Formula food of protein composition with anti-inflammatory effect and preparation method of formula food

Abstract

The invention provides a formula food of a protein composition with an anti-inflammatory effect. The protein composition with the anti-inflammatory effect comprises alpha-whey protein and beta-casein, the mass ratio of the alpha-whey protein to the beta-casein is 1: (0.17-5). Compared with the prior art, the anti-inflammatory effect of alpha-whey protein and beta-casein in a ratio of 1: (0.17-5) is superior to that of monomers of alpha-whey protein and beta-casein, and the expression of macrophage proinflammatory cytokines is reduced, so that the inflammatory reaction is reduced; the inflammatory reaction is reduced by one or more of reducing the gene expression of tumor necrosis factor-alpha, reducing the gene expression of interleukin-1alpha and reducing the activation of a transcription activator, and the application prospect is wide when the compound is applied to the infant formula food.

Description

Technical Field

[0001] The invention belongs to the technical field of food, and in particular relates to a formula food of a protein composition with anti-inflammatory effect and a preparation method thereof. Background Art

[0002] Breast milk is the gold standard for infant nutrition within 6 months. The proteins in breast milk can be roughly divided into three categories, namely casein, whey protein and mucin, among which whey protein and casein are the main ones, and mucin is mainly found in the milk fat globule membrane.

[0003] α-lactalbumin (α-LA) accounts for about 36% of the total whey protein in human milk and 22% of the total protein content, and its content gradually decreases during lactation. α-lactalbumin is an important source of bioactive peptides and essential amino acids, including tryptophan, lysine, branched-chain amino acids, and sulfur-containing amino acids (such as cysteine). In addition to providing the amino acids necessary for protein synthesis in the body, tryptophan is also a precursor of the neurotransmitter serotonin, and cysteine ​​is one of the components of glutathione and an important component of the antioxidant system. Therefore, α-lactalbumin plays an important nutritional role for newborns. Compared with other whey proteins, α-lactalbumin is relatively heat-stable when bound to calcium, and can be glycosylated with mannose, galactose, fucose, glucose, and lactose. Lactosylated α-lactalbumin can prevent infection by inhibiting the binding of pathogens to the luminal surface of intestinal epithelial cells.

[0004] β-Casein (β-CS) accounts for a large proportion of human milk casein. β-Casein is synthesized by mammary gland epithelial cells and exists in the milk of many mammals. It accounts for about 50% to 85% of the total casein content. The β-casein content shows a dynamic change pattern of first rising and then falling during lactation. After digestion and hydrolysis, β-casein produces immunostimulatory peptides and antimicrobial peptides, which stimulate immune cells, antagonize the colonization and growth of pathogenic bacteria, and participate in the intestinal immune response of infants.

[0005] The whey protein / casein ratio in human milk changes dynamically, from 80:20 in colostrum to 60:40 in transitional milk and early mature milk, and 50:50 in fully mature milk. Studies have found that casein and whey protein have different effects on milk protein structure at different ratios, such as surface hydrophobicity, free thiol content, particle size, free milk protein content, microstructure, etc. Both α-lactalbumin and β-casein have been added to infant formula as nutrients. Therefore, the combination of the two may affect the interaction of proteins and promote each other's efficacy, that is, have a synergistic effect. However, the relevant efficacy of α-lactalbumin and β-casein at different compound ratios has not yet been retrieved. Summary of the invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a formula food of a protein composition with anti-inflammatory effect and a preparation method thereof.

[0007] The present invention has been found in research that α-lactalbumin and β-casein have anti-inflammatory effects when mixed in different proportions, can reduce the expression of pro-inflammatory cytokines in macrophages to reduce inflammatory responses, reduce the gene expression of tumor necrosis factor-α, reduce the gene expression of interleukin-1α and reduce the activation of transcription activators to reduce inflammatory responses, thereby providing the following invention: a formula food of a protein composition with anti-inflammatory effects, the protein composition with anti-inflammatory effects comprising α-lactalbumin and β-casein; the mass ratio of the α-lactalbumin to the β-casein is 1:(0.17-5).

[0008] Preferably, the mass ratio of the α-lactalbumin to the β-casein is 1:(0.4-3); preferably, the mass ratio of the α-lactalbumin to the β-casein is 1:(0.65-1.35).

[0009] Preferably, the formula food comprises protein, fat and carbohydrates; the total protein content of the formula food converted into dry matter is 8 to 25 g / 100 g; the mass of whey protein is 20% to 80% of the total protein mass; the fat content of the formula food converted into dry matter is 15 to 30 g / 100 g; and the carbohydrate content of the formula food converted into dry matter is 40 to 70 g / 100 g.

[0010] Preferably, the formula food further comprises linoleic acid and α-linolenic acid; the content of linoleic acid in the formula food converted into dry matter is 2700-4500 mg / 100 g; the content of α-linolenic acid in the formula food converted into dry matter is 270-450 mg / 100 g.

[0011] Preferably, the formula food is a formula food for infants aged 0 to 6 months, and the total protein content thereof is 10 to 15 g / 100 g in terms of dry matter, the mass of whey protein is 60% to 80% of the total protein mass, the fat content is 23 to 30 g / 100 g, the linoleic acid content is 2700 to 4500 mg / 100 g, the α-linolenic acid content is 270 to 450 mg / 100 g, and the carbohydrate content is 50 to 60 g / 100 g;

[0012] The formula food is a formula food for older infants aged 6 to 12 months, and the total protein content of the formula food is 10 to 20 g / 100 g, the mass of whey protein is 40% to 70% of the total protein, the fat content is 20 to 30 g / 100 g, the linoleic acid content is 2700 to 4500 mg / 100 g, the α-linolenic acid content is 270 to 450 mg / 100 g, and the carbohydrate content is 50 to 60 g / 100 g.

[0013] The formula food is a formula food for infants aged 12 to 36 months. In terms of dry matter, the total protein content is 10 to 20 g / 100 g, the fat content is 20 to 30 g / 100 g, the linoleic acid content is 2700 to 4500 mg / 100 g, the α-linolenic acid content is 270 to 450 mg / 100 g, and the carbohydrate content is 50 to 60 g / 100 g.

[0014] Preferably, it also includes one or more of nutrients, dietary fiber, probiotics, inositol, taurine, L-carnitine, docosahexaenoic acid, eicosatetraenoic acid, lutein, nucleotides, lactoferrin, OPO, casein calcium peptide and casein phosphopeptide.

[0015] Preferably, the formula food is formula milk powder; the total protein in the formula food comes from one or more of raw milk, whole milk powder, skimmed milk powder, whey protein powder, desalted whey powder and β-casein; the fat in the formula food comes from one or more of corn oil, soybean oil, sunflower oil, 1,3-dioleyl-2-palmityl triglyceride, coconut oil, low-erucic acid rapeseed oil and linseed oil; the carbohydrates in the formula food come from lactose and / or protein raw materials containing lactose.

[0016] Preferably, the formula food reduces the inflammatory response by reducing the expression of pro-inflammatory cytokines in macrophages.

[0017] Preferably, the formula food reduces the inflammatory response by reducing one or more of the gene expression of tumor necrosis factor-α, the gene expression of interleukin-1α, and the activation of transcription activators.

[0018] The present invention also provides a method for preparing a formula food of the above protein composition with anti-inflammatory efficacy, comprising the following steps:

[0019] The protein composition with anti-inflammatory effect is mixed with the raw materials of the formula food to obtain the formula food of the protein composition with anti-inflammatory effect.

[0020] The present invention provides a formula food of a protein composition with anti-inflammatory efficacy, wherein the protein composition with anti-inflammatory efficacy comprises α-lactalbumin and β-casein; the mass ratio of the α-lactalbumin to the β-casein is 1:(0.17-5). Compared with the prior art, the present invention finds that α-lactalbumin and β-casein have an anti-inflammatory effect superior to the two monomers in the ratio range of 1:(0.17-5), reduces the expression of pro-inflammatory cytokines in macrophages, reduces the inflammatory response, reduces the gene expression of tumor necrosis factor-α, reduces the gene expression of interleukin-1α and reduces the activation of transcription activators to reduce the inflammatory response, and has a broad application prospect in the application of the formula food for children. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a graph showing the results of the formula foods obtained in Examples 1 to 7 of the present invention reducing the gene expression of the pro-inflammatory factor TNF-α;

[0022] Figure 2 This is a graph showing the results of the formula foods obtained in Examples 1 to 7 of the present invention reducing the gene expression of the pro-inflammatory factor interleukin-1α (IL-1α);

[0023] Figure 3 This is a graph showing the expression of proteins related to the STAT inflammatory pathway by the formula foods obtained in Examples 1 to 7 of the present invention;

[0024] Figure 4 The results of the formula foods obtained in Examples 1 to 7 of the present invention on the expression of proteins related to the NF-kB inflammatory pathway;

[0025] Figure 5 The results of the effects of the formula foods obtained in Examples 1 to 7 on the survival rate of macrophages are shown in the figure. DETAILED DESCRIPTION

[0026] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] The invention provides a formula food of a protein composition with anti-inflammatory effect. The protein composition with anti-inflammatory effect comprises alpha-lactalbumin and beta-casein; the mass ratio of the alpha-lactalbumin to the beta-casein is 1:(0.17-5).

[0028] The present invention finds that the ratio of alpha-lactalbumin to beta-casein in the range of 1:5 to 1:0.17 has an anti-inflammatory effect superior to that of the two monomers.

[0029] In the present invention, the α-lactalbumin and β-casein are preferably derived from cow's milk.

[0030] Preferably, the mass ratio of α-lactalbumin to β-casein is 1:(5-0.17), but does not include the endpoint ratio of 1:0.17.

[0031] Preferably, the mass ratio of α-lactalbumin to β-casein is 1:(0.4-3).

[0032] Preferably, the mass ratio of α-lactalbumin to β-casein does not include an endpoint ratio of 1:3.

[0033] More preferably, the mass ratio of α-lactalbumin to β-casein is 1:(0.65-1.35).

[0034] More preferably, the mass ratio of α-lactalbumin to β-casein is 1:(0.65-0.4).

[0035] The protein composition with anti-inflammatory efficacy provided by the present invention reduces inflammatory response by reducing the expression of pro-inflammatory cytokines in macrophages.

[0036] In the formula food of the protein composition with anti-inflammatory efficacy provided by the present invention, the content of the protein composition with anti-inflammatory efficacy in the formula food is preferably 0.1-5 mg / mL, more preferably 0.1-3 mg / mL, further preferably 0.3-1 mg / mL, and most preferably 0.14 mg / mL.

[0037] The formula food provided by the invention contains protein, fat and carbohydrate.

[0038] In the present invention, the total protein content of the formula food converted into dry matter is preferably 8-25g / 100g, more preferably 10-25g / 100g, and more preferably 10-20g / 100g; in some embodiments provided by the present invention, the total protein content of the formula food converted into dry matter is specifically 11.8g / 100g or 13.8g / 100g; the mass of whey protein is preferably 20%-80% of the total protein mass, more preferably 25%-80%, more preferably 30%-80%, and most preferably 40%-80%; the mass of α-lactalbumin is preferably 5%-20% of the total protein mass, more preferably 6%-19%; in some embodiments provided by the present invention, the mass of the α-lactalbumin is specifically 6%, 7.68%, 9.15%, 16% or 18.47% of the total protein mass.

[0039] The fat content of the formula food in terms of dry matter is preferably 15-30 g / 100 g, more preferably 18-30 g / 100 g, and even more preferably 20-30 g / 100 g; in some embodiments provided by the present invention, the fat content of the formula food in terms of dry matter is specifically 22.9 g / 100 g, 22.0 g / 100 g or 23.0 g / 100 g.

[0040] The carbohydrate content of the formula food in terms of dry matter is preferably 40-70 g / 100 g, more preferably 45-65 g / 100 g, and even more preferably 50-60 g / 100 g; in some embodiments provided by the present invention, the carbohydrate content of the formula food in terms of dry matter is specifically 57.9 g / 100 g, 55.2 g / 100 g or 52.5 g / 100 g.

[0041] The formula food provided by the present invention preferably further comprises linoleic acid and α-linolenic acid.

[0042] According to the present invention, the content of linoleic acid in the formula food converted into dry matter is preferably 2700-4500 mg / 100 g; the content of α-linolenic acid in the formula food converted into dry matter is preferably 270-450 mg / 100 g.

[0043] The formula food provided by the present invention is preferably an infant formula food and / or a children's formula food; further preferably, the infant formula food can be a formula food for infants aged 0 to 6 months, a formula food for older infants aged 6 to 12 months, or a formula food for children aged 12 to 36 months.

[0044] In the present invention, when the formula food is a formula food for infants aged 0 to 6 months, the total protein content in the dry matter is specifically 10 to 15 g / 100 g; the mass of whey protein is specifically 60% to 80% of the total protein mass; the fat content is specifically 23 to 30 g / 100 g; the linoleic acid content is specifically 2700 to 4500 mg / 100 g; the α-linolenic acid content is specifically 270 to 450 mg / 100 g; and the carbohydrate content is specifically 50 to 60 g / 100 g.

[0045] When the formula food is a formula food for older infants aged 6 to 12 months, the total protein content in the dry matter is specifically 10 to 20 g / 100 g; the mass of whey protein is specifically 40% to 70% of the total protein; the fat content is specifically 20 to 30 g / 100 g; the linoleic acid content is specifically 2700 to 4500 mg / 100 g; the α-linolenic acid content is specifically 270 to 450 mg / 100 g; and the carbohydrate content is specifically 50 to 60 g / 100 g;

[0046] When the formula food is a formula food for infants aged 12 to 36 months, the total protein content of the formula food converted into dry matter is specifically 10 to 20 g / 100 g; the fat content is specifically 20 to 30 g / 100 g; the linoleic acid content is specifically 2700 to 4500 mg / 100 g; the α-linolenic acid content is specifically 270 to 450 mg / 100 g; and the carbohydrate content is specifically 50 to 60 g / 100 g.

[0047] When the formula food is a children's formula food, the total protein content of the formula food converted into dry matter is preferably higher than 16.5 g / 100 g.

[0048] The total protein in the formula food provided by the present invention is preferably derived from one or more of raw milk, whole milk powder, skim milk powder, whey protein powder, desalted whey powder and β-casein; the raw milk is preferably raw cow's milk and / or raw goat's milk; the whole milk powder is preferably whole cow's milk powder and / or whole goat's milk powder; the skim milk powder is preferably skim milk powder and / or skim goat's milk powder; the whey protein powder is preferably one or more of bovine whey protein powder, goat whey protein powder and α-lactalbumin powder; the desalted whey powder is preferably bovine desalted whey powder and / or goat desalted whey powder; specifically, based on 1000 parts by weight of the formula food, the raw materials for the total protein include: 0 to 4000 parts by weight of raw milk, 0 to 4000 parts by weight of whole milk powder, 0 to 4000 parts by weight of the ... ~260 parts by weight, skim milk powder 0~400 parts by weight, whey protein powder 0~120 parts by weight, desalted whey powder 0~600 parts by weight and β-casein 0~40 parts by weight, and not all of them are 0 at the same time; further specifically, based on 1000 parts by weight of the formula food, the raw materials of the total protein include: raw milk 0~3500 parts by weight, whole milk powder 0~260 parts by weight, skim milk powder 0~400 parts by weight, whey protein powder 0~120 parts by weight, desalted whey powder 0~600 parts by weight and β-casein 0~40 parts by weight, and not all of them are 0 at the same time; further specifically, based on 1000 parts by weight of the formula food, the raw materials of the total protein include: raw milk 1000~3000 parts by weight , 0-260 parts by weight of whole milk powder, 0-200 parts by weight of skim milk powder, 2-80 parts by weight of whey protein powder, 100-500 parts by weight of desalted whey powder and 1-30 parts by weight of β-casein; more specifically, based on 1000 parts by weight of the formula food, the raw materials of total protein include: 1000-2500 parts by weight of raw milk, 0-260 parts by weight of whole milk powder, 0-100 parts by weight of skim milk powder, 2-35 parts by weight of whey protein powder, 150-425 parts by weight of desalted whey powder and 1-25 parts by weight of β-casein; in some embodiments provided by the present invention, based on 1000 parts by weight of the formula food, the raw materials of total protein include 2500 parts by weight of raw milk, 0-260 parts by weight of whole milk powder, 0-100 parts by weight of skim milk powder, 2-35 parts by weight of whey protein powder, 150-425 parts by weight of desalted whey powder and 1-25 parts by weight of β-casein 75 parts by weight of milk powder, 2 parts by weight of whey protein powder, 150 parts by weight of desalted whey powder and 1 part by weight of beta-casein; or 1600 parts by weight of raw milk, 8 parts by weight of whey protein powder, 250 parts by weight of desalted whey powder and 1 part by weight of beta-casein; or 1500 parts by weight of raw milk, 100 parts by weight of skimmed milk powder, 5 parts by weight of whey protein powder, 300 parts by weight of desalted whey powder and 1 part by weight of beta-casein; or 1000 parts by weight of raw milk, 27 parts by weight of whey protein powder, 400 parts by weight of desalted whey powder and 25 parts by weight of beta-casein; or 1100 parts by weight of raw milk, 35 parts by weight of whey protein powder, 425 parts by weight of desalted whey powder and 5 parts by weight of beta-casein.

[0049] The fat in the formula food provided by the present invention is preferably derived from one or more of corn oil, soybean oil, sunflower oil, 1,3-dioleyl-2-palmityl triglyceride, coconut oil, rapeseed oil and linseed oil; specifically, based on 1000 parts by weight of the formula food, the raw materials of the fat include: 0-100 parts by weight of corn oil, 0-65 parts by weight of soybean oil, 0-100 parts by weight of sunflower oil, 0-240 parts by weight of 1,3-dioleyl-2-palmityl triglyceride, 0-30 parts by weight of coconut oil, 0-85 parts by weight of rapeseed oil and 0-20 parts by weight of linseed oil, and not all of them are 0 at the same time; in the embodiments provided by the present invention, the mixed vegetable oil is specifically used as an example for explanation; in the embodiments provided by the present invention, based on 1000 parts by weight of the formula food, the raw materials of the fat include 100-200 parts by weight of the mixed vegetable oil, and further specifically include 130-185 parts by weight of the mixed vegetable oil.

[0050] The carbohydrates in the formula food provided by the present invention are preferably derived from lactose and / or protein raw materials containing lactose; the types of protein raw materials and total protein raw materials are the same, which will not be repeated here; specifically, based on 1000 parts by weight of the formula food, the carbohydrate raw materials include 0 to 500 parts by weight of lactose, which can be added according to the addition amount of the protein raw materials; further specifically, based on 1000 parts by weight of the formula food, the carbohydrate raw materials include 100 to 500 parts by weight of lactose; further specifically, based on 1000 parts by weight of the formula food, the carbohydrate raw materials include 145 to 300 parts by weight of lactose; in some embodiments provided by the present invention, based on 1000 parts by weight of the formula food, the carbohydrate raw materials specifically include 265 parts by weight, 175 parts by weight, 100 parts by weight, 175 parts by weight or 145 parts by weight of lactose.

[0051] According to the present invention, in addition to the above-mentioned components, the formula food preferably also includes other conventional components, and specifically in the present invention, it is preferred to include one or more of nutrients, dietary fiber, probiotics, inositol, taurine, L-carnitine, docosahexaenoic acid, eicosatetraenoic acid, lutein, nucleotides, lactoferrin, OPO, casein calcium peptides and casein phosphopeptides; other conventional components should comply with national standards for formula foods and relevant regulations. The nutrients can be nutrients well known to those skilled in the art, without any special restrictions, and preferably include vitamins and / or minerals in the present invention; the vitamins can be vitamins well known to those skilled in the art, without any special restrictions, and preferably include vitamin A, vitamin D, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B 12, niacin, folic acid, pantothenic acid, vitamin C, one or more of choline and biotin; the minerals may be minerals well known to those skilled in the art without particular limitation, and in the present invention, preferably are one or more of sodium, potassium, calcium, phosphorus, copper, iron, zinc, manganese, iodine, selenium, magnesium, potassium and their derivatives; in the present invention, the nutrients are preferably added via a compound nutrient package, such as a compound vitamin and / or a compound mineral; based on 1000 parts by weight of the formula food, the formula food preferably includes 0 to 20 parts by weight of nutrients, more preferably 10 to 20 parts by weight of nutrients, and even more preferably 15 to 20 parts by weight of nutrients; specifically, the source, content and usage of the vitamins and minerals should comply with the national standards for infant formula foods and relevant laws and regulations. The dietary fiber can be a dietary fiber well known to those skilled in the art, without any special restrictions, and preferably one or more of oligofructose, galacto-oligosaccharide and polyfructose in the present invention; based on 1000 parts by weight of the formula food, the formula food preferably includes 0 to 100 parts by weight of dietary fiber, more preferably 10 to 80 parts by weight of dietary fiber, more preferably 40 to 60 parts by weight of dietary fiber, and most preferably 50 to 55 parts by weight of dietary fiber. The probiotics can be a probiotic well known to those skilled in the art, without any special restrictions, and preferably Bifidobacterium lactis in the present invention; based on 1000 parts by weight of the formula food, the formula food preferably includes 0 to 3 parts by weight of probiotics, more preferably 0.01 to 2 parts by weight of probiotics, more preferably 0.1 to 1 parts by weight of probiotics, and most preferably 0.3 to 0.4 parts by weight of probiotics.

[0052] In the present invention, specifically, the formula food is formula milk powder; more specifically, the formula food is infant formula milk powder and / or children's formula milk powder.

[0053] The formula food provided by the present invention specifically reduces the inflammatory response by reducing the expression of pro-inflammatory cytokines in macrophages; more specifically, the formula food reduces the inflammatory response by reducing one or more of the gene expression of tumor necrosis factor-α, the gene expression of interleukin-1α, and the activation of transcription activators.

[0054] Tumor necrosis factor-α (TNF-α) is mainly secreted by macrophages and is a cytokine involved in systemic inflammation. The pro-inflammatory effect of TNF-α is mainly due to its ability to activate NF-kappaB (NF-kB). Almost all cell types activate NF-kB when exposed to TNF-α, leading to the expression of inflammatory genes. More than 400 genes regulated by NF-kB activation have been identified, including inflammatory cytokines, chemokines, etc. IL-1a is also an important mediator of inflammatory response and participates in cell proliferation, differentiation and apoptosis. In addition, the Janus kinase / signal transduction and activator of transcription (JAK / STAT) signaling pathway is a common pathway for many cytokine signaling, which is involved in cell proliferation, differentiation, apoptosis and inflammation to varying degrees, thereby promoting the occurrence and development of various diseases, including inflammatory bowel disease, and the activation of STAT3 is an important factor in the persistence of colonic inflammation.

[0055] The ratio of α-lactalbumin to β-casein in the formula food of the protein composition with anti-inflammatory efficacy provided by the present invention is obtained by screening the expression of inflammatory cytokines of macrophages induced by lipopolysaccharide at different ratios. The protein composition within this ratio range is superior to the anti-inflammatory effect of α-lactalbumin and β-casein monomers, and the effect is specifically reflected in reducing the gene expression of tumor necrosis factor-α and interleukin-1α, and is further confirmed in reducing NF-kappaB and STAT3 in the inflammatory pathway.

[0056] The present invention finds that alpha-lactalbumin and beta-casein have an anti-inflammatory effect superior to that of the two monomers in a ratio range of 1: (0.17-5), reduce the expression of pro-inflammatory cytokines in macrophages to reduce inflammatory response, reduce the gene expression of tumor necrosis factor-alpha, reduce the gene expression of interleukin-1alpha and reduce the activation of transcription activators to reduce inflammatory response, and the application of the present invention in pediatric formula foods has broad application prospects.

[0057] The present invention also provides a method for preparing a formula food of the above-mentioned protein composition with anti-inflammatory efficacy, comprising the following steps: mixing the protein composition with anti-inflammatory efficacy with formula food raw materials to obtain the formula food of the protein composition with anti-inflammatory efficacy.

[0058] The present invention has no particular limitation on the sources of all raw materials, and any raw materials available on the market can be used.

[0059] In the present invention, the mixing method is a method well known to those skilled in the art, and a wet method, a dry method or a wet-dry method can be used without any special limitation; specifically, in the present invention, the process flow mainly includes batching, homogenization, concentrated sterilization, spray drying, dry mixing, etc.; the protein composition with anti-inflammatory effect can be mixed together with the raw materials of the formula food when the ingredients are prepared, and can also be added during the mixing process without any special limitation.

[0060] In the embodiments provided by the present invention, the preparation is carried out specifically according to the following steps:

[0061] Mixing the other raw materials in the formula food except the spray carrier (phospholipids and anhydrous milk) and the post-mix (DHA, ARA, lactoferrin, bifidobacterium) to obtain a mixed liquid;

[0062] Homogenization: homogenize the mixed liquid;

[0063] Concentration and sterilization: The homogenized liquid is concentrated and sterilized. The concentration and sterilization conditions are: double-effect concentration, sterilization temperature ≥ 83°C, sterilization time 15 to 30 seconds, and the discharge concentration is controlled to be 48% to 52% dry matter;

[0064] Spray drying: The concentrated milk liquid is preheated to 60-70℃ in a preheater. After preheating, the material is filtered through a filter with a pore size of 0.8-1.2mm and then sent to a drying tower for spray drying. The spray drying conditions are controlled as follows: inlet air temperature 165-180℃, exhaust air temperature 75-90℃, high pressure pump pressure 160-210bar, tower negative pressure -4--2mbar;

[0065] Fluidized bed drying and cooling: The powder coming out of the drying tower is dried in the primary fluidized bed for the second time, and then cooled to 25-30°C in the secondary fluidized bed. At the same time, under the action of compressed air, a mixture of phospholipids (lecithin and / or soybean lecithin) and carrier (anhydrous butter) heated to 60-65°C is evenly dispersed on the powder surface to obtain powder particles.

[0066] Post-mixing: dry-mixing DHA, ARA, lactoferrin, bifidobacteria and the powder particles dried and cooled in a fluidized bed, and packaging to obtain a formula food.

[0067] In order to further illustrate the present invention, a formula food of a protein composition with anti-inflammatory efficacy and a preparation method thereof provided by the present invention are described in detail below in conjunction with examples.

[0068] The reagents used in the following examples are all commercially available.

[0069] Materials and Reagents

[0070] RAW264.7 macrophages, lipopolysaccharide (LPS, S11060), α-lactalbumin (α-LA), β-casein (β-CS), PBS buffer, porcine trypsin, porcine pepsin, DMEM culture medium (Gibco, 11995065); desalted whey powder was from Royal FrieslandCampina; mixed vegetable oil was from Yihai Kerry Food Industry (Tianjin) Co., Ltd.; α-lactalbumin powder was from Arla Foods Ingredients; casein powder was from Kerry Group; Bifidobacterium lactis was from The compound vitamins come from Beijing Jinkangpu Food Technology Co., Ltd.; the compound minerals come from Beijing Jinkangpu Food Technology Co., Ltd.

[0071] Example 1

[0072] The present embodiment provides a powdered formula food for older infants aged 6 to 12 months, wherein the total protein content of the powdered formula food is 11.80 g / 100 g powder, the fat content is 22.9 g / 100 g powder, the carbohydrate content is 57.9 g / 100 g powder, and the probiotic content is 1.5×10 9 CFU / 100g powder; α-lactalbumin and β-casein = 1:4.

[0073] This embodiment is realized by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight): 2500 parts of raw cow's milk, 150 parts of desalted whey powder, 75 parts of skimmed milk powder, 265 parts of lactose, 130 parts of mixed vegetable oil, 2 parts of α-lactalbumin powder, 1 part of casein powder, 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0074] The formula food for older infants of this embodiment is prepared by a dry-wet production process. The formula milk powder for older infants of this embodiment is prepared by a dry-wet production process. The main production processes and parameters include:

[0075] Mixing the other raw materials in the formula milk powder except the spray carrier (phospholipids and anhydrous milk) and the post-mix (DHA, ARA, lactoferrin, bifidobacterium) to obtain a mixed liquid;

[0076] Homogenization: homogenize the mixed liquid;

[0077] Concentration and sterilization: The homogenized liquid is concentrated and sterilized. The concentration and sterilization conditions are: double-effect concentration, sterilization temperature ≥ 83°C, sterilization time 15 to 30 seconds, and the discharge concentration is controlled to be 48% to 52% dry matter;

[0078] Spray drying: The concentrated milk liquid is preheated to 60-70℃ in a preheater. After preheating, the material is filtered through a filter with a pore size of 0.8-1.2mm and then sent to a drying tower for spray drying. The spray drying conditions are controlled as follows: inlet air temperature 165-180℃, exhaust air temperature 75-90℃, high pressure pump pressure 160-210bar, tower negative pressure -4--2mbar;

[0079] Fluidized bed drying and cooling: The powder coming out of the drying tower is dried in the primary fluidized bed for the second time, and then cooled to 25-30°C in the secondary fluidized bed. At the same time, under the action of compressed air, a mixture of phospholipids (lecithin and / or soybean lecithin) and carrier (anhydrous butter) heated to 60-65°C is evenly dispersed on the powder surface to obtain powder particles.

[0080] Post-mixing: dry-mix DHA, ARA, lactoferrin, bifidobacteria and the powder particles dried and cooled in the fluidized bed, and package to obtain the finished milk powder.

[0081] Upon testing, it was found that various indicators of the finished product met the standard requirements for formula food for older infants.

[0082] Example 2

[0083] The present embodiment provides a formula milk powder for older infants aged 6 to 12 months. The total protein content of the powdered formula food is 13.8 g / 100 g powder, the fat content is 22.0 g / 100 g powder, the carbohydrate content is 55.2 g / 100 g powder, and the probiotic content is 1×10 9 CFU / 100g powder; α-lactalbumin and β-casein = 1:3.0.

[0084] This embodiment is realized by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight): 1600 parts of raw cow's milk, 175 parts of lactose, 180 parts of mixed vegetable oil, 250 parts of desalted whey powder, 8 parts of α-lactalbumin powder, 1 part of casein powder, 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0085] The older infant formula milk powder of this embodiment is prepared by dry and wet production process, and the main process flow and parameters are the same as those of embodiment 1. After testing, the various indicators of the finished product meet the standard requirements of infant formula food.

[0086] Example 3

[0087] The present embodiment provides a formula milk powder for older infants aged 6 to 12 months. The total protein content of the powdered formula food is 11.80 g / 100 g powder, the fat content is 22.9 g / 100 g powder, the carbohydrate content is 52.5 g / 100 g powder, and the probiotic content is 1×10 9CFU / 100g powder; α-lactalbumin and β-casein = 1:2.0.

[0088] This embodiment is realized by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight): 1500 parts of raw cow's milk, 300 parts of desalted whey powder, 100 parts of skimmed milk powder, 200 parts of lactose, 185 parts of mixed vegetable oil (including OPO structured fat), 5 parts of α-lactalbumin powder, 1 part of casein powder, 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0089] The older infant formula milk powder of this embodiment is prepared by dry and wet production process, and the main process flow and parameters are the same as those of embodiment 1. After testing, the various indicators of the finished product meet the requirements of the standard of formula milk powder food.

[0090] Example 4

[0091] The present embodiment provides a formula milk powder for older infants aged 6 to 12 months. The total protein content of the powdered formula food is 11.80 g / 100 g powder, the fat content is 23.0 g / 100 g powder, the carbohydrate content is 52.5 g / 100 g powder, and the probiotic content is 1×10 9 CFU / 100g powder. α-lactalbumin and β-casein = 1:1.

[0092] This embodiment is realized by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight): 1000 parts of raw cow's milk, 400 parts of desalted whey powder, 175 parts of lactose, 185 parts of mixed vegetable oil (including OPO structured fat), 27 parts of α-lactalbumin powder, 25 parts of casein powder, 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0093] The infant formula food of this embodiment is prepared by dry and wet production process, and the main process flow and parameters are the same as those of Example 1. After testing, the various indicators of the finished product meet the standard requirements of infant formula food.

[0094] Example 5

[0095] The present embodiment provides a formula milk powder for older infants aged 6 to 12 months. The total protein content of the powdered formula food is 11.8 g / 100 g powder, the fat content is 23.0 g / 100 g powder, the carbohydrate content is 52.5 g / 100 g powder, and the probiotic content is 1×10 9 CFU / 100g powder. α-lactalbumin and β-casein = 1:0.5.

[0096] This embodiment is realized by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight): 1100 parts of raw cow's milk, 425 parts of desalted whey powder, 145 parts of lactose, 185 parts of mixed vegetable oil (including OPO structured fat), 35 parts of α-lactalbumin powder, 5 parts of casein powder, 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0097] The infant formula food of this embodiment is prepared by dry and wet production process, and the main process flow and parameters are the same as those of Example 1. After testing, the various indicators of the finished product meet the standard requirements of infant formula food.

[0098] Example 6

[0099] The present embodiment provides a powdered formula milk powder, wherein the total protein content of the powdered formula food is 11.8 g / 100 g powder, the fat content is 23.0 g / 100 g powder, the carbohydrate content is 52.5 g / 100 g powder, and the probiotic content is 1×10 9 CFU / 100g powder.

[0100] The present embodiment is realized by compounding the following raw materials in parts by weight, and the raw material composition includes (preparing 1000 parts by weight): 500 parts of desalted whey powder, 115 parts of α-lactalbumin powder, 140 parts of lactose, 220 parts of mixed vegetable oil (including OPO structured fat), 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0101] The infant formula food of this embodiment is prepared by dry and wet production process, and the main process flow and parameters are the same as those of Example 1. After testing, the various indicators of the finished product meet the standard requirements of infant formula food.

[0102] Example 7

[0103] The present embodiment provides a powdered formula milk powder, wherein the total protein content of the powdered formula food is 11.8 g / 100 g powder, the fat content is 23.0 g / 100 g powder, the carbohydrate content is 52.5 g / 100 g powder, and the probiotic content is 1×10 9 CFU / 100g powder.

[0104] This embodiment is achieved by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight): 138 parts of casein powder, 580 parts of lactose, 220 parts of mixed vegetable oil (including OPO structured fat), 30 parts of galacto-oligosaccharide, 25 parts of fructo-oligosaccharide, 0.33 parts of Bifidobacterium lactis, 3.5 parts of compound vitamins, and 13 parts of compound minerals.

[0105] The infant formula food of this embodiment is prepared by dry and wet production process, and the main process flow and parameters are the same as those of Example 1. After testing, the various indicators of the finished product meet the standard requirements of infant formula food.

[0106] Efficacy test of formula food containing α-lactalbumin and β-casein

[0107] Table 1 Design of α-lactalbumin and β-casein combination in formula food

[0108]

[0109] In vitro digestion of active proteins from bovine milk

[0110] The formula milk powder obtained in Examples 1 to 7 was dissolved in 1×PBS buffer to prepare samples with a concentration of 1 mg / mL, and 2 mL of the sample was adjusted to pH 4 with 1M HCL solution. Pepsin was added to the sample solution to make the activity of pepsin reach 200U / mL, and the sample was immediately placed on a shaker at 37°C and digested in the dark at 200rpm for 60min. Subsequently, the pH of the sample was quickly adjusted to 7 with 1M sodium bicarbonate solution, and trypsin was added to the sample solution to make the activity of trypsin reach 8.33U / mL, and digested in the dark on a shaker (200rpm) at 37°C for 60min. After digestion, the sample was immediately placed in an 85°C water bath for 3min to inactivate trypsin, cooled to room temperature, adjusted to pH 7, concentrated and freeze-dried, and stored for later use.

[0111] Detection indicators

[0112] 1) Treatment of macrophages with bovine milk-derived active protein

[0113] 2×10 5 RAW264.7 cells were inoculated into 6-well culture plates at 100 μg / well, with 3 replicates for each treatment group. The seeded cells were cultured at 37°C and 5% CO2, and after 24 hours, the serum-free and antibiotic-free DMEM medium containing bovine milk active protein (formulated milk powder of Examples 1 to 7 after in vitro digestion, concentration 0.4 mg / mL) was replaced for pretreatment for 6 hours. LPS (lipopolysaccharide) was then added to the culture medium to a concentration of 1 μg / ml, and cells were collected for RNA extraction after 2 hours of treatment, and cells were collected for protein extraction after 6 hours of treatment.

[0114] 2) Expression of inflammatory factors

[0115] RAW264.7 cells were inoculated in 12-well cell culture plates, and after 24 hours, the medium was replaced with DMEM medium containing α-LA, β-CS monomers or complexes (formulated milk powder of Examples 1 to 7 after in vitro digestion, concentration 0.4 mg / mL) and without serum and antibiotics. After 6 hours, LPS was added to a concentration of 1 μg / ml. After 2 hours, RNA was collected, and the expression levels of inflammatory factors such as IL-1α, IL-1β, and TNF-α in mRNA were quantitatively analyzed by RT-qPCR.

[0116] 3) Expression of proteins related to inflammatory pathways

[0117] RAW264.7 cells were inoculated in a 12-well cell culture plate, and after 24 hours, the medium was replaced with DMEM medium containing α-LA, β-CS monomers or complexes (formulated milk powder of Examples 1 to 7 after in vitro digestion, concentration 0.4 mg / mL) and without serum and antibiotics. After 6 hours, LPS was added to a concentration of 1 μg / ml. After another 6 hours, protein samples were collected, and the expression of proteins related to TLR4 and NF-κB pathways was analyzed by Western blot.

[0118] 4) Data Statistical Analysis

[0119] All the data obtained from the final experiment were processed using SPSS17.0 software, and the results were as follows: Figure 1 to Figure 5 As shown, Figure 1 The results of the formula foods obtained in Examples 1 to 7 reducing the gene expression of the pro-inflammatory factor TNF-α; Figure 2 The results of the formula foods obtained in Examples 1 to 7 reducing the gene expression of the pro-inflammatory factor interleukin-1α (IL-1α); Figure 3 The results of the formula foods obtained in Examples 1 to 7 on the expression of proteins related to the STAT inflammatory pathway; Figure 4 The results of the formula foods obtained in Examples 1 to 7 on the expression of proteins related to the NF-kB inflammatory pathway; Figure 5 The results of the effects of the formula foods obtained in Examples 1 to 7 on the survival rate of macrophages are shown in the figure.

Claims

1. A formula food of a protein composition having anti-inflammatory effect, characterized in that: The protein composition with anti-inflammatory effect comprises alpha-lactalbumin and beta-casein; the mass ratio of the alpha-lactalbumin to the beta-casein is 1:(0.17-5).

2. The formula food according to claim 1, characterized in that The mass ratio of the α-lactalbumin to the β-casein is 1:(0.4-3); preferably, the mass ratio of the α-lactalbumin to the β-casein is 1:(0.65-1.35).

3. The formula food according to claim 1, characterized in that The formula food comprises protein, fat and carbohydrate; the total protein content of the formula food converted into dry matter is 8-25g / 100g; the mass of whey protein is 20%-80% of the total protein mass; the fat content of the formula food converted into dry matter is 15-30g / 100g; and the carbohydrate content of the formula food converted into dry matter is 40-70g / 100g.

4. The formula food according to claim 1, characterized in that The formula food further comprises linoleic acid and α-linolenic acid; the content of linoleic acid in the formula food converted into dry matter is 2700-4500 mg / 100 g; the content of α-linolenic acid in the formula food converted into dry matter is 270-450 mg / 100 g.

5. The formula food according to claim 1, characterized in that The formula food is a formula food for infants aged 0 to 6 months, and in terms of dry matter, the total protein content is 10 to 15 g / 100 g, the mass of whey protein is 60% to 80% of the total protein mass, the fat content is 23 to 30 g / 100 g, the linoleic acid content is 2700 to 4500 mg / 100 g, the α-linolenic acid content is 270 to 450 mg / 100 g, and the carbohydrate content is 50 to 60 g / 100 g; The formula food is a formula food for older infants aged 6 to 12 months, and the total protein content of the formula food is 10 to 20 g / 100 g, the mass of whey protein is 40% to 70% of the total protein, the fat content is 20 to 30 g / 100 g, the linoleic acid content is 2700 to 4500 mg / 100 g, the α-linolenic acid content is 270 to 450 mg / 100 g, and the carbohydrate content is 50 to 60 g / 100 g. The formula food is a formula food for infants aged 12 to 36 months. In terms of dry matter, the total protein content is 10 to 20 g / 100 g, the fat content is 20 to 30 g / 100 g, the linoleic acid content is 2700 to 4500 mg / 100 g, the α-linolenic acid content is 270 to 450 mg / 100 g, and the carbohydrate content is 50 to 60 g / 100 g.

6. The formula food according to claim 1, characterized in that It also includes one or more of nutrients, dietary fiber, probiotics, inositol, taurine, L-carnitine, docosahexaenoic acid, eicosatetraenoic acid, lutein, nucleotides, lactoferrin, OPO, casein calcium peptide and casein phosphopeptide.

7. The formula food according to claim 1, characterized in that The formula food is formula milk powder; the total protein in the formula food comes from one or more of raw milk, whole milk powder, skimmed milk powder, whey protein powder, desalted whey powder and β-casein; the fat in the formula food comes from one or more of corn oil, soybean oil, sunflower oil, 1,3-dioleyl-2-palmityl triglyceride, coconut oil, low-erucic acid rapeseed oil and linseed oil; the carbohydrates in the formula food come from lactose and / or protein raw materials containing lactose.

8. The formula food according to claim 1, characterized in that The formula food reduces the inflammatory response by reducing the expression of pro-inflammatory cytokines in macrophages.

9. The formula food according to claim 1, characterized in that The formula food reduces the inflammatory response by reducing one or more of the gene expression of tumor necrosis factor-α, the gene expression of interleukin-1α and the activation of transcription activator.

10. A method for preparing a formula food of the protein composition with anti-inflammatory effect according to any one of claims 1 to 9, characterized in that: The following steps are involved: The protein composition with anti-inflammatory effect is mixed with the raw materials of the formula food to obtain the formula food of the protein composition with anti-inflammatory effect.

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