Immunity enhancing composition

Abstract

The present invention relates to a functional nutritional composition, in particular to an immunity-enhancing composition. The composition comprises: i) human milk oligosaccharides, wherein the human milk oligosaccharides at least comprise neutral fucosylated human milk oligosaccharides; ii) beta-glucans from one or more sources, wherein the ratio of the human milk oligosaccharides to the beta-glucans is 33 to 50:1, and the composition optionally comprises other nutritional ingredients.

Description

Technical Field

[0001] The invention belongs to the field of food, and relates to a functional nutritional composition, and more specifically, to a functional health care or nutritional composition with the function of improving immunity. Background Art

[0002] Under normal physiological conditions, bacteria and toxins in the intestine do not cause harm to the body. This is related to the coordination between the various systems of the body and the unique complete barrier function of the intestine. Among them, the intestinal mucosal immune barrier composed of intestinal intraepithelial lymphocytes, lamina propria lymphocytes, Peyer's lymph nodes and other intestinal-related lymphoid tissues plays a pivotal role in defending and resisting the invasion of bacteria, viruses and toxins. Intestinal mucosal immunity is the first line of defense for the body to prevent infection. In addition to the important role of IgA, intraepithelial lymphocytes (IEL) located in the small intestine, including T cells and B cells, are also indispensable for mucosal immunity. When bacteria, viruses, etc. break through the first line of defense and enter the body, the systemic immunity comes into play to identify and remove foreign invading substances, so that the human body is protected from attack. Therefore, we have developed a nutritional composition that improves the immunity of infants, children and adults, and enhances intestinal mucosal immunity and systemic immunity, including 2'-fucosyllactose (2'-FL) and yeast β-glucan.

[0003] Human milk oligosaccharides (HMOs) are the third largest solid component in breast milk after lactose and milk fat, with a content of about 5 to 12 g / L (mature breast milk). They are not destroyed by human gastric acid, nor are they broken down by digestive enzymes. They can directly reach the large intestine, stimulate the growth of beneficial bacteria (bifidobacteria and lactobacilli) in the intestine, indirectly inhibit the growth of harmful bacteria, and maintain the balance of intestinal microecology. In addition, HMOs can indirectly affect the nonspecific immune response of the human body, and their antigenic determinants play an irreplaceable role in the information transmission and immune coordination of the immune system. The intestine is the largest immune organ in the human body, with 70% of the human body's immune cells. Bifidobacteria can stimulate the intestine to produce immune substances and antibodies, enhance cell activity, and thus improve the body's immunity to pathogens and tumors. HMOs in human breast milk are composed of five monomers: D-glucose (Glc), D-galactose (Gal), N-acetylglucosamine (GlcNAc), L-fucose (Fuc) and sialic acid (Sia). Among them, 2'-fucosyllactose (2'-FL) is the HMO with the highest content in breast milk and has been shown to promote immune regulation. Although my country has not yet approved the use of HMOs in infant formula, the European Food Safety Authority concluded in 2015 that 2'-FL is safe for infants under 1 year old when added to infant formula and follow-up formula together with LNnT.

[0004] It is known that β-glucan is composed of glucose units and is widely found in microorganisms, plants and animals. It is a polysaccharide unit of glucose. Structurally, most of them are bonded by β-1,3 bonds. It is a polymer with certain biological activity. In the middle of the 20th century, foreign scholars discovered that there was a substance on the yeast cell wall that enhanced immune function. It was not until more than 20 years later that this active substance was determined to be β-1,3-glucan. Yeast glucan, one of the sources of β-glucan, comes from Saccharomyces cerevisiae. Due to its special structure, it has multiple physiological activities. In addition, the US FDA has listed it in GRAS (generally regarded as safe). As a food raw material, it is widely used in various foods.

[0005] β-glucans are thought to stimulate the immune system and modulate both humoral and cellular immunity, so they have a beneficial role in fighting infectious diseases such as bacterial, viral, fungal and parasitic diseases. They act on multiple immune receptors and activate a range of immune cells, including macrophages, neutrophils, monocytes, natural killer cells and dendritic cells. Therefore, β-glucans can modulate innate and adaptive responses and can also enhance phagocytosis. Small β-glucan fragments are eventually released by macrophages and taken up by other immune cells, resulting in various immune responses.

[0006] Although some experimental results show that 2'-fucosyllactose and β-glucan seem to have the effect of regulating immunity and improving human immunity, the research on their application in food is not sufficient. The most common use of a variety of substances with health benefits is to mix them. For example:

[0007] Cited document 1 discloses a personalized nutritional composition comprising oligosaccharides, 45% to 55% of which are α-1,2 fucosylated. The nutritional composition promotes the normal establishment of intestinal flora and intestinal immune function in the early life of infants and young children breastfed by mothers of specific genotypes, and can prevent diarrhea.

[0008] Cited document 2 proposes Cordyceps sinensis extract and its uses, studies the water-insoluble polysaccharides of Cordyceps sinensis, and obtains a water-insoluble polysaccharide extract (CEP) with good immunological activity, which is suitable for large-scale production. The content of β-glucan in the extract is 42% to 46%, and the immunological activity is high.

[0009] Reference 3 discloses a breast-milk infant formula milk powder that promotes intellectual development and intestinal health, and a preparation method and application thereof, wherein the formula milk powder is prepared by mixing cow's milk / goat's milk, desalted whey powder, composite vegetable oil, composite breast milk oligosaccharide, composite vitamins and composite minerals, etc., dissolving, homogenizing, concentrating, spray drying, and then mixing evenly with lactoferrin, sialic acid, composite fruit powder and composite probiotic powder, etc. The active ingredients of the formula milk powder are closer to breast milk by utilizing the synergistic effect between the composite components.

[0010] References:

[0011] Reference 1: CN112841316A

[0012] Reference 2: CN110511292A

[0013] Reference 3: CN112535218A Summary of the invention

[0014] Problem that the invention aims to solve

[0015] As mentioned above, various functional compositions related to improving immunity have been proposed in the art. However, in specific research practice, the inventors also found the following problems:

[0016] Reference 1 discloses a nutritional composition containing human milk oligosaccharides (HMO) to provide a precise nutritional plan for infants breastfed by Chinese mothers, but does not separately study the role of neutral fucosylated HMO in improving immunity, and does not involve β-glucan. Reference 2 discloses that the Cordyceps sinensis extract contains a large amount of β-glucan, but does not clarify the relationship between it and the effects of improving immunity, and does not involve neutral fucosylated HMO. Reference 3 discloses a formula milk powder containing HMOs and yeast β-glucan to promote intelligence and intestinal health, but does not verify the neutral fucosylated HMO through experiments, especially whether the dosage ratio of 2'-fucosyllactose (2'-FL) to yeast β-glucan can bring about a synergistic effect.

[0017] Therefore, in view of the above-mentioned deficiencies in the prior art, the problem to be solved by the present invention is to provide a composition with a simple composition and the effect of improving immunity. By scientifically designing the ratio of neutral fucosylated HMO, especially 2'-fucosyllactose and β-glucan in the composition, a synergistic effect can be obtained, thereby meeting the market demand for a composition with the function of enhancing immunity.

[0018] Solutions for solving problems

[0019] Through long-term research by the inventors of the present invention, it is found that the above technical problems can be solved by the following technical solutions:

[0020] [1]. A functional composition, characterized in that the composition comprises:

[0021] i) one or more human milk oligosaccharides (HMOs), wherein the human milk oligosaccharides include at least neutral fucosylated human milk oligosaccharides;

[0022] ii) one or more sources of β-glucan,

[0023] Wherein, in the composition, the dosage ratio of the human milk oligosaccharide to the β-glucan is 33 to 50:1, and the composition optionally includes other nutritional ingredients.

[0024] [2] The composition according to [1], characterized in that the ratio of the human milk oligosaccharide to the β-glucan is 35 to 47:1.

[0025] [3] The composition according to [1] or [2], wherein the content of the human milk oligosaccharide is 0.1% by mass or more, and the content of the β-glucan is 0.01% by mass or more.

[0026] [4] The composition according to any one of [1] to [3], characterized in that the neutral fucosylated human milk oligosaccharide comprises 2'-fucosyllactose; and the β-glucan comprises yeast β-glucan.

[0027] [5] The composition according to [4], characterized in that the 2'-fucosyllactose is selected from the group consisting of 2'-fucosyllactose obtained from fermentation medium, purified 2'-fucosyllactose separated, or artificially synthesized; and the β-glucan is selected from one or more β-glucans derived from plants or fungi.

[0028] [6] The composition according to any one of [1] to [5], characterized in that the composition is liquid, semi-solid or solid.

[0029] [7]. A composition for improving immunity, characterized in that the composition comprises the composition according to any one of [1] to [6].

[0030] [8]. A food, characterized in that the food is obtained via the composition according to any one of [1] to [7].

[0031] [9] The food according to [8], characterized in that the food comprises pasta, beverages, reconstituted food, baked goods, sauces or functional nutritional supplements.

[0032]

[10] The food according to [9], characterized in that the food is children's or baby food.

[0033] Effects of the Invention

[0034] Through the implementation of the above technical solution, the present invention has the following advantages and can obtain the following technical effects:

[0035] 1) The functional composition for improving immunity provided by the present invention has a simple and clear composition, which is more conducive to convenient and accurate compounding, and is also more conducive to convenient addition or preparation of various foods or health products;

[0036] 2) The neutral fucosylated human milk oligosaccharides in the composition of the present invention, especially 2'-fucosyllactose and β-glucan, can achieve a synergistic effect when used in a limited amount, enhance intestinal mucosal immunity and systemic immunity, and improve the body's resistance to diseases.

[0037] 3) The dosage of 2'-fucosyllactose and β-glucan in the composition of the present invention is within the safe dosage range, so it can be used for a long time and is suitable for infants, children, adults and other groups of people; DETAILED DESCRIPTION

[0038] The following is a detailed description of the present invention. The following description of the technical features is based on representative embodiments and specific examples of the present invention, but the present invention is not limited to these embodiments and specific examples. It should be noted that:

[0039] In this specification, the numerical range expressed using "a numerical value A to a numerical value B" means a range including the endpoints numerical values ​​A and B.

[0040] In the present specification, a numerical range expressed using "above" or "below" means a numerical range including the number.

[0041] In this specification, the word "may" means both performing a certain process and not performing a certain process.

[0042] In this specification, the use of "optional" or "optional" indicates that certain substances, components, execution steps, application conditions and other factors are used or not used.

[0043] In this specification, the unit names used are all international standard unit names, and unless otherwise stated, the "%" used indicates weight or mass percentage.

[0044] In the present specification, the use of "substantially" means that the standard deviation from a theoretical model or theoretical data is within a range of 5%, preferably 3%, and more preferably 1%.

[0045] In this specification, the references to "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", etc., mean that the specific elements (e.g., features, structures, properties and / or characteristics) described in connection with the embodiments are included in at least one embodiment described herein, and may or may not exist in other embodiments. In addition, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0046] <First aspect>

[0047] In the first aspect of the present invention, a functional composition is provided, comprising i) one or more human milk oligosaccharides and ii) one or more β-glucans from different sources. Furthermore, the human milk oligosaccharides at least comprise neutral fucosylated human milk oligosaccharides. In addition, the composition optionally comprises other nutritional ingredients. The above two components in the composition exhibit a significant synergistic effect, so as to enhance intestinal mucosal immunity and systemic immunity, and improve the body's resistance to disease.

[0048] The technical solution of the present invention is mainly obtained based on the following insights:

[0049] The present invention studies the effects of different contents and proportions of neutral fucosylated human milk oligosaccharides, especially the combination of 2'-fucosyllactose (2'-FL) and yeast β-glucan, on immune function through experiments. By conducting animal experiments on the immune efficacy of the combination, the functions of the two in enhancing immunity are evaluated separately and in combination with different dosages. Therefore, it is unexpectedly found that at a specific content, neutral fucosylated human milk oligosaccharides and yeast β-glucan can produce obvious synergistic effects in improving human immunity.

[0050] (Human milk oligosaccharides)

[0051] Human milk oligosaccharides (HMOs) are a general term for oligosaccharides with a degree of polymerization ≥ 3 that exist naturally in breast milk. They are based on lactose molecules and are modified by five monomers at the terminal position, namely glucose (Glc), galactose (Gal), N-acetylglucosamine (GlcNAc), fucose (Fuc) and N-acetylneuraminic acid (Neu5Ac). Each HMO molecule contains 3 to 32 monosaccharides, which are connected by different glycosidic bonds, forming the diversity and complexity of HMOs.

[0052] HMOs can be divided into three types based on their core structure: neutral fucosylated HMO (which contains fucose at the terminal position), neutral non-fucosylated HMO (containing N-acetylglucosamine at the terminal), and acidic or sialylated HMO (containing sialic acid at the terminal), and their ratios in human milk oligosaccharides are usually 35-50%, 42-55%, and 12-14%, respectively.

[0053] Fucosylated oligosaccharides are oligosaccharides with fucose residues. The fucosylated epitope of fucosylated oligosaccharides can act as a "decoy" at the mucosal surface. This epitope can use the competitive effect to prevent and / or limit the action of infectious pathogens (viral or bacterial origin) or substances secreted by pathogens (such as toxins), especially to avoid the binding of pathogens or substances secreted by pathogens to natural ligands, thereby reducing the risk of infection / inflammation. The following examples illustrate the types of fucosylated oligosaccharides: 2'-fucosyllactose (2'-FL), which is a neutral trisaccharide composed of L-fucose, D-galactose and D-glucose units, in which the monosaccharide L-fucose is connected to the disaccharide D-lactose by an α(1→2) bond. Its molecular formula is C 18 H 31 O 15 , the molecular weight is 488.439g / mol, and the molecular structure is as follows:

[0054]

[0055] 3'-fucosyllactose (3'-FL) is a neutral trisaccharide composed of L-fucose, D-galactose and D-glucose units, in which the monosaccharide L-fucose is linked to D-glucose via an α(1→3) bond. Its molecular formula is C18H32O15 and its molecular weight is 488.44 g / mol;

[0056] Lactose-N-fucopentaose I (LNFP I) is a neutral pentasaccharide composed of L-fucose, D-glucose, 2 molecules of D-galactose, and N-acetylglucosamine units, in which the monosaccharide L-fucose is linked to D-glucose via an α(1→3) bond. Its molecular formula is C 32 H 55 NO 25 , molecular weight is 853.77 g / mol;

[0057] Lactose-N-difucohexaose I (LNDFH I) is a neutral hexasaccharide composed of 2 molecules of L-fucose, D-glucose, D-galactose, and N-acetylglucosamine units, in which the monosaccharide L-fucose is connected to D-galactose and N-acetylglucosamine through α(1→2) bonds and α(1→4) bonds, respectively. Its molecular formula is C38H65NO29, and its molecular weight is 999.91g / mol;

[0058] Lactose-N-difucohexaose II (LNDFH II) is a neutral hexasaccharide composed of 2 molecules of L-fucose, D-glucose, D-galactose, and N-acetylglucosamine units, in which the monosaccharide L-fucose is connected to D-glucose and N-acetylglucosamine through α(1→3) bonds and α(1→4) bonds, respectively. Its molecular formula is C38H65NO29, and its molecular weight is 999.91g / mol.

[0059] The human milk oligosaccharide HMOs described in the present invention at least include the above-mentioned neutral fucosylated oligosaccharide, and the neutral fucosylated oligosaccharide is preferably 2'-fucosyllactose (2'-FL).

[0060] 2'-fucosyllactose (2'-FL) is a type of oligosaccharide in human milk. Its biological function has attracted widespread attention and is a hot topic in international nutrition research. Many research results have shown that oligosaccharides are important in promoting the production and development of infants and young children and regulating intestinal flora.

[0061] 2'-fucosyllactose (2'-FL) is a white to milky white powder, hygroscopic, odorless, and has a solubility of 500g / L in water. As an oligosaccharide, 2'-FL has relatively stable product properties and almost no loss occurs during the production and processing process and during the shelf life.

[0062] There is no particular limitation on the source of 2'-fucosyllactose in the present invention. In some specific embodiments of the present invention, 2'-fucosyllactose is prepared by a fermentation process, obtained from a fermentation medium using a series of filtration and chromatography steps, or separated and purified. It can also be obtained by artificial synthesis, and preferably, can be obtained by artificial synthesis.

[0063] Furthermore, there is no particular limitation on the use of 2'-fucosyllactose. 2'-fucosyllactose can be prepared as a solution before use.

[0064] In some specific embodiments of the present invention, the content of 2'-fucosyllactose in the human milk oligosaccharide is 80% by mass or more, preferably 85% by mass or more, more preferably 90% by mass or more, and even more preferably 95% by mass or more.

[0065] (β-Glucan)

[0066] Beta-glucan (abbreviated as β-glucan) belongs to fiber polysaccharides (complex sugars). Structurally, most of them are polymers linked by β-1,3 bonds and can be derived from a variety of microorganisms, animals or plants.

[0067] There is no particular limitation on the specific source of β-glucan in the present invention. The sources of β-glucan applicable to the present invention include yeast, oats, barley, fungi (mushrooms), algae, etc., or a mixture thereof. Further specific examples include oat β-glucan, kelp β-glucan, thermogel β-glucan, golden algae β-glucan, shiitake β-glucan, lichen β-glucan, oyster mushroom β-glucan, yeast polysaccharide, black fungus β-glucan, and barley polysaccharide.

[0068] In some preferred embodiments of the present invention, as for β-glucan, one of β-glucans from yeast, oats, and barley or a mixture thereof can be used, more preferably, β-glucan is selected from yeast β-glucan and / or oat β-glucan.

[0069] In some specific embodiments of the present invention, raw materials such as plants and fungi containing β-glucan can be used to give a certain content of β-glucan in the final composition. In addition, preferably, the β-glucan active ingredient with higher purity extracted from these raw materials can also be directly used.

[0070] Furthermore, there is no particular limitation on the method of using the extracted β-glucan. The β-glucan may be used directly to form a mixture, or the β-glucan may be prepared as a solution or emulsion before use.

[0071] (Synergy)

[0072] Although the existing technology has preliminary conclusions on the research of human milk oligosaccharides and β-glucan showing that both may have a certain effect in improving immunity, there is no research or report on whether the two exist or may form a synergistic immunity-improving ability.

[0073] In the present invention, it is believed that the use of neutral fucosylated human milk oligosaccharides, especially 2'-fucosyllactose and β-glucan in combination at a specific content can unexpectedly find a synergistic effect in improving immunity. Specifically, in the functional composition provided by the present invention, from the perspective of ensuring the synergistic effect and the safety of dosage, if the content of 2'-fucosyllactose and β-glucan is too low, there is a concern that the synergistic effect is not good, and if the content is too high, there is also a concern that the safety of the human body is not good.

[0074] In some specific embodiments, the ratio of neutral fucosylated human milk oligosaccharides, especially 2'-fucosyllactose to β-glucan is 33:1 or more, preferably 34:1 or more, more preferably 35:1 or more, and specifically (36, 37 or 38):1 or more. In addition, it can be 140:1 or less, preferably 50:1 or less, more preferably 48:1 or less, and specifically (47, 46, 45, 44, 43, 42 or 41):1 or less.

[0075] In addition, the present invention has no particular restrictions on the absolute content (especially the upper limit absolute content) of neutral fucosylated human milk oligosaccharides and β-glucan in the composition. On the one hand, it is related to food safety regulations, and on the other hand, it is also related to the form and use of different products. It can be cited that in some specific embodiments, in the functional nutritional composition provided by the present invention, based on the total mass of the composition, from the perspective of ensuring synergistic effects and dosage safety, the neutral fucosylated human milk oligosaccharides, especially 2'-fucosyllactose, have a content of 0.10% by mass or more, and the β-glucan content is 0.01% by mass or more. In some embodiments, the content of 2'-fucosyllactose can be 0.10-8.00 mass % (e.g., 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, 3.6, 3.8, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5 mass %); in some embodiments, β-glucan The sugar content is 0.01 to 0.3% by mass (e.g., 0.015, 0.02, 0.025, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.12, 0.14, 0.16, 0.18, 0.20, 0.22, 0.24, 0.26, 0.28, 0.30% by mass, etc.).

[0076] The present invention further found that the effect of using 2'-fucosyllactose or β-glucan alone on improving immunity is not obvious, but by matching the above contents, it is found that the two meet the above conditions of use, and a synergistic effect can be observed.

[0077] (Composition)

[0078] The functional composition of the present invention at least includes the neutral fucosylated human milk oligosaccharides described above, especially 2'-fucosyllactose (2'-FL) and β-glucan. There is no particular limitation on the formation of the composition, which can be a mixture of the plants, fungi and other substances containing these components as described above, or a mixture of two high-purity extracts.

[0079] In addition, there are no particular limitations on other components that may be used in the composition of the present invention. Without impairing the effects of the present invention, other edible ingredients, food additives or solvent components commonly used in the art may be used.

[0080] The functional composition of the present invention may be in a liquid state, or in a semi-solid state or a solid state.

[0081] Furthermore, the above-mentioned composition of the present invention has been verified through animal experiments, and it has been found that it can not only enhance intestinal mucosal immunity, but also regulate systemic immunity. Therefore, it can be used as a functional health product or functional health supplement.

[0082] <Second Aspect>

[0083] In the second aspect of the present invention, a functional nutritional supplement food is provided, especially a food for improving immunity suitable for infants, children and adults, which is obtained by processing the functional composition described in the first aspect.

[0084] There is no particular limitation on the food of the present invention, and the food may generally be pasta, beverages, reconstituted food, baked goods, sauces or functional nutritional supplements.

[0085] Wheaten food may include staple foods processed or made from flour-based raw materials, coarse grain staple foods, etc., such as steamed bread, cakes, noodles, stuffed staple foods, etc.

[0086] For baked pastries, it can be baked cakes or biscuits mainly based on cream, eggs and baking powder.

[0087] There are no particular restrictions on beverages, which may be fruit drinks, vegetable drinks, milk tea drinks, tea drinks, milk, yogurt, vitamin drinks, etc. For fruit or vegetable drinks, in addition to the two functional components of the present invention, they may also include fruit or vegetable juice, or their solid components, etc. For vitamin drinks, in addition to the two functional components of the present invention, they may also mainly include various functional vitamins and other functional ingredients, such as white sugar, sodium cyclamate, acesulfame potassium, taurine, potassium sorbate, lysine, inositol, vitamin pp, vitamin B6, vitamin B12, citric acid, etc.

[0088] The reconstituted food may typically be a reconstituted milk powder product, such as infant milk powder, adult milk powder, middle-aged and elderly milk powder, etc. In particular, the composition of the present invention is very suitable for the preparation of infant milk powder.

[0089] In addition, there are no special restrictions on functional nutritional supplement foods, which can be used as nutritional supplements or meal replacements. Such foods, in addition to the two functional components of the present invention, may also include one or more of the following components: protein, fat, essential carbohydrates, dietary fiber, supplementary elements, vitamins, plants or dietary ingredients, etc.

[0090] The protein is selected from at least one of whey protein powder, soy protein isolate, whole milk powder, whole egg powder, lactoferrin, bovine colostrum, amino acids, and protein peptides; and the amino acids are selected from at least one of L-lysine-L-glutamic acid, L-glutamic acid, L-arginine, L-tryptophan, L-glutamine, taurine, L-valine, L-isoleucine, and L-leucine; and the protein peptides are selected from one or more of soy oligopeptides, wheat protein peptides, silkworm pupa protein peptides, marine fish oligopeptide powder, kola peptides, amino peptides, and ovalbumin peptides.

[0091] The fat may include at least one of saturated fatty acids, polyunsaturated fatty acids, monounsaturated fatty acids, OPO structured lipids, DHA, EPA, ARA, and phospholipids. More specifically, the fat includes safflower seed oil, walnut oil, peanut oil, soybean oil, argan oil, olive oil, tea oil, sacha inchi oil, olive oil, coconut oil, perilla oil, deep-sea fish oil, cocoa butter, palm oil, butter, cream, lard, medium-chain triglycerides, and lecithin.

[0092] The carbohydrates include starch or modified starch. The dietary fiber includes one or more of inulin, konjac flour, galacto-oligosaccharide, fructo-oligosaccharide, isomalto-oligosaccharide, soybean polysaccharide, cyclodextrin, resistant dextrin and soybean fiber.

[0093] The supplementary elements are selected from metal ion salts of citric organic acids, such as one or more of calcium citrate, calcium L-lactate, calcium hydrogen phosphate, potassium gluconate, sodium citrate, ferrous gluconate, potassium iodide, zinc gluconate, sodium selenite, copper gluconate, chromium sulfate, manganese gluconate and magnesium gluconate.

[0094] The vitamins are selected from one or more of vitamin A, β-carotene, vitamin D3, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, pantothenic acid, folic acid, niacin, choline, inositol, and biotin.

[0095] The plants or dietary ingredients include one or more of cereals, beans, potatoes, vegetables, fungi and algae, nuts and seeds, fish and shrimp, poultry and livestock meat, and fruits.

[0096] Example

[0097] The present invention will be further described in detail below through specific examples. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0098] Animal experiment on immune efficacy of the composition

[0099] 1 Materials and methods

[0100] 1.1 Main Reagents

[0101] The main reagents used in this experiment include: RPMI-1640 culture medium (HyClone), fetal bovine serum (HyClone), Hank's solution (Beijing Solebao), anhydrous ethanol, glacial acetic acid, Cell Counting Kit-8 (CCK-8 kit, Japan Tongren), sIgA kit (Quanzhou Konodi), and transforming growth factor-β (TGF-β) kit (Quanzhou Konodi).

[0102] 1.2 Main instruments and equipment

[0103] LDZF-50KB-II vertical pressure steam sterilizer, VD-1320 clean workbench, HPG-9245 oven, full range of Eppendorf pipettes, GL-21M high-speed refrigerated centrifuge, electron microscope, PL2002 and AL104 electronic balances, vortex oscillator, HF90 CO2 incubator, AE-30 inverted biological microscope, Model 680 microplate reader.

[0104] 1.3 Experimental animals

[0105] 144 BALB / c female mice bred by Beijing Weitonglihua Experimental Animal Technology Co., Ltd. [License No.: SCXK (Beijing) 2016-0006] were selected. The experimental animal use license number is SYXK (Hei) 2019-0001. The maintenance feed was produced by Shenyang Maohua Biotechnology Co., Ltd. [License No.: SCXK (Liao) 2017-0001].

[0106] 1.4 Experimental Grouping

[0107] The experimental animal breeding room temperature was 20-25°C, the relative humidity was 40-70%, the light was alternated 24 hours a day, and the animals were allowed to drink water and eat freely. The formal experiment began after 7 days of adaptive feeding. 144 female BALB / c mice were randomly divided into 12 groups, with 12 mice in each group. Animal Experiment Examples 2-12 were gavaged every other day for 30 days, and Animal Experiment Example 1 was gavaged with an equal amount of physiological saline, and each group was fed with maintenance feed. The specific experimental doses are shown in the table below.

[0108] Comparative Examples and Experimental Examples were prepared by mixing 2′-FL and yeast β-glucan in the composition according to the ratios shown in Table 1 below.

[0109] Table 1: Animal experimental dosage

[0110]

[0111] 1.5 Detection indicators

[0112] 1.5.1 Mouse colon cytokine assay

[0113] 100 mg of colon tissue was accurately weighed, the colon sample was cut into small pieces, and then the colon and 900 μL PBS were put into a glass grinder for grinding to make a 10% suspension, centrifuged at 10000 rpm for 10 min at 4°C, and finally the colon supernatant was transferred into a centrifuge tube. The TGF-β content of the supernatant was determined using an ELISA kit (Conodi Biotechnology, Quanzhou, China) according to the instructions of the kit.

[0114] 1.5.2 Mouse fecal sIgA assay

[0115] Weigh 100 mg of feces accurately, add 1000 μL PBS, and vibrate on an oscillator for 15 seconds to make a suspension. Centrifuge at 10000 rpm for 10 minutes at 4°C, and finally transfer the fecal supernatant into a centrifuge tube. Detect the sIgA content using an ELISA kit (Conodi Biotechnology, Quanzhou, China) according to the kit instructions.

[0116] 1.5.3 Mouse splenocyte extraction

[0117] The mice were fixed, their eyeballs were removed and bled, and the mice were killed by cervical dislocation. The mice were then placed in a beaker containing 75% ethanol and soaked for 3-5 minutes. The mouse chest was cut open along the midline of the abdominal cavity on a clean bench, and the spleen was aseptically removed and placed on a 200-mesh stainless steel sieve. The center of the sieve was immersed in a plate containing Hank's solution. The spleen tissue was ground with a sterile syringe core, and the remaining tissue on the sieve was blown off with Hank's solution. The spleen tissue suspension was collected in a sterile centrifuge tube and centrifuged at 1000 r / min for 5 minutes. The supernatant was discarded, and 2-3 mL of red blood cell lysis buffer was added to the washed cells. After mixing, the cells were allowed to stand for 2-3 minutes until the red blood cells were completely broken. The cells were then centrifuged at 1000 r / min for 5 minutes and the supernatant was discarded to remove the red blood cells. The cells were then washed twice by centrifugation with RPMI-1640 incomplete culture medium. The cells were collected and resuspended in RPMI-1640 complete medium containing 10% fetal bovine serum. The cell suspension was stained with trypan blue and counted on a hemocytometer to ensure that the proportion of live cells was not less than 95%. After counting, the splenocyte concentration was adjusted to 1×10 with RPMI-1640 complete medium containing 10% fetal bovine serum. 4 pcs / mL for future use.

[0118] 1.5.4 Determination of NK cell activity

[0119] 100 μL / well of spleen cells (effector cells) and target cells were inoculated into a 96-well plate as a reaction control well (C); 100 μL of effector cells and RPMI-1640 complete culture medium were added to the effector cell well (B); 100 μL of target cells and RPMI-1640 complete culture medium were added to the target cell natural release well (A), and 200 μL of serum-free RPMI-1640 culture medium was added as a control well (zero well). The cells were cultured in a 37°C, 5% CO2 incubator for 20 hours, 15 μL / well of CCK-8 solution was added, and the culture was continued for 4 hours, and the oscillation was mixed; then the optical density of each well was measured at 450 nm using an enzyme reader. Calculate the NK cell activity as follows: NK cell activity (%) = (A+BC) / A×100%

[0120] 1.6 Statistical analysis

[0121] The experimental data were expressed as mean ± standard deviation (mean ± SD). Each experiment was repeated at least three times. GraphPad Prism 8.00, Excel 2018 and R language were used to draw charts. SPSS 16.0 software was used for statistical analysis of the experimental data. One-way analysis of variance (Duncan's multiple range test) was used. P < 0.05 indicated a significant difference.

[0122] 2. Results

[0123] 2.1 Effects of each test substance on TGF-β secretion in mouse colon

[0124] To evaluate the effects of the test substances alone or in combination on the intestinal mucosal immune function of mice, the secretion of colonic anti-inflammatory cytokine TGF-β in each group was measured, as shown in Table 2.

[0125] Table 2: Effects of each test substance on TGF-β secretion in mouse colon (mean ± SD, n = 12)

[0126]

[0127] Note: * Compared with the blank control group, P<0.05. ** Compared with the blank control group, P<0.01. # Compared with 2'-FL group (2790.0 mg / kg.bw), P<0.05, ## Compared with the 2'-FL group at 2790.0 mg / kg.bw, P<0.01; compared with the yeast β-glucan group at 117.6 mg / kg.bw, P<0.05; compared with the yeast β-glucan group at 117.6 mg / kg.bw, P<0.01.

[0128] In the above table, compared with the control group, the 2'-FL dosage of 2790.0 mg / kg.bw group and the yeast β-glucan dosage of 117.6 mg / kg.bw group increased the secretion of colon TGF-β, indicating that these two ingredients are helpful to enhance the intestinal mucosal immunity of mice.

[0129] In addition, if the values ​​of Experimental Examples 1 and 2 are directly added together, it is believed that the value in "the increase relative to the control group" should not exceed 28.95+8.32=37.27 (pg / ml). Then, it can be seen that the data of Experimental Examples 10 and 11 show that the two substances have a synergistic effect at a specific ratio.

[0130] 2.2 Effects of each test substance on sIgA secretion in mouse colon

[0131] Table 3: Effects of each test substance on sIgA secretion in mouse colon (mean±SD, n=12)

[0132]

[0133] Note: *Compared with blank control group, P<0.05; **Compared with blank control group, P<0.01; #Compared with 2'-FL group (2790.0 mg / kg.bw), P<0.05; ##Compared with 2'-FL group (2790.0 mg / kg.bw), P<0.01; ^Compared with yeast β-glucan group (117.6 mg / kg.bw), P<0.05; ^^Compared with yeast β-glucan group (117.6 mg / kg.bw), P<0.01.

[0134] Compared with the blank control group, the 2'-FL 2790.0 mg / kg.bw group and the yeast β-glucan 117.6 mg / kg.bw group could significantly increase the secretion of colon sIgA, indicating that these two components are helpful to enhance the intestinal mucosal immunity of mice.

[0135] In addition, it can be seen from the table above that animal experiment examples 1 and 2 increased the secretion of sIgA by 48.05 μg / mL and 24.43 μg / mL, respectively, relative to the blank control group. If the two are directly superimposed, it is believed that the value in "the increase relative to the control group" should not exceed 48.05+24.43=72.48 μg / mL. Animal experiment example 11 increased the secretion of sIgA by 89.11 relative to the blank control group, which is greater than the previous resin of the two directly superimposed, indicating that there is a synergistic effect between 2'-FL and yeast β-glucan, which can synergistically increase the secretion of sIgA.

[0136] 2.3 Effects of each test substance on mouse NK cell activity

[0137] Table 4: Effects of each test substance on mouse NK cell activity (mean ± SD, n = 12)

[0138]

[0139] Note: *Compared with blank control group, P<0.05; **Compared with blank control group, P<0.01; #Compared with 2'-FL group (2790.0 mg / kg.bw), P<0.05; ##Compared with 2'-FL group (2790.0 mg / kg.bw), P<0.01; ^Compared with yeast β-glucan group (117.6 mg / kg.bw), P<0.05; ^^Compared with yeast β-glucan group (117.6 mg / kg.bw), P<0.01.

[0140] Compared with the blank control group, the 2'-FL 2790.0 mg / kg.bw group significantly increased the activity of NK cells, while the yeast β-glucan 117.6 mg / kg.bw group had no effect on the activity of NK cells, indicating that only 2'-FL helps to enhance the systemic immunity of mice.

[0141] According to the comparison method in the previous text, it can be seen that Experimental Examples 8 to 11 can show a synergistic effect.

[0142] From the above experimental results, it can be seen that the neutral fucosylated human milk oligosaccharide represented by 2'-FL and yeast β-glucan in the composition of the present invention can be expected to achieve a synergistic effect in the range of 33 to 50:1, which can significantly increase the secretion of colon anti-inflammatory cytokine TGF-β and immunoglobulin sIgA, thereby enhancing the intestinal mucosal immunity of mice. In addition, it also has a significant effect on improving the activity of NK cells and enhancing the systemic immunity of mice. The composition has the effect of improving immune function, and the immunomodulatory effect is significantly better than that of the control sample.

[0143] The present invention provides a study on the immune function of a neutral fucosylated human milk oligosaccharide and yeast β-glucan composition, which provides a new idea for the development of functional foods in the future. In particular, 2'-FL and yeast β-glucan have broad prospects in regulating the body's immunity. The combination of the two can not only enhance intestinal mucosal immunity, but also regulate systemic immunity.

[0144] <Application Examples>

[0145] In the following application examples, “parts” are all parts by weight, and the % content of components are all % content by weight.

[0146] In addition, in the following application examples, unless otherwise specified, the raw materials used are from the following sources:

[0147] Raw milk: farm (P06-11)

[0148] Whole milk powder: Heilongjiang Feihe Dairy Co., Ltd.

[0149] Skim milk powder: Kerry, Ireland

[0150] Demineralized whey powder (D90): Valio, Finland

[0151] Concentrated whey protein powder: Hilmar, USA

[0152] Edible vegetable blended oil: Cargill Grain and Oil (Nantong) Co., Ltd.

[0153] Anhydrous butter: New Zealand Tatua

[0154] 2'-fucosyllactose (2'-FL 93%): Royal FrieslandCampina

[0155] Yeast β-glucan (soluble, 98%): Kerry, USA

[0156] Lactose: Hilmar, USA

[0157] Corn syrup solids: Bowling Biotech Co., Ltd.

[0158] Fructo-oligosaccharide: Baolingbao Biotechnology Co., Ltd.

[0159] Isomaltooligosaccharide: Baolingbao Biotechnology Co., Ltd.

[0160] Calcium carbonate: Nantong Licheng Bioengineering Co., Ltd.

[0161] Calcium hydrogen phosphate: Zhengzhou Ruipu Bioengineering Co., Ltd.

[0162] Potassium chloride: Zhengzhou Ruipu Bioengineering Co., Ltd.

[0163] Sodium citrate: Rizhao Jinhe Biochemical Group Co., Ltd.

[0164] Compound choline chloride: Beijing Jinkangpu Food Technology Co., Ltd.

[0165] Compound vitamins: DSM Vitamins (Shanghai) Co., Ltd.

[0166] Compound minerals: DSM Vitamins (Shanghai) Co., Ltd.

[0167] Compound nutrients: DSM Vitamins (Shanghai) Co., Ltd.

[0168] Application Example 1

[0169] Infant formula milk powder containing 2'-FL and yeast beta-glucan, wherein every 1000 parts of infant formula milk powder is prepared from the following components in parts by weight:

[0170] The raw materials used in the milk powder of the present invention are: 470 parts of desalted whey powder (D90), 207 parts of edible vegetable blending oil, 181 parts of raw cow milk (in terms of dry matter), 97 parts of lactose, 21.5 parts of 2'-FL (the content of 2'-FL in each part is 93%), 5 parts of sodium citrate, 3 parts of calcium carbonate, 3 parts of calcium hydrogen phosphate, 2.8 parts of compound choline chloride, 2 parts of potassium chloride, 1.7 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%), 3 parts of compound vitamins, 2 parts of compound minerals and 1 part of compound nutrients. The content of 2'-FL in the product is 2.00%, and the content of yeast beta-glucan is 0.17%.

[0171] Application Example 2

[0172] Infant formula milk powder containing 2'-FL and yeast beta-glucan, wherein every 1000 parts of infant formula milk powder is prepared from the following components in parts by weight:

[0173] The raw materials used in the milk powder of the present invention are: 470 parts of desalted whey powder (D90), 207 parts of edible vegetable blending oil, 181 parts of raw cow milk (based on dry matter), 75.5 parts of lactose, 43 parts of 2'-FL (the content of 2'-FL in each part is 93%), 5 parts of sodium citrate, 3 parts of calcium carbonate, 3 parts of calcium hydrogen phosphate, 2.8 parts of compound choline chloride, 2 parts of potassium chloride, 1.7 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%), 3 parts of compound vitamins, 2 parts of compound minerals, and 1 part of compound nutrients. The content of 2'-FL in the product is 4.00%, and the content of yeast beta-glucan is 0.17%.

[0174] Application Example 3

[0175] An infant formula milk powder containing 2'-FL and yeast beta-glucan is prepared from the following components in parts by weight per 1000 parts of the infant formula milk powder:

[0176] The raw materials used in the milk powder of the present invention are: 318 parts of raw cow milk (in terms of dry matter), 300 parts of desalted whey powder (D90), 241.2 parts of lactose, 115 parts of edible vegetable blending oil, 4.3 parts of 2'-FL (the content of 2'-FL in each part is 93%), 4 parts of sodium citrate, 3 parts of calcium carbonate, 2.8 parts of compound choline chloride, 2 parts of calcium hydrogen phosphate, 2 parts of potassium chloride, 1.7 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%), 3 parts of compound vitamins, 2 parts of compound minerals and 1 part of compound nutrients. The content of 2'-FL in the product is 0.40%, and the content of yeast beta-glucan is 0.17%.

[0177] Application Example 4

[0178] Infant formula milk powder containing 2'-FL and yeast beta-glucan, each 1000 parts of infant formula milk powder is prepared from the following components in parts by weight:

[0179] The raw materials used in the milk powder of the present invention are: 400 parts of desalted whey powder (D90), 372 parts of raw cow milk (in terms of dry matter), 131 parts of lactose, 80 parts of edible vegetable blending oil, 4.3 parts of 2'-FL (the content of 2'-FL in each part is 93%), 4 parts of calcium carbonate, 2.8 parts of compound choline chloride, 0.9 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%), 3 parts of compound vitamins, 1 part of compound minerals, and 1 part of compound nutrients. The content of 2'-FL in the product is 0.40%, and the content of yeast beta-glucan is 0.09%.

[0180] Application Example 5

[0181] The modulated milk powder containing 2'-FL and yeast β-glucan is suitable for children to eat. Each 1000 parts of the modulated milk powder is prepared from the following components in parts by weight:

[0182] The raw materials used in the milk powder of the present invention are: 267 parts of raw cow milk (in terms of dry matter), 265 parts of whole milk powder, 214 parts of desalted whey powder (D90), 155 parts of skimmed milk powder, 80 parts of oligofructose, 5 parts of concentrated whey protein powder, 5 parts of calcium carbonate, 4.3 parts of 2'-FL (the content of 2'-FL in each part is 93%), 1.4 parts of compound choline chloride, 2 parts of compound vitamins, 1 part of compound minerals, and 0.3 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%). The content of 2'-FL in the product is 0.40%, and the content of yeast beta-glucan is 0.03%.

[0183] Application Example 6

[0184] The modulated milk powder containing 2'-FL and yeast β-glucan is suitable for consumption by adults and middle-aged and elderly people. Each 1000 parts of the modulated milk powder is prepared from the following components in parts by weight:

[0185] The raw materials used in the milk powder of the present invention are: 700 parts of whole milk powder, 112.8 parts of lactose, 110 parts of solid corn syrup, 40 parts of isomaltooligosaccharide, 21.5 parts of 2'-FL (the content of 2'-FL in each part is 93%), 12 parts of calcium carbonate, 1.4 parts of compound choline chloride, 1 part of compound vitamins, 1 part of compound minerals, and 0.3 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%). The content of 2'-FL in the product is 2.00%, and the content of yeast beta-glucan is 0.03%.

[0186] Application Example 7

[0187] The modulated milk powder containing 2'-FL and yeast β-glucan is suitable for consumption by adults and middle-aged and elderly people. Each 1000 parts of the modulated milk powder is prepared from the following components in parts by weight:

[0188] The raw materials used in the milk powder of the present invention are: 700 parts of whole milk powder, 112.2 parts of lactose, 110 parts of solid corn syrup, 40 parts of isomaltooligosaccharide, 21.5 parts of 2'-FL (the content of 2'-FL in each part is 93%), 12 parts of calcium carbonate, 1.4 parts of compound choline chloride, 1 part of compound vitamins, 1 part of compound minerals, and 0.9 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%). The content of 2'-FL in the product is 2.00%, and the content of yeast beta-glucan is 0.09%.

[0189] Application Example 8

[0190] The modulated milk powder containing 2'-FL and yeast β-glucan is suitable for consumption by adults and middle-aged and elderly people. Each 1000 parts of the modulated milk powder is prepared from the following components in parts by weight:

[0191] The raw materials used in the milk powder of the present invention are: 700 parts of whole milk powder, 110 parts of solid corn syrup, 91.3 parts of lactose, 43 parts of 2'-FL (the content of 2'-FL in each part is 93%), 40 parts of isomaltooligosaccharide, 12 parts of calcium carbonate, 1.4 parts of compound choline chloride, 1 part of compound vitamins, 1 part of compound minerals, and 0.3 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%). The content of 2'-FL in the product is 4.00%, and the content of yeast beta-glucan is 0.03%.

[0192] Application Example 9

[0193] The modulated milk powder containing 2'-FL and yeast β-glucan is suitable for consumption by adults and middle-aged and elderly people. Each 1000 parts of the modulated milk powder is prepared from the following components in parts by weight:

[0194] The raw materials used in the milk powder of the present invention are: 700 parts of whole milk powder, 110 parts of solid corn syrup, 90.7 parts of lactose, 43 parts of 2'-FL (the content of 2'-FL in each part is 93%), 40 parts of isomaltooligosaccharide, 12 parts of calcium carbonate, 1.4 parts of compound choline chloride, 1 part of compound vitamins, 1 part of compound minerals, and 0.9 parts of yeast beta-glucan (the content of yeast beta-glucan in each part is 98%). The content of 2'-FL in the product is 4.00%, and the content of yeast beta-glucan is 0.09%.

[0195] Industrial Applicability

[0196] The functional composition provided by the present invention can be prepared industrially and is suitable for improving immunity.

Claims

1. Use of a functional composition in preparing a food for improving immunity, characterized in that: The composition comprises: i) one or more human milk oligosaccharides (HMOs), wherein the human milk oligosaccharides include at least 2'-fucosyllactose; ii) β-glucan from one or more sources, the β-glucan comprising yeast β-glucan; Wherein, in the composition, the ratio of the human milk oligosaccharide to the β-glucan is 35 to 47:1, and the composition optionally includes other nutritional ingredients. The improvement of immunity is caused by the synergistic effect of component i) and component ii); In the composition, the content of the human milk oligosaccharide is 0.1% by mass or more, and the content of the β-glucan is 0.01% by mass or more.

2. The use according to claim 1, characterized in that: The 2'-fucosyllactose is obtained from fermentation medium or separated pure 2'-fucosyllactose; the β-glucan is selected from one or more β-glucans derived from plants or fungi.

3. The use according to claim 1 or 2, characterized in that: The composition is liquid, semi-solid or solid.

4. The use according to claim 1 or 2, characterized in that: The food includes pasta, beverages, reconstituted food, baked cakes, sauces or functional nutritional supplement foods.

5. The use according to claim 1 or 2, characterized in that: The food is children's or baby food.

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