Composition for regulating intestinal flora and lipid absorption in early life and application thereof

Abstract

The invention discloses a composition for regulating intestinal flora and lipid absorption in the early stage of life and application. The composition for regulating intestinal flora and lipid absorption in the early stage of life comprises galactooligosaccharide and a milk fat globule membrane. Compared with the prior art, the composition has the advantages that the milk fat globule membrane and the galactooligosaccharide coordinate to regulate the intestinal flora in the early stage of life and promote intestinal barrier development and lipid absorption.

Description

Technical Field

[0001] The present invention relates to the field of infant formula food, and in particular to a composition and application thereof for regulating intestinal flora and lipid absorption in early life. Background Art

[0002] The intestinal flora of infants in the early stages of life (0 to 3 years old) has strong plasticity and is a critical period for the development of the flora. At this stage, under the action of microorganisms and oligosaccharides, the intestinal flora of breastfed infants usually shows a high abundance of aerobic bacteria and facultative anaerobes. After weaning, due to the intake of solid food and more indigestible polysaccharides, the intestinal flora of infants will undergo a phased change. With the improvement of neonatal development, the intestinal flora shows diversity and stability, forming an intestinal flora structure dominated by anaerobic bacteria such as Bacteroidetes, Bifidobacterium and Clostridium in the Firmicutes. Since the intestinal barrier function and nutritional metabolism of infants are easily affected by the intestinal microbiota, healthy intervention of the intestinal flora of infants in early life plays an important role in promoting the integrity of the infant's intestine and the growth and development of the body. Furthermore, the intestinal barrier of infants in the early stages of life (0 to 6 months) is not fully developed, there are large gaps between intestinal epithelial cells, and the intestinal permeability is high, which easily increases the absorption of harmful metabolites and thus causes intestinal inflammatory diseases. Therefore, active intervention in the integrity of the intestinal barrier has an important impact on preventing infant intestinal diseases and promoting healthy development. In addition, infants' fat digestion and absorption abilities are immature in the early stages of life. Gastric acid, lipase, and bile salts can only reach the level of effective fat digestion at 6 to 9 months, and fat absorption ability can only reach adult levels at 2 years old. Therefore, active intervention in infants' fat absorption ability can improve their absorption of bioactive lipids and energy substances in the early stages of life and promote their growth and development.

[0003] Milk fat globule membrane (MFGM) is a biological membrane with a complex three-layer structure that encapsulates milk fat in milk. It is a natural component of milk and is mainly composed of glycerophospholipids (phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylinositol (PI)), sphingomyelin (SM) and various milk fat globule membrane proteins. A large number of clinical and animal studies have shown that MFGM has a variety of positive effects on the immune system, neurodevelopment and cognitive ability, intestinal function and lipid metabolism of infants and young children. Literature has reported that the intestinal flora composition of rat pups fed with milk powder containing MFGM is more similar to that of pups fed with breast milk, and can promote intestinal proliferation and differentiation.

[0004] Galacto-oligosaccharides (GOS) are the third most abundant solid component in breast milk. In infant formula, GOS is usually used to simulate oligosaccharides in breast milk. The direct effects of GOS on infants mainly include two points. First, it can regulate the intestinal flora and promote the growth of beneficial bacteria. This effect is due to the competitive adhesion and proliferation of beneficial bacteria and pathogens, which increases the relative abundance of beneficial bacteria and reduces the relative abundance of pathogens. Many studies have found that infant formula with GOS can increase the abundance of infant bifidobacteria and lactobacilli, and reduce the abundance of pathogens such as Escherichia coli and Salmonella. Second, it produces short-chain fatty acids, thereby reducing the occurrence of colon inflammation. During the development of the infant's intestinal flora, Enterobacteriaceae dominates the distribution of flora in the gastrointestinal tract. By fermenting oligosaccharides, short-chain fatty acids are produced, the pH value in the gastrointestinal tract is lowered, and the growth of harmful pathogens is inhibited.

[0005] Most of the research on MFGM and GOS focuses on exploring their health effects under independent action.

[0006] In addition, breast milk lipids are an important source of energy for infants, accounting for 3-5% of breast milk components and providing 45-55% of the total energy for infants. Breast milk lipids are mainly composed of triglycerides (mainly 1,3-dioleic acid-2-palmitic acid triglyceride (OPO) and 1-oleic acid-2-palmitic acid-3-linoleic acid triglyceride (OPL)), lipids and fatty acids, among which fatty acids are mainly composed of saturated fatty acids (average value: 36.4%), monounsaturated fatty acids (average value: 32.2%) and polyunsaturated fatty acids (average value: 22.4%). The fat composition used in the current commercially available infant formula milk powder is single, which can only meet the requirement that the sn-2 structural lipid or fatty acid composition is similar to that of breast milk, but ignores the composition of OPL and medium-and long-chain fatty acids (MLCT). In infant formula milk powder, raw cow's milk must be used as a raw material to achieve an effect similar to that of breast milk lipid composition, which has high production costs and complicated production processes.

[0007] Therefore, it is necessary to design a suitable composition for regulating the intestinal flora, lipid absorption, and suitable fatty acid composition in early life and apply it to infant formula. Summary of the invention

[0008] The first technical problem to be solved by the present invention is to provide a composition for regulating intestinal flora and lipid absorption in early life in view of the current status of the prior art.

[0009] The second technical problem to be solved by the present invention is to provide the use of the above composition in the preparation of infant formula milk powder.

[0010] The technical solution adopted by the present invention to solve the above first technical problem is: a composition for regulating intestinal flora and lipid absorption in early life, characterized by comprising galacto-oligosaccharide and milk fat globule membrane. The technical solution adopted by the present invention to solve the above second technical problem is: use of the composition in the preparation of infant formula milk powder.

[0011] Preferably, in the infant formula milk powder, the galacto-oligosaccharide accounts for 4.1% by mass, and the whey protein powder with milk fat globule membrane accounts for 1.1% by mass.

[0012] Preferably, in the infant formula milk powder, the galacto-oligosaccharide accounts for 4.1% by mass, and the whey protein powder with milk fat globule membrane accounts for 3.5% by mass.

[0013] Preferably, the infant formula milk powder further comprises a fat composition, which comprises the following components by mass percentage:

[0014] 1,3-dioleyl-2-palmitoyl triglyceride 32%;

[0015] Sunflower oil 26%;

[0016] High oleic sunflower oil 17%, of which oleic acid accounts for 84.6% according to the gas chromatography peak area ratio;

[0017] Breast milk-like fat 15%;

[0018] Coconut oil 7%;

[0019] Linseed oil 3%.

[0020] In this way, the following effects are achieved: first, there are rich types of fatty acids, among which coconut oil is the main source of saturated fatty acids, high oleic sunflower oil is an important source of monounsaturated fatty acids, sunflower oil and linseed oil are the main sources of polyunsaturated fatty acids, breast milk-like oils are the main source of OPL and MLCT, and are also a partial source of sn-2 structural lipids, and OPO is the main source of sn-2 structural lipids; second, infant formula milk powder adopts the above-mentioned oil ingredients and can achieve an effect similar to that of breast milk lipid composition without using raw cow's milk as raw material, which is beneficial to reducing production costs. In addition to satisfying that the sn-2 structural lipid or fatty acid composition is similar to that of breast milk, OPL and MLCT are also considered, so that the OPL and MLCT in the oil composition are closer to the breast milk lipid composition.

[0021] Preferably, the infant formula milk powder further comprises a fat composition, which comprises the following components by mass percentage:

[0022] Saturated fatty acids 33.17 ± 1.04%;

[0023] Monounsaturated fatty acids 43.63 ± 0.05%;

[0024] Polyunsaturated fatty acids 23.20 ± 0.99%;

[0025] Preferably, the infant formula milk powder comprises the following ingredients by mass percentage:

[0026] 3.5% whey protein powder with milk fat globule membrane;

[0027] Whey protein isolate powder 10.7%;

[0028] The oil and fat composition 28.3%;

[0029] Lactose 52.2%;

[0030] The galacto-oligosaccharide 4.8%;

[0031] 1 stage complex macrovitamin iodine premix 0.1%;

[0032] Stage 1 complex macrovitamin premix 0.1%;

[0033] 1st stage composite macro mineral premix 0.25%;

[0034] Stage 1 Complex Trace Mineral Premix 0.1%.

[0035] Preferably, the first stage composite macro-vitamin iodine premix and the first stage composite macro-vitamin premix comprise the following components by mass percentage:

[0036] Choline bitartrate 42.612%;

[0037] Sodium L-ascorbate 35.713%;

[0038] Taurine 7.102%;

[0039] Inositol 7.102%;

[0040] L-Carnitine tartrate 2.130%;

[0041] dl-α-tocopherol acetate 1.758%;

[0042] Niacinamide 1.711%;

[0043] D-calcium pantothenate 0.89%;

[0044] Riboflavin 0.325%;

[0045] Thiamine nitrate 0.271%;

[0046] Vitamin A acetate 0.17%;

[0047] Pyridoxine hydrochloride 0.153%;

[0048] Folic acid 0.035%;

[0049] Phytomenadione 0.015%;

[0050] D-biotin 0.009%;

[0051] Vitamin D3 0.005%;

[0052] Cyanocobalamin 0.0005%.

[0053] Preferably, the first stage composite macro-mineral premix and the first stage composite trace mineral premix comprise the following components by mass percentage:

[0054] Potassium chloride 60.483%;

[0055] Tricalcium phosphate 21.793%;

[0056] Potassium hydrogen phosphate 11.871%;

[0057] Magnesium sulfate 4.824%;

[0058] Zinc sulfate 0.506%;

[0059] Ferrous sulfate 0.422%;

[0060] Copper sulfate 0.076%;

[0061] Potassium iodide 0.018%;

[0062] Manganese sulfate 0.004%;

[0063] Sodium selenite 0.003%.

[0064] In order to facilitate the storage of milk powder, the moisture content of the infant formula milk powder is controlled below 3wt%. Thus, although GOS has a certain hygroscopicity, MFGM contains phospholipids that are more easily oxidized, and the stable storage of milk powder is facilitated by controlling the moisture content.

[0065] Compared with the prior art, the present invention has the following beneficial effects:

[0066] (1) The present invention provides a milk fat globule membrane and galacto-oligosaccharide composition, which has been verified by experimental studies to have the ability to synergistically regulate early intestinal flora, promote intestinal barrier development and lipid absorption.

[0067] (2) In the improved scheme, by designing different components and ratios, the OPO, OPL, MLCT and fatty acid composition in the oil composition is made closer to the lipid composition of breast milk; in addition, the use of the oil composition can achieve a lipid composition similar to that of breast milk without adding raw cow's milk, which is conducive to simplifying the production process of infant formula milk powder and reducing production costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] Figure 1 Design illustrations for animal experiments;

[0069] Figure 2 The graphs show the periodic changes of Shannon (a), Simpson (b), ACE (c) and Chao 1 (d) indexes. The asterisk “*” indicates p < 0.05.

[0070] Figure 3 Figure 1 shows the nonmetric multidimensional scaling (NMDS) analysis of the GOS+High MFGM group and the Control, GOS, MFGM, and GOS+Low MFGM groups at 0 (a), 8 (b), 15 (c), and 24 (d). The table shows the results of the NMDS analysis. 2 The closer it is to 1, the stronger the reliability of NMDS analysis is, and p < 0.05 indicates that there is a significant difference between the groups;

[0071] Figure 4 The effect of GOS and MFGM on the composition of the top 100 absolute abundance of intestinal flora in SD rats at the genus level and phylum level;

[0072] Figure 5Wilcoxon test plots of GOS+High MFGM and Control (a), GOS (b), MFGM (c), and GOS+Low MFGM (d);

[0073] Figure 6 The continuous changes of the abundance of Enterococcus (a), Roseburia (b), Lachnospiracea_ND3007_group (c) and Desulfovibrio (d) in the GOS+High MFGM group on days 0, 8, 15 and 24;

[0074] Figure 7 Continuous changes in the abundance of Prevotellaceae_NK3B31_group (a) in the GOS+Low MFGM and GOS+High MFGM groups, Alistipes (b) in the GOS, GOS+Low MFGM and GOS+High MFGM groups, and unclassified_Clostridia_UCG_014 (c), UCG_005 (d) and Marvinbryantia (e) in the five different diet groups on days 0, 8, 15 and 24;

[0075] Figure 8 The contents of fecal short-chain fatty acids in the Control, GOS, MFGM, GOS+Low MFGM and GOS+High MFGM groups on the 24th day (a); the continuous changes of short-chain fatty acids in the Control (b), GOS (c), MFGM (d), GOS+Low MFGM (e) and GOS+High MFGM groups (f) on the 0th, 8th, 15th and 24th days; the expression levels of short-chain fatty acid G protein-coupled receptors (GPR43 and GPR109a) in the jejunum (g) and colon (h);

[0076] Fig. 9 The figure shows the effects of GOS and MFGM on the expression of intestinal barrier (MUC2 and ZO-1) and lipid absorption-related genes (SLC27A, FABP4 and Dgat1) in SD rats;

[0077] Fig.10 The average Zeta potential (a) and particle size distribution (b) of infant formula emulsion; the average Zeta potential (c) and particle size distribution (d) of reconstituted infant formula emulsion. Different lowercase letters (a, b, c) represent significant differences between groups (p<0.05);

[0078] Fig.11Microscopic morphology (a), XRD (b) and DSC curve (c) of infant formula milk powder;

[0079] Fig.12 The graph shows the effects of GOS and MFGM on the acid value (a), peroxide value (b) and malondialdehyde content (c) of infant formula during storage. DETAILED DESCRIPTION

[0080] The present invention is further described in detail below with reference to the accompanying drawings.

[0081] Example 1 Effect verification of the composition

[0082] 1. Test methods

[0083] 1.1 Experimental design and sample collection

[0084] Experimental Materials:

[0085] Galacto-oligosaccharide (GOS) was purchased from Yunfu Xinjinshan Biotechnology Co., Ltd. (model GOS-700). The purity of GOS in this batch of products was 83.7%.

[0086] Milk fat globule membrane MFGM, using whey protein powder with milk fat globule membrane (MFGM), the whey protein powder was purchased from Arla Foods Ingredients MFGM-10, in which the phospholipid content is 7% of the total substance content;

[0087] In this study, a total of 50 male SD rats were raised and randomly divided into 5 groups after adaptive feeding, with 10 rats in each group. Five dose groups were set:

[0088] ①Control group: normal diet, pure water gavage;

[0089] ②GOS group: normal diet, oral administration of GOS solution (i.e., oral administration of the above-mentioned GOS-700 dissolved in water), the dose is 900 mg GOS-700 / kg body weight / day;

[0090] ③MFGM group: normal diet, gavage of MFGM solution (i.e., gavage of the above-mentioned whey protein powder with milk fat globule membrane (MFGM), dissolved in water), gavage dose of 650 mg whey protein powder / kg body weight / day;

[0091] ④GOS+Low MFGM group: normal diet, oral administration of GOS and MFGM mixed solution, the dosage was 900 mg GOS-700 / kg body weight / day, 210 mg whey protein powder / kg body weight / day;

[0092] ⑤GOS+High MFGM group: normal diet, oral administration of GOS and MFGM mixed solution, the dose was 900 mg GOS-700 / kg body weight / day, 650 mg whey protein powder / kg body weight / day, the oral administration volume was 1 mL for all groups.

[0093] The above-mentioned oral gavage dose mainly takes into account the average infant intake of about 150 mL of formula milk / kg body weight / day, as well as the oral gavage range that animal experimental rats can tolerate.

[0094] The experimental design is detailed in Figure 1 The body weight and food intake of rats were weighed and calculated in a 3-day cycle, and oral gavage samples were prepared on the basis of body weight for the last 3 days. During the experimental cycle, feces of rats were collected on days 0, 8, 15, and 24 for subsequent analysis.

[0095] 1.2 Establishment of sample detection method based on rat experiment

[0096] 1.2.1 High-throughput 16S rRNA sequencing of rat feces

[0097] The full-length 16S microbial diversity was used to quantitatively analyze the structural composition of the rat intestinal flora. In brief, nucleic acids were extracted from fecal samples using a DNA extraction kit. The nucleic acid concentration was determined using the Qubit assay, and the nucleic acid samples were diluted with 0.1×TE to a concentration of 1 ng / μL. Subsequently, a quantitative Spike-in internal reference was added, and specific primers with barcodes were synthesized according to the full-length primer sequence for PCR amplification. The amplified PCR products were mixed, and the library construction kit provided by PacBio was used to repair damage, end repair, and connect sequencing adapters to the amplified products. The reaction process was carried out on a PCR instrument, and finally the AMpure PB magnetic bead purification kit was used to recover the PCR-free library for the machine. The library that passed the concentration and size test was sequenced on the machine.

[0098] 1.2.2 Analysis of fecal short-chain fatty acids

[0099] 3 mL of saturated NaCl solution was added to the fecal sample and allowed to stand for 30 min; 120 μL of sulfuric acid (sulfuric acid: water = 1:10, v / v) was added to acidify; 4.5 mL of n-hexane was taken to extract short-chain fatty acids and centrifuged at 4 °C, 10000 rpm for 20 min. The supernatant was added with anhydrous sodium sulfate and centrifuged to remove water. Finally, the supernatant was filtered into a gas phase vial with a 0.22 μm organic syringe filter until the sample was analyzed. Sample preparation was carried out under ice bath conditions to prevent the volatilization of short-chain fatty acids. At the same time, the short-chain fatty acid standard was gradiently diluted to 500 μg / mL, 400 μg / mL, 300 μg / mL, 200 μg / mL, 100 μg / mL, 50 μg / mL, and 25 μg / mL, and the short-chain fatty acids were determined by gas chromatography and a standard curve was drawn.

[0100] 1.2.3 Jejunum / colon RT-qPCR

[0101] Intestinal tissue samples were ground into powder on liquid nitrogen, and RNA from jejunum and colon samples was extracted using an RNA extraction kit and reverse transcribed into cDNA. Specific primers for mucin2 (MUC2), tight junction protein-1 (Zonulaoccluden-1, ZO-1), solute carrier family 27 member 4 (SLC27A4), fatty acid-binding protein 4 (FABP4), diacylglycerol O-acyltransferase-1 (Dgat1), G protein-coupled receptor 43 (GPR43) and G protein-coupled receptor 109a (GPR109a) were designed and synthesized, and the primers had good specificity. RT-qPCR was performed on PerfectStart Green for real-time quantitative polymerase chain reaction, and the cycle parameters were: 95℃ for 15min and 40 cycles, including 94℃ for 30s, 60℃ for 30s, and 72℃ for 30s. The linearity of the dissociation curve was determined using a fluorescent quantitative PCR instrument and analyzed using iCycler software. The average cycle time of the linear part of the curve was designated as Ct. The following formula was used for normalization: relative expression level change fold of gene = 2 (Ct 基因 -Ct 内参(GAPDH) ), and the results are shown as the mean fold change ± standard deviation relative to the Control group (set to 1).

[0102] Table 1 Primer design for RT-qPCR reaction of SD rats

[0103]

[0104]

[0105] 1.2.4 Intestinal histopathological observation

[0106] About 1 cm of rat jejunum and colon tissues were taken and fixed in 10% neutral formalin fixative. After paraffin embedding, sectioning (5 μm), and hematoxylin and eosin staining, the villus height and crypt depth were observed and measured using a digital slice scanner.

[0107] 2. Results

[0108] 2.1 Effects of GOS and MFGM on the diversity and structural composition of rat intestinal flora

[0109] like Figure 2 As shown in (a, b), on the 24th day of intervention, the Shannon index and Simpson index of the GOS+High MFGM group were significantly increased compared with the GOS+Low MFGM group (p<0.05), indicating that the synergistic effect of GOS and MFGM on intestinal flora was related to the dose of MFGM.

[0110] like Figure 2 As shown in (a, b, c, d), on the 8th day of GOS+High MFGM intervention, the Shannon, Simpson, ACE and Chao 1 indices increased from significantly lower than those in the MFGM group on the 0th day to no significant difference with the MFGM group, and the Shannon, ACE and Chao 1 indices showed a continuous increasing trend over time, indicating that GOS+High MFGM intervention can improve the microbial diversity of intestinal flora, and is proportional to the intervention time.

[0111] like Figure 3 As shown in (a, b, c, d), NMDS analysis was performed using binary-jaccard metric distance, and there were significant differences in β-diversity between samples on days 8, 15, and 24, indicating that GOS+High MFGM intervention can affect cyclical changes in intestinal flora.

[0112] like Figure 4 As shown in the figure, all samples had similar taxonomic communities and showed relatively high abundances of Firmicutes, Bacteroidota, Proteobacteria, and Acidobacteriota. In addition, on the 8th day of intervention, the species abundance of all samples showed an upward trend compared with that on the 0th day, especially in the GOS+Low MFGM and GOS+High MFGM groups, indicating that the synergistic effect of GOS and MFGM can significantly increase the species abundance of the intestinal flora in early life.

[0113] The results showed that GOS+High MFGM intervention for 24 days significantly increased the abundance of Ligilactobacillus, Prevotellaceae_NK3B31_group, Marvinbryantia, and Blautia (p<0.05) ( Figure 5 ). This indicates that GOS+High MFGM intervention can increase the abundance of beneficial microorganisms in the early intestinal flora that promote intestinal barrier development, improve immune regulation, anti-inflammation, and produce short-chain fatty acids. Figure 6 As shown in the figure, GOS+High MFGM intervention can reduce the abundance of Enterococcus, which is a risk of infection, and increase the abundance of beneficial bacteria such as Roseburia, Lachnospiracea_ND3007_group and Desulfovibrio. Figure 7 As shown in the figure, GOS+High MFGM can increase the abundance of beneficial bacteria Previotellaceae_NK3B31_group and Alistipes. As beneficial intestinal bacteria, the abundance of unclassified_Clostridia_UCG_014, UCG_005 and Marvinbryantia increased significantly under the five intervention modes, and the GOS+High MFGM intervention showed a more significant increase trend.

[0114] 2.2 Effects of GOS and MFGM on short-chain fatty acids in rat feces

[0115] After 24 days of feeding, the total short-chain fatty acid content in the MFGM (1.172±0.001mg / g), GOS+Low MFGM (1.162±0.006mg / g) and GOS+High MFGM (1.165±0.007mg / g) groups was significantly lower than that in the Control (1.698±0.02mg / g) and GOS (1.697±0.016mg / g) groups ( Figure 8 ). By analyzing the effects of GOS and MFGM on G protein-coupled receptor genes in the jejunum and colon, it was found that the expression of GPR109a gene was significantly upregulated in the colon of rats in the MFGM, GOS+Low MFGM and GOS+High MFGM groups. It is speculated that the high expression of GPR109a may be related to the absorption of short-chain fatty acids. This is mainly because MFGM, as the main component of biofilm, plays a positive role in promoting the development of intestinal epithelial cells and the absorption of nutrients.

[0116] 2.3 Effects of GOS and MFGM on intestinal barrier and lipid absorption in rats

[0117] like Fig. 9 As shown, GOS+High MFGM significantly increased the expression of tight junction proteins MUC2 and ZO-1 in jejunal epithelial cells, indicating that GOS+High MFGM can effectively consolidate the safety of the intestinal barrier and control the occurrence of intestinal diseases by promoting intestinal mucosal development and reducing cell gaps.

[0118] In addition, GOS+High MFGM also significantly increased the expression of SLC27A4, FABP4 and Dgat1, and played a positive role in promoting the absorption of long-chain fatty acids, free fatty acids and the synthesis of triglycerides in the jejunum and colon. Through the measurement of jejunal villus height (Table 2), GOS+High (or Low) MFGM can significantly improve the development of jejunal villi, thereby improving the ability to absorb substances. The results of this study show that GOS+High MFGM can promote the absorption and synthesis of lipids by upregulating the expression of SLC27A4, FABP4 and Dgat1 genes and promoting the development of intestinal villi.

[0119] Table 2 Effects of GOS and MFGM on the length of villi and crypt depth in rat jejunum and colon

[0120]

[0121]

[0122] Note: Different lowercase letters (a, b, c) represent significant differences among different groups (p<0.05).

[0123] Example 2 Composition rich in regulating early intestinal flora and infant formula milk powder with lipid composition close to breast milk

[0124] 1. Formula milk powder composition

[0125] According to the experimental calculation of GOS+High MFGM group in Example 1, the intake of GOS-700 for infants needs to meet 900

[0126] mg / kg body weight / day, the intake of whey protein powder with milk fat globule membrane (MFGM) needs to meet 650mg / kg body weight

[0127] / day. Based on the average infant intake of about 150mL formula milk / kg body weight / day, this is equivalent to 900mg GOS-700 / 150mL

[0128] Formula milk and 650 mg of whey protein powder with MFGM / 150 mL of formula milk. The formula milk powder is prepared as follows:

[0129] 90 mL of warm water is used to mix 12.4 g of milk powder to obtain 100 mL of formula milk. 150 mL of formula milk should contain about 18.6 g of milk powder. The GOS-700 content is 900 mg / 18.6 g of milk powder. Considering that the purity of GOS in GOS-700 is 83.7%

[0130] (Example 1), the content of GOS in milk powder is about (900mg*83.7%) / 18.6g, that is, by mass fraction, GOS

[0131] The content of whey protein powder with MFGM is: 650mg / 18.6g milk powder, that is, by mass fraction, the whey protein powder with MFGM accounts for 3.5% of the mass of milk powder.

[0132] Similarly, corresponding to the GOS+LOW MFGM group experiment, the content of GOS in milk powder is the same as above, and the content of whey protein powder with MFGM is: 210mg / 18.6g milk powder, that is, by mass fraction, the whey protein powder with MFGM is

[0133] White powder accounts for 1.1% of the weight of milk powder.

[0134] Specifically, the infant formula milk powder comprises the following components by weight percentage: whey egg white with milk fat globule membrane

[0135] White powder 3.5%, isolated whey protein powder 10.7%, oil and fat composition 28.3%, lactose 52.2%, galacto-oligosaccharide 4.8%,

[0136] 1 stage composite macrovitamin iodine premix 0.1%, 1 stage composite macrovitamin premix 0.1%, 1 stage composite macromineral premix 0.25%, 1 stage composite trace mineral premix 0.1%. According to the weight percentage in the oil composition

[0137] The oil composition includes: 1,3-dioleyl-2-palmitoyl triglyceride: 32%, sunflower oil: 26%, high oleic sunflower oil: 17%, breast milk-like oil (including OPO, OPL and MLCT): 15%, coconut oil: 7%, linseed oil: 3%. Among them, 1,3-dioleyl-2-palmitoyl triglyceride, high oleic sunflower oil (according to the gas chromatographic peak area ratio, oleic acid accounts for 84.6%), and breast milk-like oil (MoMiFAT brand) are produced and sold publicly by Wilmar (Shanghai) Biotechnology Research and Development Center Co., Ltd. The first stage compound macrovitamin iodine premix and the first stage compound macrovitamin premix include the following ingredients by mass percentage: 42.612% choline bitartrate, 35.713% L-sodium ascorbate, 7.102% taurine, 7.102% inositol, 2.130% L-carnitine tartrate, 1.758% dl-α-tocopheryl acetate, 1.711% niacinamide, 0.89% D-calcium pantothenate, 0.325% riboflavin, 0.271% thiamine nitrate, 0.17% vitamin A acetate, 0.153% pyridoxine hydrochloride, 0.035% folic acid, 0.015% phytonadione, 0.009% D-biotin, vitamin D 3 The grease composition comprises the following components by mass percentage: 33.17±1.04% saturated fatty acid; 43.63±0.05% monounsaturated fatty acid; 23.20±0.99% polyunsaturated fatty acid; the first stage composite macro-mineral premix and the first stage composite trace mineral premix comprise the following components by mass percentage: 60.483% potassium chloride, 21.793% tricalcium phosphate, 11.871% dipotassium hydrogen phosphate, 4.824% magnesium sulfate, 0.506% zinc sulfate, 0.422% ferrous sulfate, 0.076% copper sulfate, 0.018% potassium iodide, 0.004% manganese sulfate, and 0.003% sodium selenite.

[0138] The milk powder is beneficial for regulating the intestinal flora in early life, promoting the development of the intestinal barrier and lipid absorption; the oil and fat composition can achieve a lipid composition similar to that of breast milk without adding raw cow's milk, which is beneficial for simplifying the production process of infant formula milk powder and reducing production costs.

[0139] 2. Preparation method of formula milk powder

[0140] The preparation method of infant formula comprises the following steps:

[0141] (1) Ingredients: weigh whey protein powder rich in milk fat globule membrane, isolated whey protein powder, lactose and galacto-oligosaccharide according to a predetermined ratio, mix them evenly, and completely hydrate them;

[0142] (2) Emulsification and sterilization: emulsify the mixed ingredients and the oil composition at 55°C for 30 minutes and sterilize;

[0143] (3) Shearing and homogenization: The mixture was sheared at 8000 rpm for 1 min in a high-speed shearing machine, and then homogenized three times in a high-pressure homogenizer at a homogenization pressure of 150 bar;

[0144] (4) Spray drying: the air inlet temperature is 180°C and the outlet temperature is 90°C for drying;

[0145] (5) Dry mixing: mixing the vitamin and mineral premix with the spray-dried milk powder;

[0146] (6) Packaging: Nitrogen-filled packaging.

[0147] 3. Characterization of infant milk emulsion and milk powder includes the following:

[0148] (1) Effects of MFGM and GOS on particle size distribution and zeta potential of infant formula emulsions and infant formula milk powder reconstituted emulsions

[0149] like Fig.10 As shown in the figure, the Zeta potential of the infant formula milk powder containing GOS+High MFGM is -34mV, the average particle size is about 400nm, and the particle size is between 200 and 1000nm ( Fig.10 (a, b)). After the spray-dried milk powder was reconstituted, the Zeta potential was -35 mV, the average particle size distribution was between 400 and 500 nm (the first peak), and the particle size distribution of a small part (the second peak) of the emulsion was between 4000 and 6000 nm ( Fig.10 (c, d)). This indicates that the milk powder has good resolubility, with only a small amount of particles with poor solubility existing, which can ensure the stability of the reconstituted emulsion.

[0150] (2) Analysis of milk powder moisture content, water activity, surface oil content, surface microstructure, lactose crystallinity, and glass transition temperature

[0151] In Table 2, the sources of galacto-oligosaccharide GOS and milk fat globule membrane MFGM are the same as those in Example 1. That is, galacto-oligosaccharide GOS was purchased from Yunfu Xinjinshan Biotechnology Co., Ltd. (model GOS-700), and the purity of GOS in this batch of products was 83.7%. Milk fat globule membrane MFGM was obtained from whey protein powder with milk fat globule membrane (MFGM), which was purchased from Arla Foods Ingredients. MFGM-10, in which the phospholipid content is 7% of the total substance content.

[0152] Specifically, in Table 2, GOS group (GOS-700 accounts for 4.1% of the weight of milk powder); MFGM group (whey protein powder with MFGM accounts for 3.5% of the weight of milk powder); GOS+low MFGM group (GOS-700 accounts for 4.1% of the weight of milk powder, whey protein powder with MFGM accounts for 1.1% of the weight of milk powder), GOS+high MFGM group (GOS-700 accounts for 4.1% of the weight of milk powder, whey protein powder with MFGM accounts for 3.5% of the weight of milk powder). As shown in Table 2, the moisture content of infant formula milk powder containing GOS+High MFGM is about 3.52%, the water activity is about 0.244, and the surface oil content is 1.84%. The moisture content of the three infant formula milk powders containing GOS (GOS, GOS+Low MFGM and GOS+High MFGM) increased significantly, and all were >3%, indicating that the addition of GOS with strong hygroscopicity is more likely to lead to an increase in the viscosity of the milk powder. Therefore, it is recommended that milk powder containing GOS be dried to a lower moisture content to increase the shelf life of the product.

[0153] Table 2 Moisture content, water activity and surface oil content of infant formula milk powder

[0154]

[0155] Note: Different lowercase letters (a, b, c) represent significant differences among the groups (p<0.05).

[0156] The microstructure of the infant formula powder containing GOS+High MFGM is between 4 and 14 μm, and is smooth and spherical, with no obvious agglomeration ( Fig.11 (a)). The milk powder did not show crystallization peaks at 19.1° and 20.1° ( Fig.11 (b)), indicating that there is no α-lactose monohydrate and anhydrous α:β lactose crystals in the milk powder, and the lactose in the milk powder is in an amorphous state. The glass transition temperature (T g ) is 47.72℃, which is between MFGM and GOS milk powder ( Fig.11 (c)) indicates that GOS supplementation has the effect of reducing T g trend, thereby reducing the physical stability of milk powder.

[0157] (3) Effects of MFGM and GOS on storage stability of infant formula

[0158] like Fig.12As shown in the figure, after the milk powder was stored at 40°C for 15 days, the acid value, peroxide value and malondialdehyde content increased significantly. Among them, the acid value and malondialdehyde content of the infant formula milk powder containing GOS+High MFGM had a higher growth trend, indicating that the contribution of GOS to the hygroscopicity of milk powder and the highly oxidizable phospholipids contained in MFGM may accelerate the oxidation of milk powder. In addition, the b* value of the infant formula milk powder containing GOS+High MFGM increased significantly with the increase of storage time (Table 3), indicating that this milk powder has an obvious browning trend during the pro-oxidation process. In summary, infant formula milk powder containing GOS+High MFGM should be stored at a lower temperature while controlling the moisture content (below 3wt%) to reduce the occurrence of lipid oxidation and Maillard reaction.

[0159] Table 3 Effect of GOS and MFGM on color values ​​L and b of infant formula during storage

[0160]

[0161] Note: Different capital letters (A, B, C) represent that different groups have significant differences (p<0.05) at a certain sampling time (Day 5, 10 or 15); different lowercase letters (a, b, c) represent that a group has significant differences (p<0.05) at different sampling times (Day 5, 10 and 15).

[0162] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A composition for regulating intestinal flora and lipid absorption in early life, characterized in that: Including galacto-oligosaccharides and milk fat globule membrane.

2. Use of the composition according to claim 1 in preparing infant formula milk powder.

3. The use according to claim 2, characterized in that: In the infant formula milk powder, the galacto-oligosaccharide accounts for 4.1% by mass, and the whey protein powder with milk fat globule membrane accounts for 1.1% by mass.

4. The use according to claim 2, characterized in that: In the infant formula milk powder, the galacto-oligosaccharide accounts for 4.1% by mass, and the whey protein powder with milk fat globule membrane accounts for 3.5% by mass.

5. The use according to claim 2, characterized in that: The infant formula milk powder further comprises a fat composition, which comprises the following components by mass percentage: 1,3-dioleyl-2-palmitoyl triglyceride 32%; Sunflower oil 26%; High oleic sunflower oil 17%, of which oleic acid accounts for 84.6% according to the gas chromatography peak area ratio; Breast milk-like fat 15%; Coconut oil 7%; Linseed oil 3%.

6. The use according to claim 2, characterized in that: The infant formula milk powder further comprises a fat composition, which comprises the following components by mass percentage: Saturated fatty acids 33.17 ± 1.04%; Monounsaturated fatty acids 43.63 ± 0.05%; Polyunsaturated fatty acids 23.20±0.99%.

7. The use according to claim 5 or 6, characterized in that: The infant formula milk powder comprises the following ingredients by mass percentage: 3.5% whey protein powder with milk fat globule membrane; Whey protein isolate powder 10.7%; The oil and fat composition 28.3%; Lactose 52.2%; The galacto-oligosaccharide 4.8%; 1 stage complex macrovitamin iodine premix 0.1%; Stage 1 complex macrovitamin premix 0.1%; 1st stage composite macro mineral premix 0.25%; Stage 1 Complex Trace Mineral Premix 0.1%.

8. The use according to claim 7, characterized in that: The first stage composite macro-vitamin iodine premix and the first stage composite macro-vitamin premix comprise the following components by mass percentage: Choline bitartrate 42.612%; Sodium L-ascorbate 35.713%; Taurine 7.102%; Inositol 7.102%; L-Carnitine tartrate 2.130%; dl-α-tocopherol acetate 1.758%; Niacinamide 1.711%; D-calcium pantothenate 0.89%; Riboflavin 0.325%; Thiamine nitrate 0.271%; Vitamin A acetate 0.17%; Pyridoxine hydrochloride 0.153%; Folic acid 0.035%; Phytomenadione 0.015%; D-biotin 0.009%; Vitamin D3 0.005%; Cyanocobalamin 0.0005%.

9. The use according to claim 7, characterized in that: The first stage composite macro-mineral premix and the first stage composite trace mineral premix comprise the following components by mass percentage: Potassium chloride 60.483%; Tricalcium phosphate 21.793%; Potassium hydrogen phosphate 11.871%; Magnesium sulfate 4.824%; Zinc sulfate 0.506%; Ferrous sulfate 0.422%; Copper sulfate 0.076%; Potassium iodide 0.018%; Manganese sulfate 0.004%; Sodium selenite 0.003%.

10. The use according to claim 2, characterized in that: The moisture content of the infant formula milk powder is controlled below 3wt%.

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