WO2024052406A1 - Compositions nutritionnelles cuites comprenant des oligosaccharides de lait humain - Google Patents

Compositions nutritionnelles cuites comprenant des oligosaccharides de lait humain Download PDF

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Publication number
WO2024052406A1
WO2024052406A1 PCT/EP2023/074447 EP2023074447W WO2024052406A1 WO 2024052406 A1 WO2024052406 A1 WO 2024052406A1 EP 2023074447 W EP2023074447 W EP 2023074447W WO 2024052406 A1 WO2024052406 A1 WO 2024052406A1
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Prior art keywords
milk
mixture
nutritional composition
lacto
less
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PCT/EP2023/074447
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English (en)
Inventor
Joeri Beauprez
Ut VAN NGUYEN
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Inbiose N.V.
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Publication of WO2024052406A1 publication Critical patent/WO2024052406A1/fr

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    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/14—Organic oxygen compounds
    • A21D2/18—Carbohydrates
    • A21D2/181—Sugars or sugar alcohols
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/20—Animal feeding-stuffs from material of animal origin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/153—Nucleic acids; Hydrolysis products or derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
    • A23K50/42—Dry feed

Definitions

  • the present invention relates to a heated nutritional composition comprising a milk saccharide or a mixture of at least two different milk saccharides. More specifically, the present invention relates to a heated nutritional composition comprising a mammalian milk saccharide or a mixture of at least two different mammalian milk saccharides. Even more specifically, the present invention relates to a heated nutritional composition comprising a mammalian milk oligosaccharide or a mixture of at least two different mammalian milk oligosaccharides Even more specifically, the present invention relates to a heated nutritional composition comprising a human milk oligosaccharide or a mixture of at least two different human milk oligosaccharides.
  • Milk saccharides i.e. saccharides which are found in milk of animals such as mammals and humans, in particular mammalian milk oligosaccharides (MMOs) and human milk oligosaccharides (Urashima T. et al., 2011, Milk oligosaccharides, Nova Biomedical books, New York ISBN 978-1-61122-831-1; Coppa et al, 2013, Ital. J. Pediatr. 2013, 39(2): p. Datsenko and Wanner, PNAS 2000, 97(12): p. 6640-6645), are gaining more and more attention as a replete amount of beneficial effects have been demonstrated. A replete amount of milk saccharide structures have been elucidated so far.
  • milk oligosaccharides found in animals such as mammals and humans comprise lactose at the reducing end (Urashima et al, 2011).
  • Other milk oligosaccharides comprise N-acetyllactosamine (Gal-pi,4-GlcNAc) or lacto-N-biose (Gal- pi,3-GlcNAc) at the reducing end (Urashima et al, 2011; Wrigglesworth et al, 2020, PLoS ONE 15(12); Urashima et al, 2013, Biosci. Biotechnol. Biochem 77(3): p. 455-466; Wei et al, 2018, Sci. Rep. 8:4688).
  • Examples hereof are 3-FLN (Gal-pi,4-(Fuc-al,3-)GlcNAc; also known as Lewis x antigen), 3'-SLN (Neu5Ac- a2,3-Gal-pi,4-GlcNAc), 6'-SLN (Neu5Ac-a2,6-Gal-pi,4-GlcNAc) (Urashima et al, 2011; Wrigglesworth et al, 2020; Wei et al, 2018). Further, milk saccharides comprise milk glycosaminoglycans (GAGs; Coppa et al, 2013; Rai et al, 2021, Int. J. biol.
  • Macromolecules 193(A) : p. 137-144) Antigens of the human ABO blood group system such as the A determinant (GalNAc-alphal,3(Fuc-alphal,2)-Gal), B determinant (Gal- alphal,3(Fuc-alphal,2)-Gal) and H-determinant (Fuc-alphal,2-Gal) have been shown to be present in saccharides of the milk of mammals (e.g. A tetrasaccharide in a lion and leopard; Wrigglesworth et al, 2020, PLoS ONE 15(12)).
  • a determinant GalNAc-alphal,3(Fuc-alphal,2)-Gal
  • B determinant Gal- alphal,3(Fuc-alphal,2)-Gal
  • H-determinant Fuc-alphal,2-Gal
  • milk saccharides in particular milk oligosaccharides such as MMOs and HMOs
  • MMOs and HMOs milk oligosaccharides
  • HMOs and MMOs are further known to act as decoys to reduce the risk of infections by bacterial and viral pathogens which adhere to human cells by binding to these cells' surface glycoproteins.
  • various HMOs and MMOs possess an anti-inflammatory effect and act as immunomodulators (e.g. reducing the risk of developing food allergies).
  • (oligo)saccharides in particular sialylated mammalian/human milk (oligo)saccharides, in the presence of other constituents such as proteins, amino acids etc, which are present in common food and feed products, could react and/or are labile (e.g. sialylated oligosaccharides in an acidic environment), especially under conditions of high temperature (i.e. at a temperature of above 100°C, in particular above 150°C) and/or in the presence of moisture (e.g. water, milk).
  • labile e.g. sialylated oligosaccharides in an acidic environment
  • high temperature i.e. at a temperature of above 100°C, in particular above 150°C
  • moisture e.g. water, milk
  • the invention is directed to the surprising finding that milk saccharides, even sialylated milk saccharides which are known to be even more sensitive to the environmental conditions, retain more than 85.0 % of their concentration (expressed as % of the total weight of dry matter) in a nutritional composition upon being heated at temperature of 100°C or higher, even up to 225°C.
  • the invention provides a nutritional composition comprising a milk saccharide or a mixture of at least two different milk saccharides, wherein said nutritional composition was heated at a temperature of 100°C or higher.
  • the invention provides a method for the production of a heated nutritional composition, preferably a baked nutritional composition.
  • the invention provides a nutritional composition comprising a milk saccharide or a mixture of at least two different milk saccharides, preferably a milk oligosaccharide or a mixture of at least two different milk oligosaccharides, wherein said nutritional composition was heated at a temperature of 100°C or higher.
  • the nutritional composition according to the invention is for human and/or animal consumption, preferably for human consumption, more preferably for animal consumption.
  • said nutritional composition is an animal food or animal feed.
  • said nutritional composition is a pet food or pet feed.
  • said nutritional composition is a companion animal food or companion animal feed.
  • the expression “food and/or feed” and “food/feed” is preferably replaced with “food” or is preferably replaced with “feed”, more preferably replaced with “feed”.
  • “food” preferably refers to a nutritional composition for human consumption
  • “feed” preferably refers to a nutritional composition for animal consumption.
  • the nutritional composition according to the invention is edible.
  • the nutritional composition according to the invention is not a liquid product (i.e. is not a liquid nutritional composition).
  • the nutritional composition according to the invention is not a powder (i.e. is not a powdered nutritional composition).
  • a powdered nutritional composition can be obtained by various drying techniques as known in the art (spray drying and freeze drying are widely used examples hereof).
  • the nutritional composition according to the invention is not a milk replacer.
  • milk replacer refers to a nutritional composition which serves as a substitute for mother's milk.
  • the nutritional composition according to the invention is a baked nutritional composition.
  • the terms “bake”, “baking” and “baked” are well- understood by the skilled person (it is referred to the book entitled “Food processing technology: principles and practice” (Fellows; second edition; Woodhead Publishing Limited; 2000), chapter 16, i.e. p. 341-352, which is incorporated by reference).
  • the terms “bake”, “baking” and “baked” preferably refer to the process of cooking of a product, preferably a food/feed by heat, preferably dry heat, without direct exposure of said product, preferably food/feed, to a flame, preferably in an oven.
  • baking heat is transferred into the product, preferably food/feed, (from e.g. hot surface and/or hot air) while moisture is transferred from the product, preferably food/feed, to the air that surrounds said food/feed.
  • “baking” does not encompass (hot) extrusion. The latter is for example a typical high- temperature-short-time (HTST) method in contrast to baking.
  • HTST high- temperature-short-time
  • said nutritional composition according to the invention is selected from a list consisting of a cookie, pet food (preferably dry pet food), a pet treat, a bread, a cake, a pie, a tart, a pastry, a candy bar, an energy bar, granola, a granola bar, a quiche, cereal, a pizza, a corn chip, a tortilla chip, a potato chip and a baked cracker; preferably selected from a list consisting of a cookie, pet food (preferably dry pet food), a pet treat, a bread, a cake, a pie, a tart, a pastry, a candy bar, an energy bar, granola, a granola bar and cereal; more preferably selected from a list consisting of a cookie, pet food (preferably dry pet food), a pet treat, a pastry, a candy bar, an energy bar, granola, a granola bar and cereal
  • Said cookie is for human and/or animal consumption, preferably for animal consumption as described herein.
  • Said pet food is preferably food for a companion animal.
  • a particular preferred example of food for a companion animal is a treat for a companion animal (such as kibbles and biscuits).
  • the nutritional composition is a treat or toy for a companion animal or is part of a treat or toy for a companion animal.
  • a treat or toy for the treatment of dental problems such as inhibition of plaque formation and/or removal of tartar, is particularly preferred.
  • said nutritional composition according to the invention is a pet food, more preferably a dry pet food, even more preferably a dry pet food for a companion animal (i.e. dry companion animal food).
  • a dry pet food typically contains at least 85.0 % (w/w), preferably at least 90.0 % (w/w), of dry matter (in other words, the moisture content is 15.0 % (w/w) or less, preferably 10.0 % (w/w) or less).
  • a dry pet food comprises kibbles, biscuits, meals or extruded product.
  • a dry pet food is preferably selected from kibble, biscuit, meal and an extruded product, more preferably selected from kibble, biscuit and meal, even more preferably selected from kibble and biscuit.
  • Kibbles and biscuits are usually prepared by mixing all the ingredients into a homogeneous dough which is then baked.
  • the dough is formed into the desired shape before baking (which is similar to the baking of cookies and crackers).
  • the dough is usually spread onto large sheets before baking. After cooling, the sheets are broken into bite-size pieces.
  • a meal is produced by mixing dried, flaked or granular ingredients.
  • Extruded products have become more and more popular wherein the ingredients are mixed together as to form a dough which is then cooked/baked under conditions of high pressure and temperature (100°C or above). Extrusion results in rapid cooking/baking, resulting in an increased digestibility and palatability. Extrusion is nowadays frequently used to make biscuits and kibbles for example.
  • a total moisture content of 10% (w/w) or less can be achieved through for example hot-air drying as known in the art.
  • said nutritional composition comprises at least 85.0 %, preferably at least 87.0 %, more preferably at least 90.0 %, most preferably at least 92.0 % dry matter.
  • said nutritional composition comprises 15.0 % (w/w) or less, preferably 13.0 % (w/w) or less, more preferably 10.0 % (w/w) or less, most preferably 8.0 % (w/w) or less moisture.
  • said moisture is liquid, more preferably is water and/or milk, most preferably is water.
  • the % of dry matter i.e.
  • dry extract can be determined by a thermogravimetric analysis or a Karl Fisher titration analysis, preferably by a Karl Fisher titration analysis (preferably as described herein); preferably wherein said % dry extract is calculated as (mass of nutritional composition after 24 hours at 105°C / initial mass of the nutritional composition) x 100.
  • moisture can be preferably replaced with “liquid” and vice versa, unless explicitly stated otherwise.
  • the moisture content (% w/w) of a cake is 15-30% (w/w)) and of a bread is 35-45% (w/w), while biscuits/cookies typically have a moisture content of 5-10% (w/w), pastries have a moisture content of ⁇ 15% (w/w) (Figoni, 2011, How baking works : exploring the fundamentals of baking science, 3th edition, John Wiley & Sons Inc).
  • said nutritional composition before being heated at a temperature of 100°C or higher according to the invention is a dough or a batter, preferably a dough.
  • said unheated nutritional composition comprises 3.0-70.0 % w/w, preferably 3.0-60.0 % w/w, more preferably 3.0-50.0 % w/w, even more preferably 5.0-50.0 % w/w, most preferably 10.0-50.0 % w/w, moisture; preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • a product that comprises a-b % of a component A means that the product can contain further components besides component A but component A can only be present in an amount falling in the range a-b.
  • said nutritional composition before being heated at a temperature of 100°C or higher according to the invention i.e. unheated nutritional composition, preferably unbaked nutritional composition
  • said unheated nutritional composition comprises 10.0-70.0 % w/w, preferably 15.0-70.0 % w/w, more preferably 20.0-70.0 % w/w, even more preferably 25.0-70.0 % w/w, even more preferably 30.0-70.0 % w/w, even more preferably 35.0-70.0 % w/w, even more preferably 35.0-60.0 % w/w, most preferably 40.0-60.0 % w/w, moisture; preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • the term “dough” preferably refers to a mixture of (i) flour and/or meal and (ii) other ingredients such that said mixture is firm enough to knead or roll. Dough may be fresh or frozen, preferably fresh.
  • the preparation of frozen dough is described in the book entitled “Frozen and Refrigerated doughs and batters” (Kulp et al, 1995, Frozen and refrigerated doughs and batters, Amer Assn of Cereal Chemists) which is incorporated by reference.
  • the term “batter” preferably refers to a mixture of (i) flour and/or meal and (ii) other ingredients such that said mixture is thinner and more liquid than dough. Batter is consequently not firm enough to knead (mixing is usually done by a hand mixer or electric mixer), in other words batter it is not malleable in contrast to dough.
  • said unheated nutritional composition has a pH of 2.0-8.0, preferably 2.5-7.5, more preferably 2.5-7.0, even more preferably 3.0-7.0, even more preferably 3.0-6.5, most preferably 3.0-6.0.
  • said nutritional composition is a synthetic nutritional composition.
  • a synthetic nutritional composition refers to a composition which is artificially prepared and preferably refers to a nutritional composition comprising at least one component that is produced ex vivo, either chemically and/or biologically, e.g. by means of chemical reaction, enzymatic reaction or recombinantly, or purified by humans. It is preferred that a synthetic nutritional composition of the invention is not identical with a naturally occurring nutritional composition.
  • said nutritional composition according to the invention comprises flour and/or meal, preferably flour.
  • flour as understood by the skilled person is a powder which is obtained by grinding grains, roots, beans, nuts or seeds.
  • Meal as understood by the skilled person is identical to flour, except that the particle size is slightly more coarse than that of flour.
  • said nutritional composition according to the invention comprises flour obtained from any one or more selected from the list consisting of wheat, buckwheat, barley, oat, spelt, rye, sorghum, maize, triticale, millet, teff, sunflower, sesame, linseed, amaranth, flaxseed, pumpkin, rapeseed, soybean, farro, kamut, quinoa, beans, peas, chick peas, nuts (e.g.
  • said nutritional composition according to the invention comprises a lipid ingredient, preferably any one or more selected from a list consisting of an oil, a fat and a fat substitute, more preferably an oil and/or a fat.
  • said oil is selected from the list consisting of sunflower oil, olive oil, corn oil, peanut oil, coconut oil, soy oil, safflower oil, palm oil, fish oil and rapeseed oil.
  • said fat is butter, margarine or shortening, more preferably said fat is butter or margarine, even more preferably said fat is butter.
  • said nutritional composition according to the invention comprises any one or more selected from the list consisting of an egg, egg white, egg yolk and egg replacer.
  • said nutritional composition according to the invention comprises a liquid, preferably milk and/or water, more preferably water. It is particularly preferred that said liquid is not milk.
  • said nutritional composition preferably comprises 15.0 % (w/w) or less, preferably 13.0 % (w/w) or less, more preferably 10.0 % (w/w) or less, most preferably 8.0 % (w/w) or less of said liquid.
  • said nutritional composition before being heated at a temperature of 100°C or higher according to the invention (i.e.
  • unheated nutritional composition preferably unbaked nutritional composition
  • said nutritional composition according to the invention comprises sugar, preferably sugar which is different from the milk saccharide(s), preferably milk oligosaccharide(s), according to the invention, and/or a sweetener.
  • said nutritional composition according to the invention comprises one or more food and/or feed ingredients, preferably one or more functional ingredients, more preferably selected from the list consisting of salt, salty-like ingredient, sugar, sweetener, ethanol, bicarbonate, baking powder, diacetyl, natural aromas ingredient, synthetic aromas ingredient, flavor ingredient, flavor improvement ingredient, enzyme, oxidizing agent, reducing agent, emulsifier, vitamin, coloring agent, antimicrobial agent and food-grade microorganism.
  • a “functional ingredient” refers to an ingredient which provides a desired function to the nutritional composition according to the invention.
  • Such function can be taste (e.g. addition of salt, salty-like ingredient, sugar, sweetener, natural/synthetic aromas ingredient, diacetyl, flavor ingredient and flavor improvement ingredient), appearance (e.g. coloring agent), retention of flavor (natural/synthetic aromas ingredient, flavor ingredient and especially flavor improvement ingredient), shelf life (e.g. antimicrobial agent), stability (e.g. enzyme), structure/texture (e.g. bicarbonate, emulsifier, food-grade microorganism, enzyme), leavening (e.g. food-grade microorganism) and health (e.g. foodgrade microorganism) among other functions as known by the skilled person.
  • taste e.g. addition of salt, salty-like ingredient, sugar, sweetener, natural/synthetic aromas ingredient, diacetyl, flavor ingredient and flavor improvement ingredient
  • appearance e.g. coloring agent
  • said nutritional composition according to the invention comprises a leavening agent, preferably chosen from the list consisting of baking soda, baking powder, whipped egg white, cream and a food-grade microorganism, preferably chosen from the list consisting of baking soda, baking powder and a food-grade microorganism, more preferably a foodgrade microorganism.
  • a leavening agent preferably chosen from the list consisting of baking soda, baking powder and a food-grade microorganism, more preferably a foodgrade microorganism.
  • said food-grade microorganism is a yeast.
  • the nutritional composition according to the invention comprises (i) a milk saccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (it is referred to the section "milk saccharide”), preferably a milk oligosaccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk oligosaccharides, and (ii) one or more conventional ingredients, for example the ingredients described in the present section.
  • milk saccharide preferably a milk oligosaccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk oligosaccharides
  • one or more conventional ingredients for example the ingredients described in the present section.
  • said nutritional composition comprises a milk saccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides.
  • said nutritional composition comprises a milk oligosaccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk oligosaccharides.
  • saccharide refers to a molecule comprising at least one monosaccharide.
  • saccharide refers to a molecule that comprises at least one monosaccharide and does not comprise a moiety which is different from a sugar such as a nucleobase, a nucleotide or an amino acid.
  • saccharide refers to a molecule consisting of one or more monosaccharides, i.e. "saccharide” refers to a monosaccharide, disaccharide, oligosaccharide and polysaccharide, preferably as defined herein.
  • saccharide refers to an oligosaccharide and polysaccharide, preferably as defined herein. Most preferably, “saccharide” refers to an oligosaccharide, preferably as defined herein.
  • milk saccharide refers to a saccharide which is found in milk of an animal, preferably a mammal and/or human.
  • said milk saccharide and said milk saccharides are a milk oligosaccharide and milk oligosaccharides, respectively.
  • said nutritional composition comprises a milk oligosaccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk oligosaccharides.
  • saccharides/oligosaccharides preferably means "structurally different” or “structurally distinct”. These terms are hence preferably interchangeably used in the context of the present invention.
  • oligosaccharide as used in the context of the present invention preferably refers to a saccharide containing 2 up to and including 20 monosaccharides, i.e. the degree of polymerization (DP) is 2-20.
  • said oligosaccharide according to the invention contains at least 3 monosaccharides, i.e. the degree of polymerization is preferably 3-20. More preferably, said oligosaccharide according to the invention consists of 3-9, preferably 3-8, more preferably 3-7 monosaccharides.
  • said oligosaccharide according to the invention is not lactose.
  • said oligosaccharide according to the invention is not lacto-N-triose II.
  • An oligosaccharide can be a linear structure or can include branches. The linkage (e.g. glycosidic linkage, galactosidic linkage, glucosidic linkage, etc.) between two sugar units can be expressed, for example, as 1,4, l->4, or (1-4), used interchangeably herein. Each monosaccharide can be in the cyclic form (e.g. pyranose or furanose form).
  • An oligosaccharide can contain both alpha- and beta-glycosidic bonds or can contain only beta-glycosidic bonds.
  • polysaccharide refers to a saccharide containing a plurality of repeating units comprised of simple sugars.
  • said polysaccharide preferably has a degree of polymerization which is at least 40 (and preferably ⁇ 3000).
  • said milk saccharide or any one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides in said mixture is/are a mammalian milk saccharide, more preferably a mammalian milk oligosaccharide (MMO).
  • said milk saccharide (preferably milk oligosaccharide), or any one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides (preferably milk oligosaccharides) in said mixture is/are a human milk saccharide (preferably a human milk oligosaccharide (HMO)).
  • said nutritional composition according to the invention comprises a human milk oligosaccharide or a mixture of at least two , preferably at least three, more preferably at least 4, most preferably at least 5, different human milk oligosaccharides.
  • milk saccharide is preferably replaced with the term “mammalian milk saccharide”, even more preferably replaced with the term “human milk saccharide”.
  • milk oligosaccharide is preferably replaced with the term “mammalian milk oligosaccharide”, even more preferably replaced with the term "human milk oligosaccharide”.
  • Mammalian milk (oligo)saccharides comprise (oligo)saccharides present in milk found in any phase during lactation including colostrum milk from humans (i.e. human milk (oligo)saccharides) and mammals including but not limited to cows (Bos Taurus), sheep (Ovis aries), goats (Capra aegagrus hircus), bactrian camels (Camelus bactrianus), horses (Eguus ferus caballus), pigs (Sus scropha), dogs (Canis lupus familiaris), ezo brown bears (Ursus arctos yesoensis), polar bear (Ursus maritimus), Japanese black bears (Ursus thibetanus japonicus), striped skunks (Mephitis mephitis), hooded seals (Cystophora cristata), Asian elephants (Elephas maximus), African elephant (Loxodont
  • said milk saccharide/oligosaccharide is selected from the list consisting of (i) oligosaccharide comprising lactose (Gal-bl,4-Glc) at its reducing end; (ii) oligosaccharide comprising N-acetyllactosamine (LacNAc) at its reducing end; (iii) oligosaccharide comprising lacto-N-biose (LNB) at its reducing end; (iv) milk glycosaminoglycan; (v) antigen of the human ABO blood group system, optionally bound to a monosaccharide selected from the list consisting of betal,3-GlcNAc, betal,4-GlcNAc, betal,3-GalNAc and
  • said milk saccharide/oligosaccharide is selected from the list consisting of (i) oligosaccharide comprising lactose (Gal-bl,4-Glc) at its reducing end; (ii) oligosaccharide comprising N- acetyllactosamine (LacNAc) at its reducing end; (iii) oligosaccharide comprising lacto-N-biose (LNB) at its reducing end; and (iv) antigen of the human ABO blood group system, optionally bound to a monosaccharide selected from the list consisting of betal,3-GlcNAc, betal,4-GlcNAc, betal,3-GalNAc and -betal,4-Glc.
  • said milk saccharide/oligosaccharide is selected from the list consisting of (i) oligosaccharide comprising lactose (Gal-bl,4-Glc) at its reducing end; (ii) oligosaccharide comprising N- acetyllactosamine (LacNAc) at its reducing end; and (iii) oligosaccharide comprising lacto-N-biose (LNB) at its reducing end.
  • said milk saccharide/oligosaccharide is an oligosaccharide comprising lactose (Gal-bl,4- Glc) at its reducing end.
  • Said oligosaccharide comprising lactose (Gal-bl,4-Glc) at its reducing end refers to a replete amount of oligosaccharide structures as found in the milk of animals such as mammals and humans (e.g. Table 2 of
  • Examples include: neutral, fucosylated oligosaccharides: e.g. 2'-fucosyllactose (2'FL), 3-fucosyllactose (3-FL), difucosyllactose (diFL), Lacto-N-fucopentaose I (LNFP-I; Fuc-al,2-Gal-bl,3-GlcNAc-bl,3-Gal- bl,4-Glc), GalNAc-LNFP-l (GalNAc-al,3-(Fuc-al,2)-Gal-bl,3-GlcNAc-bl,3-Gal-bl,4-Glc), Lacto-
  • 2'-fucosyllactose 2'-fucosyllactose (2'FL), 3-fucosyllactose (3-FL), difucosyllactose (diFL), Lacto-N-fucopentaos
  • N-fucopentaose (LNFP-II; Gal-bl,3-(Fuc-al,4)-GlcNAc-bl,3-Gal-bl,4-Glc), Lacto-N- fucopentaose III (LNFP III; Gal-bl,4-(Fuc-al,3)-GlcNAc-bl,3-Gal-bl,4-Glc), Lacto-N- fucopentaose V (LNFP-V; Gal-bl,3-GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc), Lacto-N- fucopentaose VI (LNFP-VI; Gal-bl,4-GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc), lacto-N- neofucopentaose (LNnFP
  • Monofucosyllacto-N-hexaose III Difucosyllacto-N-hexaose, difucosyl-lacto-N-neohexaose, LNnDFH (Gal-bl,4-(Fuc-al,3)-GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc), A-tetrasaccharide (GalNAc-al,3-(Fuc-al,2)-Gal-bl,4-Glc),... neutral, non-fucosylated oligosaccharides: e.g.
  • Lacto-N-triose II (LN3), Lacto-N-neotetraose (LNnT), Lacto-N-tetraose (LNT), para-Lacto-N-neopentaose, para-Lacto-N-pentaose, para- Lacto-N-neohexaose, para-Lacto-N-hexaose, GalNAc-bl,3-Gal-al,4-Gal-bl,4-Glc (globo-N- tetraose),... charged oligosaccharides: e.g.
  • Said oligosaccharide comprising N-acetyllactosamine (LacNAc) at its reducing end refers to oligosaccharides such as but not limited to 3'-sialyllactosamine (3'SLacNAc; 3'-SLN; Neu5Ac-a2,3-Gal- pi,4-GlcNAc), 6'-sialyllactosamine (6'SLacNAc; 6'-SLN; Neu5Ac-a2,6-Gal-pi,4-GlcNAc), sialyl Lex (sialyl Lewis x; Neu5Ac-a2,3-Gal-pi,4-[Fuc-al,3]-GlcNAc), Neu5Gc-a2,3-Gal-bl,4-GlcNAc, 2'FLacNAc, 3-FLacNAc (lewis x; 3-FLN; Gaip-l,4-[Fuc-
  • Said oligosaccharide comprising lacto-N-biose (LNB) at its reducing end refers to oligosaccharides such as but not limited to 3'-sialyllacto-N-biose (3'SLNB), 6'-sialyllacto-N-biose (6'SLNB), sialyl Lea (sialyl Lewis a; Neu5Ac-a2,3-Gal-pi,3-[Fuc-al,4]-GlcNAc), 2'FLNB, 4-FLNB, Leb (Lewis b; Fuc-al,2-Gal-bl,3-(Fuc-al,4)- GIcNAc),... .
  • Said glycosaminoglycan is preferably selected from the list consisting of chondroitin sulfate, heparan sulfate, hyaluronic acid, dermatan sulfate and heparin.
  • Said antigen of the human ABO blood group system refers to A determinant (GalNAc-alphal,3(Fuc- alphal,2)-Gal), B determinant (Gal-alphal,3(Fuc-alphal,2)-Gal) and H-determinant (Fuc-alphal,2-Gal), which can optionally be bound to a monosaccharide selected from the list consisting of betal,3-GlcNAc, betal,4-GlcNAc, betal,3-GalNAc and -betal,4-Glc.
  • Said lewis-type antigen oligosaccharide preferably refers to Hl antigen, which is Fuc-al,2-Gal-pi,3- GIcNAc, or in short 2'FLNB; Lewisa (or Lea), which is the trisaccharide Gal-pi,3-[Fuc-al,4]-GlcNAc, or in short 4-FLNB; Lewisb (or Leb), which is the tetrasaccharide Fuc-al,2-Gal-pi,3-[Fuc-al,4]-GlcNAc, or in short DiF-LNB; sialyl Lewisa (or sialyl Lea) which is 5-acetylneuraminyl-(2-3)-galactosyl-(l-3)- (fucopyranosyl-(l-4))-N-acetylglucosamine, or written in short Neu5Ac-a2,3-Gal-pi,3-[Fuc
  • said milk saccharide (preferably said milk oligosaccharide) is not lactose and none of the milk saccharides (preferably said milk oligosaccharides) of said mixture according to the invention are lactose.
  • said milk saccharide (preferably oligosaccharide) is not a disaccharide and none of the milk saccharides (preferably oligosaccharides) of said mixture according to the invention are a disaccharide.
  • said milk saccharide (preferably milk oligosaccharide) or any one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides (preferably milk oligosaccharides) of said mixture according to the invention is/are (has/have been) isolated from a microbial cultivation or fermentation, cell culture, enzymatic reaction or chemical reaction, preferably isolated from a microbial cultivation or fermentation, or cell culture, more preferably isolated from a microbial cultivation or fermentation, most preferably isolated from a microbial fermentation.
  • said milk saccharide (preferably milk oligosaccharide) or any one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides (preferably milk oligosaccharides) of said mixture according to the invention is/are obtained from an in vitro and/or ex vivo culture of cells, wherein said cells are preferably chosen from the list consisting of a microorganism, said microorganism is preferably a bacterium, a yeast or a fungus; a plant cell; an animal cell or a protozoan cell.
  • the latter bacterium preferably is an Escherichia coli strain, more preferably an Escherichia coli strain which is a K-12 strain, even more preferably the Escherichia coli K-12 strain is E. coli MG1655.
  • the latter yeast is preferably selected from Pichia pastoris, Yarrowia lipolitica, Saccharomyces cerevisiae and Kluyveromyces lactis.
  • the latter fungus belongs preferably to the genus Rhizopus, Dictyostelium, Penicillium, Mucor or Aspergillus.
  • the latter plant cell includes cells of flowering and non-flowering plants, as well as algal cells.
  • said plant cell is a tobacco, alfalfa, rice, cotton, rapeseed, tomato, corn, maize or soybean cell.
  • the latter animal cell is preferably derived from a non-human animal or is a genetically modified cell line derived from human cells excluding embryonic stem cells.
  • the latter protozoan cell preferably is a Leishmania tarentolae cell.
  • two or more different cells produce the milk saccharides of the mixture according to the invention, wherein each cell produces a different milk saccharide and/or a different mixture of milk saccharides.
  • said milk saccharide (preferably milk oligosaccharide) or any one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides (preferably milk oligosaccharides) of said mixture according to the invention is/are obtained from an in vitro and/or ex vivo culture of mammary epithelial cells, mammary myoepithelial cells and/or mammary progenitor cells, preferably wherein said cells are generated from non-mammary adult stem cells, more preferably wherein said cells are generated from mesenchymal stem cells.
  • WO2021/067641 and WO2021/242866 mimmary epithelial cells derived from non-mammary adult stem cells, preferably from mesenchymal stem cells
  • WO2021/142241 mimmary epithelial cells, mammary myoepithelial cells, mammary progenitor cells.
  • said milk saccharide (preferably milk oligosaccharide) or any one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides (preferably milk oligosaccharides) of said mixture according to the invention is obtained from an in vitro and/or ex vivo culture of microorganism cells, preferably said microorganism is a bacterium or a yeast, more preferably said microorganism is a bacterium, even more preferably said microorganism is Escherichia coli.
  • said nutritional composition comprises a mammalian milk oligosaccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different mammalian milk oligosaccharides, wherein said mammalian milk oligosaccharide(s) or wherein at least one, preferably at least two, more preferably at least three, most preferably all, oligosaccharides of said mixture is/are selected from a list consisting of a neutral MMO and a sialylated MMO, preferably a sialylated MMO.
  • said mammalian milk oligosaccharide(s) or wherein at least one, preferably at least two, more preferably at least three, most preferably all, oligosaccharides of said mixture is/are selected from a list consisting of lacto-N-triose II, 3-fucosyllactose, 2'-fucosyllactose, 6-fucosyllactose, 2',3-difucosyllactose, 2',2-difucosyllactose, 3,4- difucosyllactose, 6'-sialyllactose, 3'-sialyllactose, 3,6-disialyllactose, 6,6'-disialyllactose, 8,3- disialyllactose, 3,6-disialyllacto-N-tetraose, lactodifucotetraose, lactodifucot
  • a neutral oligosaccharide as used herein and as generally understood in the state of the art is an oligosaccharide that has no negative charge originating from a carboxylic acid group.
  • a neutral MMO is preferably selected from a list consisting of lacto-N-triose II, 3-fucosyllactose, 2'- fucosyllactose, 6-fucosyllactose, 2',3-difucosyllactose, 2',2-difucosyllactose, 3,4-difucosyllactose, lactodifucotetraose, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose II, lacto-N- fucopentaose I, lacto-N-fucopentaose III, lacto-N-fucopentaose V,
  • a sialylated IVIMO is preferably selected from a list consisting of 6'-sialyllactose, 3'-sialyllactose, 3,6- disialyllactose, 6,6'-disialyllactose, 8,3-disialyllactose, 3,6-disialyllacto-N-tetraose, sialyllacto-N- neotetraose d, sialyllacto-N-neotetraose c, sialyllacto-N-tetraose b, sialyllacto-N-tetraose a, monofucosylmonosialyllacto-N-neotetraose c, sialyllacto-N-hexaose and sialyllacto-N-neohexaose II; more
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.001 to 15.0 % (w/w), preferably 0.001 to 10.0 % (w/w), more preferably 0.001 to 5.0 % (w/w), even more preferably 0.001 to 3.0 % (w/w), even more preferably 0.001 to 1.0 % (w/w), most preferably 0.010 to 1.0 % (w/w), of the total weight, i.e.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.001 to 15.0 % (w/w), preferably 0.001 to 10.0 % (w/w), more preferably 0.001 to 5.0 % (w/w), even more preferably 0.001 to 3.0 % (w/w), even more preferably 0.001 to 1.0 % (w/w), most preferably 0.010 to 1.0% (w/w), of the total weight of the unheated nutritional composition (preferably unbaked nutritional composition).
  • the nutritional composition before being heated according to the invention comprises 0.001 to 15.0 % (w/w), preferably 0.001 to 10.0 % (w/w), more preferably 0.001 to 5.0 % (w/w), even more preferably 0.001 to 3.0 % (w/w), even more preferably 0.001 to 1.0 % (w/w), most preferably 0.010 to 1.0 % (w/w), i.e. % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • said unheated nutritional composition is a dough or batter, preferably a dough.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.05 to 15.0 % (w/w), preferably 0.05 to 10.0 % (w/w), more preferably 0.05 to 5.0 % (w/w), even more preferably 0.05 to 3.0 % (w/w), even more preferably 0.05 to 1.0 % (w/w), most preferably 0.10 to 1.0 % (w/w), of the total weight, i.e. % (w/w), of said nutritional composition before being heated at a temperature of 100°C or higher according to the invention as described herein.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.05 to 15.0 % (w/w), preferably 0.05 to 10.0 % (w/w), more preferably 0.05 to 5.0 % (w/w), even more preferably 0.05 to 3.0 % (w/w), even more preferably 0.05 to 1.0 % (w/w), most preferably 0.10 to 1.0 % (w/w), of the total weight of the unheated nutritional composition (preferably unbaked nutritional composition).
  • the nutritional composition before being heated according to the invention i.e.
  • unheated nutritional composition comprises 0.05 to 15.0 % (w/w), preferably 0.05 to 10.0 % (w/w), more preferably 0.05 to 5.0 % (w/w), even more preferably 0.05 to 3.0 % (w/w), even more preferably 0.05 to 1.0% (w/w), most preferably 0.10 to 1.0% (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.01 to 60.0 % (w/w), preferably 0.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.01 to 60.0 % (w/w), preferably 0.
  • the total weight of dry matter of the unheated nutritional composition comprises 0.01 to 60.0 % (w/w), preferably 0.
  • said unheated nutritional composition is a dough or batter, preferably a dough.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of the total weight of dry matter of said nutritional composition before being heated at a temperature of 100°C or higher according to the invention as described herein.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.1 to 40.0% (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0% (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of the total weight of dry matter of the unheated nutritional composition (preferably unbaked nutritional composition).
  • the total weight of dry matter of the nutritional composition before being heated according to the invention i.e.
  • unheated nutritional composition comprises 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides constitutes 0.01 to 60.0 % (w/w), preferably 0.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.01 to 60.0 % (w/w), preferably 0.
  • the total weight of dry matter of the heated nutritional composition comprises 0.01 to 60.0 %, preferably 0.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides constitutes 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of the total weight of dry matter of said nutritional composition after being heated at a temperature of 100°C or higher according to the invention as described herein.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) according to the invention constitutes 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of the total weight of dry matter of the heated nutritional composition, preferably baked nutritional composition as described herein.
  • the total weight of dry matter of the nutritional composition after being heated according to the invention comprises 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) in said nutritional composition after being heated at a temperature of 100°C or higher according to the invention constitutes at least 85.0%, preferably at least 87.5%, more preferably at least 90.0%, even more preferably at least 92.0%, of the amount of said milk oligosaccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) in said nutritional composition before being heated at said temperature of 100°C or higher according to the invention.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) in said heated nutritional composition according to the invention comprises at least 85.0%, preferably at least 87.5%, more preferably at least 90.0%, even more preferably at least 92.0%, of the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) in the unheated nutritional composition (preferably unbaked nutritional composition).
  • said unheated nutritional composition is a dough or batter, preferably a dough.
  • said nutritional composition comprises a milk saccharide or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides, wherein said nutritional composition was heated at a temperature of 100°C or higher.
  • milk saccharide and “milk saccharides” are as described in the section "milk saccharide”.
  • Pasteurization is a treatment using mild heat in order to eliminate pathogens and hence extend the shelf life of a product. As pasteurization typically occurs at a temperature which is lower than 100°C, it is not encompassed by the scope of the present invention. Hence, in a preferred embodiment of the present invention, said heating at a temperature of 100°C or higher does not encompass pasteurization.
  • said nutritional composition was heated at a temperature of at least 100°C, preferably at least 125°C, more preferably at least 135°C, even more preferably at least 150°C, even more preferably at least 160°C, even more preferably at least 170°C, most preferably at least 180°C.
  • said nutritional composition was heated at a temperature of 225°C or lower, preferably at 220°C or lower.
  • said nutritional composition was heated at a temperature of 100-225°C, preferably 125-225°C, more preferably 135-225°C, even more preferably 150-225°C, even more preferably 160-225°C, even more preferably 170-225°C, even more preferably 180-225°C, most preferably 180-220°C.
  • a preferred heating method comprises baking and/or extrusion.
  • Said extrusion can be dry extrusion or wet extrusion (wherein steam injection is applied), preferably said extrusion is dry extrusion.
  • said extrusion is not wet extrusion.
  • a more preferred heating method comprises baking.
  • baking of a nutritional composition can be achieved by subjecting it to heat (e.g. hot air), preferably dry heat.
  • the nutritional composition is steam-heated.
  • said nutritional composition according to the invention was heated by baking, more preferably by subjecting the nutritional composition to heat, preferably dry heat.
  • Said oven can be a direct heating oven or an indirect heating oven.
  • Said oven can be a continuous oven or a semi- continuous oven.
  • Such types of ovens are known to the skilled person. Examples can be found in said book entitled “"Food processing technology: principles and practice” on p. 343-348.
  • the feature "oven” as applied in the field and the present application does not encompass a barrel (which can be heated/cooled) used during extrusion.
  • said nutritional composition was heated for at least 2 minutes, preferably at least 3 minutes, more preferably at least 5 minutes, even more preferably at least 7 minutes, most preferably at least 10 minutes. In an additional and/or alternative preferred embodiment of the invention, said nutritional composition was heated for 60 minutes or less, preferably 45 minutes or less, more preferably 30 minutes or less, even more preferably 25 minutes or less, most 20 minutes or less.
  • the skilled person understands that the preferred heat and duration will vary depending upon the particular nutritional composition, preferably baked nutritional composition. Depending on the properties of the nutritional composition, thickness of said composition and desired doneness, the skilled person will be able to routinely determine the appropriate temperature and duration of the baking process.
  • said nutritional composition is heated until the desired doneness has been reached, more preferably heated until said nutritional composition is cooked.
  • said nutritional composition according to the invention is obtainable by a method according to the second aspect of the invention.
  • the invention provides a method for the production of a heated nutritional composition, said method comprising the steps of: providing a solution or a powder, preferably a powder, comprising a milk saccharide (preferably a milk oligosaccharide) or comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides), providing one or more food and/or feed ingredients, preferably one or more functional ingredients, mixing said solution or powder with said food and/or feed ingredient(s) to obtain a mixture, preferably a homogenous mixture, and heating said obtained mixture at a temperature of 100°C or higher, preferably until said mixture has reached the desired doneness, more preferably until said mixture is cooked.
  • a solution or a powder preferably a powder
  • a milk saccharide preferably a milk oligosaccharide
  • said nutritional composition is according to the first aspect of the present invention (it is referred to the section "Nutritional composition").
  • the term “cooked” is preferably replaced with “baked”.
  • milk saccharide "milk oligosaccharide”, “milk saccharides”, “milk oligosaccharides”, “food ingredient”, “feed ingredient”, “functional ingredient”, “heating” as described throughout the second aspect of the invention and claims are preferably as described in the first aspect of the invention.
  • said solution comprising a milk saccharide (preferably a milk oligosaccharide) or comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides) is not milk.
  • said solution is water.
  • said powder comprising a milk saccharide (preferably a milk oligosaccharide) or comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides) is not milk powder.
  • a milk saccharide preferably a milk oligosaccharide
  • said powder comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides) is not milk powder.
  • the mixture obtained by mixing said solution or powder with said food and/or feed ingredients is a dough or batter, preferably a dough.
  • said mixing is preferably done by kneading.
  • said method according to the invention further comprises a step wherein the obtained mixture, preferably said homogenous mixture, is provided a desired shape.
  • the obtained dough can optionally be divided in small portions and each small portion is subsequently formed into the shape of a biscuit.
  • said method according to the invention further comprises a leavening step, said step is preferably conducted after obtaining said mixture, preferably said homogenous mixture.
  • a leavening agent preferably as described in the first aspect of the invention
  • the solution or powder comprising said saccharide(s) and one or more food and/or feed ingredients.
  • leavening causes the mixture (preferably homogenous mixture) to expand by gas which is released when a leavening agent comes into contact with a liquid, acid or heat.
  • said method according to the invention further comprises a drying step, preferably at a temperature > 50°C and/or preferably ⁇ 100°C, more preferably ⁇ 80°C, even more preferably ⁇ 70 °C, most preferably ⁇ 60°C.
  • said drying step is conducted in an oven.
  • said drying step occurs after obtaining said heated mixture.
  • said drying step occurs before heating said mixture.
  • heating said mixture comprises baking and/or extrusion.
  • Said extrusion can be dry extrusion or wet extrusion (wherein steam injection is applied), preferably said extrusion is dry extrusion.
  • heating said mixture comprises baking, preferably in an oven. Accordingly, such a method results in the production of a baked nutritional composition, preferably a baked nutritional composition as described in the first aspect of the present invention (it is referred to the section "Nutritional composition").
  • said food and/or feed ingredient(s) is as described in the section "Nutritional composition ingredients" of the first aspect of the invention.
  • said mixture obtained by mixing said solution or powder with said food and/or feed ingredient(s) comprises 3.0-70.0 % w/w, preferably 3.0-60.0 % w/w, more preferably 3.0-50.0 % w/w, even more preferably 5.0-50.0 % w/w, of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • said mixture obtained by mixing said solution or powder with said food and/or feed ingredient(s) comprises 10.0-70.0 % w/w, preferably 15.0-70.0 % w/w, more preferably 20.0-70.0 % w/w, even more preferably 25.0-70.0 % w/w, even more preferably 30.0- 70.0 % w/w, even more preferably 35.0-70.0 % w/w, even more preferably 35.0-60.0 % w/w, most preferably 40.0-60.0 % w/w, moisture; preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • said heated nutritional composition according to the invention comprises 15.0 % (w/w) or less, preferably 13.0 % (w/w) or less, more preferably 10.0 % (w/w) or less, most preferably 8.0 % (w/w) or less of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • the method for the production of a heated nutritional composition comprises a step of providing flour and/or meal, preferably flour.
  • said flour and/or meal is provided before mixing all ingredients to obtain a mixture.
  • Said flour and said meal are preferably as described in the section "Nutritional composition ingredients" of the first aspect of the invention.
  • the method for the production of a heated nutritional composition comprises a step of providing a liquid.
  • a liquid is provided before mixing all ingredients to obtain a mixture.
  • Said liquid is preferably as described in the section "Nutritional composition ingredients" of the first aspect of the invention.
  • the method for the production of a heated nutritional composition comprises a step of providing one or more functional ingredients.
  • said functional ingredient is provided before mixing all ingredients to obtain a mixture.
  • Said functional ingredient is preferably as described in the section "Nutritional composition ingredients" of the first aspect of the invention.
  • the method for the production of a heated nutritional composition comprises a leavening step. Said step is preferably conducted after obtaining said mixture, preferably said homogenous mixture.
  • a leavening agent preferably as described in the first aspect of the invention
  • a leavening agent is mixed together with the solution or powder comprising said saccharide(s) and one or more food and/or feed ingredients.
  • the invention provides a method for the production of a heated nutritional composition, said method comprising the steps of: providing a solution or a powder, preferably a powder, comprising a milk saccharide (preferably milk oligosaccharide) or comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides), providing flour and/or meal, preferably flour, providing a liquid, providing one or more food and/or feed ingredients, preferably one or more functional ingredients, mixing said solution or powder with all ingredients to obtain a mixture, preferably a homogenous mixture, optionally leaven said mixture, optionally providing a desired shape to said mixture, optionally leaven said mixture, and heating said obtained mixture at a temperature of 100°C or higher, preferably until said mixture has reached the desired doneness, more preferably until said mixture is cooked.
  • a solution or a powder preferably a powder, comprising a milk saccharide (preferably milk oligosacc
  • said method according to the invention further comprises a drying step, preferably at a temperature > 50°C and/or preferably ⁇ 100°C, more preferably ⁇ 80°C, even more preferably ⁇ 70 °C, most preferably ⁇ 60°C ).
  • said drying step is conducted in an oven.
  • said drying step occurs after obtaining said heated mixture.
  • said drying step occurs before heating said mixture.
  • the step of "providing one or more food and/or feed ingredients” is optional in the disclosed methods (in other words, said step can be omitted) if said methods comprise the step of "providing flour and/or meal, preferably flour” and/or the step of "providing a liquid”.
  • said nutritional composition is according to the first aspect of the present invention (it is referred to the section "Nutritional composition”).
  • said food and/or feed ingredients is as described in the section "Nutritional composition ingredients" of the first aspect of the invention.
  • said flour and said meal are as described in the section "Nutritional composition ingredients” of the first aspect of the invention.
  • said liquid is as described in the section "Nutritional composition ingredients” of the first aspect of the invention.
  • the invention provides a method for the production of a baked nutritional composition, said method comprising the steps of: providing a solution or a powder, preferably a powder, comprising a mammalian milk oligosaccharide or comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different mammalian milk oligosaccharides, providing flour and/or meal, preferably flour, providing a liquid, providing one or more food and/or feed ingredients, preferably one or more functional ingredients, mixing said solution or powder with all ingredients to obtain a mixture, preferably a homogenous mixture, optionally leaven said mixture, optionally providing a desired shape to said mixture, optionally leaven said mixture, and baking said obtained mixture at a temperature of 100°C or higher, preferably until said mixture has reached the desired doneness, more preferably until said mixture is baked.
  • said method according to the invention further comprises a drying step, preferably at a temperature > 50°C and/or preferably ⁇ 100°C, more preferably ⁇ 80°C, even more preferably ⁇ 70 °C, most preferably ⁇ 60°C ).
  • said drying step is conducted in an oven.
  • said drying step occurs after obtaining said heated mixture.
  • said drying step occurs before heating said mixture.
  • said nutritional composition is according to the first aspect of the present invention (it is referred to the section "Nutritional composition”).
  • said food and/or feed ingredients is as described in the section "Nutritional composition ingredients" of the first aspect of the invention.
  • said flour and said meal are as described in the section "Nutritional composition ingredients” of the first aspect of the invention.
  • said liquid is as described in the section "Nutritional composition ingredients” of the first aspect of the invention.
  • the mixture obtained by mixing said solution or powder with the other ingredients is a dough or batter, preferably a dough.
  • said mixing is preferably done by kneading.
  • heating said mixture comprises baking and/or extrusion.
  • Said extrusion can be dry extrusion or wet extrusion (wherein steam injection is applied), preferably said extrusion is dry extrusion.
  • heating said mixture comprises baking, preferably in an oven. Accordingly, such a method results in the production of a baked nutritional composition, preferably a baked nutritional composition as described in the first aspect of the present invention (it is referred to the section "Nutritional composition").
  • said obtained mixture (before heating/baking) comprises 0.001 to 15.0 %, preferably 0.001 to 10.0 %, more preferably 0.001 to 5.0 %, even more preferably 0.001 to 3.0 %, even more preferably 0.001 to 1.0 %, most preferably 0.010 to 1.0 %, i.e. % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • said obtained mixture (before heating/baking) comprises 0.05 to 15.0 % (w/w), preferably 0.05 to 10.0 % (w/w), more preferably 0.05 to 5.0 % (w/w), even more preferably 0.05 to 3.0 % (w/w), even more preferably 0.05 to 1.0 % (w/w), most preferably 0.10 to 1.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the total weight of dry matter of said obtained mixture comprises 0.01 to 60.0 % (w/w), preferably 0. 1 to 60.0 % (w/w), more preferably 1.0 to 60.0 % (w/w), even more preferably 1.0 to 50.0 % (w/w), even more preferably 5.0 to 50.0 % (w/w), even more preferably 10.0 to 50.0 % (w/w), even more preferably 10.0 to 40.0 % (w/w), most preferably 10.0 to 40.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the total weight of dry matter of said obtained mixture comprises 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the total weight of dry matter of said heated (preferably baked) nutritional composition comprises 0.01 to 60.0 % (w/w), preferably 0. 1 to 60.0 % (w/w), more preferably 1.0 to 60.0 % (w/w), even more preferably 1.0 to 50.0 % (w/w), even more preferably 5.0 to 50.0 % (w/w), even more preferably 10.0 to 50.0 % (w/w), even more preferably 10.0 to 40.0 % (w/w), most preferably 10.0 to 40.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the total weight of dry matter of said heated (preferably baked) nutritional composition comprises 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the amount of said milk saccharide (preferably milk oligosaccharide) or the amount of said mixture of at least two different milk saccharides (preferably milk oligosaccharides) in the obtained heated nutritional composition is at least 85.0%, preferably at least 87.5%, more preferably at least 90.0%, even more preferably at least 92.0%, of the amount of said mammalian milk oligosaccharide or the amount of said mixture of at least two different mammalian milk oligosaccharides in the mixture before heating said mixture.
  • said mixture obtained by mixing said solution or powder with the other ingredients comprises 3.0-70.0 % w/w, preferably 3.0-60.0 % w/w, more preferably 3.0-50.0 % w/w, even more preferably 5.0-50.0 % w/w, most preferably 10.0-50.0 % w/w, of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • said mixture obtained by mixing said solution or powder with the other ingredients comprises 10.0-70.0 % w/w, preferably 15.0-70.0 % w/w, more preferably 20.0-70.0 % w/w, even more preferably 25.0-70.0 % w/w, even more preferably 30.0- 70.0 % w/w, even more preferably 35.0-70.0 % w/w, even more preferably 35.0-60.0 % w/w, most preferably 40.0-60.0 % w/w, of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • said heated nutritional composition comprises 15.0 % (w/w) or less, preferably 13.0 % (w/w) or less, more preferably 10.0 % (w/w) or less, most preferably 8.0 % (w/w) or less of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • a solution or a powder comprising a milk saccharide (preferably milk oligosaccharide) or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides) is provided.
  • a milk saccharide preferably milk oligosaccharide
  • a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides) is provided.
  • said solution or powder comprising a milk saccharide (preferably milk oligosaccharide) or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides), is typically isolated (preferably obtained) from a microbial cultivation or fermentation, cell culture, enzymatic reaction or chemical reaction, preferably isolated from a microbial cultivation or fermentation, or cell culture, more preferably isolated from a microbial cultivation or fermentation, most preferably isolated from a microbial fermentation, as described in the present application, resulting in a solution containing said milk saccharide or said mixture of different milk saccharides.
  • Such a solution can for example be obtained by a method comprising the steps of:
  • said solution or powder comprising a milk saccharide (preferably milk oligosaccharide) or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides), is obtained from an in vitro and/or ex vivo culture of cells, wherein said cells are preferably chosen from the list consisting of a microorganism, said microorganism is preferably a bacterium, a yeast or a fungus; a plant cell; an animal cell or a protozoan cell.
  • the latter bacterium preferably is an Escherichia coli strain, more preferably an Escherichia coli strain which is a K-12 strain, even more preferably the Escherichia coli K-12 strain is E. coli MG1655.
  • the latter yeast is preferably selected from Pichia pastoris, Yarrowia lipolitica, Saccharomyces cerevisiae and Kluyveromyces lactis.
  • the latter fungus belongs preferably to the genus Rhizopus, Dictyostelium, Penicillium, Mucor or Aspergillus.
  • the latter plant cell includes cells of flowering and non-flowering plants, as well as algal cells.
  • said plant cell is a tobacco, alfalfa, rice, cotton, rapeseed, tomato, corn, maize or soybean cell.
  • the latter animal cell is preferably derived from a non-human animal or is a genetically modified cell line derived from human cells excluding embryonic stem cells.
  • the latter protozoan cell preferably is a Leishmania tarentolae cell.
  • two or more different cells produce the milk saccharides of the mixture according to the invention, wherein each cell produces a different milk saccharide and/or a different mixture of milk saccharides.
  • said solution or powder comprising a milk saccharide preferably milk oligosaccharide
  • a milk saccharide preferably milk oligosaccharide
  • a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides) is obtained from an in vitro and/or ex vivo culture of mammary epithelial cells, mammary myoepithelial cells and/or mammary progenitor cells, preferably wherein said cells are generated from non-mammary adult stem cells, more preferably wherein said cells are generated from mesenchymal stem cells.
  • WO2021/067641 and WO2021/242866 mimmary epithelial cells derived from non-mammary adult stem cells, preferably from mesenchymal stem cells
  • WO2021/142241 mimmary epithelial cells, mammary myoepithelial cells, mammary progenitor cells.
  • said solution or powder comprising a milk saccharide (preferably milk oligosaccharide) or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides (preferably milk oligosaccharides), is obtained from an in vitro and/or ex vivo culture of microorganism cells, preferably said microorganism is a bacterium or a yeast, more preferably said microorganism is a bacterium, even more preferably said microorganism is Escherichia coli.
  • slurry and powder are known to the skilled person which can be used to obtain a slurry (i.e. concentrated solution) or powder from a solution containing said milk saccharide or said mixture of at least two different milk saccharides.
  • the slurry and powder according to the invention are preferably obtained by spray drying, freeze drying, spray freeze-drying, crystallization, lyophilization, band or belt drying, drum or roller drying, and/or agitated thin film drying, preferably by spray drying, drum or roller drying, or agitated thin film drying, of a solution containing said milk saccharide or said mixture of milk saccharides.
  • said obtained powder contains ⁇ 15 wt. %, preferably ⁇ 10 wt. %, more preferably ⁇ 9 wt. %, more preferably ⁇ 8 wt. %, more preferably ⁇ 7 wt. %, even more preferably ⁇ 5 wt. %, even more preferably ⁇ 4 wt. % of liquid, even more preferably ⁇ 3 wt. % of liquid, even more preferably ⁇ 2 wt. % of liquid, most preferably ⁇ 1 wt. %, preferably wherein said liquid is water.
  • the liquid content is preferably determined by a thermogravimetric analysis or Karl Fisher titration analysis, more preferably by Karl Fisher titration analysis (preferably as described herein).
  • said milk saccharide and said mixture of different milk saccharides is as described in the section "Milk saccharide" of the first aspect of the invention.
  • a method for the production of a heated nutritional composition comprising the steps of: providing a solution or a powder, preferably a powder, comprising a milk saccharide, preferably milk oligosaccharide, or comprising a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different milk saccharides, preferably milk oligosaccharides, providing one or more food and/or feed ingredients, preferably one or more functional ingredients, mixing said solution or powder with said food and/or feed ingredients to obtain a mixture, preferably a homogenous mixture, and preferably wherein said mixture is a dough or batter, more preferably a dough, optionally leaven said mixture, optionally providing a desired shape to said mixture, optionally leaven said mixture, and heating, preferably comprising baking and/or extrusion, more preferably comprising baking, said mixture at a temperature of 100°C or higher preferably until said mixture has reached the desired doneness, more preferably until said mixture
  • a method according to embodiment 1 or 2, wherein said milk saccharide or at least one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides in said mixture is obtained from an in vitro and/or ex vivo culture of cells, wherein said cells are preferably chosen from the list consisting of a microorganism, a plant cell, an animal cell or a protozoan cell, more preferably wherein said cells are a bacterium, a yeast or a fungus.
  • a method according to any one of embodiments 1 to 3, said method further comprises a drying step, preferably at a temperature > 50°C and/or preferably ⁇ 100°C, more preferably ⁇ 80°C, even more preferably ⁇ 70 °C, most preferably ⁇ 60°C.
  • heating comprises baking and/or extrusion, preferably baking and/or dry extrusion, more preferably baking.
  • heating is for at least 2 minutes, preferably at least 3 minutes, more preferably at least 5 minutes, even more preferably at least 7 minutes, most preferably at least 10 minutes and/or 60 minutes or less, preferably 45 minutes or less, more preferably 30 minutes or less, even more preferably 25 minutes or less, most 20 minutes or less.
  • said milk saccharide or at least one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides in said mixture is a milk oligosaccharide, preferably having a degree of polymerization of 2-20, more preferably 3-20, even more preferably 3-9, even more preferably 3-8, most preferably 3-7.
  • said milk saccharide or at least one, preferably at least two, more preferably at least three, even more preferably at least four, most preferably all, of said milk saccharides in said mixture is selected from the list consisting of (i) oligosaccharide comprising lactose (Gal-bl,4-Glc) at its reducing end; (ii) oligosaccharide comprising N-acetyllactosamine (LacNAc) at its reducing end; (iii) oligosaccharide comprising lacto-N-biose (LNB) at its reducing end; (iv) milk glycosaminoglycan; (v) antigen of the human ABO blood group system, optionally bound to a monosaccharide selected from the list consisting of betal,3-GlcNAc, betal,4- GIcNAc, betal,3-GalNAc and -betal,4-
  • a method according to any one of embodiments 1 to 11, wherein said milk saccharide and at least one, preferably at least two, more preferably at least three, most preferably all, oligosaccharides of said mixture is selected from a list consisting of a neutral mammalian milk oligosaccharide and a sialylated mammalian milk oligosaccharide, preferably a sialylated mammalian milk oligosaccharide.
  • a method according to any one of embodiments 1 to 12, wherein said obtained mixture comprises, preferably consists of, 3.0-70.0 % w/w, preferably 3.0-60.0 % w/w, more preferably 3.0-50.0 % w/w, even more preferably 5.0-50.0 % w/w, most preferably 10.0-50.0 % w/w, of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • a method according to any one of embodiments 1 to 14, wherein said obtained mixture comprises, preferably consists of, 0.001 to 15.0 %, preferably 0.001 to 10.0 %, more preferably 0.001 to 5.0 %, even more preferably 0.001 to 3.0 %, even more preferably 0.001 to 1.0 %, most preferably 0.010 to 1.0 %, i.e. % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • the total weight of dry matter of said obtained mixture comprises, preferably consists of, 0.01 to 60.0 % (w/w), preferably 0. 1 to 60.0 % (w/w), more preferably 1.0 to 60.0 % (w/w), even more preferably 1.0 to 50.0 % (w/w), even more preferably 5.0 to 50.0 % (w/w), even more preferably 10.0 to 50.0 % (w/w), even more preferably 10.0 to 40.0 % (w/w), most preferably 10.0 to 40.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • said heated nutritional composition comprises, preferably consists of, 15.0 % (w/w) or less, preferably 13.0 % (w/w) or less, more preferably 10.0 % (w/w) or less, most preferably 8.0 % (w/w) or less of moisture, preferably wherein said moisture is liquid, more preferably said moisture is milk and/or water, even more preferably said moisture is water.
  • the total weight of dry matter of said heated nutritional composition comprises, preferably consists of, 0.01 to 60.0 % (w/w), preferably 0. 1 to 60.0 % (w/w), more preferably 1.0 to 60.0 % (w/w), even more preferably 1.0 to 50.0 % (w/w), even more preferably 5.0 to 50.0 % (w/w), even more preferably 10.0 to 50.0 % (w/w), even more preferably 10.0 to 40.0 % (w/w), most preferably 10.0 to 40.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • a nutritional composition comprising a milk saccharide or mixture of at least two different milk saccharides, preferably a milk oligosaccharide or mixture of at least two different milk oligosaccharides, wherein said nutritional composition was heated at a temperature of 100°C or higher.
  • a nutritional composition according to embodiment 20 which is obtainable by a method according to any one of embodiments 1 to 19.
  • a nutritional composition according to any one of embodiments 20 to 32, wherein the total weight of dry matter of said nutritional composition comprises, preferably consists of, 0.01 to 60.0 % (w/w), preferably 0. 1 to 60.0 % (w/w), more preferably 1.0 to 60.0 % (w/w), even more preferably 1.0 to 50.0 % (w/w), even more preferably 5.0 to 50.0 % (w/w), even more preferably 10.0 to 50.0 % (w/w), even more preferably 10.0 to 40.0 % (w/w), most preferably 10.0 to 40.0 % (w/w), of said milk saccharide or said mixture of milk saccharides.
  • a method for the production of a baked nutritional composition comprising the steps of: providing a solution or a powder, preferably a powder, comprising a human milk oligosaccharide (HMO) or a mixture of at least two, preferably at least three, more preferably at least 4, most preferably at least 5, different human milk oligosaccharides (HMOs), providing one or more food and/or feed ingredients, preferably one or more functional ingredients, mixing said solution or powder with said food and/or feed ingredients to obtain a mixture, preferably a homogenous mixture, and preferably wherein said mixture is a dough or batter, more preferably a dough, optionally leaven said mixture, optionally providing a desired shape to said mixture, optionally leaven said mixture, and baking said obtained mixture at a temperature of at least 150°C until said mixture has reached the desired doneness, preferably until said mixture is cooked.
  • HMO human milk oligosaccharide
  • HMOs human milk oligosaccharide
  • said obtained mixture comprises 0.05 to 15.0 % (w/w), preferably 0.05 to 10.0 % (w/w), more preferably 0.05 to 5.0 % (w/w), even more preferably 0.05 to 3.0 % (w/w), even more preferably 0.05 to 1.0 % (w/w), most preferably 0.10 to 1.0 % (w/w), of said HMO or said mixture of HMOs.
  • a method according to any one of embodiments 1 to 4, wherein the total weight of dry matter of said obtained mixture comprises 0.1 to 40.0% (w/w), preferably 0.1 to 30.0% (w/w), more preferably 0.1 to 20.0 % (w/w), even more preferably 0.1 to 10.0 % (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of said HMO or said mixture of HMOs.
  • said obtained mixture comprises 15.0-70.0 % w/w, preferably 20.0-70.0 % w/w, more preferably 25.0-70.0 % w/w, even more preferably 30.0-70.0 % w/w, even more preferably 35.0-70.0 % w/w, even more preferably 35.0-60.0 % w/w, most preferably 40.0-60.0 % w/w, of moisture.
  • said baked nutritional composition comprises 15.0 % (w/w) or less, preferably 13.0 % (w/w) or less, more preferably 10.0 % (w/w) or less, most preferably 8.0 % (w/w) or less of moisture.
  • a method according to any one of embodiments 1 to 9, said method further comprises a drying step, preferably at a temperature > 50°C and/or preferably ⁇ 100°C, more preferably ⁇ 80°C, even more preferably ⁇ 70 °C, most preferably ⁇ 60°C.
  • HMO and at least one, preferably at least two, more preferably at least three, most preferably all, HMOs of said mixture is selected from a list consisting of a neutral HMO and a sialylated HMO, preferably a sialylated HMO.
  • the total weight of dry matter of said baked nutritional composition comprises 0.1 to 40.0 % (w/w), preferably 0.1 to 30.0 % (w/w), more preferably 0.1 to 20.0% (w/w), even more preferably 0.1 to 10.0% (w/w), even more preferably 0.1 to 5.0 % (w/w), most preferably 0.2 to 5.0 % (w/w), of said milk HMO or said mixture of HMOs.
  • a nutritional composition obtainable by a method according to any one of embodiments 1 to 16.
  • the verbs "to comprise”, “to have” and “to contain”, and their conjugations are used in their non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded.
  • the verb "to consist essentially of” means that a solution as defined herein may comprise additional component(s) than the ones specifically identified, said additional component(s) not altering the unique characteristic of the invention.
  • the verbs "to comprise”, “to have” and “to contain”, and their conjugations may be preferably replaced by "to consist” (and its conjugations) or “to consist essentially of” (and its conjugations) and vice versa.
  • indefinite article “a” or “an” does not exclude the possibility that more than one of the element is present, unless the context clearly requires that there be one and only one of the elements.
  • the indefinite article “a” or “an” thus usually means “at least one”.
  • the word “about” or “approximately” or “around” when used in association with a numerical value preferably means that the value may be the given value (of 10) more or less 10%, preferably 5%, more preferably 1% of the value.
  • monosaccharide refers to a sugar that is not decomposable into simpler sugars by hydrolysis, is classed either an aldose or ketose, and contains one or more hydroxyl groups per molecule. Monosaccharides are saccharides containing only one simple sugar.
  • LNT II LNT-II
  • LN3 lacto-N-triose II
  • lacto-N-triose II lacto-N-triose
  • lacto-N-triose lacto-N-triose
  • GlcNAcpi-3Gaipi-4Glc as used in the present invention, are used interchangeably.
  • LNT lacto-N-tetraose
  • lacto-/V-tetraose lacto-/V-tetraose
  • Gaipi-3GlcNAcpi-3Gaipi-4Glc as used in the present invention, are used interchangeably.
  • LNnT lacto-N-neotetraose
  • lacto-/V-neotetraose lacto-/V-neotetraose
  • Gaipi-4GlcNAcpi- 3Gaipi-4Glc as used in the present invention, are used interchangeably.
  • LSTa LS-Tetrasaccharide a
  • Sialyl-lacto-N-tetraose a sialyllacto-N-tetraose a
  • Neu5Ac-a2,3-Gal-bl,3-GlcNAc-bl,3-Gal-bl,4-Glc as used in the present invention, are used interchangeably.
  • LSTb LS-Tetrasaccharide b
  • Sialyl-lacto-N-tetraose b sialyllacto-N-tetraose b
  • Gal- bl,3-(Neu5Ac-a2,6)-GlcNAc-bl,3-Gal-bl,4-Glc as used in the present invention, are used interchangeably.
  • LSTc "LS-Tetrasaccharide c", "Sialyl-lacto-N-tetraose c", “sialyllacto-N-tetraose c”, “sialyllacto-N-neotetraose c" or "Neu5Ac-a2,6-Gal-bl,4-GlcNAc-bl,3-Gal-bl,4-Glc" as used in the present invention, are used interchangeably.
  • LSTd "LS-Tetrasaccharide d"
  • Sialyl-lacto-N-tetraose d "sialyllacto-N-tetraose d”
  • sialyllacto-N-neotetraose d or "Neu5Ac-a2,3-Gal-bl,4-GlcNAc-bl,3-Gal-bl,4-Glc".
  • DSLNnT and “Disialyllacto-N-neotetraose” are used interchangeably and refer to Neu5Ac- a2,6-[Neu5Ac-a2,6-Gal-bl,4-GlcNAc-bl,3]-Gal-bl,4-Glc.
  • DSLNT and “Disialyllacto-N-tetraose” are used interchangeably and refer to Neu5Ac-a2,6- [Neu5Ac-a2,3-Gal-bl,3-GlcNAc-bl,3]-Gal-bl,4-Glc.
  • LNFP-I lacto-N-fucopentaose I
  • LNFP I lacto-N-fucopentaose I
  • LNF I LNF I
  • LNF I OH type I determinant "LNF I", “LNF1", “LNF 1” and “Blood group H antigen pentaose type 1" are used interchangeably and refer to Fuc-al,2-Gal-bl,3-GlcNAc-bl,3-Gal-bl,4-Glc.
  • GalNAc-LNFP-l and "blood group A antigen hexaose type I" are used interchangeably and refer to GalNAc-al,3-(Fuc-al,2)-Gal-bl,3-GlcNAc-bl,3-Gal-bl,4-Glc.
  • LNFP-II lacto-N-fucopentaose II
  • lacto-N-fucopentaose II are used interchangeably and refer to Gal-bl,3-(Fuc- al,4)-GlcNAc-bl,3-Gal-bl,4-Glc.
  • LNFP-III and "lacto-N-fucopentaose III” are used interchangeably and refer to Gal-bl,4-(Fuc- al,3)-GlcNAc-bl,3-Gal-bl,4-Glc.
  • LNFP-V lacto-N-fucopentaose V
  • lacto-N-fucopentaose V are used interchangeably and refer to Gal-bl,3- GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc.
  • LNFP-VI LNnFP V
  • lacto-N-neofucopentaose V are used interchangeably and refer to Gal-bl,4-GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc.
  • LNnFP I and “Lacto-N-neofucopentaose I” are used interchangeably and refer to Fuc-al,2- Gal-bl,4-GlcNAc-bl,3-Gal-bl,4-Glc.
  • LNDFH I Lacto-N-difucohexaose I
  • LNDFH-I LNDFH I
  • LNDFH I LNDFH I
  • Le b -lactose LNDFH I
  • Lewis-b hexasaccharide are used interchangeably and refer to Fuc-al,2-Gal-bl,3-[Fuc-al,4]-GlcNAc-bl,3-Gal- bl,4-Glc.
  • LNDFH II Lacto-N-difucohexaose II
  • Lewis a-Lewis x and “LDFH II” are used interchangeably and refer to Fuc-al,4-(Gal-bl,3)-GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc.
  • LNnDFH Lacto-N-neoDiFucohexaose
  • Lewis x hexaose are used interchangeably and refer to Gal-bl,4-(Fuc-al,3)-GlcNAc-bl,3-Gal-bl,4-(Fuc-al,3)-Glc.
  • alpha-tetrasaccharide and “A-tetrasaccharide” are used interchangeably and refer to Gal N Acai, 3-(Fuc-al,2)-Gal-bl,4-Glc. Examples
  • 3'sialyllactose (3'SL) and 6'sialyllactose (6'SL) were recombinantly produced in E. coli and purified as described in WO 2022/034079 (Examples 11, 13 and 14; the E. coli strains are described in Example 3 of WO 2018/122225 for 6'SL and Example 7 of WO 2018/122225 for 3'SL wherein an alpha-2, 3- sialyltransferase from Pasteurella multocida, i.e.
  • the milk saccharide composition in pre-baking and post-baking dog cookies/biscuits samples were determined by mixing 4 g of said sample into 20 ml of citrate buffer (0.02M, pH 5.0). An internal standard (galactoronic acid) was added until a final concentration of 3 mg/ml. The mixture was incubated at 70°C for 10 min before it was blended at 20000 rpm for 5 min using an ultra turrax (IKA T 18), followed by shaking a 37°C for 30 min. Protein precipitation was performed using MeOH at the ratio of 2:1 volume of MeOH : sample. The mixture was kept at -20°C for lh, followed by centrifugation at 14000 rpm for 5 min.
  • the saccharides were separated in 60 min while maintaining a constant ratio of 25 % of eluent B using the following gradient: an initial isocratic step maintained for 10 min of 75 % of eluent A, an initial increase from 0 to 4 % of eluent C over 8 min, a second isocratic step maintained for 6 min of 71 % of eluent A and
  • Example 2 Materials and Methods (production of dog cookies/biscuits by baking)
  • Oligosaccharides 3'SL and 6'SL were mixed into 1 kg of dog food meal (moisture content of 5.5 % w/w) from United Petfood (major components shown in Table 1) at the inclusion ratio of 12 g oligosaccharides per kg dog food. Before subjecting the mixture to the baking process, said mixture was first homogenized for 15 min. A portion of 100 g of the homogenized mixture was preserved at -80°C for analytics ("pre-baked sample").
  • the homogenized dog food (860 g) was added to 640 mL water and mixed by hand to obtain a dough.
  • the dough was shaped manually (approximately 10 g per cookie/biscuit) before baking for 11 min at 220°C in a pre-heated oven. Subsequently, the baked cookies/biscuits were dried in the oven for 50 min at 60°C. A portion of 100 g was preserved at -80°C for analytics ("post-baked sample").
  • Dog food cookies/biscuits (water content ⁇ 10.0 % w/w) containing spray-dried 3'SL and spray-dried 6'SL were prepared according to the baking procedure described in Example 2.
  • the concentrations of 3'SL and 6'SL were determined before baking ("pre-baking") and after baking ("post-baking") and expressed as mg/g dried matter (it is referred to Example 1) as shown in Table 2.

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Abstract

La présente invention concerne une composition nutritionnelle chauffée, plus spécifiquement cuite, comprenant un saccharide de lait ou un mélange d'au moins deux saccharides de lait différents.
PCT/EP2023/074447 2022-09-06 2023-09-06 Compositions nutritionnelles cuites comprenant des oligosaccharides de lait humain WO2024052406A1 (fr)

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