0042] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0043] In the present application, "about" is used to qualify the numerical ranges and parameters of the present application are approximate values, the relevant values in the specific examples have been presented as precisely as possible. Unless otherwise expressly stated, it is to be understood that all ranges, numbers, values and percentages used herein are to be "about" modified. Herein, "about" generally means within ± 1% or ± 0.5% of a specified value or range, depending on what is typical in the art. Further, the numerical values, numerical ranges, appearing in the present application are to be understood to be inclusive of the inevitable system errors in industrial production.

[0044] As used herein, the term "and / or" encompasses all combinations of the items linked by the term "and / or". It is to be understood that each combination is individually contemplated herein. For example, "A and / or B" encompasses "A", "A and B", and "B". For example, "A, B, and / or C" encompasses "A", "B", "C", "A and B", "A and C", "B and C", and "A and B and C".

[0045] In the present application, the unit names used are international standard unit names, and if not specifically stated, the "%" used means the weight or mass percentage content.

[0046] In the present application, "infants" means the human group under 3 years of age, which includes infants from 0 to 6 months of age, older infants from 6 to 12 months of age, and toddlers from 12 to 36 months of age.

[0047] In the present application, "children" means the human group from 3 to 6 years of age.

[0048] In the present application, "adolescents" means the human group from 7 to 18 years of age.

[0049] In the present application, "pregnant and lactating women" includes pregnant women and lactating women.

[0050] In the present application, "elderly" means the human group over 41 years of age.

[0051] In the present application, "animal milk" means the liquid obtained from the mammary glands of mammals in the lactation period. The term "animal milk" should be interpreted broadly and encompasses both raw milk (i.e. the liquid obtained directly from the mammary glands) and standardized dairy products (such as, for example, skimmed milk or full-fat milk).

[0052] Unless otherwise defined, other technical and scientific terms used in the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0053] < Nutritional composition >

[0054] The present application provides a nutritional composition having an effect of promoting absorption of amino acids and minerals, which comprises essential active ingredients as shown in (a)~(b) below.

[0055] (a) α-lactalbumin;

[0056] (b) human milk oligosaccharide.

[0057] The present application does not particularly limit the source of α-lactalbumin, which can be derived from cow's milk, goat's milk, goat's milk, etc. The α-lactalbumin in natural milk can be obtained by conventional separation and purification techniques, such as thermal agglomeration-enzymatic hydrolysis, membrane separation-chromatography combination, selective precipitation, etc. In addition, the above-mentioned α-lactalbumin can be obtained directly by commercially available products in addition to being obtained by the above methods.

[0058] The present application does not particularly limit the human milk oligosaccharide, and one or more human milk oligosaccharides allowed to be used in food can be used. In some preferred exemplary embodiments, the human milk oligosaccharide includes 3-fucosyllactose and / or 6'-sialyllactose.

[0059] The 6'-sialyllactose (6'-SL) described in the present application is a disaccharide containing sialic acid, which is mainly composed of lactose (D-galactose and D-glucose) and sialic acid (N-acetylneuraminic acid) connected by α-2,6 glycosidic bond, C 23 H 39 O 19 , and the molecular weight is 633.55 g / mol.

[0060] The present application does not particularly limit the source of the 6'-sialyllactose, which can be of natural origin, synthetic origin, microbial fermentation origin, etc. Typically, for 6'-sialyllactose, sialic acid (N-acetylneuraminic acid, Neu5Ac) can be connected to lactose by chemical reaction and the like steps to synthesize; lactose + CMP-Neu5Ac (cytidine-5'-monophosphate-sialic acid) as a substrate, catalyzed by sialic acid synthase (such as NeuA) and CMP-sialic acid synthase (such as CSS) to generate CMP-Neu5Ac, under the action of sialic acid transferase, the sialic acid of CMP-Neu5Ac is transferred to the 6'-OH position of lactose to generate 6'-sialyllactose. In addition, 6'-FL can also be derived from animal milk such as cow's milk, etc.

[0061] 3-fucosyllactose (3-FL) is a neutral trisaccharide composed of a lactose (Galβ1-4Glc) molecule core and an L-fucose (L-Fucose) group connected to the glucose unit of lactose by α-1,3-glycosidic bond, with a molecular formula of C 18 H 32 O15 The source of 3-FL is not particularly limited in the present application, and typically, it can be obtained by common chemical synthesis methods, microbial fermentation, etc. In addition, 3-FL can also be derived from animal milk such as cow milk, etc.

[0062] The present application finds that, compared with α-lactalbumin alone, the nutritional composition provided by the present application can more effectively promote the absorption of amino acids and minerals, i.e., there is a synergistic effect between the above two or three.

[0063] In some embodiments, in the nutritional composition described in the present application, the (a) α-lactalbumin, (b) human milk oligosaccharides, especially 6'-SL and / or 3-FL are the main active ingredients, i.e., the nutritional composition described in the present application mainly relies on the (a) α-lactalbumin, (b) human milk oligosaccharides, especially 6'-SL and / or 3-FL contained therein to exert specific physiological activity functions, such as helping to promote the absorption of amino acids and minerals. In other words, in some embodiments, the active ingredients (ingredients for exerting specific physiological activity functions, i.e., ingredients for exerting the effect of helping to promote the absorption of amino acids and minerals) of the nutritional composition consist of the following (a) and (b): (a) α-lactalbumin, (b) human milk oligosaccharides, especially 6'-SL and / or 3-FL.

[0064] In some embodiments, the nutritional composition comprises active ingredients (ingredients for exerting specific physiological activity functions, i.e., ingredients for exerting the effect of helping to promote the absorption of amino acids and minerals) and non-active ingredients (substances that do not exert the effect of helping to promote the absorption of amino acids and minerals). Exemplarily, the non-active ingredients can be other nutrients, any food acceptable adjuvant, and / or substances that are accompanied in the production or acquisition process of the active ingredients and cannot be effectively separated from the active ingredients or do not need to be separated. In some embodiments, the nutritional composition consists of the active ingredients and the non-active ingredients.

[0065] In some embodiments, the nutritional composition further comprises other species of whey protein other than a-lactalbumin, and other species of human milk oligosaccharides other than 3-fucosyllactose and 6'-sialyllactose. Exemplarily, the other species of whey protein other than a-lactalbumin include b-lactoglobulin, immunoglobulin, serum albumin, lactoferrin, etc.; the other species of human milk oligosaccharides other than 3-fucosyllactose and 6'-sialyllactose include 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lacto-N-tetraose, lacto-N-neotetraose, and lacto-N-fucopentaose I, etc., as described above, which are used in compliance with the requirements of laws and regulations.

[0066] In some embodiments, in the nutritional composition, the mass ratio of (a) the essential active ingredient shown to (b) the essential active ingredient shown can be (10~20):(1~25), preferably 15:(1~20), for example, can be 15:1, 15:2, 15:3, 15:4, 15:5, 15:6, 15:7, 15:8, 15:9, 15:10, 15:11, 15:12, 15:13, 15:14, 15:15, 15:16, 15:17, 15:18, 15:19, 15:20.

[0067] In some exemplary embodiments, the human milk oligosaccharides comprise 3-fucosyllactose and / or 6'-sialyllactose.

[0068] In some embodiments, in the nutritional composition, the three active ingredients of the a-lactalbumin, the 6'-sialyllactose and the 3-fucosyllactose are included, and the mass ratio of the a-lactalbumin, the 6'-sialyllactose and the 3-fucosyllactose can be (10-20):(1-5):(5-20), preferably 15:(1-5):(5-15); for example, it can be 15:1:5, 15:1:6, 15:1:7, 15:1:8, 15:1:9, 15:1:10, 15:1:11, 15:1:12, 15:1:13, 15:1:14, 15:1:15, 15:2:5, 15:2:6, 15:2:7, 15:2:8, 15:2:9, 15:2:10, 15:2:11, 15:2:12, 15:2:13, 15:2:14, 15:2:15, 15:3:5, 15:3:6, 15:3:7, 15:3:8, 15:3:9, 15:3:10, 15:3:11, 15:3:12, 15:3:13, 15:3:14, 15:3:15, 15:4:5, 15:4:6, 15:4:7, 15:4:8, 15:4:9, 15:4:10, 15:4:11, 15:4:12, 15:4:13, 15:4:14, 15:4:15, 15:5:5, 15:5:6, 15:5:7, 15:5:8, 15:5:9, 15:5:10, 15:5:11, 15:5:12, 15:5:13, 15:5:14, 15:5:15, etc.

[0069] The present application does not make special limitation to the form of the nutritional composition, and typically, it can be a liquid or a solid, etc. From the perspective of production, transportation, storage and use convenience, the nutritional composition of the present application is preferably a powdery solid.

[0070] <Use of the nutritional composition>

[0071] The present application proposes that (a) a-lactalbumin, (b) human milk oligosaccharides, especially 3-fucosyllactose and / or 6'-sialyllactose, are compounded in a certain ratio, which can help to promote the absorption of amino acids and minerals, and there is a synergistic effect between a-lactalbumin and human milk oligosaccharides, specifically, there is a synergistic effect between a-lactalbumin and human milk oligosaccharides, especially 3-fucosyllactose or 6'-sialyllactose, or there is a synergistic effect among a-lactalbumin, 3-fucosyllactose and 6'-sialyllactose.

[0072] In some embodiments, the amino acids include free amino acids, preferably, the free amino acids include 20 amino acids obtained by protein hydrolysis, including glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), aspartic acid (Asp), glutamic acid (Glu), lysine (Lys), arginine (Arg), histidine (His), cysteine (Cys), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), serine (Ser), and threonine (Thr).

[0073] In some embodiments, the minerals include calcium and / or zinc; preferably, the minerals include calcium and zinc.

[0074] In some embodiments, the facilitating absorption of amino acids and minerals includes facilitating absorption of free amino acids and minerals, the free amino acids including 20 amino acids obtained by protein hydrolysis, the minerals including calcium and / or zinc.

[0075] In some embodiments, the facilitating absorption of amino acids and minerals includes facilitating transmembrane transport of amino acids and minerals by intestinal cells.

[0076] In some embodiments, the facilitating absorption of amino acids and minerals includes facilitating expression of amino acid transporters, calcium transporters, and / or zinc transporters. Preferably, the facilitating absorption of amino acids and minerals includes facilitating expression of amino acid transporters, calcium transporters, and zinc transporters.

[0077] In some exemplary embodiments, the amino acid transporters include SLC6A14; the calcium transporters include TRPV6; and the zinc transporters include ZnT1.

[0078] The present application is not particularly limited to specific food products containing or prepared using the nutritional composition described above.

[0079] In some embodiments, the food product described herein is in a liquid or solid form at room temperature.

[0080] In some embodiments, the food product is a baby food product, a child food product, an adolescent food product, a pregnant or lactating woman food product, an adult food product, or a senior food product.

[0081] In some embodiments, the food product described herein is a confectionery, such as a hard candy, a gummy candy, a crisp candy, a pressed candy, an aerated candy, and the like.

[0082] In some embodiments, the food of the present application is a beverage, such as a carbonated beverage, a tea-based beverage, a coffee-based beverage, a fruit / vegetable juice beverage, a lactic acid bacteria beverage, and the like.

[0083] In some embodiments, the food of the present application is a dairy product, such as powdered milk, cheese, yogurt, liquid milk, and the like.

[0084] In some embodiments, the food of the present application is a baked food, such as bread, cake, cookie, and the like.

[0085] In some embodiments, the food of the present application is a dietary supplement, such as a hard capsule, a soft capsule, a tablet, an oral solution, a pill, a granule, a powder, and the like.

[0086] In some embodiments, in the food of the present application, the mass ratio of (a) the essential active ingredient to (b) the essential active ingredient can be (10-20):(1-25), preferably 15:(1-20), such as 15:1, 15:2, 15:3, 15:4, 15:5, 15:6, 15:7, 15:8, 15:9, 15:10, 15:11, 15:12, 15:13, 15:14, 15:15, 15:16, 15:17, 15:18, 15:19, 15:20. In some exemplary embodiments, the human milk oligosaccharide includes 3-fucosyl lactose and / or 6'-sialyllactose.

[0087] In some embodiments, the food product described in the present application comprises α-lactalbumin, 6'-sialyllactose and 3-fucosyllactose. The mass ratio of the α-lactalbumin, the 6'-sialyllactose and the 3-fucosyllactose can be (10~20):(1~5):(5~20), preferably 15:(1~5):(5~15); for example, it can be 15:1:5, 15:1:6, 15:1:7, 15:1:8, 15:1:9, 15:1:10, 15:1:11, 15:1:12, 15:1:13, 15:1:14, 15:1:15, 15:2:5, 15:2:6, 15:2:7, 15:2:8, 15:2:9, 15:2:10, 15:2:11, 15:2:12, 15:2:13, 15:2:14, 15:2:15, 15:3:5, 15:3:6, 15:3:7, 15:3:8, 15:3:9, 15:3:10, 15:3:11, 15:3:12, 15:3:13, 15:3:14, 15:3:15, 15:4:5, 15:4:6, 15:4:7, 15:4:8, 15:4:9, 15:4:10, 15:4:11, 15:4:12, 15:4:13, 15:4:14, 15:4:15, 15:5:5, 15:5:6, 15:5:7, 15:5:8, 15:5:9, 15:5:10, 15:5:11, 15:5:12, 15:5:13, 15:5:14, 15:5:15, etc.

[0088] The present application does not particularly limit the specific absolute content of the α-lactalbumin, the human milk oligosaccharide, especially 3-fucosyllactose and 6'-sialyllactose in the food product, which meets the requirements of local food-related laws and regulations.

[0089] In some embodiments, in the food product, the content of the α-lactalbumin is 0.5%-4.5%, the content of the human milk oligosaccharide is 0.01%~2.5%, preferably, the content of the 3-fucosyllactose is 0.1%~2%, and the content of the 6'-sialyllactose is 0.01%~0.5%, based on the total dry matter content of the food product.

[0090] In addition to the above-described components in the nutritional composition, the food product can also comprise other ingredients, such as other proteins / amino acids, carbohydrates, fats, vitamins, minerals, and other common food ingredients.

[0091] In addition, depending on the kind of food and the final needs of the target, in some embodiments, any one or more of the following ingredients are contained in the food: plant product ingredients, animal milk product ingredients, animal meat product ingredients, functional additive ingredients, and any acceptable adjuvants.

[0092] For the plant product ingredients, examples include fruits such as figs, pomegranates, kiwis, oranges, tangerines, pineapples, strawberries, apples, bananas, grapes, pears, cherries, blueberries, blackberries, blackcurrants, cranberries, raspberries, melons, emblics, and mulberries, or extracts thereof; fruit and vegetable materials such as onions, cucumbers, tomatoes, cauliflowers, red beetroots, spinach, kohlrabi, Brussels sprouts, garlic, basil, Oregon grass, or extracts thereof; cereals such as rice (indica rice, japonica rice, glutinous rice), wheat (wheat, barley, oat, rye), corn, sorghum, millet, foxtail millet, japonica, buckwheat, soybeans, beans, peas, mung beans, adzuki beans, kidney beans, or extracts thereof; nut materials such as walnuts, pistachios, cashews, hazelnuts, almonds, apricot kernels, pine nuts, peanuts, melon seeds, chestnuts, macadamia nuts, ginkgo nuts, or extracts thereof; coffee or extracts thereof; and some medicinal and edible plant materials or extracts thereof.

[0093] For the animal milk product ingredients, examples include fresh milk derived from cows, sheep, and the like, and reprocessed milk products such as whole milk powder, skim milk powder, concentrated whey protein powder, desalted whey powder, whey protein powder, hydrolyzed whey protein powder, casein powder, and the like.

[0094] For the animal meat product ingredients, examples include meat product ingredients of pigs, cows, sheep, aquatic products, or birds.

[0095] For the functional additive ingredients, examples include vitamin supplements (e.g., vitamin A, beta-carotene, vitamin D3, vitamin E, vitamin Kl, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, pantothenic acid, folic acid, niacin, biotin, and the like), mineral supplements (e.g., iron, copper, manganese, zinc, cobalt, nickel, chromium, vanadium, fluorine, selenium, iodine, silicon, tin, and the like), nucleotide supplements (e.g., inosine), dietary fibers (e.g., inulin, konjac powder, galactooligosaccharides, fructooligosaccharides, isomaltooligosaccharides, soybean polysaccharides, cyclodextrins, resistant dextrins, soybean fibers, and the like), functional polyunsaturated fatty acid supplements (e.g., arachidonic acid oil powder, docosahexaenoic acid oil powder, and the like), and the like.

[0096] For the any acceptable adjuvants, examples include solvents, antioxidants, antibacterial agents, thickening agents, diluents, co-solvents, stabilizers, emulsifiers, fillers, disintegrants, lubricants, coating materials, anti-caking agents, flavoring agents, sweetening agents, food essences, food colorants, and the like.

[0097] Examples

[0098] Embodiments of the present application will be described in detail below with reference to Examples, but those skilled in the art will understand that the following Examples are for illustrative purposes only and should not be construed as limiting the scope of the present application. The specific conditions not mentioned in the Examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not specified by the manufacturer, but are conventional products that can be obtained commercially.

[0099] Example 1: Effect of nutritional compositions on free total amino acid and mineral absorption rate

[0100] Caco-2 cells have been shown to be a good model for studying the absorption of substances. Under in vitro culture conditions, the Caco-2 cell model can be used to study the absorption mechanism of substances in the small intestine, the interaction of nutrients during the absorption process, and the relationship between the chemical structure of substances and transport in vivo. Therefore, the present application evaluates the absorption rate of free amino acids and minerals after digestion of each sample by Caco-2 cells by in vitro simulated digestion of different nutritional compositions, to analyze the bioavailability of different samples.

[0101] I. Experimental methods

[0102] 1. Raw materials:

[0103] Alpha-lactalbumin: Arla, Alpha-10;

[0104] 6'-SL: DSM, GlyCare TM 6SL 9001;

[0105] 3-FL: DSM, GlyCare 3-FL 9001.

[0106] 2. Cells: Caco-2 (human colorectal adenocarcinoma cells).

[0107] 3. Preparation of milk protein mineral-containing compositions

[0108] According to the protein content of the sample, the total protein concentration was adjusted to 10 mg / mL with deionized water to prepare 50 mL of milk protein sample. The amount of minerals added was based on the mineral addition level in infant formula, i.e. 32.7 mg of calcium, 2.8 mg of magnesium, 0.45 mg of iron, and 0.27 mg of zinc per g of protein. The proportion of the content of the nutritional compositions in each experimental example is shown in Table 1.

[0109] Table 1: Content of nutritional compositions in each experimental example

[0110]

[0111] 4. In vitro gastrointestinal digestion

[0112] The basic scheme of the experiment uses a dynamic biomimetic digestion system provided by Xiaodong Yijian Instrument Equipment Co., Ltd. to carry out in vitro dynamic simulation digestion scheme. The digestion solution mainly includes the following:

[0113] Preparation method of simulated gastric fluid (SGF) electrolyte stock solution: dissolve the SGF powder in the product with a clean beaker, add 0.825 mL of MgCl2(H2O)6 solution, completely dissolve and transfer to a 1L volumetric flask, add 3.25 mL of 6M HCl, and dilute to 1L with distilled water, then transfer to a glass reagent bottle for storage. Preparation of digestion solution simulation solution: take 98.5 mL of SGF electrolyte stock solution, add 50 μL of CaCl2·2H2O solution, adjust the pH with 6M HCl, and adjust the dynamic in vitro digestion to 3.2. Take a portion of Pepsin + Lipase and dissolve in the above solution (when dissolving the enzyme, pay attention to minimize the loss of enzyme, otherwise it will affect the enzyme concentration of the digestion solution simulation solution, it is recommended to rinse the reagent bottle with the above solution).

[0114] Preparation method of simulated intestinal fluid (SIF) electrolyte stock solution: dissolve the SIF powder in the product with a clean beaker, add 2.75 mL of MgCl2(H2O)6 solution, completely dissolve and transfer to a 1L volumetric flask, add 1.75 mL of 6M HCl, and dilute to 1L with distilled water, then transfer to a glass reagent bottle for storage. Preparation of digestion solution simulation solution: take 98.3 mL of SIF electrolyte stock solution, add 200 μL of CaCl2(H2O)2 solution, and adjust the pH to 6.5 with 6M HCl. Take a portion of Pancreatin and Bile salt and dissolve in the above solution (when dissolving the enzyme, pay attention to minimize the loss of enzyme, otherwise it will affect the enzyme concentration of the digestion solution simulation solution).

[0115] Fourteen 100 mL centrifuge tubes were prepared for gastric digestion stage, 30 mL gastric juice was added respectively, and incubated at 37°C for 10 min. Fourteen 30 mL different compounded compositions were incubated at 37°C for 10 min, and added to the incubated digestion liquid respectively, and incubated at 37°C on a shaking table (180 r / min), and the timing was started. Each centrifuge tube was controlled to take sample at 60 min of digestion time, and 1 M NaHCO3 was added to adjust the pH to 7.0, and placed on ice to stop the reaction. In the simulated intestinal digestion stage, 30 mL digestion product was mixed with 30 mL SIF, and incubated at 37°C on a shaking table. Each centrifuge tube was sampled at 120 min, and 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride was added to a final concentration of 4 mM, and placed on ice to stop the reaction. The digestion product was aliquoted, sealed, and placed at -20°C for subsequent digestion product determination.

[0116] 5. In vitro Caco-2 cell absorption experiment

[0117] The recovered Caco-2 cells were inoculated in culture bottles and placed in a 37°C, 5% CO2 incubator for culture. When the Caco-2 cells in the culture bottles grew to the logarithmic phase, they were inoculated in Transwell chambers, and complete culture medium was added to the top side and the base side of the filter membrane, respectively, and then placed in the incubator for culture. The cell morphology was observed daily under a microscope.

[0118] 5.1 Amino acid and mineral transport absorption ratio determination

[0119] A bidirectional transmembrane transport experiment was performed using the Caco-2 cell monolayer model formed after 21 days of culture. The cell culture medium on both sides of the Transwell chamber was discarded, and the cells were washed twice with calcium and magnesium-free Hanks buffer for standby. The 0.22 μm filter was used to filter the final digestion product of each group. 0.25 mL of the digestion product sample and 0.15 mL of calcium and magnesium-free Hanks buffer were added to the AP side of the Caco-2 cell Transwell chamber, and 1.5 mL of calcium and magnesium-free Hanks buffer was added to the BL side. The chamber was placed in the incubator for 2 h, then 0.1 mL of BL side liquid was aspirated, the inner chamber was removed, and the cells on the polycarbonate membrane were carefully scraped off and ultrasonically broken. The content of total amino acids in each group of initial AP side addition and final BL side transfer and cells was determined by ninhydrin colorimetry, and the content of calcium, magnesium, iron, and zinc elements in each group of initial AP side addition and final BL side transfer and cells was determined by flame atomic absorption spectrometry. The absorption ratios of amino acids, calcium, magnesium, iron, and zinc elements in each group were calculated, respectively, according to the following formula:

[0120] Amino acid, calcium and zinc element absorption ratio = content of amino acid, calcium and zinc element in cells / content of initial added amino acid, calcium and zinc element on AP side x 100%.

[0121] II. Experimental results

[0122] 1. Free total amino acid and mineral absorption rate

[0123] The total protein and mineral concentrations of each experimental example were the same, and after digestion by the in vitro gastrointestinal digestion model, an equal amount of the digestion solution was taken to the in vitro absorption model of Caco-2 cells, and the transport ratio of nutrients transported to the lower chamber was as shown in Table 2. Compared with the blank group, the content of α-lactalbumin was increased in Comparative Example 1, and the results showed that the free total amino acid transport ratio, calcium transport ratio and zinc transport ratio were increased (P <0.05), indicating that increasing the proportion of α-lactalbumin in the protein can improve the absorption of amino acids and minerals; based on Comparative Example 1, Experimental Examples 1, 2 and 3 added 6’-SL, and the results showed that the free total amino acid transport ratio, calcium transport ratio and zinc transport ratio were increased (P <0.05), indicating that the addition of 6’-SL can further improve the absorption of amino acids and minerals; based on Comparative Example 1, Experimental Examples 4, 5 and 6 added 3-FL, and the results showed that the free total amino acid transport ratio, calcium transport ratio and zinc transport ratio were increased (P <0.05), indicating that the addition of 3-FL can further improve the absorption of amino acids and minerals. Based on Comparative Example 1, Experimental Examples 1 to 6, Experimental Examples 7 to 12 added α-lactalbumin, 6’-SL and 3-FL in combination, and the results showed that the free total amino acid transport ratio, calcium transport ratio and zinc transport ratio were further improved (P <0.05).

[0124] Specifically, taking Experimental Example 2, Experimental Example 4 and Experimental Example 9 as examples, compared with the blank group, Experimental Examples 2 and 4 increased the free total amino acid transport ratio by 4.9 and 6.6 respectively, while Experimental Example 9 increased the free total amino acid transport ratio by 14.5, which was greater than the sum of the first two (4.9+6.6); Experimental Examples 2 and 4 increased the calcium transport ratio by 6.2 and 5.8 respectively, while Experimental Example 9 increased the calcium transport ratio by 17.7, which was greater than the sum of the first two (6.2+5.8); Experimental Examples 2 and 4 increased the zinc transport ratio by 6 and 6.1 respectively, while Experimental Example 9 increased the zinc transport ratio by 16.4, which was greater than the sum of the first two (6+6.1). The above results show that there is a synergistic effect between the nutritional composition of α-lactalbumin, 6’-SL and 3-FL, which can promote the increase of free total amino acid, calcium and zinc absorbed by intestinal cells.

[0125] Table 2: Free total amino acid transport ratio, calcium transport ratio and zinc transport ratio

[0126]

[0127] Note: Different letters indicate significant differences (P < 0.05) between treatments.

[0128] Example 2: Effect of nutritional composition on the expression of amino acid and mineral transport carrier proteins

[0129] I. Experimental methods

[0130] 1. Raw materials: same as in Example 1.

[0131] 2. Cells: same as in Example 1.

[0132] 3. Preparation of milk protein mineral-containing composition: same as in Example 1.

[0133] 4. In vitro gastrointestinal digestion: same as in Example 1.

[0134] 5. In vitro Caco-2 cell absorption experiment: same as in Example 1.

[0135] 5.1. Determination of the expression of amino acid and mineral transport carrier proteins

[0136] The main absorption mode of amino acids and minerals by small intestinal epithelial cells is active transport, i.e. into the body using transport carriers. Amino acid transport carrier proteins are an important class of proteins on the cell membrane, responsible for assisting the entry and exit of amino acids from cells, maintaining the homeostasis of amino acids in cells. SLC6A14 is a neutral / alkaline amino acid transport protein in the SLC superfamily 6, which has broad substrate specificity, transporting all amino acids except glutamic acid and aspartic acid, and has a typical neurotransmitter transport protein-like folding structure. TRPV6 is the main ion channel for calcium ion absorption in the small intestine, responsible for transporting calcium ions in the intestinal lumen to the cytoplasm. ZnT1 is a key transport protein for zinc absorption in the intestine.

[0137] Caco-2 cells were lysed, washed with PBS for 3 times, then 0.6 mL of culture medium and 0.6 mL of sample were added, after 24 h of intervention, the liquid was poured out, washed with PBS for 3 times, then 200 μL of cell lysis solution (containing 1 mM PMSF (phenylmethylsulfonyl fluoride)) was added, after 1-2 s of treatment, the liquid was sucked into a centrifuge tube. The whole process was carried out on ice, 4°C, 5000g centrifugation for 5 min, and the supernatant was taken. The protein concentration in the supernatant was measured by BCA method, and the transport protein content in the supernatant was determined according to the operation instruction of ELISA kit.

[0138] II. Experimental results

[0139] 1. Expression of amino acid and mineral transport carrier proteins

[0140] SLC6A14 is a neutral / aromatic amino acid transporter protein, which is widely expressed in tissues such as the intestine, and is involved in amino acid absorption and metabolic regulation through a sodium / chloride-dependent transport mechanism. TRPV6 is a calcium transporter protein, a calcium-selective ion channel, which is mainly involved in calcium absorption in the intestine and kidney, and regulates calcium homeostasis. Its abnormal expression is related to diseases such as osteoporosis, and is the focus of research on calcium metabolism disorders. ZnT1 is a zinc transporter protein, which maintains intracellular zinc homeostasis by effluxing zinc, and is essential for cell antioxidant, signal transduction and apoptosis regulation. It plays a core role in intestinal zinc absorption and neuron protection, and its dysfunction is related to zinc deficiency and neurodegenerative diseases. The expression amount of the amino acid and mineral transport carrier protein is shown in Table 3. Compared with the blank group, the content of α-lactalbumin in Comparative Example 1 was increased, and the results showed that the expression amounts of SLC6A14, TRPV6 and ZnT1 were increased (P < 0.05); on the basis of Comparative Example 1, 6'-SL was added in Experimental Examples 1, 2 and 3, and the results showed that the expression of SLC6A14, TRPV6 and ZnT1 was increased (P < 0.05); on the basis of Comparative Example 1, 3-FL was added in Experimental Examples 4, 5 and 6, and the results showed that the expression of SLC6A14, TRPV6 and ZnT1 was increased (P < 0.05). Compared with Comparative Example 1, Experimental Examples 1 to 6, Experimental Examples 7 to 12 synchronously added α-lactalbumin, 6'-SL and 3-FL, and the results of each group showed that the expression of SLC6A14, TRPV6 and ZnT1 was significantly increased (P < 0.05).

[0141] Specifically, compared with Experimental Example 3, Experimental Example 5 and Experimental Example 8, relative to the blank group, Experimental Examples 3 and 5 increased the expression amount of SLC6A14 by 0.018 and 0.023 respectively, and Experimental Example 8 increased the expression amount of SLC6A14 by 0.048, which was greater than the sum of the first two (0.018+0.023); Experimental Examples 3 and 5 increased the expression amount of TRPV6 by 21.04 and 33.59 respectively, and Experimental Example 8 increased the expression amount of TRPV6 by 63.68, which was greater than the sum of the first two (21.04+33.59); Experimental Examples 3 and 5 increased the expression amount of ZnT1 by 1.59 and 1.85 respectively, and Experimental Example 8 increased the expression amount of ZnT1 by 3.55, which was greater than the sum of the first two (1.59+1.85). The above results show that there is a synergistic effect between the nutritional composition of α-lactalbumin, 6'-SL and 3-FL, which can promote the expression of SLC6A14, TRPV6 and ZnT1 transporters, and improve the transport and absorption of cells to amino acids, calcium and zinc.

[0142] Table 3: Expression amount of amino acid and mineral transport carrier protein

[0143]

[0144] Note: Different letters indicate significant differences (P < 0.05) among treatments.

Claims

1. A nutritional composition for promoting absorption of amino acids and minerals, characterized in that, The nutritional composition comprises essential active ingredients shown in (a) and (b) below: (a) α-lactalbumin; (b) human milk oligosaccharide; wherein, in the nutritional composition, the mass ratio of the essential active ingredient shown in (a) to the essential active ingredient shown in (b) is (10-20):(1-25).

2. The nutritional composition according to claim 1, characterized in that, The human milk oligosaccharide includes 3-fucosyllactose and / or 6'-sialyllactose; provided that, if the human milk oligosaccharide includes 3-fucosyllactose and 6'-sialyllactose, the mass ratio of the α-lactalbumin, the 6'-sialyllactose and the 3-fucosyllactose is (10-20):(1-5):(5-20).

3. Nutritional composition according to claim 1 or 2, characterized in that, The nutritional composition further comprises whey protein other than α-lactalbumin.

4. Use of the nutritional composition according to any one of claims 1 to 3 in the manufacture of a foodstuff for facilitating absorption of amino acid and mineral.

5. Use according to claim 4, characterized in that, The amino acid includes free amino acid.

6. Use according to claim 4 or 5, characterized in that, The mineral includes calcium and / or zinc.

7. Use according to any one of claims 4 to 6, characterized in that, The facilitating absorption of amino acid and mineral includes facilitating expression of amino acid transporter protein, calcium transporter protein and / or zinc transporter protein.

8. Use according to any one of claims 4 to 7, characterized in that, The foodstuff is selected from the group consisting of infant food, child food, adolescent food, pregnant and lying-in woman food, adult food and elderly food.

9. Use according to any one of claims 4 to 8, characterized in that, The foodstuff contains any one or more of the following ingredients: plant product ingredient, animal milk product ingredient, animal meat product ingredient, functional additive ingredient and any acceptable adjuvant.

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