[go: up one dir, main page]

EP4539679A1 - Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de bifidobactéries - Google Patents

Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de bifidobactéries

Info

Publication number
EP4539679A1
EP4539679A1 EP23734576.4A EP23734576A EP4539679A1 EP 4539679 A1 EP4539679 A1 EP 4539679A1 EP 23734576 A EP23734576 A EP 23734576A EP 4539679 A1 EP4539679 A1 EP 4539679A1
Authority
EP
European Patent Office
Prior art keywords
lst
lacto
lnfp
fucopentaose
bifidobacteria
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
EP23734576.4A
Other languages
German (de)
English (en)
Inventor
Nicole Seifert
Wilbert SYBESMA
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.)
DSM IP Assets BV
Original Assignee
DSM IP Assets 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 DSM IP Assets BV filed Critical DSM IP Assets BV
Publication of EP4539679A1 publication Critical patent/EP4539679A1/fr
Pending legal-status Critical Current

Links

Classifications

    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • 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
    • 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
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
    • 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
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • 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
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/28—Oligosaccharides
    • A23V2250/284—Oligosaccharides, non digestible
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/145—Gasseri
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/149—Jensenii
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/165—Paracasei
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/173—Reuteri
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/175—Rhamnosus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/513—Adolescentes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/515—Animalis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/517—Bifidum
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/519—Breve
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/529—Infantis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/531—Lactis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/533—Longum
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the present invention relates to the use of human milk oligosaccharides (HMOs) for improving the regeneration and/or viability of Bifidobacteria in acidic environments.
  • HMOs human milk oligosaccharides
  • HMOs were found to increase the number of viable Bifidobacteria upon their rehydration (regeneration) in acidic liquids. This improves probiotic potential of these bacteria in food, beverages, dietary supplements, and oral pharmaceuticals, as their viability during preparation for consumption, after ingestion, and/or along the path through the gastrointestinal tract increases.
  • probiotic is a term used to describe live bacteria which, when ingested in adequate amounts, provide a benefit to the human or animal host. The viability of a probiotic is therefore of crucial importance for its efficacy.
  • Bifidobacterium is a genus of gram-positive, nonmotile, anaerobic bacteria. They are ubiquitous inhabitants of the gastrointestinal tract, but the strains have also been isolated from the vagina and mouth of mammals, including humans. Bifidobacteria are one of the major genera of bacteria that make up the gastrointestinal tract microbiota in mammals. Some bifidobacterial ae known as probiotics, such as, for example, Bifidobacterium (B.) bifidum, B. longum (B. longum ssp. longum), B. animalis (B. animalis ssp. animalis), B. lactis (B. animalis ssp. lactis), B. breve, B. infantis (B. longum ssp. infantis), B adolescentis , and B. thermacidophilum.
  • B. animalis ssp. animalis and B. animalis ssp. lactis were previously described as two distinct species. Presently, both are considered B. animalis, with the subspecies (abbreviated “subsp.” or “ssp.”) Bifidobacterium animals ssp. animalis and Bifidobacterium animals ssp. lactis.
  • Dried product forms include capsules, beadles, tablets, sachets, powders, and the like. They can be directly swallowed or dissolved in a liquid before swallowing. These products depend on their ability to regenerate (rehydrate) and deliver viable, functional bacteria in amounts which result in a health benefit.
  • Rehydration involves an important step in the recovery of dehydrated bacteria; an inadequate rehydration/ regeneration step may lead to poor cell viability and a low final survival rate. Rehydration is therefore a highly critical step in the revitalization of a lyophilized culture.
  • Dried probiotics need to regenerate upon reconstitution/ rehydration, which is a very harsh process, dependent upon pH, temperature, osmolarity and other variables. Reconstitution is usually with excessive water, more than that removed during the dehydration process, thereby resulting in osmotic shock.
  • Many bacteria simply do not "revive”: 99% of probiotic bacteria can be killed prior to reaching their destination in the intestine when they are dissolved in an acidic liquid (such as a juice or a carbonated soft drink), or when they encounter the acidic environment of the stomach.
  • Live probiotics which are delivered e.g. in food, such as yoghurt, also need to survive the stomach acid before reaching the intestine.
  • probiotic bacteria which can be delivered in a safe, reliable form, can facilitate a smooth regeneration and improved viability, and is accessible to all consumers.
  • the present invention relates to the following items:
  • HMO human milk oligosaccharide(s)
  • Bifidobacteria are one or more selected from the following group of bacteria: Bifidobacterium (B.) bifidum, B. longum (B. longum ssp. longum), B. animalis (B. animalis ssp. animalis), B. lactis (B. animalis ssp. lactis), B. breve, B. infantis (B. longum ssp. infantis), B. adolescentis , and B. thermacidophilum. 5.
  • Bifidobacterium (B.) bifidum B. longum (B. longum ssp. longum)
  • B. animalis B. animalis ssp. animalis
  • B. lactis B. animalis ssp. lactis
  • B. breve B. infantis (B. longum ssp. infantis)
  • B. adolescentis B. thermacidophilum. 5.
  • any one of items 1-4, wherein the one or more HMO(s) is selected from the group consisting of: lacto-N-fucopentaose I (LNFP-I), lacto-N-fucopentaose II (LNFP-II), lacto-N-fucopentaose III (LNFP- III), lacto-N-fucopentaose V (LNFP-V), lacto-N-fucopentaose VI (LNFP-VI), 3’sialyllacto-N-tetraose a (LST a), 6’-sialyllacto-N-tetraose b (LST b), 6’-sialyllacto-N-neotetraose (LST c), Lacto-N- difucohexaose I (LNDFH-I), Lacto-N-difucohexaose
  • a method of improving the regeneration and/or viability of Bifidobacteria in an acidic environment, wherein the Bifidobacteria are contained in a powder composition comprising mixing the powder composition with one or more HMO(s) prior to or during the process of adding the powder composition to a dietary supplement, medicament, foodstuff or beverage, wherein the one or more HMO(s) is a sialylated and/or fucosylated HMO with at least five monosaccharide units.
  • a composition comprising Bifidobacteria and one or more HMO(s), wherein the HMO is a sialylated or fucosylated HMO and has at least five monosaccharide units.
  • composition of item 8 wherein the Bifidobacteria are dried, preferably wherein the Bifidobacteria are lyophilized.
  • Bifidobacteria are one or more selected from the following group of bacteria: Bifidobacterium (B.) bifidum, B. longum (B. longum ssp. longum), B. animalis (B. animalis ssp. animalis), B. lactis (B. animalis ssp. lactis), B. breve, B. infantis (B. longum ssp. infantis), B. adolescentis , and B. thermacidophilum.
  • Bifidobacterium B.
  • B. longum B. longum ssp. longum
  • B. animalis B. animalis ssp. animalis
  • B. lactis B. animalis ssp. lactis
  • B. breve B. infantis (B. longum ssp. infantis)
  • B. adolescentis B. thermacidophilum.
  • composition of any one of items 8-10, wherein the one or more HMO(s) is selected from the group consisting of: lacto-N-fucopentaose I (LNFP-I), lacto-N-fucopentaose II (LNFP-II), lacto-N- fucopentaose III (LNFP-III), lacto-N-fucopentaose V (LNFP-V), lacto-N-fucopentaose VI (LNFP- VI), 3’sialyllacto-N-tetraose a (LST a), 6’-sialyllacto-N-tetraose b (LST b), 6’-sialyllacto-N-neotetraose (LST c), Lacto-N-difucohexaose I (LNDFH-I), Lacto-N-difucohexaose
  • composition of any one of items 8-11 wherein the one or more HMO(s) is selected from the group consisting of: LNFP-I, LNFP-III, LST a, and LST c; preferably the HMO is LST c.
  • An acidic composition comprising the composition of any one of items 8-12.
  • beverage selected from the group consisting of: carbonated mineral water, sports drinks, carbonated soft drinks, fruit juices, fruit drinks, sodas, energy drinks, and cold teas, and coffee.
  • FIGURE 1 Shows the experimental setup of the regeneration and viability assessment of lyophilized probiotics under pH 3.0 acidic conditions.
  • FIGURE 2 Shows the regeneration and viability of lyophilized Bifidobacterium breve incubated for 30 minutes at pH 3.0 without HMOs (control). The original sample was plated on agar plates and incubated for 48 h in anaerobic chamber.
  • FIGURE 3 A) Shows the regeneration and viability of lyophilized Bifidobacterium breve incubated for 30 minutes at pH 3.0 in combination with LNFP-III. The original sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates. B) Shows the viability of lyophilized Bifidobacterium breve incubated for 30 minutes at pH 3.0 in combination with LNFP-III, compared to the control (B. breve only).
  • FIGURE 5 A) Shows the regeneration and viability of lyophilized Bifidobacterium breve incubated for 30 minutes at pH 3.0 in combination with LST c. 1 :10 dilutions and the original sample were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution steps 1 :10 and 1 :100 in duplicates. B) Shows the viability of lyophilized Bifidobacterium breve incubated for 30 minutes at pH 3.0 in combination with LST c, compared to the control (B. breve only).
  • FIGURE 6 Shows the regeneration and viability of lyophilized Bifidobacterium longum incubated for 30 minutes at pH 3.0 without HMOs (control). The original sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates.
  • FIGURE 7 A) Shows the regeneration and viability of lyophilized Bifidobacterium longum incubated for 30 minutes at pH 3.0 in combination with LNFP-III. The original sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates. B) Shows the viability of lyophilized Bifidobacterium longum incubated for 30 minutes at pH 3.0 in combination with LNFP-III, compared to the control (B. longum only).
  • FIGURE 8 A) Shows the regeneration and viability of lyophilized Bifidobacterium longum incubated for 30 minutes at pH 3.0 in combination with LST c. The original sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates. B) Shows the viability of lyophilized Bifidobacterium longum incubated for 30 minutes at pH 3.0 in combination with LST c, compared to the control (B. longum only).
  • FIGURE 9 Shows the regeneration and viability of lyophilized Bifidobacterium bifidum incubated for 30 minutes at pH 3.0 without HMOs (control). 1 :10 dilutions of the original sample were plated on agar plates and incubated for 5 days in anaerobic chamber. In the picture from left to right: Dilution steps 1 :100, 1 :1000 (E-2 - E-3) in duplicates.
  • FIGURE 10 A) Shows the regeneration and viability of lyophilized Bifidobacterium bifidum incubated for 30 minutes at pH 3.0 in combination with LST c. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 5 days in anaerobic chamber. In the picture from left to right: E-2 - E-3 in duplicates. B) Shows the viability of lyophilized Bifidobacterium bifidum incubated for 30 minutes at pH 3.0 in combination with LST c, compared to the control (B. bifidum only).
  • FIGURE 11 Shows the regeneration and viability of lyophilized Bifidobacterium infantis incubated for 30 minutes at pH 3.0 without HMOs (control). The sample was plated on agar plates in duplicates and incubated for 72 h in anaerobic chamber.
  • FIGURE 14 A) Shows the regeneration and viability of lyophilized Bifidobacterium infantis incubated for 30 minutes at pH 3.0 in combination with LST c. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 72 h in anaerobic chamber. In the picture from left to right: Undiluted, 1 :10 - 1 :100 - 1 :1000 in duplicates. B) Shows the viability of lyophilized Bifidobacterium infantis incubated for 30 minutes at pH 3.0 in combination with LST c, compared to the control (B. infantis only).
  • FIGURE 15 Shows the regeneration and viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 without HMOs (control). 1 :10 dilutions of the original sample were plated on agar plates and incubated for 5 days in anaerobic chamber. In the picture from left to right: Dilution steps 1 :1000, 1 :10’000 (E-3 - E-4) in duplicates.
  • FIGURE 16 A) Shows the regeneration and viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 in combination with LNFP-I. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 5 days in anaerobic chamber. In the picture from left to right: E-3 - E-4 in duplicates.
  • B) Shows the viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 in combination with LNFP-I, compared to the control (B. lactis only). Results are expressed as mean values (n 2) with standard deviation (SD) of colony-forming units (CFU) per milliliter calculated from B. lactis colonies on agar plates when plated at dilution step E-4 ( Figure 16A). * indicates statistically significant difference relative to control, p 0.0439.
  • FIGURE 17 A) Shows the regeneration and viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 in combination with LST a. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 5 days in anaerobic chamber. In the picture from left to right: E-3 - E-4 in duplicates.
  • B) Shows the viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 in combination with LST a, compared to the control (B. lactis only). Results are expressed as mean values (n 2) with standard deviation (SD) of colonyforming units (CFU) per milliliter calculated from B. lactis colonies on agar plates when plated at dilution step E-4 ( Figure 17A). ** indicates statistically significant difference relative to control, p 0.007.
  • FIGURE 18 A) Shows the regeneration and viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 in combination with LST c. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 5 days in anaerobic chamber. In the picture from left to right: E-3 - E-4 in duplicates.
  • B) Shows the viability of lyophilized Bifidobacterium lactis incubated for 1 h at pH 2.0 in combination with LST c, compared to the control (B. lactis only). Results are expressed as mean values (n 2) with standard deviation (SD) of colonyforming units (CFU) per milliliter calculated from B. lactis colonies on agar plates when plated at dilution step E-4 ( Figure 18A). ** indicates statistically significant difference relative to control, p 0.0054.
  • FIGURE 19 Shows the regeneration and viability of lyophilized Bifidobacterium animalis incubated for 2 h at pH 2.0 without HMOs (control). 1 :10 dilutions of the original sample were plated on agar plates and incubated for 4 days in anaerobic chamber. In the picture from left to right: Dilution steps 1 :100, 1 :1000, 1 :10’000 (E-2 - E-3 - E-4) in duplicates.
  • FIGURE 20 A) Shows the regeneration and viability of lyophilized Bifidobacterium animalis incubated for 2 h at pH 2.0 in combination with LST a. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 4 days in anaerobic chamber. In the picture from left to right: E-2 - E-3 - E-4 in duplicates.
  • B) Shows the viability of lyophilized Bifidobacterium animalis incubated for 2 h at pH 2.0 in combination with LST a, compared to the control (B. animalis only). Results are expressed as mean values (n 2) with standard deviation (SD) of colony-forming units (CFU) per milliliter calculated from B. animalis colonies on agar plates when plated at dilution step E-3 ( Figure 20A). * indicates statistically significant difference relative to control, p 0.0150.
  • FIGURE 21 A) Shows the regeneration and viability of lyophilized Bifidobacterium animalis incubated for 2 h at pH 2.0 in combination with LST c. 1 :10 dilutions of the original sample were plated on agar plates and incubated for 4 days in anaerobic chamber. In the picture from left to right: E-2 - E-3 - E-4 in duplicates.
  • B) Shows the viability of lyophilized Bifidobacterium animalis incubated for 2 h at pH 2.0 in combination with LST c, compared to the control (B. animalis only). Results are expressed as mean values (n 2) with standard deviation (SD) of colony-forming units (CFU) per milliliter calculated from B. animalis colonies on agar plates when plated at dilution step E-4 ( Figure 21 A). ** indicates statistically significant difference relative to control, p 0.0020.
  • FIGURE 22 Shows the regeneration and viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 without HMOs (control). The undiluted sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates.
  • FIGURE 23 A) Shows the regeneration and viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 in combination with LNFP-III. The undiluted sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates. B) Shows the viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 in combination with LNFP-III, compared to the control (B. adolescentis only).
  • FIGURE 24 A) Shows the regeneration and viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 in combination with LST a. The undiluted sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates. B) Shows the viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 in combination with LST a, compared to the control (B. adolescentis only).
  • FIGURE 25 A) Shows the regeneration and viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 in combination with LST c. The undiluted sample and a 1 :10 dilution were plated on agar plates and incubated for 48 h in anaerobic chamber. In the picture from left to right: Undiluted sample and dilution step 1 :10 in duplicates. B) Shows the viability of lyophilized Bifidobacterium adolescentis incubated for 3 h at pH 3.0 in combination with LST c, compared to the control (B. adolescentis only).
  • HMOs Human milk oligosaccharides
  • HMOs have a core structure comprising a lactose unit at the reducing end that can be elongated by one or more p-N-acetyl-lactosaminyl and/or one or p-more lacto- N-biosyl units, and which core structure can be substituted by an a-L-fucopyranosyl (“fucosyl”) and/or an a-N- acetyl-neuraminyl (“sialyl”) moiety. See, e.g., Urashima et al.: Milk Oligosaccharides. Nova Science Publisher (2011); and Chen, Adv. Carbohydr. Chem. Biochem. 72, 113 (2015)).
  • HMOs can be isolated or enriched by well-known processes from milk(s) secreted by mammals including, but not limited to human, bovine, ovine, porcine, or caprine species.
  • the HMOs can also be produced by well- known processes using microbial fermentation, enzymatic processes, chemical synthesis, or combinations of these technologies.
  • sialylated oligosaccharides can be made as described in WO 2012/113404, and mixtures of human milk oligosaccharides can be made as described in WO 2012/113405.
  • sialylated oligosaccharides can be made as described in WO 2012/007588
  • fucosylated oligosaccharides can be made as described in WO 2012/127410
  • diversified blends of human milk oligosaccharides can be made as described in WO 2012/156897 and WO 2012/156898.
  • W02001/04341 and WO 2007/101862 describe how to make core human milk oligosaccharides optionally substituted by fucose or sialic acid using genetically modified E. coli.
  • HMOs with five or more monosaccharide units produced by fermentation is described, for example, in WO2016/040531 , WO2019/008133, W02022/034067, WO2019/020707, W02020/115671 , WO2022/243312 and EP 3 848 471 .
  • EP22209675 describes the combination of fermentation and enzymatic processes to produce HMOs with five or more monosaccharide units.
  • fucosylated HMOs with five monosaccharide units include lacto-N-fucopentaose (a1-2)-Gal-(b1-3)-GlcNAc-(b1-3)-Gal-(b1-4)-Glc), lacto-N-fucopentaose (b1- GlcNAc-(b1-3)-Gal-(b1-4)-Glc), lacto-N-fucopentaose III (LNFP-III, Gal-(b GlcN (b1-4)-Glc), lacto-N-fucopentaose V (LNFP-V, Gal-(b1-3)-GlcNAc-(b1- c-(a lacto-N-fucopentaose VI (LNFP-VI, Gal-(b1-4)-GlcNAc-(b1-3)-Gal-(b1 lc).
  • sialylated HMOs with five monosaccharide units examples include 3’-sialyllacto-N-tetraose a (LST a, Neu5Ac-(a2-3)-Gal- (b1-3)-GlcNAc-(b1-3)-Gal-(b1-4)-Glc), 6’-sialyllacto-N-tetraose b (LST b, Gal-(b1-3)-[Neu5Ac-(a2-6)]-GlcNAc- (b1-3)-Gal-(b1-4)-Glc), and 6’-sialyllacto-N-neotetraose( LST c, Neu5Ac-(a2-6)-Gal-(b1-4)-GlcNAc-(b1-3)-Gal- (b1-4)-Glc).
  • fucosylated HMOs with six monosaccharide units examples include Lacto-N-difucohexaose I (LNDFH-I, Fuc- (a1-2)-Gal-(b1-3)-[Fuc-(a1-4)]-GlcNAc-(b1-3)-Gal-(b1-4)-Glc), Lacto-N-difucohexaose II (LNDFH-II, Gal-(b1-
  • sialylated HMOs with six monosaccharide units include 3’,6-Disialyllacto-N-tetraose (DSLNT, Neu5Ac-(a2-3)-Gal-(b1-3)-[Neu5Ac-(a2- 6)]-GlcNAc-(b1-3)-Gal-(b1-4)-Glc).
  • Examples of fucosylated and sialylated HMOs with six monosaccharide units include Sialyl-lacto-N-fucopentaose I (F-LST b, Fuc-(a1-2)-Gal-(b1-3)-[Neu5Ac-(a2-6)]-GlcNAc-(b1-3)- Gal-(b1-4)-Glc), Sialyl-lacto-N-fucopentaose II (F-LST a, Neu5Ac-(a2-3)-Gal-(b1-3)-[Fuc-(a1-4)]-GlcNAc-(b1-
  • Regeneration means the process of regaining/ restoring a dried bacteria’s viability (i.e., “reviving” the bacterial cells by rehydration, wherein “rehydration” means restoring fluid). This process is also sometimes referred to as “reconstitution”.
  • “Viability” is the ability of a bacterial cell to live and function as a living cell.
  • One way of determining the viability of bacterial cells is by spreading them on an agar plate with suitable growth medium and counting the number of colonies formed after incubation for a predefined time (plate counting). Alternatively, FACS analysis may be used.
  • “Improving the regeneration” of Bifidobacteria means to increase the amount (number) of Bifidobacteria successfully regenerating/ reviving compared to the respective control (i.e., the amount/ number of Bifidobacteria without the addition of HMO).
  • “Improving the viability” of Bifidobacteria means to increase the amount (number) of viable Bifidobacteria compared to the respective control (i.e., the amount/ number of Bifidobacteria without the addition of HMO).
  • “Acidic” means having a pH below 7.0 (for example, having a ⁇ 6.0, or ⁇ 5.0, or ⁇ 4.0, or ⁇ 3.0, or in the range of 2.0-6.0, etc.).
  • the pH measured in the stomach is in the range of about 1.5-3.5.
  • the pH of fruit juices is in the range of about 2.0-4.5.
  • “Dried” means that the probiotic has been subjected to any of the following processes: lyophilization (freeze-drying), fluidized bed drying, atmospheric air drying, spray-drying, liquid-drying (L-drying), or vacuum drying. These processes are generally known in the art. A dried probiotic may be rehydrated by restoring its water content.
  • the present inventors have found that Bifidobacteria, when admixed with a human milk oligosaccharide (HMO), have a significantly increased regeneration and viability when coming into contact with a low pH (acidic) environment, such as stomach acid or an acidic beverage.
  • HMO human milk oligosaccharide
  • this combination can offer a reliable way of delivering an adequate amount of Bifidobacteria to a host (human or animal), either in pharmaceutical-like forms, or in food-based forms.
  • the present invention relates to the use of one or more human milk oligosaccharide(s) (HMO(s)) for improving the regeneration and/or viability of Bifidobacteria in an acidic environment.
  • HMO(s) human milk oligosaccharide(s)
  • the HMDs of the invention are sialylated and/or fucosylated HMDs with at least five monosaccharide units.
  • the acidic environment is or contains an acidic liquid.
  • the acidic environment is a beverage or the stomach.
  • the acidic environment has a pH below 7.0.
  • the pH is below 6.0. More preferably, the pH is below 5.0, or even below 4.0.
  • the pH is in the range of 1 .0-6.0.
  • the pH is in the range of 1 .0-5.0.
  • the pH is in the range of 1 .0-4.0.
  • the pH range corresponds to the pH usually measured in the stomach (1 .5-3.5).
  • Other pH ranges specifically considered are those of beverages (2.0-6.0):
  • the pH of fruit and vegetable juices is in the range of 2.0-4.5, that of coffee in the range of 4.5-6.0.
  • Most sodas have a pH in the range of 2.5-4.0.
  • the Bifidobacteria are dried.
  • the dried bacteria may be the result of any known dehydration process, including freeze-drying (lyophilization), spray-drying, and liquid-drying.
  • the Bifidobacteria are lyophilized.
  • Dried product forms include capsules, beadles, tablets, sachets, powders, and the like. They can be directly swallowed or dissolved in a liquid before swallowing.
  • the HMDs are used to improve the regeneration/ rehydration of such dried bacteria.
  • the Bifidobacteria may be live bacteria which are contained, for example, in probiotic drinks or food.
  • the HMDs are used to improve the viability of such live bacteria, for example by helping them survive contact with stomach acid.
  • the Bifidobacteria used may be any type of Bifidobacteria.
  • the Bifidobacteria are probiotics, including probiotics known to have beneficial effects in the gut, such as, for example, Bifidobacterium (B.) bifidum, B. longum, B. animalis, B. animalis ssp. lactis, B. breve, B. infantis, B. adolescentis , and B. thermacidophilum.
  • the Bifidobacteria are selected from the following group of bacteria: Bifidobacterium (B.) bifidum, B. longum, B. animalis, B.
  • the Bifidobacteria are selected from the group consisting of: B. bifidum, B. longum, B. animalis ssp. lactis, B. breve, and B. infantis.
  • B. animalis ssp. lactis is particularly preferred.
  • Bifidobacteria Preferred strains of Bifidobacteria are any of the following: Bifidobacterium breve DSM 33789, Bifidobacterium longum DSM 32946, Bifidobacterium bifidum DSM 32403, Bifidobacterium infantis DSM 32687, and Bifidobacterium animalis ssp. lactis DSM 32269.
  • the HMO used may be any HMO which is sialylated and/or fucosylated and has at least five monosaccharide units.
  • fucosylated HMDs with five monosaccharide units which may be used are: lacto-N- fucopentaose I (LNFP-I), lacto-N-fucopentaose II (LNFP-II), lacto-N-fucopentaose III (LNFP-III), lacto-N- fucopentaose V (LNFP-V), and lacto-N-fucopentaose VI (LNFP-VI).
  • sialylated HMDs with five monosaccharide units which may be used are: 3’-O-sialyllacto-N-tetraose a (LST a), 6’-O-sialyllacto-N-tetraose b (LST b), and 6’-O-sialyllacto-N-neotetraose (LST c).
  • the HMO is one or more selected from the group consisting of: LNFP-I, LNFP-III, LST a, and LST c. LST c is particularly preferred.
  • the HMO used may further be any HMO which is sialylated and/or fucosylated and has at least six monosaccharide units.
  • fucosylated HMOs with six monosaccharide units include Lacto-N- difucohexaose I (LNDFH-I), Lacto-N-difucohexaose II (LNDFH-II) and Lacto-N-difucohexaose III (LNDFH-III.
  • Examples of sialylated HMOs with six monosaccharide units include 3’,6-Disialyllacto-N-tetraose (DSLNT).
  • Examples of fucosylated and sialylated HMOs with six monosaccharide units include Sialyl-lacto-N- fucopentaose I (F-LST b), Sialyl-lacto-N-fucopentaose II (F-LST a) and Sialyl-lacto-N-fucopentaose III (F-LST c).
  • the present invention relates to a method of improving the regeneration and/or viability of Bifidobacteria in an acidic environment, wherein the Bifidobacteria are contained in a powder composition, said method comprising mixing the powder composition with one or more HMO(s) prior to or during the process of adding the powder composition to a dietary supplement, medicament, foodstuff or beverage, wherein the HMO is a sialylated and/or fucosylated HMO with at least five monosaccharide units.
  • the powder composition may further optionally comprise lyoprotection agents and/or processing aids.
  • the method is for improving the regeneration of probiotic blends upon reconstitution in liquid, wherein the probiotic blend comprises probiotic culture powders which are blended with carrier material and or other functional material aimed to dilute the number of probiotics and/or make the probiotic blend more functional, said method comprising adding an HMO to the liquid.
  • the HMO may be added to the liquid prior to introduction of the probiotic culture powders to the liquid; substantially simultaneously to the introduction of the probiotic culture to the liquid; or after the introduction of the probiotic culture to the liquid.
  • the HMO can be added to the probiotic culture powders and the resultant mixture added to the liquid.
  • the HMO is preferably added in an effective/protective amount.
  • the effective/protective amount of the one or more HMO(s) may be from 0.5 g to 15 g, more preferably 1 g to 10 g.
  • the effective amount is from 2 g to 7.5 g of the one or more HMO(s).
  • the probiotic comes in direct contact with the stomach acid without prior mixing with another liquid. It has been found that the HMOs will protect the Bifidobacteria from the harsh effects of stomach acid, and allow a better regeneration and greater survival rate.
  • the present invention relates to compositions comprising Bifidobacteria and one or more HMO(s), wherein the one or more HMO(s) is a sialylated and/or fucosylated HMO with at least five monosaccharide units.
  • the Bifidobacteria of the inventive compositions may be any type of Bifidobacteria.
  • the Bifidobacteria are probiotics, including probiotics known to have beneficial effects in the gut, such as, for example, Bifidobacterium (B.) bifidum, B. longum, B. animalis, B. animalis ssp. lactis, B. breve, B. infantis, B. adolescentis, and B. thermacidophilum.
  • the Bifidobacteria are selected from the following group of bacteria: Bifidobacterium (B.) bifidum, B. longum, B. animalis, B.
  • the Bifidobacteria are selected from the group consisting of: B. bifidum, B. longum, B. animalis ssp. lactis, B. breve, and B. infantis; B. animalis ssp. lactis is particularly preferred.
  • Bifidobacteria Preferred strains of Bifidobacteria are any of the following: Bifidobacterium breve DSM 33789, Bifidobacterium longum DSM 32946, Bifidobacterium bifidum DSM 32403, Bifidobacterium infantis DSM 32687, and Bifidobacterium animalis ssp. lactis DSM 32269.
  • the HMO comprised in the inventive compositions may be any HMO which is sialylated and/or fucosylated and has at least five monosaccharide units.
  • fucosylated HMDs with five monosaccharide units include: lacto-N-fucopentaose I (LNFP-I), lacto-N-fucopentaose II (LNFP-II), lacto-N-fucopentaose III (LNFP-II I), lacto- N-fucopentaose V (LNFP-V), and lacto-N-fucopentaose VI (LNFP-VI).
  • sialylated HMDs with five monosaccharide units examples include: 3’-O-sialyllacto-N-tetraose a (LST a), 6’-O-sialyllacto-N-tetraose b (LST b), and 6’-O-sialyllacto-N-neotetraose (LST c).
  • the HMO may further be any HMO which is sialylated and/or fucosylated and has at least six monosaccharide units.
  • fucosylated HMOs with six monosaccharide units include: Lacto-N-difucohexaose I (LNDFH-I), Lacto-N-difucohexaose II (LNDFH-II) and Lacto-N-difucohexaose III (LNDFH-III.
  • Examples of sialylated HMOs with six monosaccharide units include: 3’,6-Disialyllacto-N-tetraose (DSLNT).
  • Examples of fucosylated and sialylated HMOs with six monosaccharide units include Sialyl-lacto-N-fucopentaose I (F-LST b), Sialyl-lacto-N-fucopentaose II (F-LST a) and Sialyl-lacto-N-fucopentaose III (F-LST c).
  • the HMO is one or more selected from the group consisting of: LNFP-I, LNFP-I 11 , LST a, and LST c.
  • LST c is particularly preferred.
  • composition comprising the probiotic and HMO may optionally contain other ingredients such as vitamins, minerals, flavorings, and further nutritional supplementation.
  • the composition of the invention may comprise a probiotic dose between 1 E+08 and 1 E+12 cfu.
  • the probiotic dose is at least 1 E+08, 2E+08, 3E+08, 4E+08, 5E+08, 6E+08, 7E+08, 8E+08, 9E+08, 1 E+09, 2E+09, 3E+09, 4E+09, 5E+09, 6E+09, 7E+09, 8E+09, 9E+09, 1 E+10, 2E+10, 3E+10, 4E+10, 5E+10, 6E+10, 7E+10, 8E+10, 9E+10, 1 E+11 , 2E+11 , 3E+11 , 4E+11 , 5E+11 , 6E+11 , 7E+11 , 8 E+11 , 9E+11 , or 1 E+12 cfu.
  • the composition of the invention preferably comprises an effective/protective amount of one or more sialylated and/or fucosylated HMO(s) with at least five monosaccharide units from 0.5 g to 15 g, more preferably 1 g to 10 g.
  • the effective amount is from 2 g to 7.5 g of the one or more human milk oligosaccharides (amount per HMO if a single HMO is used, and total HMOs if several HMOs are used, respectively).
  • compositions comprising the probiotic and the HMO(s) can be in the form of a nutritional composition.
  • the nutritional composition can be a food composition, a rehydration solution, a medical food or food for special medical purposes, a nutritional supplement, an early life nutrition product and the like.
  • the nutritional composition can contain sources of protein, lipids and/or digestible carbohydrates and can be in powdered or liquid forms.
  • the Bifidobacteria of the inventive compositions are dried.
  • the dried bacteria may be the result of any known dehydration process, including freeze-drying (lyophilization), spray-drying, and liquid-drying.
  • the Bifidobacteria are lyophilized.
  • the composition comprising the probiotic and the HMO(s) is in a powdery form, such as in a sachet, a dissolvable capsule or tablet, or any other convenient dry formulation.
  • the composition may also be in a liquid form, such as a liquid concentrate.
  • compositions of the invention may be used as a starter culture for fermented foods and drinks, such as spoonable dairy yoghurt, drinkable yoghurt or other fermented beverages, and spoonable non-dairy yoghurt.
  • Starter cultures obtained from probiotic providers typically contain so-called lyoprotection agents and/or processing aids added during their production. These are often proprietary to the provider and may include: disaccharides (saccharose, lactose, trehalose), polyols (mannitol, sorbitol), and polymers (maltodextrin, dextran, inulin), as well as others.
  • lyoprotection agents and/or processing aids added during their production. These are often proprietary to the provider and may include: disaccharides (saccharose, lactose, trehalose), polyols (mannitol, sorbitol), and polymers (maltodextrin, dextran, inulin), as well as others.
  • the inventive composition is consisting essentially of Bifidobacteria and one or more HMO(s), wherein the HMO is a sialylated and/or fucosylated HMO with at least five monosaccharide units.
  • these two elements are the only bioactive ingredients; other ingredients such as binders, fillers, etc. may also be present.
  • the present invention relates to an acidic composition comprising the compositions of the present invention.
  • the acidic composition is a liquid composition.
  • acidic liquids contemplated in this invention include: carbonated mineral water, sports drinks, carbonated soft drinks (such as coca cola), fruit juices (such as orange juice or apple juice), fruit drinks, sodas, energy drinks, cold teas, and coffee.
  • one embodiment of this invention is an acidic drink comprising a reconstituted Bifidobacteria probiotic, and a protective amount of an HMO.
  • the composition of the invention has a pH below 7.0.
  • the pH is below 6.0. More preferably, the pH is below 5.0, or even below 4.0.
  • the pH is in the range of 1 .0-6.0.
  • the pH is in the range of 1 .0-5.0.
  • the pH is in the range of 1 .0-4.0.
  • the preferred pH range for beverages is 2.0-6.0; for fruit and vegetable juices it is in the range of 2.0-4.5, for coffee in the range of 4.5-6.0, for sodas in the range of 2.5-4.0.
  • the following combinations of HMOs and Bifidobacteria are preferred in the uses, methods, and compositions of the present invention:
  • the above Bifidobacterium breve is the strain Bifidobacterium breve DSM 33789 which was deposited at the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany, according to the Budapest Treaty on 1 Feb 2021 , and has the accession number DSM 33789.
  • the above Bifidobacterium longum is the strain Bifidobacterium longum ssp. longum DSM 32946 which was deposited at the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany, according to the Budapest Treaty on 7 Nov 2018, and has the accession number DSM 32946.
  • the above Bifidobacterium bifidum is the strain Bifidobacterium bifidum DSM 32403 which was deposited at the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany, according to the Budapest Treaty on 15 Dec 2016, and has the accession number DSM 32403.
  • the above Bifidobacterium infantis is the strain Bifidobacterium infantis DSM 32687 which was deposited at the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany, according to the Budapest Treaty on 15 Nov 2017, and has the accession number DSM 32687.
  • the above Bifidobacterium animalis ssp. lactis is the strain Bifidobacterium animalis ssp. lactis DSM 32269 which was deposited at the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany, according to the Budapest Treaty on 26 Feb 2016, and has the accession number DSM 32269.
  • Example 1 Bifidobacterium breve DSM 33789;
  • Example 2 Bifidobacterium longum DSM 32946;
  • Example 3 ( Figures 9-10): Bifidobacterium bifidum DSM 32403;
  • Example 4 Bifidobacterium infantis DSM 32687;
  • Example 5 Bifidobacterium animalis ssp. lactis DSM 32269;
  • Example 6 ( Figures 19-21): Bifidobacterium animalis ssp. animalis DSM 16284 (DSM Austria GmbH);
  • Example 7 ( Figures 22-25): Bifidobacterium adolescentis DSM 33750 (DSM Austria GmbH).
  • Lyophilized probiotics (0.4 mg/ml), alone or in combination with HMOs (5% w/v), were dissolved into sterile pH 3.0 (or pH 2.0) water or PBS, warmed to 37°C, and vigorously mixed for about 30 seconds until no visible clumps remained. The tubes were incubated at 37°C for the times indicated (see description of drawings). The samples were further diluted, and 100 pl were spread in duplicates onto MRS agar plates which were incubated for at least 48 h at 37°C in anaerobic chambers. The regeneration and viability of the probiotics were determined by counting the colonies on the plates after the indicated times of incubation. For the experimental setup, see Figure 1 .
  • Regeneration and viability of lyophilized Bifidobacterium animal is alone under pH 2.0 acidic conditions: When the lyophilized Bifidobacterium animalis bacteria are dissolved without HMOs, only very few bacterial colonies resulting from viable cells are observed ( Figure 19). Regeneration and viability of lyophilized Bifidobacterium animalis under pH 2.0 acidic conditions, in combination with LST a: When the lyophilized Bifidobacterium animalis bacteria are simultaneously dissolved with 5% LST a, the regeneration and viability of the bacteria in acidic conditions is significantly increased compared to the control ( Figures 20 A and B).

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Nutrition Science (AREA)
  • Mycology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Saccharide Compounds (AREA)

Abstract

La présente invention concerne l'utilisation d'oligosaccharides de lait humain (HMO) pour améliorer la régénération et/ou la viabilité de bifidobactéries dans des environnements acides. On a découvert que les HMO augmentaient le nombre de bifidobactéries viables lors de leur réhydratation (régénération) dans des liquides acides. Ceci améliore le potentiel probiotique de ces bactéries dans des aliments, des boissons, des compléments alimentaires et des produits pharmaceutiques oraux, comme leur viabilité pendant la préparation pour la consommation, après ingestion et/ou le long du trajet à travers le tractus gastro-intestinal augmente.
EP23734576.4A 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de bifidobactéries Pending EP4539679A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA202200588A DK202200588A1 (en) 2022-06-20 2022-06-20 Mixture of fucosylated HMOs
PCT/EP2023/066702 WO2023247578A1 (fr) 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de bifidobactéries

Publications (1)

Publication Number Publication Date
EP4539679A1 true EP4539679A1 (fr) 2025-04-23

Family

ID=87059739

Family Applications (4)

Application Number Title Priority Date Filing Date
EP23734577.2A Pending EP4539680A1 (fr) 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de lactobacillus rhamnosus
EP23734576.4A Pending EP4539679A1 (fr) 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de bifidobactéries
EP23744674.5A Pending EP4539681A1 (fr) 2022-06-20 2023-06-20 Mélange de hmo fucosylés
EP23734575.6A Pending EP4539678A1 (fr) 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de lactobacilles

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP23734577.2A Pending EP4539680A1 (fr) 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de lactobacillus rhamnosus

Family Applications After (2)

Application Number Title Priority Date Filing Date
EP23744674.5A Pending EP4539681A1 (fr) 2022-06-20 2023-06-20 Mélange de hmo fucosylés
EP23734575.6A Pending EP4539678A1 (fr) 2022-06-20 2023-06-20 Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de lactobacilles

Country Status (9)

Country Link
EP (4) EP4539680A1 (fr)
KR (1) KR20250024846A (fr)
CN (1) CN119403457A (fr)
AU (1) AU2023286780A1 (fr)
DE (1) DE202023103382U1 (fr)
DK (2) DK202200588A1 (fr)
ES (1) ES1317021Y (fr)
FR (1) FR3136650B3 (fr)
WO (4) WO2023247577A1 (fr)

Family Cites Families (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2796082B1 (fr) 1999-07-07 2003-06-27 Centre Nat Rech Scient Procede de production d'oligosaccharides
ES2456292T3 (es) 2006-03-09 2014-04-21 Centre National De La Recherche Scientifique (Cnrs) Procedimiento de producción de oligosacáridos sialilados
WO2008033520A2 (fr) 2006-09-15 2008-03-20 The Regents Of The University Of California Séquences géniques de bifidobactéries et leur utilisation
CA2805501A1 (fr) 2010-07-16 2012-01-19 Glycom A/S Synthese de nouveaux derives sialo-oligosaccharide
DE12746649T1 (de) 2011-02-16 2016-09-29 Glycosyn LLC Biosynthese menschlicher milcholigosaccaride in manipulierten Bakterien
WO2012113405A1 (fr) 2011-02-21 2012-08-30 Glycom A/S Hydrogénolyse catalytique d'une composition d'un mélange de précurseurs d'oligosaccharides et ses utilisations
RU2013146524A (ru) 2011-03-18 2015-04-27 Глюком А/С Синтез новых фукозо-содержащих производных углеводородов
AU2012257395A1 (en) * 2011-05-13 2013-12-12 Glycom A/S Method for generating human milk oligosaccharides (HMOs) or precursors thereof
US9382564B2 (en) 2011-05-13 2016-07-05 Glycom A/S Diversification of human milk oligosaccharides (HMOs) or precursors thereof
JP6129821B2 (ja) * 2011-05-13 2017-05-17 グリコシン リミテッド ライアビリティー カンパニー プレバイオティクスとしての、精製された２’−フコシルラクトース、３−フコシルラクトース、およびラクトジフコテトラオースの使用
US20150265661A1 (en) 2012-04-13 2015-09-24 Trustees Of Boston College Prebiotic effect of sialyllactose
EP3191499A4 (fr) 2014-09-09 2018-06-06 Glycosyn LLC Alpha (1,3) fucosyltransférases destinées à être utilisées dans la production d'oligosaccharides fucosylés
US10415021B2 (en) * 2014-10-24 2019-09-17 Glycom A/S Mutated fucosidase
US10314852B2 (en) * 2014-10-24 2019-06-11 Glycom A/S Mixtures of HMOs
EP3425052A1 (fr) 2017-07-07 2019-01-09 Jennewein Biotechnologie GmbH Fucosyltransférases et leur utilisation dans la production d'oligosaccharides fucosylés
WO2019020707A1 (fr) 2017-07-26 2019-01-31 Jennewein Biotechnologie Gmbh Sialyl-transférases et leur utilisation dans la production d'oligosaccharides sialylés
US20220282262A1 (en) 2018-12-04 2022-09-08 Glycom A/S Synthesis of the fucosylated oligosaccharide lnfp-v
EP3848471A1 (fr) 2020-01-10 2021-07-14 Chr. Hansen HMO GmbH Production fermentative séquentielle d'oligosaccharides
EP4192944A1 (fr) 2020-08-10 2023-06-14 Inbiose N.V. Production de mélanges d'oligosaccharides par une cellule
US20240035055A1 (en) * 2020-08-10 2024-02-01 Inbiose N.V. Process for purification of an oligosaccharide solution produced by cell cultivation or microbial fermentation
CN112189850A (zh) * 2020-09-17 2021-01-08 合生元（广州）健康产品有限公司 益生元在促进胃肠道中益生菌存活的应用
DK181242B1 (en) 2021-05-17 2023-05-30 Dsm Ip Assets Bv GENETICALLY ENGINEERED CELLS COMPRISING A RECOMBINANT NUCLEIC ACID SEQUNCE ENCODING AN α-1,2-FUCOSYLTRANSFERASE CAPABLE OF PRODUCING LNFP-I, NUCLEIC ACID SEQUENCES ENCODING SAME AND METHODS FOR USE OF SAME

Also Published As

Publication number Publication date
EP4539678A1 (fr) 2025-04-23
DE202023103382U1 (de) 2023-11-29
WO2023247578A1 (fr) 2023-12-28
CN119403457A (zh) 2025-02-07
WO2023247577A1 (fr) 2023-12-28
KR20250024846A (ko) 2025-02-19
AU2023286780A1 (en) 2025-01-02
EP4539681A1 (fr) 2025-04-23
ES1317021U (es) 2025-04-10
DK202300035U3 (da) 2023-09-22
DK202300035U9 (da) 2024-07-05
DK202200588A1 (en) 2024-02-23
FR3136650B3 (fr) 2024-06-28
EP4539680A1 (fr) 2025-04-23
WO2023247483A1 (fr) 2023-12-28
WO2023247579A1 (fr) 2023-12-28
ES1317021Y (es) 2025-07-01
FR3136650A3 (fr) 2023-12-22

Similar Documents

Publication Publication Date Title
AU2010311325B2 (en) Nutritional compositions comprising lactoferrin and probiotics and kits of parts thereof
CN102006787B (zh) 延长保质期的益生菌食品
Soccol et al. The potential of probiotics: a review.
RU2521500C2 (ru) ПРИМЕНЕНИЕ ШТАММА Bifidobacterium lactis CNCM I-3446 У ДЕТЕЙ РОЖДЕННЫХ, ПУТЕМ КЕСАРЕВА СЕЧЕНИЯ (ВАРИАНТЫ) И ПРОБИОТИЧЕСКАЯ КОМПОЗИЦИЯ, СОДЕРЖАЩАЯ УКАЗАННЫЙ ШТАММ (ВАРИАНТЫ)
WO2011096808A1 (fr) Utilisation de sialyl-oligosaccharides pour gestion du poids
TW201100537A (en) Short-time high temperature treatment generates microbial preparations with anti-inflammatory profiles
Misra et al. Applications of probiotics as a functional ingredient in food and gut health
CN102065867A (zh) 预防感染的营养物
CA3017371A1 (fr) Compositions alimentaires pour le sevrage
JP7474596B2 (ja) 新規ビフィドバクテリウム属細菌、および当該細菌を含む組成物、並びに当該細菌の増殖促進用の組成物
US20240108668A1 (en) Improving protein digestion and amino acid bioavailability by probiotic strains
TW201105252A (en) Prevention and treatment of allergic diarrhoea
CN116709935A (zh) 改善氨基酸和的用益生细菌的蛋白质生物加工
WO2019182160A1 (fr) Composition pour améliorer un constituant de lait maternel
EP1997880A1 (fr) Bifidobactérie du lait maternel, composition correspondante, leur utilisation et milieu de culture pour les obtenir
EP4539679A1 (fr) Utilisation d'oligosaccharides de lait humain pour améliorer la viabilité de bifidobactéries
WO2024121103A1 (fr) Probiotiques pour le traitement et/ou la prévention d'états associés à la colonisation par helicobacter pylory
JP7787936B2 (ja) 新規ビフィドバクテリウム属細菌、および当該細菌を含む組成物、並びに当該細菌の増殖促進用の組成物
Kumar et al. CHAPTER: 16 ROLE OF PROBIOTIC MILK IN HUMAN HEALTH
CN115137074A (zh) 婴幼儿用排便促进剂
Sreeja et al. Bifidus milks
TW201130429A (en) Nutritional compositions comprising lactoferrin and probiotics and kits of parts thereof
MANDAL MICROENCAPSULATION OF PROSIOTICS AND
Soccol et al. Mogućnosti primjene probiotika
L Harvey et al. Development of the LAFTI range of probiotic cultures

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20241219

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC ME MK MT NL NO PL PT RO RS SE SI SK SM TR

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)