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WO2025224241A1 - Préparation de galacto-oligosaccharide enrichie en sialyllactose - Google Patents

Préparation de galacto-oligosaccharide enrichie en sialyllactose

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Publication number
WO2025224241A1
WO2025224241A1 PCT/EP2025/061218 EP2025061218W WO2025224241A1 WO 2025224241 A1 WO2025224241 A1 WO 2025224241A1 EP 2025061218 W EP2025061218 W EP 2025061218W WO 2025224241 A1 WO2025224241 A1 WO 2025224241A1
Authority
WO
WIPO (PCT)
Prior art keywords
sialyllactose
lactose
process according
galacto
anion exchange
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2025/061218
Other languages
English (en)
Inventor
Linqiu Cao
Trinath PATHAPATI
Alfred Willy Bonte
Johannes Petronella Maria NIEDERER
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
FrieslandCampina Nederland BV
Original Assignee
FrieslandCampina Nederland BV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by FrieslandCampina Nederland BV filed Critical FrieslandCampina Nederland BV
Publication of WO2025224241A1 publication Critical patent/WO2025224241A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
    • A23C9/1206—Lactose hydrolysing enzymes, e.g. lactase, beta-galactosidase
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/14—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment
    • A23C9/142—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration
    • A23C9/1425—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration by ultrafiltration, microfiltration or diafiltration of whey, e.g. treatment of the UF permeate
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula

Definitions

  • the invention relates to the field of nutritional ingredients. More in particular, it relates to a method for producing a galacto-oligosaccharides (GOS) preparation with a relatively high content of oligosaccharides that naturally occur in milk.
  • GOS galacto-oligosaccharides
  • GOS Various physiological functions of GOS have been reported, including the capacity to stimulate the growth of bifidogenic bacteria in the gut, to support normal gut transit, to contribute to natural defenses and to enhance mineral absorption. GOS has received particular attention for their prebiotic effects that promote the growth of Bifidobacterium, Lactobacillus, and other enteric bacteria. Therefore, GOS is commonly used in formula feeding (infant formula, follow-on formula, and young child formula).
  • GOS Galacto-oligosaccharides
  • Typical GOS preparations mainly comprise di- to hexa-saccharides.
  • GOS is produced from lactose by a transglycosylation reaction with a betagalactosidase enzyme (enzyme class EC.3.2.1 .23).
  • Beta-galactosidase enzymes are produced in many microorganisms such as Bacillus circulans, Aspergillus oryzae, Kluyveromyces marxianus, Kluyveromyces fragilis, Sporobolomyces singularis, Papiliotrema terrestris, and Lactobacillus fermentum. Beta-galactosidases differ in their three-dimensional structures, resulting in stereo- and regioselectivity of the glycosidic bonds that are formed.
  • a fungal beta-galactosidase derived from Aspergillus predominantly produces (31 -6 bonds (thus resulting in a GOS preparation that predominantly comprises [31-6 bonds, which may be referred to as “6’- GOS”), while a bacterial beta-galactosidase derived from Bacillus predominantly produce [31-4 bonds (resulting in a GOS preparation that predominantly comprises (31 - 4 bonds, which may also be referred to as “4’-GOS”).
  • beta-galactosidase produced by B. circulans possesses particularly strong transgalactosylation activity.
  • GOS prepared by B. circulans beta-galactosidase is sold worldwide.
  • lactose The starting material for GOS production, lactose, is generally sourced from milk and obtained by crystallization from whey ultrafiltration permeate (herein referred to as “whey permeate”). This whey permeate results from ultrafiltration of whey.
  • the lactose concentration in this whey permeate is generally in the range 75-90 wt% on dry matter.
  • Whey permeate as such can also be used as the lactose source for GOS production. And so can delactosed whey permeate (conventionally referred to as DLP or OPL), i.e. the mother liquor from the lactose crystallization, as disclosed in WO 2006/087391 , WO 2018/020473, and WO 2020/141032.
  • DLP delactosed whey permeate
  • OPL the mother liquor from the lactose crystallization
  • the process disclosed in WO 2006/087391 involves the enzymatic treatment with a beta-galactosidase of a lactose source obtained by concentration, lactose removal (e.g. crystallization), and demineralization (by reverse osmosis, nanofiltration, or ion exchange) of whey permeate.
  • a beta-galactosidase of a lactose source obtained by concentration, lactose removal (e.g. crystallization), and demineralization (by reverse osmosis, nanofiltration, or ion exchange) of whey permeate.
  • WO 2020/141032 discloses GOS production starting from demineralized (e.g. by electrodialysis) delactosed whey permeate.
  • demineralized e.g. by electrodialysis
  • this document proposes to add calcium in order to precipitate the corresponding calcium salts.
  • WO 2018/020473 involves the precipitation of minerals from delactosed whey permeate by way of heat treatment, followed by ultrafiltration and nanofiltration.
  • whey permeate as the lactose source is that it allows valorization of one of the most abundant waste streams in the dairy industry.
  • this permeate contains a significant amount of oligosaccharides naturally present in milk, such as the sialyllactoses 3’-sialyllactose (3’-SL) and 6’- sialyllactose (6’-SL), thereby enabling the preparation of GOS containing such oligosaccharides.
  • oligosaccharides are also known to have significant health benefits by supporting resistance to pathogens, gut maturation, immune function, and cognitive development.
  • the commercial production of such sialyllactoses currently involves fermentation reactions with genetically engineered microorganisms.
  • GOS can be added to formula feeding in concentrations of 2.4-8.0 g/L, preferably 3.0-
  • the main sialyllactose in bovine milk can be added to formula feeding in concentrations up to about 0.3 g/L.
  • GOS preparation comprising, based on dry matter, 0.2-3.0 wt% sialyllactose, thereby allowing the direct introduction of this GOS preparation in formula feeding.
  • sialyllactose content be significantly higher than 3.0 wt%, direct introduction would run the risk of introducing more sialyllactose than allowed.
  • a disadvantage of delactosed whey permeate is its high mineral content, which requires extensive demineralization and concentration steps. Furthermore, the sialyllactose content of the final GOS is difficult to control.
  • a further disadvantage of using delactosed whey permeate is that it contains nitrogencontaining compounds and compounds resulting in color formation, which cannot be removed with a conventional anion exchange column without losing sialyllactose.
  • SL sialyllactose
  • the present invention relates to a process for the production of a galactooligosaccharide preparation with, based on dry matter, at least 35 wt% galactooligosaccharides other than lactose and a sialyllactose content in the range 0.2-3.0 wt%, comprising the steps of: i) providing a whey permeate with a lactose concentration in the range 75-90 wt% on dry matter, ii) removing mono- and multivalent ions from said whey permeate, thereby obtaining a demineralized whey permeate, iii) isolating sialyllactose by submitting the demineralised whey permeate to a macroporous or gel-type anion exchange resin and subsequently eluting the sialyllactose from said macroporous or gel-type anion exchange resin with an acidic salt solution, iv) subjecting the isolated sialyll
  • the process of the present invention starts with whey permeate, i.e. the lactose-rich ultrafiltration permeate remaining after protein concentration from whey.
  • Whey results from separating (skimmed) milk into a casein-rich and a whey proteinrich fraction; either by renneting (i.e. cheese making), acidification, or microfiltration.
  • Whey resulting from cheese making is referred to as cheese whey;
  • whey resulting from acidification is referred to as acid whey;
  • the permeate resulting from the microfiltration of milk is referred to as ideal whey.
  • Whey proteins are conventionally concentrated by submitting whey to ultrafiltration.
  • This ultrafiltration conventionally uses a membrane with a molecular weight cut-off (MWCO) in the range 5-10 kDa.
  • MWCO molecular weight cut-off
  • a major part of the water, lactose, sialyllactose, minerals, and vitamins passes the membrane and forms the whey permeate, whereas the proteins are retained by the membrane as a whey protein concentrate.
  • the whey permeate to be used in the process of the present invention can be a cheese whey permeate, an acid whey permeate, or an ideal whey permeate.
  • the whey permeate is a cheese whey permeate (CWP) or an acid whey permeate. Most preferably, it is a cheese whey permeate.
  • the lactose content of the whey permeate is in the range 75-90 wt%.
  • the sialyllactose content of whey permeate is generally around 0.1 -0.3 wt%, based on dry solids.
  • the whey permeate is submitted to one or more demineralization steps in order to remove monovalent and multivalent ions.
  • the main multivalent ions are calcium and phosphate ions.
  • This demineralisation can be conducted with electrodialysis, precipitation of the minerals, ion exchange, and/or nanofiltration.
  • the demineralisation step should be conducted in a way that prevents removal of significant amounts of sialyllactose from the whey permeate.
  • Demineralisation preferably results in a whey permeate that is demineralized to an ash content of about 0.1 -5.0 wt% , more preferably 0.1 -4.0 wt%, even more preferably 0.1- 3.0 wt%, more preferably 0.1 -2.0 wt%, and most preferably 0.1 -1 .0 wt.%, based on dry matter.
  • the demineralisation is performed by electrodialysis, thereby removing both monovalent and multivalent ions.
  • multivalent ions are replaced with monovalent ions in at least one cation exchange step in order to replace calcium ions with monovalent cations, such as sodium or potassium, and at least one anion exchange step in order to replace phosphate ions with monovalent anion, such as chloride, after which the monovalent ions can be removed by membrane filtration or further ion exchange steps.
  • monovalent ions such as sodium or potassium
  • anion exchange step in order to replace phosphate ions with monovalent anion, such as chloride
  • anion exchange step it should be ensured that this step does not result in significant removal of sialyllactose.
  • This can be achieved by performing the anion exchange with an anion exchange resin in the chloride form, preferably a geltype styrene-divinylbenzene or gel-type crosslinked acrylic anion exchange resin with tertiary amine functional groups, more preferably dimethylamine groups.
  • the ion exchange steps are preferably followed by a membrane filtration step using a membrane that permeates monovalent ions, such as sodium, potassium, and chloride, thereby further demineralizing and at the same time concentrating the whey permeate.
  • Sialyllactose should remain in the membrane filtration retentate.
  • the whey permeate is preferably concentrated to a dry matter content in the range 10-25 wt%, preferably 15-20 wt%.
  • any membrane suitable for desalination purposes may be used for this step.
  • a nanofiltration membrane with a molecular weight cut-off in the range 200-300 Da is used.
  • the membrane is a polypiperazine amide membrane. Said nanofiltration is preferably combined with diafiltration.
  • a membrane with a molecular weight cut-off in the range 400-2500, preferably 400-2000, more preferably 400-1500, even more preferably 400-1000, even more preferably 400-700, more preferably 400-600, and most preferably 400-500 Dalton, is used.
  • Such membrane not only results in demineralization, but additionally results in removal of part of the lactose, thereby increasing the sialyllactose/lactose ratio.
  • Such membrane filtration is preferably performed until the resulting retentate has a lactose content, based on dry matter, in the range 90-99.8 wt% and the sialyllactose content, relative to lactose, in the range 0.2-3.0 wt%.
  • This membrane filtration may be combined with diafiltration. Diafiltration serves in a further reduction of multivalent ions (e.g. phosphate) and non-protein nitrogen (NPN).
  • This membrane filtration is preferably performed at a temperature in the range 5-60°C, preferably 10-50°C, most preferably 40-50°C. Since the solubility of lactose increases with temperature, higher filtration temperatures allow higher lactose concentrations. On the other hand, temperatures in the range 5-15°C, more preferably 5-10°C, may be desired for reducing the growth of microorganisms.
  • monovalent ions may also be removed by cation and anion exchange steps.
  • cation exchange is performed prior to anion exchange. Apart from removing minerals, this ion exchange may also reduce the non-protein nitrogen (NPN) content of the demineralised whey permeate.
  • NPN non-protein nitrogen
  • This cation exchange step is preferably performed using a strong cation exchange resin in the free acid form (H + form).
  • the resin is preferably a styrene-divinylbenzene cation exchange resin, more preferably macroporous or gel-type styrene- divinylbenzene cation exchange resin.
  • the cationexchange material comprises sulfonic acid functional groups.
  • the cation-exchange material is a strong acid cation exchange resin having a styrene/divinylbenzene gel-type matrix and sulfonic acid functional groups.
  • the anion exchange resin should be a gel-type anion exchange resin (e.g. cross-linked polystyrene-divinylbenzene gel) with a moisture content of 30-48%, preferably 35-45%.
  • a gel-type anion exchange resin e.g. cross-linked polystyrene-divinylbenzene gel
  • the demineralization capacity becomes too low; at high moisture contents, the affinity for sialyllactose increases. Due to the low moisture content and the dense network of a gel-type resin, the mobility of sialyllactose is lower than that of the abundantly present chloride anions. As a result, sialyllactose will remain in the liquid phase, whereas chloride anions are preferentially bound to the resin.
  • the anion exchange resin preferably has strong anion exchange groups, preferably type II anion exchange groups.
  • a column of absorbent material This can be a conventional absorbent material such as activated carbon, but preferably is a cation-exchange type of material.
  • This cation exchange material preferably has a macroporous structure with a higher porosity and a lower density of ionic groups than the cation exchange resin referred to above.
  • the porosity of this cation exchange material is preferably in the range 0.8 to 1.2 ml/g, more preferably 0.9 to 1 .1 ml/g, and most preferably 0.95 to 1 .05 ml/g.
  • the BET surface area is preferably > 600 m 2 /g, more preferably > 650 m 2 /g, even more preferably > 670 m 2 /g, and most preferably > 700 m 2 /g.
  • the absorbent material preferably is a styrene/divinyl benzene copolymer matrix of which the hydrophilicity is increased by the presence of sulphonic acid groups.
  • it is used to absorb cationic and non-cationic components, typically organic components, in particular non-cationic components, more in particular components that are neutral at the pH of the solution.
  • An advantage of such material over absorbent materials like activated carbon is its inertness towards absorption of acidic oligosaccharides.
  • the demineralised whey permeate is contacted with a macroporous or gel-type anion exchange resin that is capable of capturing and binding sialyllactose.
  • a macroporous or gel-type anion exchange resin that is capable of capturing and binding sialyllactose.
  • the moisture content of the macroporous or gel-type anion exchange resin is greater than 45% and more preferably is between 50 and 70%. In such resins, large pores are present.
  • the resin is preferably in the free base form.
  • the temperature during this step is preferably lower than 20°C and more preferably lower than 10°C.
  • the sialyllactose can subsequently be eluted from said resin with a small volume of a an acidic salt solution, preferably a NaCI and/or KCI solution acidified with HCI.
  • a an acidic salt solution preferably a NaCI and/or KCI solution acidified with HCI.
  • the acidity of this solution ensures that the pH of the resulting sialyllactose-containing eluate will not rise above levels that negatively affect the stability of sialyllactose.
  • the sialyllactose-containing eluate preferably has a pH in the range 3-9, more preferably 4-8, most preferably 5-6.
  • the eluate is then demineralized, preferably by nanofiltration, preferably in combination with diafiltration.
  • the pH of the resulting demineralized eluate is preferably adjusted to a value in the range 3-9, more preferably 4-8, most preferably 5-6.
  • the isolated and demineralised sialyllactose is then added to a galacto-oligosaccharide composition, thereby forming the galactooligosaccharide preparation.
  • the galacto-oligosaccharide preparation is obtained by adding the isolated and demineralised sialyllactose to a lactose source, followed by converting said lactose source into galacto-oligosaccharide using a beta-galactosidase enzyme.
  • the resulting GOS preparation contains, based on dry matter, at least 35 wt% preferably 40-90 wt%, most preferably 50-70 wt% galactooligosaccharides other than lactose, and 0.2-3.0 wt%, preferably 0.2-2.0 wt%, and most preferably 0.4-1.0 wt% sialyllactose.
  • the GOS preparation preferably contains 0.2-5.0 wt%, preferably 0.3-4.0 wt%, more preferably 0.3-3.0 wt%, even more preferably 0.4-2.0 wt%, and most preferably 0.5-1 .0 wt% sialyllactose.
  • This sialyllactose content manly consist of 3’-sialyllactose and 6’-sialyllactose and can be determined with the aid of a chromatographic technique using an Acquity premier glycan BEH amide 130A 1.7 pm column and a fluorescence detector.
  • the sialyllactose content is calculated as the sum of the content of 3'-sialyllactose and 6'-sialyllactose.
  • the dry matter content of the lactose source containing the isolated and demineralised sialyllactose is preferably adjusted to at least 35 wt%, preferably at least 40 wt%, more preferably at least 45 wt%, and most preferably at least 50 wt% prior to conversion into galactooligosaccharide. This can be achieved by conventional techniques, such as membrane filtration or evaporation. If the dry matter content is already in this range, no action is required.
  • the pH of the lactose source containing the isolated and demineralised sialyllactose is preferably adjusted to a value in the range 5-7, preferably 5.5-6.5, most preferably 6.0-6.3. If the pH is already in this range, no action is required. If the pH is outside this range, it can be adjusted by the addition of acid or base, such as citric acid or NaOH.
  • Suitable [3-galactosidase enzymes for converting the lactose into galactooligosaccharide include [3-galactosidase enzymes selected from the group consisting of Bacillus circulans, Aspergillus oryzae, Kluyveromyces marxianus, Kluyveromyces fragilis, Sporobolomyces singularis, Lactobacillus fermentum, and Papiliotrema terrestris (Cryptococcus Papiliotrema terrestris).
  • a preferred enzyme is [3- galactosidase produced by Bacillus circulans.
  • the enzymatic conversion is preferably conducted at a temperature of about 20-60°C, more preferably 40-60°C, most preferably 45-55°C.
  • the enzyme is preferably used in an amount of 0.60-2.5 LU/gram, preferably of 0.65- 2.0 LU/gram, most preferably 0.65-1.1 LU/gram.
  • a lactase unit (LU) is defined as the quantity of enzyme that liberates 1 micromole glucose per minute at the early stage of the reaction at 40°, pH 6.0.
  • the enzymatic reaction may take 6-50 hours, preferably 10-48 hours, even more preferably 18-24 hours, after which the enzyme may be denatured (e.g. by heating at about 95-100°C or adjusting the pH to about 3.5 or less) and the GOS preparation can be subjected to additional purification and concentration steps to obtain a syrup, which may optionally be dried, e.g. spray-dried, to form a powder.
  • the enzyme may be denatured (e.g. by heating at about 95-100°C or adjusting the pH to about 3.5 or less) and the GOS preparation can be subjected to additional purification and concentration steps to obtain a syrup, which may optionally be dried, e.g. spray-dried, to form a powder.
  • the enzyme may be used in powder form (e.g. freeze dried, vacuum dried, or spray dried) or liquid form (e.g. dissolved or dispersed in water, a phosphoric acid buffer solution, a tri-ethanol amine buffer solution, a tris-hydrochloric acid buffer solution, or a GOOD buffer solution).
  • powder form e.g. freeze dried, vacuum dried, or spray dried
  • liquid form e.g. dissolved or dispersed in water, a phosphoric acid buffer solution, a tri-ethanol amine buffer solution, a tris-hydrochloric acid buffer solution, or a GOOD buffer solution.
  • the enzyme is used in immobilized form.
  • Various ways of enzyme immobilization are known in the art. They typically comprise a porous carrier onto which the beta-galactosidase is immobilized via covalent binding, via physical adsorption (charge-charge or van der Waals interaction), via gel encapsulation, or a combination thereof.
  • carrier-free immobilized enzymes such as CLEC (crosslinked enzyme crystals) or CLEA (crosslinked enzyme aggregates) may also be applied.
  • Carriers that can promote direct covalent binding of the enzyme are preferred, in view of their ease of operation and absence of leakage into the reaction mixture.
  • An example of a solid carrier is an activated acrylic polymer, preferably a functionalized polymethacrylate matrix.
  • a hexamethylenamino-functionalized polymethacrylate matrix (Sepabeads) or a microporous acrylic epoxy-activated resin, like Eupergit C 250L, can be used.
  • immobilized enzyme allows a repeated batch operating system involving several consecutive batches (‘cycles’) of GOS purification. It also allows for recycling of enzyme, which enables semi-continuous operation and multiple reuse of the enzyme.
  • the reaction is conducted in an enzymatic membrane reactor.
  • the resulting product may be submitted to purification steps, evaporation, and/or (spray) drying.
  • one of the purification steps involves a decolorization step using a cation exchange resin in the free acid form and an absorbent resin as defined above. Apart from removing colour, this step also adds to a further reduction of the ash content.
  • binding to an anion exchange resin should be prevented, either by not conducting an anion exchange step, or by applying an anion exchange step with a resin that does not bind sialyllactose as discussed above.
  • no anion exchange step is performed at this stage; not only does that prevent the introduction of additional anions in the product, the use of cation exchange in the absence of anion exchange also results in a pH reduction of the product that improves microbial stability. The latter thus discards the need for acid addition for achieving microbial stability.
  • the sialyllactose-enriched galacto-oligosaccharide preparation resulting from the process of the present invention can be used in nutritional compositions for human subjects of any age.
  • the subject is an adolescent or an adult.
  • An adolescent is herein defined as a person having an age of from 13 to 20 years.
  • An adult is herein defined as a person having an age of 20 years or higher.
  • the subject is a child having an age of 3 years (36 months) to 13 years.
  • the subject is child having an age of 0 to 3 years, preferably having an age of 24 months or below, more preferably having an age of 18 months or below.
  • the nutritional composition is a MUM composition for pregnant women, a growing up milk (GUM), a follow-up formula, or an infant formula.
  • GUM growing up milk
  • Such nutritional compositions may further comprise a protein source, a digestible carbohydrate source, and/or a lipid source.
  • Protein sources are known in the art, particularly for employment in infant formula, and include dairy proteins (whey proteins, casein) and/or plant protein sources (e.g. pea, faba bean, canola, soy proteins).
  • dairy proteins whey proteins, casein
  • plant protein sources e.g. pea, faba bean, canola, soy proteins
  • Examples of digestible carbohydrate sources are disaccharides such as lactose and saccharose, monosaccharides, such as glucose, and maltodextrins, starch, and carbohydrate sources having a prebiotic effect.
  • Suitable lipid sources include mono-, di-, and triglycerides, phospholipids, sphingolipids, fatty acids, and esters or salts thereof.
  • the lipids may have an animal, vegetable, microbial or synthetic origin.
  • polyunsaturated fatty acids such as gamma linolenic acid (GLA), dihomo gamma linolenic acid (DHGLA), arachidonic acid (AA), stearidonic acid (SA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), docosapentaenoic acid (DPA) and conjugated linoleic acid (CLA).
  • suitable vegetable lipid sources include sun flower oil, high oleic sun flower oil, coconut oil, palm oil, palm kernel oil, soy bean oil, etc.
  • suitable lipid sources of animal origin include milkfat, for example anhydrous milkfat (AMF), cream, etc. In a preferred embodiment, a combination of milkfat and lipids of vegetable origin are used.
  • the nutritional composition may further comprise probiotic bacteria, such as include bacteria of the genus Bifidobacteria (e.g. B. breve, B. longum, B. infantis, B. bifidum), Lactobacillus (e.g. L Acidophilus, L paracasei, L johnsonii, L plantarum, L reuteri, L rhamnosus, L casei, L lactis), and Streptococcus (e.g. S. thermophilus).
  • B. breve and B. longum are especially suitable probiotics.
  • composition may contain one or more conventional micro ingredients, such as vitamins, antioxidants, minerals, free amino acids, nucleotides, taurine, carnitine and polyamines.
  • suitable antioxidants are BHT, ascorbyl palmitate, vitamin E, alpha and beta carotene, lutein, zeaxanthin, lycopene and phospholipids.
  • This cheese whey permeate was demineralized by removing mono- and multivalent ions.
  • multivalent ions were removed by passing the whey permeate at 10°C over a weak base anion exchange resin in the chloride form (XA3112; having a crosslinked acrylic gel structure matrix, tertiary amine functional groups, and a moisture holding capacity of 56-64%), followed by passing it over a strong acid cation exchange resin in the sodium form (XA 2033; having a styrene divinylbenzene matrix, sulphonate functional groups, and a moisture holding capacity of 43-45%).
  • XA3112 having a crosslinked acrylic gel structure matrix, tertiary amine functional groups, and a moisture holding capacity of 56-64%
  • XA 2033 having a styrene divinylbenzene matrix, sulphonate functional groups, and a moisture holding capacity of 43-45%.
  • the resulting demineralised product was then submitted to nanofiltration using a Dow FilmtecTM NF-2540 membrane (DOW; a thin-film poly(piperazine-amide) composite membrane rejecting organics with a molecular weight above 200 Da), in order to remove monovalent ions.
  • a concentrate was obtained through continuous recirculation of the demineralised product over the membrane at 10°C and a trans membrane pressure of 20-25 bar. As a result, the dry matter content in the concentrate increased to 17% and the lactose concentration increased from 83% to 93% on dry matter.
  • the nanofiltration retentate (10°C) was subsequently passed over a cation exchange resin in the free acid form (XA2041 Na; a macroporous strong cation resin with a styrene-divinylbenzene copolymer matrix, SO3 functional groups, and a moisture holding capacity of 48 ⁇ 6 %), an anion exchange resin in the free base form (UBA150; a strong anion exchange resin with styrene-divinylbenzene matrix, trimethyl ammonium groups functional groups, and a moisture holding capacity of 39-45%), and an absorbent resin (XA5072 H; a cation exchange resin with a styrene divinylbenzene copolymer matrix, SO3 functional groups, an average surface area of 700 m 2 /g, and a moisture content 54 ⁇ 3 %).
  • a cation exchange resin in the free acid form XA2041 Na; a macroporous strong cation resin with a s
  • the demineralized product with a pH of 8.9, was subjected to membrane filtration using a 1000 Damembrane at a controlled temperature of 10°C and a pressure of 8 bar.
  • This membrane filtration increased the dry matter content from 11.4 to 14.2 wt%, reduced the ash content from 0.74 to 0.03 wt% on dry matter, increased the sialyllactose content from 0.18 to 0.6 wt% on dry matter, and increased the lactose content from 95 to 96 wt% on dry matter.
  • the retentate was submitted to another anion exchanger in the free base form (XA90 CL; a strong base anionic resin with a styrene divinylbenzene copolymer matrix, - N + (CH 3 )2C 2 H 4 OH functional groups and a moisture holding capacity 54 ⁇ 6 %), which bound substantially all the sialyllactose.
  • XA90 CL a strong base anionic resin with a styrene divinylbenzene copolymer matrix, - N + (CH 3 )2C 2 H 4 OH functional groups and a moisture holding capacity 54 ⁇ 6 %), which bound substantially all the sialyllactose.
  • the bound sialyllactose was recovered by eluting the resin with a 0.2M NaCI solution. During elution, the pH was neutralized using hydrochloric acid to a pH of about 5.5.
  • the sialyllactose-rich eluate was demineralized and concentrated by nanofiltration using a FilmtecTM NF-2540 membrane (DOW; a thin-film poly(piperazine-amide) composite membrane rejecting organics with a molecular weight above 200 Da), thereby transmitting monovalent ions.
  • a concentrate was obtained through continuous recirculation of the demineralised product over the membrane at 10°C and a trans membrane pressure of 20-25 bar. As a result, the dry matter content in the concentrate increased to 19%.
  • sialyllactose-rich NF retentate was blended with a 51 wt% demineralized lactose solution of in a weight ratio 7:100 in order to standardize to a siallyllactose content on dry matter of 0.41 wt%.
  • This concentrated solution was used to make GOS via an enzymatic reaction.
  • the pH of the solution was set to 6.3, kept constant during the reaction by adding a citrate buffer, and a Bacillus c/rcu/ans-originating beta-galactosidase enzyme (2.7 Lll/g) was added. After 42 h reaction at 58°C, the solution was heated to 130°C to inactivate the enzyme and the pH of the resulting GOS syrup was adjusted to 3.8 with citric acid.
  • the GOS syrup was concentrated to 75 wt% dry matter by evaporation.

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Abstract

L'invention concerne un procédé de production d'une préparation de galacto-oligosaccharide enrichie en sialyllactose à partir d'un perméat de lactosérum, ledit procédé impliquant l'élimination d'ions, et une chromatographie.
PCT/EP2025/061218 2024-04-25 2025-04-24 Préparation de galacto-oligosaccharide enrichie en sialyllactose Pending WO2025224241A1 (fr)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006087391A1 (fr) 2005-02-21 2006-08-24 Nestec S.A. Melange d'oligosaccharides
US20110098244A1 (en) * 2008-03-14 2011-04-28 Alfred Willy Bonte Process for Isolating Sialic Acid Containing Oligosaccharides, and the Compositions Containing Sialic Acid Containing Oligosaccharides Obtainable Thereby
US20140087021A1 (en) * 2011-05-24 2014-03-27 Nestec S.A 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
US20150174179A1 (en) * 2006-03-07 2015-06-25 Nestec S.A. Synbiotic mixture
WO2018020473A1 (fr) 2016-07-28 2018-02-01 Fonterra Co-Operative Group Limited Produit laitier et procédé associé
WO2020141032A1 (fr) 2019-01-02 2020-07-09 Frieslandcampina Nederland B.V. Procédé pour la préparation de préparation de gos avec de la bêta-galactosidase provenant de cryptococcus terrestrisis, préparations de gos pouvant être obtenues de cette manière et leurs utilisations
US20210330687A1 (en) * 2018-09-06 2021-10-28 Frieslandcampina Nederland B.V. Bifidogenic hypoallergenic gos compositions and methods for providing the same involving beta-galactosidase from a strain of lactobacillus delbrueckii ssp bulgaricus

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006087391A1 (fr) 2005-02-21 2006-08-24 Nestec S.A. Melange d'oligosaccharides
US20150174179A1 (en) * 2006-03-07 2015-06-25 Nestec S.A. Synbiotic mixture
US20110098244A1 (en) * 2008-03-14 2011-04-28 Alfred Willy Bonte Process for Isolating Sialic Acid Containing Oligosaccharides, and the Compositions Containing Sialic Acid Containing Oligosaccharides Obtainable Thereby
US20140087021A1 (en) * 2011-05-24 2014-03-27 Nestec S.A 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
WO2018020473A1 (fr) 2016-07-28 2018-02-01 Fonterra Co-Operative Group Limited Produit laitier et procédé associé
US20210330687A1 (en) * 2018-09-06 2021-10-28 Frieslandcampina Nederland B.V. Bifidogenic hypoallergenic gos compositions and methods for providing the same involving beta-galactosidase from a strain of lactobacillus delbrueckii ssp bulgaricus
WO2020141032A1 (fr) 2019-01-02 2020-07-09 Frieslandcampina Nederland B.V. Procédé pour la préparation de préparation de gos avec de la bêta-galactosidase provenant de cryptococcus terrestrisis, préparations de gos pouvant être obtenues de cette manière et leurs utilisations
US20220087298A1 (en) * 2019-01-02 2022-03-24 Frieslandcampina Nederland B.V. Method for preparing gos-preparation with beta-galactosidase from cryptococcus terrestris, gos preparations obtainable thereby and uses thereof

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