TW201249349A - Milk oligosaccharide-galactooligosaccharide composition for infant formula containing the soluble oligosaccharide fraction present in milk, and having a low level of monosaccharides, and a process to produce the composition - Google Patents

Abstract

The invention discloses an oligosaccharide mixture derived from cow's milk that can be easily spray dried comprising (a) a soluble oligosaccharide population which is the same as that of soluble oligosaccharides found in cow's milk and (b) β -galactooligosaccharides formed by the action of β -galacotsidase on lactose and the milk oligosaccharides. The mixture having a total monosaccharide content of less than 5% w/v and a lactose: oligosaccharide ratio of less than 20. A process for obtaining such a mixture, which includes a nanofiltration step, is disclosed. Nutritional compositions, especially infant formula, comprising said oligosaccharide mixture are also disclosed.

Description

201249349 VI. Description of the Invention: [Technical Field] The present invention relates to a mixture of oligosaccharides derived from cow's milk and a food product comprising the oligo (os) mixture (especially an infant formula), and a preparation of the oligosaccharide mixture Methods. [Prior Art] A large number of bacteria having favorable and adverse effects on visceral physiology and having other systemic effects survive in the human colon. The dominant bacterial population found in the colon includes anaerobic bacilli (Bifidobacteria, Eubacteria, Clostridium (CVoiinWa), and Botrytis bacillus). Matrix availability, redox potential, pH, 〇2 tension and its distribution in the colon have fluctuating activity. In general, intestinal bacteria can be classified as species that exert potentially harmful or beneficial effects on their host. Effects (which may be caused by, for example, Clostridium or anaerobic bacilli) include dysentery, infection 'liver damage, cancer and intestinal rot. It can inhibit the growth of harmful bacteria, stimulate immune function, improve the digestion and absorption of essential nutrients and vitamins. Synthetic, causing a health-promoting effect. It is necessary to increase the number and/or activity of bacterial populations (such as bifidobacteria and lactobacilli) which may have health promoting properties. Such "beneficial bacteria" which have a beneficial effect on the host are named " Probiotics. Probiotics include many types of bacteria but are generally selected from four bacterial species: Lactobacillus acidophilus ac/i/ Op/i/Z/w·?), Bifidobacterium, Lactobacillus (Zac/ococcwi) and Pediococcus (/»eshan.wcw). Health benefits associated with probiotics such as lactobacilli or bifidobacteria include 164518.doc 201249349 enhanced systemic cellular immune responses, such as improved antibody production and phagocytosis (swallowing) of white blood cells. Certain probiotic strains have been associated with boosting the immune system and thus prevent or reduce the extent of infection. Some probiotic strains are associated with preventing allergies and alleviating the severity of allergies. Several strains have been documented to be effective in improving intestinal diseases, particularly diseases. Regarding the specific situation of the baby, the baby's gastrointestinal tract is considered to be sterile prior to birth. During the birth process, its exposure to bacteria and bacteria from the mother's digestive tract and skin begins to grow there. There is a big difference in the composition of the visceral microbial phase in response to infant fakes. The fecal flora of human-fed infants includes a large number of Bifidobacterium flora containing some Lactobacillus species. However, formula-fed infants have a more complex microbial phase, usually present in the genus Bifidobacterium and Bacteroides. (7) "I", Clostridium and Streptococcus (汾repMcoaz·) » After weaning, we began to establish a type of visceral microbial phase similar to adult type. It is recommended to breastfeed all babies. However, in some cases, for medical reasons The milk is not enough or unsuccessful, or the mother chooses not to be breast-friendly. For such cases, the infant formula has been developed. In order to establish a healthy intestinal flora similar to human-fed infants, it is necessary to supplement the baby with probiotics. Formulations. Examples of probiotics currently used in infant formula include Lactobacillus rhamnosus, available from Valio 〇y, Finland under the trademark LGG (Zaciokc/Z/wi ATCC 53103, by BioGaia

Α·Β sold under the trademark Reuteri, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus lactis CNCM 1-2116, Rodrigues rewieri), BLIS I64518.doc 201249349 from New Zealand

Technologies Limited sells Lactobacillus johnsonii CNCM 1-1225 under the trademark KI2, S. salivarius {Streptococcus DSM 13084, especially from Denmark

Bifidobacterium sold by Christian Hansen company under the trademark Bb 12 (Bz/zWoiacierz’ww CNCM 1-3446, by Morinaga, Japan

Milk Industry Co. Ltd. sold under the trademark BB536, Bifidobacterium breve (owgwm) ATCC BAA-999, sold by Danisco under the trade Bb-03 short Bifidobacterium Z> reve), by Morinaga Short Bifidobacterium sold under the trademark M-16V, Bifidobacterium infantis sold by Procter & Gamble Co. under the trademark Bifantis, by Institut Rosell (Lallemand) under the trademark R0070 Short-type Bifidobacterium for sale. Such bacteria can be administered in an amount of about 10 to about 20 billion colony forming units (CFU) per day to maintain intestinal microflora healthily, preferably about 5 billion to about 1 〇〇 亿活菌/天. Another factor affecting healthy intestinal flora is the beneficial bacteria in the intestine. Probiotics help selectively stimulate the growth and/or activity of probiotics in the colon. Thus improving the healthy indigestible food component of the host. The probiotic is not broken down or absorbed in the stomach or small intestine, and thus completely enters the colon which is selectively fermented by beneficial bacteria, in the sense that Indigestible Examples of helper biomass include certain oligosaccharides, such as oligosaccharides (FOS) and galactooligosaccharides (GOS). Human milk is known to contain higher indigestible oligosaccharides than most other animal milk-containing children. Digestive oligosaccharides represent the third largest solid in human milk. 164518.doc 201249349 Knife (after lactose and lipids)' is 丨2 to 丨5 g/丨 in colostrum and 5-8 in cooked milk. The concentration of g/l exists. Human milk oligosaccharide (HM〇) is extremely resistant to enzymatic hydrolysis, indicating that this type of oligosaccharide can exhibit basic functions not directly related to its calorie value. Therefore, probiotics and probiotics Synergistically provides an important health benefit for infants. Probiotics not only selectively promote the growth of probiotics added to infant formula, but also promote the growth of endogenous probiotics that work synergistically with the added probiotics. Therefore, the health benefits provided by the probiotics described above are enhanced by the presence of beneficial bacteria. Therefore, as the understanding of the composition of human milk is increased, it is also proposed to add beneficial bacteria to the infant formula. Bacterial helper A sufficient amount to stimulate the healthy microflora in the viscera and to cause the proliferation of such "good" bacteria. Typical levels are from about 1 to about 1 〇g per serving of food or from about 5% to about 4 〇〇/0. The recommended daily intake of fiber for babies. Therefore, it is possible to purchase various infant formulas that supplement the beneficial bacteria to help the mixture of oligosaccharides and galactooligosaccharides. However, such mixtures are only provided in human milk. A mixture of oligosaccharides present. More than one oligosaccharide component has been detected in human milk, some of which are not detected in animal milk such as cow's milk, or only a small amount is detected. Some sialylated oligosaccharides and fucosylated oligosaccharides are present in cow's milk and colostrum, but only in very small amounts. EP 0 975 23 5 B1 describes a synthetic nutritional composition comprising one or more human milk oligosaccharides, wherein the HMO in the composition is selected from 8 HMOs (3-164518.doc 201249349 fucosyllactose, milk -N-rock thin pentose in, milk-N-fucoyl pentose π, difucosyllactose, 2·-fucosyllactose, milk-Ν-fucoid pentose I, milk-Ν-new The group of tetrasaccharides and milk-Ν-fucose pentose V), the European patent shows: - The oligosaccharide protects babies from viral and bacterial infections in the respiratory, gastrointestinal and urethra. U.S. Patent Application Serial No. 2003/0129278 describes an oligosaccharide mixture based on oligosaccharides produced by biting several animal milks, characterized in that it comprises at least two oligosaccharide notes, each consisting of at least two Different oligarchy compositions. The difference in the oligosaccharide population in the oligosaccharide mixture is the extraction of animal milk from the oligodomain. ΕΡ 0 458 358 relates to a method for preparing a skim milk powder comprising from 1 to 15% by weight of galactooligosaccharides, comprising: (0 concentrated skim milk to obtain a concentrated milk having a solid content of 20 to 50% by weight, (Π) adding β-galactosidase to the concentrated milk to cause an enzymatic reaction, heating the resulting reaction mixture for 30 seconds to 15 minutes to 70 to 85 ° C to end the enzyme reaction, and (iv) Spray-drying the mixture of the end reaction. From the inventors 2W〇2〇〇6/087391 discloses a mixture of oligosaccharides derived from animal milk and a method for preparing the oligo-mix. The oligo-purine mixture is used as a specific person. The beneficial bacteria in the viscera are effective and have a distribution of oligosaccharides closer to human milk than those provided by the mixture of sugar and galactooligosaccharides. In order to develop a probiotic-promoting infant formula, it is necessary to provide a kind of 164518.doc

S 201249349 ::=, the distribution of the near vinegar distribution (at least in qualitative terms) can be varied in the presence of milk. This means that the ugly mixture should contain the intermediate oligobiliary (four) points. "Soluble pride" (4) The content is not the same as oligosaccharides. The relative set of different soluble oligosaccharides in the source milk does not have to remain in the oligosaccharide mixture. Moreover, it is desirable to maintain a low lactose/oligosaccharide ratio to avoid introducing unnecessary high amounts of milk:' and to reduce the amount of the two required to achieve the desired amount of oligosaccharide in the infant formula. Moreover, the sugar-supplying mixture should have a very low protein content so that the amino acid distribution of the infant formula is not strongly affected. It is an object of the present invention to provide an oligosaccharide mixture which is effective as a probiotic-promoting substance in a human viscera and which has a human milk that is closer to human milk than a mixture of a fruit-supplying sugar and a galactooligosaccharide. Sugar distribution and very low monosaccharide concentration. Accordingly, it is an object of the present invention to provide an oligosaccharide mixture comprising an oligosaccharide soluble fraction present in the milk from which the oligosaccharide mixture is derived. In the context of the present invention, "a mixture of oligosaccharides derived from cow's milk" means that the oligosaccharide is obtained from cow's milk. Thus, the different soluble oligosaccharides present in the source milk are also present in the final oligosaccharide mixture of the invention, but are not necessarily in the same proportions. Another object of the present invention is to provide an oligosaccharide mixture having a relatively high concentration of oligosaccharides (generally 20 to 50% by weight). There is a need for a food that is targeted especially to young children, infants and/or newborn babies' which helps to ensure a normal immune or inflammatory state or to reduce or reduce the effects of food allergies. There is a need to provide a food that is specifically effective as a probiotic-promoting substance in human viscera. 164518.doc 201249349 There is a need for a food 0 that provides the above benefits while maintaining an individual's balanced normal metabolism requiring an improvement in human visceral conditions achieved by non-drug-based interventions that are compatible with vulnerable individuals such as infants or young children. There is a need for a food that is orally tolerant to allergens. There is a need for a milk oligosaccharide component having a very low protein which can be in the form of a slurry or powder without the need for any carrier to be added to the infant formula. SUMMARY OF THE INVENTION The present invention relates to a sugar-supplying mixture derived from cow's milk, which comprises: a. a soluble sugar-collecting group comprising a soluble oligosaccharide fraction (bovine milk oligosaccharide or CMOS) found in cow's milk; b. a β-galactooligosaccharide formed by β-galactosidase on lactose present in bovine milk oligosaccharide and, if necessary, also in other cow's milk, the mixture having less than 5% w/v, preferably less than 3% of the total monosaccharide content 'and less than 10, preferably less than 3 lactose: total oligosaccharide ratio. The mixture has the following monosaccharide, disaccharide and oligosaccharide composition as a percentage of dry matter: c. 30 to 60% lactose d. 0.5 to 2.5% glucose e. 0.5 to 2.5% galactose f. 20 to 50 % oligosaccharide and β-semi-milk sugar g. 0.2 to 2% sialic galactose. Another aspect of the invention is a method of making an OS mixture. The steps include: 164518.doc •10· 201249349 a) Concentration of the deproteinized milk material to 50 to 75% of total solids (TS); b) subjecting the concentrated milk material to a lactose removal step to produce lactose having less than 100: a liquid of oligosaccharide ratio; c) purifying the liquid as needed; d) treating the liquid to be purified as needed by using β-galactosidase to produce a liquid comprising β-semi-milk sugar; e) as needed, for example The liquid is demineralized by a weak cation column and optionally a mixed bed column and/or an anion exchange column; f) performing a nanofiltration step which can be carried out after or after the optional demineralization step It must be carried out after or at the same time as being treated with his galactosidase. In another aspect, the invention provides a product obtainable by the method of the invention. The OS mixture of the invention has advantageous properties. When the mixture has a moisture content of 2, it has a glass transition temperature (Tg) in the range of -85t. This physical property allows the mixture to be easily spray dried without agglomeration or adhesion. Moreover, compared to the (10) mixture of the (10) mixture of the non-nano materials, the mixture of the 〇s of the present invention is less inclined & is less prone to the Maillard reaction (the magical Uard react-. Better physical stability 0 ^ The OS mixture of the present invention can be incorporated into a nutritional composition formulation, an infant formula, a baby food, an infant, a baby formula, a larger formula or a growing milk, preferably an initial Infant Formulations Accordingly, this nutritional composition is an object of the present invention. 164518.doc 201249349 Another aspect of the present invention resides in the use of a nutritional composition comprising a mixture of the present invention for enhancing immune protection and/or Or reduce the risk of infection and / or reduce the impact of food allergies and related food allergies on health. [Embodiment] Definition: The following terms used herein have the following meanings. The term "baby" means a child less than 12 months. "Child" means a child between the ages of 1 and 3. The "infant formula" means a specific nutritional use intended for a newborn baby of 4 to 6 months old. Its food to meet the nutritional needs of these infants (22 December 2006 concerning the larger infant formula and infant formula of

European Commission Directive 2006/141/EC Article 2). The term "larger infant formula" means a food intended for use in a particular nutritional use of an infant over 4 months of age and as a major liquid element in a progressively diverse diet of such infants. The "initial infant formula" indicates food intended for the specific nutritional use of infants during the first 4 months of life. The term "child food" means food intended for a particular nutritional use of an infant during the first trimester. The term "infant sputum composition" means a food intended for a particular nutritional use in infants during the first year of life. The term "growth milk" means a lotion-based beverage suitable for the specific nutritional needs of a child. The term "increased oral tolerance to allergens" means reduced sensitivity to allergens when orally administered 164518.doc 201249349. The term "nutritional composition" means a composition that provides nutrition to an individual. The nutritional composition is usually administered orally or intravenously, and it usually comprises a source of lipids or fats and a source of protein. The term "synthetic composition" means a composition obtained by a chemical and/or biological (e.g., enzymatic) route which is chemically identical to a mixture naturally occurring in mammalian milk. The term "hypoallergenic composition" means a composition which is less likely to cause an allergic reaction. The term "oligosaccharide" means a sugar polymer comprising a small amount (generally 2 to 1 unit) of a monosaccharide component. The term "sialylated" refers to sugar collection with sialic acid residues. The term "probiotic-promoting" means that the host's non-digestible carbohydrates are advantageously affected by selectively stimulating the growth and/or activity of beneficial bacteria such as bifidobacteria in the human colon (Gibson GR, Roberfroid MB. Dietary modulation). Of the human colonic microbiota: introducing the concept of prebiotics. J. Nutr. 1995; 125: 1401-12). The term "probiotic" means a component of a microbial cell preparation or microbial cell that has a beneficial effect on the health or well-being of the host. (Salminen S, Ouwehand A., Benno Y. et al., "Probiotics: how should they be defined" Trends Food Sci. Technol· 1999: 10 107-10). 164518.doc 201249349 Allergy is an allergy that is detected by a doctor and can be treated intermittently or in a more persistent manner. "Food allergies" are allergies associated with nutritional compositions. The term "oligosaccharide distribution" means the identity of an oligosaccharide population. All percentages are by weight unless otherwise stated. Oligosaccharides: Oligosaccharides (os) are defined herein as those naturally occurring in animal milk and having a degree of polymerization (DP) of 3 to 20. "The oligosaccharides are soluble in milk. Unless otherwise stated, in the text All further cited oligosaccharides refer to soluble (soluble in milk) oligosaccharides. The present invention provides a sugar-supplying mixture derived from cow's milk wherein the mixture has a lactose (DP = 2): oligosaccharide ratio of less than 20' is less than, and preferably between 1.25 and 8. This corresponds to a 4 to 200-fold decrease in lactose content in the oligosaccharide mixture, which is equivalent to an increase of 4 to 200 times the ratio between oligosaccharide and lactose. Thus, the mixture has the same oligosaccharide distribution as found in the milk from which the oligosaccharide (i.e., its source) is obtained, but the oligosaccharide is at a higher concentration. The oligosaccharide mixture of the present invention also comprises a galacto-oligosaccharide (G〇s) derived from the sero-galactosidase mainly for lactose and, if desired, to a much lower extent, in the presence of milk. Some soluble oligosaccharides (bovine milk oligosaccharides or CM〇S). Thus, in one embodiment, the sputum-galacto-oligosaccharide is derived from the action of the galactosidase on lactose and bovine milk oligosaccharides, and the enzyme is added during the preparation of the sugar-supplying mixture. The inventors have not yet accurately determined the type of enzyme and the extent of action that contribute to CM〇S. Thus, the relative proportions of the different oligosaccharides in the oligobiliary complex of the present invention may differ from the source of the milk from which they provide the mixture. The non-galactosidase has dual activities: it decomposes lactose into monosaccharides, galactose and glucose 'and the second' by transferase activity, which catalyzes the formation of subsequent galacogenic sugar (GOS). Such oligosaccharides are composed of glucose and galactose monomers, 164518.doc

S 201249349 DP is 3 to 1 〇 and is known to have beneficial bacteria-promoting activity. There are more than 2 different 〇(10) constitutive structures in the oligosaccharide mixture of the present invention. The oligosaccharide distribution can be characterized by HPLC, mass spectrometry and other methods. Roots The preferred HPLC method for the presence of the sample is the extraction in water. By reacting with 2 o-aminobenzamide, by forming a Schiff test--, the light-extracted extraction of 0Se, followed by reduction of the double bond with the cyano-halogenation reaction to produce a stable oligo Glycosyl benzamide (〇8_8) derivative. The labeled extract is diluted with acetonitrile and then injected into an HPLC-fluorimeter equipped with a trap column. Separation was carried out in μιη, 4 6χΐ5〇mm column, and the labeled 0S was detected in a fluorescent meter under 33〇11111, £11142〇11111. By using maltotriose as an external standard and using Khumbu III The sugar is used as an internal standard to measure the reaction of 〇s_2ab, and the quantification of different OS is carried out. Figure 1 shows the total soluble oligosaccharide fraction in the three consecutive steps in the preparation of the mixture of the 丨8 mixture as determined by the above HPLC method. (1) 〇s powder after partial removal of lactose and demineralization by crystallization, (2) hydrolysis by p_galactosidase to produce GOS, and (3) after nanofiltration (corresponding to Example 2) Nano OS-GOS). This is the sugar distribution. So 'can be in the same drawing ratio Three chromatograms were viewed under the ruler, samples (2) 〇s-GOS and (3) diluted 20 and 10 times by NE-filtered by nanofiltration. The list of oligos identified from HPLC data is listed in Table 1. Not all sugar collections have been identified, but the distributions in the three chromatograms are extremely similar. This confirms that the CMOS population of the original milk is indeed present in the 〇s mixture of the present invention. The inventors have noted the 3'SL peak (corresponding to Sialyl oligosaccharide 3, sialic galactose) is mainly visible in chromatogram (1) rather than chromatograms (2) and (3). It is believed to be hydrolyzed in CMOS by 164518.doc 201249349 β-galactose (tetra) Step (4), which results in the retention time of (10) close to (10), and this will affect the separation of the peaks. The inventors have confirmed that the peaks at 32.29 minutes (1) and 32 2 minutes (7) are not 3'SL, but represent G. 〇s structure (data not shown). Because of the dilution factor applied to CMOS-GOS samples, in the chromatogram (1) with sialyl oligosaccharide 6'L (Neu5Ac(a2-6)Gal(pi-4)Glc The relevant peaks are no longer visible in the chromatogram (2). Table 1 (1) OS powder derived from crystallization and demineralization No. retention time min Peak name concentration g/lOOg 1 21.68 Maltose n*a. 2 22.51 Hex2 0.0489 3 23.00 Hex2 0.0330 4 23.59 Lactose n*a. 5 25.32 Hex2 1.0309 6 28.02 Laminaria triose (int· std) 0.0000 7 30.12 HexNAc-Hex2 0.1950 8 31.36 Hex3 0.0382 9 31.94 Hex3 1.0213 10 32.80 3'SL 0.2843 11 33.47 Hex3 0.0072 12 34.03 Hex3 0.1100 13 34.58 Hex3 0.0126 14 34.73 Hex3 0.0058 15 35.73 6'SL 0.0781 16 44.00 Hex6 0.0072 -16 · 164518.doc 201249349 (2)Using β-half Os-GOS after lactosidase treatment retention time peak concentration min g/lOOg 1 22.04 Hex2 0.2134 2 22.51 nana 3 23.22 lactose na 4 24.88 Hex2 0.6273 5 25.11 Hex2 0.7422 7 29.67 HexNAc-Hex2 0.0318 8 30.15 Hex3 0.2734 9 30.9 Hex3 0.0403 10 31.49 Hex3 0.5278 11 31.81 Hex3 0.1499 12 32.29 GOS 0.1065 13 32.69 Hex3 0.0966 14 33.02 Hex3 0.3952 15 33.57 Hex3 2.2039 16 34.15 Hex3 0.1860 17 35.27 Hex4 0.0136 18 36.53 Hex4 0.0206 19 37.85 Hex4 0.1200 20 38.54 Hex4 0.1499 21 39.08 Hex4 0.0667 22 40.5 Hex4 0.4755 23 45.94 Hex6 0.0812 164518.doc 17 20 1249349 (3) OS-GOS finally filtered by nanometer No. Retention time peak name concentration min g/lOOg 1 21.89 Maltose na 2 22.38 Hex2 0.4295 3 23.12 Lactose na 4 24.74 Hex2 1.3178 5 24.92 Hex2 1.5554 7 29.52 HexNAc-Hex2 0.2429 8 30.00 Hex3 2.3388 9 30.74 Hex3 0.1065 10 31.34 Hex3 4.4451 11 32.2 GOS 0.4389 12 32.55 Hex3 0.2517 13 32.89 Hex3 1.6449 14 33.44 Hex3 17.2782 15 34.01 Hex3 0.7043 16 35.18 6'SL 0.1053 17 35.67 Hex4 0.0206 18 36.39 Hex4 0.2057 19 37.71 Hex4 0.9629 20 38.42 Hex4 1.2078 21 38.94 Hex4 0.7056 22 39.7 Hex5 0.4053 23 40.37 Hex5 4.5556 24 43.39 Hex6 0.0828 25 43.68 Hex6 0.1968 26 44.41 Hex6 0.1536 27 45.82 Hex6 0.3741

Hex = hexose Hex 2-6 = number of hexoses equivalent to disaccharide, tri, tetra, penta, hexasaccharide 164518.doc • 18 - 201249349 The os mixture of the invention has a total monodomain of less than 5% w/v The content w/v' is preferably less than 3%. The mixture can be incorporated into the mouth of an infant or adult A _ mouth and confers probiotic, immunomodulatory and protective effects. Although the sugar-supplying mixture of the present invention is derived from cow's milk, milk can also be obtained from any type of animal, specifically cow (milk), goat, buffalo, horse, camel or sheep. '"' A method for preparing the oligosaccharide mixture. Starting material: The starting material in the method for preparing the oligo-mixed mixture of the present invention is a deproteinized milk material, such as milk from which protein has been removed, and whey protein removed. Whey or any prepared or modified whey material. Such materials include acid whey and sweet whey. Preferred starting materials are milk ultrafiltration permeate and whey ultrafiltration permeate. Alternatively, the starting material may be Reconstituted powder, such as powdered ultrafiltration permeate. Because the protein causes undesirable Maillard reactions and browning during concentration, the starting material must be deproteinized. It can be by any known means (eg acid precipitation, Heat treatment, ion exchange), the starting material is deproteinized. Preferably, however, the protein can be removed by ultrafiltration which also removes the lipid from the starting material. The pH of the starting material can be between 3. Between 7.5 and 5, but a pH range of 5 to 6 is preferred to prevent oligosaccharide hydrolysis, such as the sialic acid galactose, and to help reduce browning. a) Thickness of the starting material 164518.doc 201249349 Eight temperatures are not increased to the extent that sugar will be hydrolyzed (eg, desialylated), and the protein is concentrated to 5 to 75% total solids (TS) by any known method, preferably § 55 to 60 〇/〇 TS. It is preferably concentrated at a temperature of 50 to 90 ° C, more preferably 50 to 75 ° C. Evaporation is a preferred technique, which can be carried out at a pressure of 80 to 200 mbar. In this method, the temperature does not rise above 60 C, which ensures that the sugar collection is not adversely affected. Or, if the starting material is a powder, it can be concentrated by appropriate rehydration of the powder. Appropriate degree b) Preferably, the removal of lactose is carried out by crystallization and removal of lactose crystals. The step of removing the lactose can be carried out in a concentrated starting material, for example with or without seeding. The concentrated material is cooled to crystallize the lactose. Next, the lactose crystals are removed by any known method (e.g., centrifugation, filtration, and/or decantation). An alternative method of isolating lactose from vinegar is to utilize differential solubility. Spray drying and then adding water to change The lactose is crystallized. / The liquid obtained is highly rich in oligosaccharides, and the oligosaccharide: lactose is 2 to 2 times higher than the oligosaccharide-lactose ratio found in the milk from which the liquid is derived. The re-illusionation and the step of transferring the lactose can be repeated as needed. The final ratio of lactose:oligosaccharide is: 250, preferably less than 125, more preferably less than ι〇〇, In particular, less than 20, preferably less than 1 〇. ° This step can be carried out according to known methods. C) Purification of the liquid This step is optional and can be known by anyone skilled in the art. 164518.doc -20- 201249349 Method (eg centrifugation). d) Treatment of deproteinized liquid with P-galactosidase to produce a liquid comprising p-galactooligosaccharide (GOS) so 'can be concentrated in the milk substance ( The liquid is treated before step (a)) and/or after the lactose removal step (step (b)). This preferably takes place after the completion of the lactose removal step. The beta-galactosidase used is preferably derived from Aspergillus oryzae (hpergi/Zw. This enzyme may be from Aman®, Japan as lactase F or from Enzyme Development Corporation (EDC), New Y〇rk, USA as Enzeco The fungal lactase concentrate is commercially available, and the enzyme activity measured according to the FCCIv method can be between 1, 〇〇〇 and 30,000 u/kg of lactose. The pH can be in the range of 3 to 7, between 4 and 7 〇t. Enzymatic treatment of the starting material of lactose ingredient containing 5 to 70 g/1 〇〇g total solids (TS) at a temperature between 〇·5 to 1 g/kg of sugar-supplying mixture Preferably, the dry matter per kilogram of oligosaccharide mixture uses about 15 grams of enzyme and the incubation time is between 20 and 7 (TC is between 1 and 8 hours. The enzyme is inactivated by heating after use. After treatment with 0.5 to 6 mg of β-galactose/g TS having a TS concentration of 25 to 50% and a liquid of about 15 to 40% lactose, the resulting solution may comprise about 1 to 4% oligosaccharide, about 9 to 25% GOS, about 15 to 30% lactose, about 5 to 15% galactose and about 2 to 15 angstroms/glucose. Oligosaccharide: galactooligosaccharide ((}〇3) is better at 1: 2 to 1:25 Within the circumference, preferably 1:5 to 1:2. e) Demineralization step: This step is optional. It can be by any known method (such as ion exchange, electrodialysis, ultrafiltration or the like). A combination of methods) demineralizing a liquid. Example 164518.doc 201249349 If the substance can be passed through a weakly cation column and a mixed bed column and/or an anion column, followed by electrodialysis or nanofiltration, which can be neutral The demineralization step is carried out at acidic pH. It can be carried out before or after the hydrolysis step (d). It can also be carried out partially before hydrolysis and after hydrolysis. 0 nm filtration step This step is a method of the invention It is necessary. Liquid nanofiltration can remove monovalent cations and anions and monosaccharides. It is necessary to remove monosaccharides from the mixture because (1) it does not have beneficial bacteria-promoting activity and (π) when it is a beneficial bacteria-promoting ingredient. Used in the preparation of infant formulas, which induce undesired reactions with proteins. For example, 'monosaccharides can cause the reactivity of Maillard reactions, in which the amino acid residues of proteins are blocked, thus reducing the nutrition of infant formulas. Amino acid is an essential amino acid that must be provided in the diet, and blocked lysine cannot be utilized by the body. Therefore, it is necessary to reduce the amount of Maillard reaction occurring during the manufacture of the infant formula. The lower the monosaccharide concentration, the higher the oligosaccharide concentration. Unexpectedly, the inventors have also discovered that a lower monosaccharide content allows the oligosaccharide mixture to be dried without the addition of a carrier. CMOS-GOS mixtures typically require 25 to 35%. A carrier such as maltodextrin or lactose to form a powder. The carrier is not required to be a powder in the manufacture of powders and is a great advantage in the preparation and use of the CMOS-GOS mixture of the present invention. After filtration through the nanoparticle, the resulting liquid should have a monosaccharide concentration of less than 5% w/v, preferably less than 3% w/v, and preferably less than 20, preferably less than 1 Å of lactose: oligosaccharide ratio . Nanofiltration was carried out by passing the liquid through a membrane having a pore size small enough to retain the oligosaccharide but large enough to allow the single 164518.doc-22-201249349 sugar to pass. For this purpose, commercially available crucibles having a range of from 2 Torr to 1 Dalton can be used as known in the art. A non-limiting example of a membrane that can be used, inter alia, as

Nadyr DS NP030 and MMS-LD-3838. The nanosteps can be carried out before or after the optional demineralization step. However, the 'nano pass' step can also be used to demineralize liquids. This filtration step must be carried out after or at the same time as the hydrolysis step. ^Nano = 遽 The step is carried out simultaneously with the hydrolysis step, and P_galactose (4) is freely dissolved or fixed in the retentate container on the nanofiltration membrane. / Nano can be combined with the dialysis (four) step to achieve the desired single enzyme content. 'Using an equal volume of demineralized water to refine the retentate from the nanofiltration several times during the diafiltration step' then re-pass the nano Filter membrane. In general, the diafiltration step is repeated using 1 to 5 parts, preferably 3 to 5 parts by volume of water. After the nanofiltration step,? The retentate may comprise about 15 to 30 〇 / 〇 TS ' of which 40 to 6 % is lactose, 〇 5 to 25% is glucose, 05 to 2 ^ is galactose and about 10 to 50% is for recruitment. The ratio of oligo-β•galactooligosaccharides (g〇s) does not change significantly due to the nanofiltration step. The resulting nano-transition liquid system is in the form of a slurry and it can be used directly in the form of a slurry or it can be concentrated by evaporation to 6 to 85%, preferably 74 to 85% TS and more preferably 74. To 嶋ts to make it stable to store, or it can then be dried (for example by spray drying) to produce a powder. Mouth mist drying methods known in the art of I can be used. Preferably, less than 5% of the carrier, more preferably 0% of the carrier, may be added during the drying step. 164518.doc -23· 201249349

Carrier (generally 15 to 30%) so that the mixture can be spray dried. The powder may adhere to spray drying and thus 'in order to form a powder, generally glucan, maltodextrin, or arabic will occasionally agglomerate. And adhesion. For example, the wall on the tower' thus blocks the dryer. Due to the addition of carrier molecules such as proteins, glucosamine, glutinous starch and glucose or lactose syrup. Thus, the increase in Tg observed for the nanofiltration CMOS-GOS mixture of the present invention allows for the successful successful spray drying of such mixtures without agglomeration in the absence of such carriers. In addition, the increase in Tg for a given moisture content has the advantage of improving the physical stability of the powder during storage. The OS mixture of the present invention therefore has the key advantage of being easily spray dried without the addition of a carrier. Because &, the amount of oligobiliary in the final infant formula can be increased without introducing an undesirable amount of carrier molecule. This creates the flexibility to deliver ingredients to the plant in liquid or powder type I (depending on the plant's configuration). The powder preparation process can be simplified. The inventors have tested a similar I64518.doc •24·201249349 nanofiltration method for removing monosaccharides for commercially available galactooligosaccharide mixtures. The economy. These tests indicate (data not shown)

To spray dry successfully. The enzyme-enzyme mixture of the present invention may be in the form of a powder additionally comprising less than 5% of the carrier molecules or even no carrier molecules. Moreover, the oligosaccharide of the present invention causes a significant decrease in the reactivity of the Maillard reaction. It is confirmed in Fig. 3 (Example 3) that the ring shows the liquid shrinkage and powder of the present invention and the fraction containing (4) the nano-filtered milk oligosaccharide and (9) the commercially available semi-milk sugar component (by Fries Und Campina) The concentration of blocked effluent acid compared to the Vivina 丨8 bovine milk oligosaccharide mixture supplied. The formulations tested are described in Table 2 (Example 3). The mixture without nano-passing was finally spray dried. In the infant formula, extremely significant off-amino acid capping is caused. Conversely, if the CMOS-GOS fraction has been nanofiltered prior to addition to the formulation (whether added, wet or dry), the amine acid capping is much lower. This is because the removal of a large amount of monosaccharides causes a Maillard reaction. After spray drying, the resulting powder contains about 5% lactose and the remainder is an oligosaccharide mixture (about 20 to 40%, including saliva). Acidified oligosaccharide), less than 3% monosaccharide (such as glucose and galactose), about 10% non-protein nitrogen-containing compound, 3% residual protein, and some residual salts. Use of the OS mixture of the present invention: In the present invention In a preferred aspect, it will be The oligosaccharide mixture is incorporated into a food product. Within the scope of the present invention, the term "food" is intended to cover any edible material. Therefore, it may be a food intended for human consumption, specifically a baby 164518.doc • 25· 201249349 Formula, dehydrated milk powder 'which includes a mixture of growing milk or mash. The infant formula can be prepared in any suitable manner. For example, by blending together a suitable source of protein, any carbohydrate other than lactose and knowing Fat source 'Manufacture baby formula'»Add emulsifier if needed. Vitamins and minerals can be added at this time, but usually added later to avoid thermal degradation. Any lipophilic vitamins, emulsifiers, etc. can be dissolved in fat source. The 're-blending' can then be mixed with water (preferably water that has been subjected to reverse osmosis) to form a liquid mixture. The liquid mixture is then heat treated to reduce the amount of bacteria. For example, the liquid mixture is rapidly heated to about 80. 〇 to a temperature in the range of about i°°°°C for about 5 seconds to about 5 minutes. This can be done by steam injection or by heat exchange. The device (for example, a plate heat exchanger) is carried out. The liquid mixture can then be cooled to about 60 ° C to about 85 ° C by, for example, rapid cooling. Then, for example, it can be divided into two stages (in the first stage at about 7) The liquid mixture is homogenized from MPa to about 40 MPa and from about 2 MPa to about M Μρ & in the second stage. Then, the homogenized mixture is further cooled to add any heat sensitive components such as vitamins and minerals. The pH and total solids (TS) content can be conveniently standardized at this point. The homogenized mixture is transferred to a suitable drying apparatus (such as a spray dryer or freeze dryer) and converted to a powder. The powder should have a weight of less than about 5 % moisture content. The oligosaccharide mixture of the invention may be added to an infant formula or other food product by wet mixing or by dry mixing at an appropriate stage in the manufacturing process, but preferably prior to heat treatment and evaporation. Immediately added by wet mixing. 1645I8.doc •26· 201249349 However, it is clear to those skilled in the art that the amount of carbohydrates in the infant formula needs to be adjusted in consideration of the other carbohydrates provided in the oligosaccharide mixture. The final concentration of the oligosaccharide mixture in the infant or infant food or formula is preferably between 2 and 20 g/丨, more preferably about 5 g/i of the edible formulation. However, such levels should not be considered limiting and should be adjusted to the target population (eg based on the weight and age or health of the child or infant). Preferably, the formulation comprising the oligosaccharide mixture of the invention is fed to the infant each time it is fed. Alternatively, the oligosaccharide mixture can be added to the infant or adult food by dry mixing. The mixture may be added to the infant or infant formula at a concentration of from about 1 to 15 g oligosaccharide per gram of dry formulation without causing an abnormally high lactose content in the formulation' however, such amounts should not be considered limiting. It should be adjusted according to the target population (for example, based on the weight and age of young children or babies, or the health of a specific group of people). Although it is preferred to supplement the specific nutrient and nourishment specific to the infant or to advantageously supplement no specific target or, for example, the oligosaccharide mixture of the present invention. Such cheeses, fermented milk, milk-based fermented products, products or dairy-based products. Nutritional composition Foods that target children's nutrition, but target adults. The health care products for incorporation into the elderly may include, in particular, milk, yoghurt, milk ice cream, fermented probiotics based on the present invention, and more preferably the digestive tract or intestinal compound. Sugar mixture. It preferably also includes, wherein the stimulating mixture promotes the probiotics to grow or multiply. Infant formula, infant food The nutritional composition is preferably an initial infant formula 164518.doc 27· 201249349 The product 'baby cereal composition, larger infant formula or growing milk, preferably an initial infant formula. The invention will now be further described with reference to the following examples. Example 1 Method for preparing a CMOS-GOS mixture of the invention: 207,000 kg of whey ultrafiltration permeate was pre-concentrated to 29% (w/w) total solids (TS) 'pasteurized at about 75 ° C for about 30 The seconds were then concentrated by evaporation at 6 ° C to reach 58 ° /. TS (w/w). The liquid was dispensed into 3 crystallizers and each crystallizer was cooled at a rate of 2 / hr for 24 hours to crystallize the lactose. The crystallized lactose is washed and then removed by a wringer. The remaining liquid is purified by a decanter. In a manner known per se, 114,000 kg of liquid demineralization with 23% TS obtained from the purifier, resulting in 109,000 kg with 90% of 14.4% TS, by a combination of a weak cation column and a mixed bed column Demineralized liquid. 5,400 kg of this demineralized oligosaccharide mixture was concentrated to 52% TS by evaporation' and then heated to 6 °C in a standard vessel at a pH of 5.5 to 6.5. The concentration of lactose, glucose 'galactose, galactooligosaccharides and other oligosaccharides in the mixture was measured. 0.5 g Enzeco fungal lactase concentrate (Enzyme Development Corporation, New Y〇rk, USA) was added per kg of TS and the mixture was maintained at 60 ° C until a 15% glucose concentration was obtained. The temperature was then raised to 90 °c by direct steam injection for 3 seconds to inactivate the enzyme. A mixture of 1,500 kg of 52.4% TS hydrolyzed oligosaccharide was diluted with soft water to obtain a 25% TS solution of 645, 8d 〇C -28-sr 201249349. The solution was normalized at pH 2.5 to 6.5 and at 36 ° C to 25 ° C and 25-30 bar at 36.25 m2 of MMS-LD-3838 membrane with a nominal molecular weight retention of 300 Daltons. The rice in the rice filter line is too low. The retentate from the nanotransition was filtered by dialysis against 1 to 5 times its volume of soft water. The nano-transitioned G〇s-containing oligosaccharide mixture with 22% TS was heat treated at 108 C for 5 seconds and evaporated to 55 〇/0 TS. The concentrate was spray dried in an Egron column using conditions known in the art. In a second variant of the method, the same nano-filtered mixture of oligosaccharides containing G 〇s is heat treated at 108 ° C for 5 seconds, but evaporated to 74% TS to achieve a water activity of less than 0.86 (aw) 'It gives the concentrated product a storage stability of at least 3 months. The viscosity of a concentrated product with 74% TS and the ease of use of the evaporator to achieve this high concentration level indicate that higher concentrations can be achieved. The concentrations of lactose, glucose, galactose, galactooligosaccharide and other sugar-supplying sugars in the mixture were again measured and the results are shown in Example 2 below. Example 2 This example shows the total solid oligosaccharide, monosaccharide, ash substance 'lactic acid, citric acid change' during the process of the present invention in Example 1. CMOS corresponds to the mixture before hydrolysis step d) 'CMOS-GOS corresponds to hydrolysis The mixture after step d), nano CMOS-GOS corresponds to the mixture after nanofiltration step f). These samples were also used to generate the data in the HPLC chromatograms (1), (2) and (3) and Table 1 of Figure 1. I64518.doc -29- 201249349 w/v % CMOS CMOS-GOS Nano CMOS-GOS Total Solids 98.1 52.4 97.2 Gray Matter 1.83 - 2.46 Lactic Acid - 0.52 Citric Acid 0.34 - 0.50 Lactose 82.7 44.05 51.14 Glucose 1.2 13.6 1.52 Galactose 4.08 6.59 1.01 OS+GOS 1.3 17.87 36.20 Sialyl lactose 0.29 0.29 0.55 Example 3 Reduced reactivity of Maillard reaction after nanofiltration: Formulation of the infant formula of Table 2 below and the reactivity of the Maillard reaction in the table MS W indicates modified sweet whey, which is sweet whey from which casein glycopeptide (CGMP) has been removed. Infant formulations of the CMOS-GOS blend of the present invention added as a liquid concentrate or in the form of a dry powder were tested as compared to infant formulas including non-nanofiltered CMOS and commercially available GOS (Vivinal® GOS). Standard reaction conditions were used to test the reactivity of the Maillard reaction. The results are shown in Figure 3. The sugar-supplying mixture of the present invention, which is added to the infant formula in liquid form or in powder form, results in a significant decrease in the reactivity of the Maillard reaction in the final spray-dried product. 164518.doc •30·

S 201249349 Table 2 Nano CMOS-GOS Liquid Nano CMOS-GOS Dry Matter CMOS Vivinal® GOS Wet Mix Weight % Wet Mix Weight % Wet Mix Weight % Oil Mix 25 Oil Mix 25 Oil Mixture 25 MSW 30 MSW 45 MSW 15 nm CMOS-GOS 16 CMOS 20 Vivinal® GOS3 7.9 Skim Milk Skim 10 Skim Milk 10 LC-PUFAs 1 LC-PUFAs 1.3 LC-PUFAs 1.3 Salt Salts 1.5 Vitamin Premix Vitamin Premix Vitamin Premix 0.3 Lecithin Lecithin Lecithin 0.2 Dry Mixture Dry Mixture Lactose 14 Nano CMOS-GOS 2 16 Lactose 14 Trace Element Premix 0.3 Trace Element Premix 0.3 Trace Element Premix 0.3 Probiotics 0.1 Probiotics 0.1 Probiotics 0.1 1 Long-chain polyunsaturated fatty acids 2 Example 1 3 Vivinal® GOS supplied by Friesland Campina (NL) Example 4 An example of an infant formula containing an oligosaccharide mixture according to the present invention is given below. This example is based on the addition of a premium whey-based infant formula (from Nestis) of the 164518.doc 31 201249349 specific oligosaccharide of the present invention at a level as indicated below. Nutrients per 100 kcal per liter of energy (kcal) 100 670 Protein (g) 1.83 12.3 Fat (g) 5.3 35.7 Linoleic acid (g) 0.79 5.3 α-linolenic acid (mg) 101 675 lactose (g) 11.2 74.7 Minerals (g) 0.37 2.5 Na (mg) 23 150 K (mg) 89 590 Cl (mg) 64 430 Ca (mg) 62 410 P (mg) 31 210 Mg (mg) 7 50 Mn(pg) 8 50 Se (pg 2 13 Vitamin A (μδ RE*) 105 700 Vitamin Dbg) 1.5 10 Vitamin E (mgTE") 0.8 5.4 Vitamin ΚΙ (μ8) 8 54 Vitamin C (mg) 10 67 164518.doc -32- 201249349 Vitamin B1 (mg 0.07 0.47 Vitamin B2 (mg) 0.15 1.0 Niacin (mg) 1 6.7 Vitamin B6 (mg) 0.075 0.50 Folic acid (pg) 9 60 Pantothenic acid (mg) 0.45 3 Vitamin Β12 (μ8) 0.3 2 Biotin (pg) 2.2 15 Bile test (mg) 10 67 Fe (mg) 1.2 8 i (pg) 15 100 Cu (mg) 0.06 0.4 Zn (mg) 0.75 5 oligosaccharide CMOS + GOS nano ***, λ (g) 2.07 13.9 * RE The retinol equivalent **TE is the tocopherol equivalent * * * The oligosaccharide mixture of the present invention [Simple description] @ 1 : Determination of bovine milk oligosaccharide (CMOS) and bovine milk oligosaccharide/β-half-oligosaccharide by HPLC Sugar (CMOS-GOS) mixture in the treatment period of Example 1. Total soluble sugars raised points of the three fractions. The HPLC chromatogram corresponds to data obtained from the following samples: (1) OS powder after removal of part of lactose and demineralization by crystallization (2) production of GOS by hydrolysis using β-galactosidase The resulting OS-GOS (3) nano-filtered nano-filtered OS-GOS (corresponding to the nano OS-GOS in Example 2). Thus, the 6.5518.doc •33-201249349 chromatogram can be viewed at approximately the same scale, and the samples (2) OS-GOS and (3) are diluted 20 and 10 times by the NE-GOS filtered by nanometer. Figure 2: Measured powder of CMOS-GOS containing 30% maltodextrin carrier (black), CMOS-GOS (dashed line) containing 30% lactose, and nano-filtered CMOS-GOS mixture according to the present invention ( Gray) The glass transition temperature (Tg) of the powder. The water content of the sample was determined by the Karl Fischer method. Since the sample cannot be dried to the desired humidity level without the addition of a carrier, it is not possible to obtain a CMOS-GOS glass transition temperature value without a carrier. However, the data seems to indicate that, under no carrier, the Tg value of the CMOS-GOS without nanofiltration is close to 35 to 4 (TC. Tg measurement by differential scanning calorimetry (DSC). Circle 3: The percentage of blocked amide acid (In Maillard reaction) was determined in the three infant formulas detailed in 2. Measurements were made before (grey) and after (black) spray drying. Spray drying was carried out at 92t. In the table, the "CMOS Vivinal" block indicates the addition of a nano-filtered GOS fraction (Vivinal® GOS from FrieslandCampina) and a liquid concentrate (gray) and powder (black) baby without a nano-passed CMOS fraction. Data for the formulation (see Table 2). The column "Nano-CMOS-GOS Wet Additive" indicates the addition of an infant formula (liquid concentrate and powder) of the nanofiltration-filtered CMOS-GOS mixture according to the present invention in the form of a wet mixture. The data of "Nano-CMOS-GOS Dry Additive" column indicates the data of the infant formula added with the nano-filtered CMOS-GOS mixture according to the present invention in the form of a dry powder.

164S18.doc -34- S

Claims (1)

201249349 VII. Scope of application: 1. A mixture of oligosaccharides derived from cow's milk, comprising: a. a soluble oligosaccharide population comprising a soluble oligosaccharide fraction found in cow's milk, b. by β-galactoside A beta-semi-fragrance sugar formed by the action of lactose in the milk oligosaccharide, the mixture having a total monosaccharide content of less than 5% w/v and a lactose ratio of less than 1 :: total sugar-receiving ratio. 2. The sugar-supplying mixture of claim 1 wherein the mixture has a total monosaccharide content of less than 3% w/v. 3. The oligosaccharide mixture of claim 1 or 2 wherein the mixture has a total sugar ratio of less than 3: total ratio. 4. The oligosaccharide mixture of claim 1 or 2 wherein the mixture has the following monosaccharide, disaccharide and oligosaccharide percentages expressed as a percentage of dry matter: a. 30 to 60% lactose b. 0.5 to 2.5% glucose c. 5.5 to 2.5% galactose d. 20 to 50% oligosaccharide and β·galactooligosaccharide* e. 0.2 to 2% sialic galactose. 5. The oligosaccharide mixture of claim 1 or 2, wherein the mixture has a glass transition temperature in the range of from 7 Torr to 85 ° C when the mixture has a moisture content of 25%. 6. The oligosaccharide mixture of claim 1 or 2, wherein it is in powder form and further comprises less than 5% of carrier molecules. 164518.doc 201249349 7. The oligosaccharide mixture of claim 1 or 2, wherein the mixture is in the form of a concentrated slurry having 6 to 85% total solids (TS)', preferably greater than 74% TS. A nutritional composition comprising the ugly mixture of any one of claims 1 to 7. 9. The nutritional composition of claim 8 wherein it further comprises a probiotic, preferably wherein the oligosaccharide mixture promotes growth or reproduction of the probiotic in the digestive tract or intestine. 10. The nutritional composition according to item 8 or 9, which is an initial infant formula, an infant formula, a baby food, a baby babies composition, a larger infant formula or a creche, preferably an initial infant formula. 11. Use of a nutritional composition according to claim 8 or 9 for improving the immune protection and/or reducing the risk of infection and/or reducing the incidence of food allergies and the health effects of related food allergies. 12. A method of preparing an oligosaccharide mixture derived from cow's milk, comprising the steps of: a) concentrating the deproteinized milk material to 50 to 75% of total solids (TS); b) removing the concentrated milk material to remove lactose a step of producing lactose: a liquid having a oligobiliary ratio of less than 100; 0 purifying the liquid as needed; d) treating the optionally purified liquid with β-galactosidase to produce a liquid comprising β-semi-milk sugar; e) demineralizing, for example, by passing the liquid through a weak cation column and optionally a mixed bed column and/or an anion exchange column; f) performing a nanofiltration step 'which can be optionally demineralized Substance step S 164518.doc 201249349 before or after 'and must be carried out after treatment with β-galactosidase or simultaneously. 13. The method of claim 12, wherein the step comprises: a lactose crystallization step and a concentration step to remove lactose crystals, and to produce lactose: oligosaccharide ratio less than 100' 14. The method of claim 12 or 13, wherein the step of adjusting is ambiguous to obtain a total monosaccharide content of less than 5% w/v, preferably less than 3% w/v and lactose: oligosaccharide ratio less than 20' Preferably, it is less than a mixture of cerium. 15. The method of claim 12 or 13, wherein the method is adjusted to obtain a mixture of any one of claims 1 to 7. 16. The method of claim 12 or 13, wherein the deproteinized milk material is a milk ultrafiltration permeate or whey ultrafiltration permeate. 17. The method of claim 12 or 13, wherein the beta galactosidase is derived from M. oryzae. 18. The method of claim 12 or 13, wherein the nanofiltration step is combined with hydrazine to 5 diafiltration steps. 19. The method of claim 12 or 13, further comprising the step of spray drying the liquid product to produce a powder. 20. The method of claim 20 wherein less than 5% of the carrier, preferably 〇%, of the carrier is added to the mixture during the spray drying step. A product obtained by the method of any one of claims 12 to 2. I645l8.doc