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EP3703721A1 - Composition et procédé pour des composés germinatifs dans des produits alimentaires et des boissons probiotiques pour la consommation humaine - Google Patents

Composition et procédé pour des composés germinatifs dans des produits alimentaires et des boissons probiotiques pour la consommation humaine

Info

Publication number
EP3703721A1
EP3703721A1 EP18873222.6A EP18873222A EP3703721A1 EP 3703721 A1 EP3703721 A1 EP 3703721A1 EP 18873222 A EP18873222 A EP 18873222A EP 3703721 A1 EP3703721 A1 EP 3703721A1
Authority
EP
European Patent Office
Prior art keywords
bacillus
food
tea
beverage
probiotic
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
EP18873222.6A
Other languages
German (de)
English (en)
Other versions
EP3703721A4 (fr
Inventor
Jordan E. CHURCH
Gabriel F.K. EVERETT
Charles J. Greenwald
Jr. Lester LEVY
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.)
NCH Corp
Original Assignee
NCH Corp
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 NCH Corp filed Critical NCH Corp
Publication of EP3703721A1 publication Critical patent/EP3703721A1/fr
Publication of EP3703721A4 publication Critical patent/EP3703721A4/fr
Pending legal-status Critical Current

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
    • A23F3/00—Tea; Tea substitutes; Preparations thereof
    • A23F3/06—Treating tea before extraction; Preparations produced thereby
    • A23F3/14—Tea preparations, e.g. using additives
    • 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
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • 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
    • A23K20/142—Amino acids; Derivatives thereof
    • 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
    • A23L2/52—Adding ingredients
    • 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
    • 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
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/175—Amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
    • 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
    • C12N1/205—Bacterial isolates
    • 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
    • C12N3/00—Spore forming or isolating processes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • A61K2035/115—Probiotics
    • 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
    • C12R2001/07—Bacillus
    • C12R2001/10—Bacillus licheniformis
    • 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
    • C12R2001/07—Bacillus
    • C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus

Definitions

  • the human microbiome is a product of our genetics, our diets, and our environment.
  • Probiotics may be used to alter the human microbiome.
  • Probiotics for human consumption are available as capsules and as food products (e.g. yogurt, beverages, etc.).
  • probiotic tea there are examples of probiotic tea on the market, there are none that include an activating component to germinate probiotic spores for improved efficacy.
  • an invention is described that includes the provision of activating compounds with Bacillus bacterial spores for human probiotic consumption in tea, soups, gravies, sauces, hot chocolate, and other food and beverage products, particularly heated liquids and fluids.
  • FIG. 1 shows photographs of Bacillus species in a nutrient germinant composition according to a preferred embodiment at time zero (when hot water is initially added) and after steeping in hot water for 60 minutes at various temperatures according to preferred methods of the invention, the darker spots indicate germinated bacteria and the lighter/brighter spot indicate non-germinated spores;
  • FIG. 2 shows photographs of Bacillus species and a nutrient germinant composition according to a preferred embodiment in steeped tea (black and green tea) according to a preferred method of the invention, the darker spots indicate germinated bacteria and the lighter/brighter spots indicate non-germinated spores.
  • probiotics may be used to alter the human microbiome in order to regulate the digestive system and bolster the immune system.
  • probiotics are non-pathogenic bacteria that may include species of lactic acid bacteria (Lactobacillus), Bifidobacterium, and Propionibacterium. Species of Bacillus are also common as probiotics. Probiotics have been widely used in animals as an alternative to hormones and antibiotics, in plants to enhance certain growth characteristics, and in humans as a digestive supplement.
  • Probiotics for human consumption are available as capsules and as food products (e.g. yogurt, juice, etc.).
  • Probiotics delivered in tea are also on the market.
  • Probiotic tea products contain probiotic bacteria packaged with tea in a porous bag. Tea bags are steeped for a period of time in hot water and are consumed.
  • Kombucha tea is a type of probiotic drink in which a symbiotic culture of bacteria and yeast (SCOBY) is added to tea containing sugar. The SCOBY then acts to ferment the sugar in the tea to produce vitamins and acids. In this case, the sugar acts as a substrate and the drink does not contain amino acids that are known to activate bacteria.
  • SCOBY symbiotic culture of bacteria and yeast
  • Bacillus species as probiotics (e.g. B. coagulans)
  • the bacteria are supplied as spores.
  • Spores represent a unique life stage of specific species of bacteria in that these species have the ability to transition into a dormant spore stage if environmental conditions are unfavorable. Spores are naturally dehydrated, are stable for extended periods of time, and are resistant to environmental stresses (e.g. heat, salt, pH, etc.).
  • the spores are preferably more resistant to the high temperatures of steeping water.
  • spores In order for spores to return to a live, vegitatively growing state, they must first germinate. Spore germination is a multistep process in which spores are revived from the dormant state. The first step is one by which spores are activated and are induced to germinate, typically by an environmental signal called a germinant.
  • This signal can be a nutrient such as an L-amino acid, potassium ions (e.g. KCI), and/or natural osmoprotectant compounds (e.g. ectoine). Nutrient germinants bind to receptors in the inner-membrane of the spore to initiate germination. Additionally, sugars have been shown to increase the binding affinity of L-amino acids for their cognate receptors.
  • the germinant signal initiates an irreversible cascade of events, also known as commitment. Commitment is fast and is typically >90% complete in 2 minutes. As germination progresses, the spore rapidly takes on water. As the spore absorbs water it consequently loses its refractivity. This loss of refractivity towards the end of the first phase in the germination process allows spore germination to be observed via phase-contrast microscopy. The initial phase is typically complete within 30 to 60 minutes depending on the temperature.
  • the second phase of germination is an outgrowth step in which the spore's metabolic, biosynthetic, and DNA replication/repair pathways initiate.
  • the outgrowth period includes a ripening period in which no morphological changes (such as cell growth) occur, but the spore's molecular machinery (e.g. transcription factors, translation machinery, biosynthesis machinery, etc.) is activated. This period can vary in length based on the initial resources that are packaged with the spore during the process of sporulation.
  • a ripening period in which no morphological changes (such as cell growth) occur, but the spore's molecular machinery (e.g. transcription factors, translation machinery, biosynthesis machinery, etc.) is activated. This period can vary in length based on the initial resources that are packaged with the spore during the process of sporulation.
  • spores can be induced to germinate via heat-activation. Spores of various Bacillus species have been heat-activated at strain specific temperatures (e.g. B. subtilis spores can be heat-activated at 75°C for 30 minutes while B. licheniformis spores can be heat-activated at 65°C for 20 minutes). The heat- activation is believed to cause a transient, reversible unfolding of spore proteins. If the spore proteins are not presented with a nutrient germinant, such as L-alanine, they will refold and effectively reverse the germination process.
  • a nutrient germinant such as L-alanine
  • the present invention describes a composition and method for activating probiotic spores in in food and beverage products, such as steeped teas, soups, gravies, sauces, hot chocolate, coffee, and other products, particularly heated liquids and fluids.
  • the method includes the provision of nutrient germinants with Bacillus spores that will be heat activated and subsequently germinated with nutrient germinants while the food or beverage product is being heated, such as while tea is steeping in hot water.
  • the two probiotic components bacterial spores and nutrient germinants
  • the ingredients may be provided as mixed ingredients (nutrient germinant composition and spores) in a single serve packet to be added to tea or another food or beverage product, mixed components (nutrient germinant composition and spores) in a single bag containing tea or another food or beverage product, or as separated components in separate tea bags (e.g. two bags containing any combination of the nutrient germinant composition, spores, and tea) or other container for other food or beverage products.
  • spores can be heat-activated in the presence of nutrient germinants in a method that is described in U.S. Patent Application No 15/479,773, which is incorporated herein by reference.
  • nutrient compositions may contain one or more L-amino acids, D-glucose, D-fructose, a biological buffer, a potassium ion source, and/or a natural osmoprotectant. All components in the nutrient formulation must be Generally Regarded as Safe (GRAS) for human consumption by the United States Food and Drug Administration. At the time of this writing, most, although not all, of the components described herein have been deemed GRAS.
  • GRAS Generally Regarded as Safe
  • the nutrient germinant composition can include one or more L-amino acids.
  • Preferred L-amino acid(s) included in the nutrient formulation are L-alanine, L-asparagine, L- valine, and/or L-cysteine.
  • the choice of L-amino acids is determined by the species of Bacillus used in the probiotic formulation.
  • the L-amino acids can be provided in the form of any suitable source, such as their pure forms and/or a hydrolysate of soy protein.
  • the nutrient germinant composition can optionally contain an amount of D-glucose and/or D-fructose.
  • the choice of sugar is determined by the species of Bacillus used in the probiotic formulation.
  • the nutrient germinant composition can optionally contain one or more sources of potassium ions.
  • KCI can be included as a source of potassium ions in the nutrient formulation.
  • the inclusion of potassium ions is determined by the species of Bacillus used in the probiotic formulation.
  • the nutrient germinant composition can optionally contain one or more biological buffers.
  • the biologic buffer is a buffer that can buffer the nutrient formulation and/or nutrient-spore formulation, to maintain the formulation at the proper pH for spore germination (about pH 6-8).
  • Preferred biologic buffers include, but are not limited to, a phosphate buffer or a HEPES sodium salt.
  • monosodium phosphate and disodium phosphate, preferably used together, can be included in the nutrient formulation as buffers.
  • a HEPES buffer may be used.
  • the nutrient solution can optionally contain an osmoprotectant compound.
  • an osmoprotectant compound Preferably, ectoine, a natural osmoprotectant produced by some species of bacteria, may be included. The inclusion of an osmoprotectant is determined by the species of Bacillus used in the probiotic formulation.
  • the nutrient composition is any one of the nutrient compositions described in U.S. Patent Application No 15/479,773.
  • a nutrient composition may contain ingredients in one or more of the above-reference categories of ingredients and may contain one or more ingredients from within those categories.
  • Preferred embodiments of spore formulations include one or more Bacillus species of spores, including but not limited to, Bacillus licheniformis, Bacillus subtillis, Bacillus amyloliquiefaciens, Bacillus polymyxa, Bacillus thuringiensis, Bacillus megaterium, Bacillus coagulans, Bacillus lentus, Bacillus clausii, Bacillus circulans, Bacillus firmus, Bacillus lactis, Bacillus laterosporus, Bacillus laevolacticus, Bacillus polymyxa, Bacillus pumilus, Bacillus simplex, Bacillus sphaericus, Bacillus sonorensis, Bacillus, horneckiae, Bacillus axarquiensis, Bacillus mucilaginosus, Bacillus olivae, and any combinations thereof.
  • Bacillus species of spores including but not limited to, Bacillus licheniform
  • Preferred spore formulations can contain 1 -20 or more species of Bacillus spores.
  • a spore formulation can contain 3 strains of Bacillus bacteria.
  • 2 strains of the Bacillus bacteria can each be a different strain of the species Bacillus Iicheniformis and the third strain is a species of Bacillus subtilis.
  • about 80% of the formulation can be Bacillus Iicheniformis (40% of each strain) and 20% of the spores in the spore formulation can be is Bacillus subtilis.
  • the spores of the strain(s) included in the spore formulation can be mixed with water or other suitable carrier and/or organic salts.
  • the Bacillus species that can be contained in the spore formulations can produce and/or be capable of producing one or more enzymes including, but not limited to, proteases, amylases, lipases, glycosidases, cellulases, esterases, and xylanases. Tests and assays for determining the production of such enzymes from a Bacillus species are generally known in the art and to one of ordinary skill in the art.
  • the spore formulation can contain about 0.1 % to 90% by weight spores, along with salt or other suitable carrier, such as sodium bicarbonate, or maltodextrin, or a combination thereof. In some embodiments, the spore formulation contains about 5% by weight spores.
  • the spore formulation can be and/or include a powder or other dry form (e.g. spray-dried form of a liquid spore concentrate, or lyophilized spore formulation) containing spores.
  • the total concentration of spores in the spore formulation can range from about 1 x 10 5 CFU/mL or spores/g to 1 x 10 14 CFU/mL or spores/g or any specific concentration or range therein.
  • the total preferred concentration of spores in the spore formulation can be about 1 , 1 .125, 1 .5, 1 .75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, or 9.75 x 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 ,
  • any one specific spore species can be present in the spore formulation at a concentration that can range from about 1 x 10 5 CFU/mL to 1 x 10 14 CFU/mL or any specific range therein.
  • the preferred concentration of any one specific spore species present in the preferred embodiments of spore formulations according to the invention can be about 1 , 1 .125, 1.5, 1 .75, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6, 6.25, 6.5, 6.75, 7, 7.25, 7.5, 7.75, 8, 8.25, 8.5, 8.75, 9, 9.25, 9.5, or 9.75 x 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 ,
  • the spore formulation is biodegradable.
  • the concentrated spore formulation can contain about 1 -9 x 10 9 or 10 10 CFU/mL or spores/g.
  • the concentrated spore formulation can contain about 10 10 CFU/mL or spores/g.
  • the composition comprises a nutrient composition, a spore formulation, and tea.
  • the composition comprises a nutrient composition, a spore formulation, and a food or beverage product, such as soups, gravies, sauces, hot chocolate.
  • the nutrient composition and spore formulation in these embodiments are one of the above described embodiments, both in dry powdered or other dry solid form. Any variety of tea or other food or beverage product may be used.
  • the food or beverage product is one that is heated before being consumed.
  • a nutrient composition and spore formulation are added to any variety of tea.
  • the nutrient composition and spore formulation are one of the above described embodiments.
  • the nutrient composition, spore formulation, and tea or other food or beverage product are combined into a mixture prior to steeping in hot water or otherwise heating the food or beverage product.
  • the nutrient composition, spore formulation, and tea may be separate components that are added together to hot water to prepare the probiotic tea or added to the other food or beverage product to be heated or already heated, or any combination of two of the components may be pre- mixed together and added to the third component prior to or after adding hot water to prepare the probiotic tea or prior to or after heating the other food or beverage product.
  • the order of addition of the three components and the hot water or hot food or beverage is not critical and any order may be used.
  • the mixture is allowed to steep, preferably for 2 to 10 minutes prior to being consumed by a human.
  • Two minutes is the preferred minimum amount of steeping time, but the total time may vary based on the type of tea used and the strength of the tea desired by the person who will consume the probiotic tea.
  • green tea is typically steeped for 2-4 minutes while Oolong tea is typically steeped for 5-8 minutes. Consumption is preferably by drinking or otherwise orally ingesting the probiotic tea.
  • the hot water is at a temperature in the range of 42 to 100°C when initially added to the other components.
  • the hot water temperature is preferably not below 42°C, and the range may vary based on the type of tea being used.
  • green tea is typically steeped in the range of 65-80°C and black tea is usually steeped in the range of 80-100°C.
  • compositions according to preferred embodiments of the invention were tested according to preferred methods of the invention.
  • the compositions, methods, and results are described below.
  • EXAMPLE 1 A probiotic blend of Bacillus spores (B. subtilis and B. licheniformis) at a final concentration of approx. 4 x 10 6 CFU/mL, where CFU stands for colony forming unit, and a nutrient germinant composition according to a preferred embodiment of the invention comprising L-alanine (0.21 g), monosodium phosphate (0.14g), and disodium phosphate (0.5g), were added to approx. 8 fluid ounces of tap water at various temperatures.
  • the amount of spores added represents a "dose" of 10 9 CFU of probiotic bacteria, as recommended by a Harvard Health study.
  • the amount of nutrient germinants used represents the minimum concentration of germinants necessary to initiate germination.
  • the temperatures used represent a range of tea steeping temperatures.
  • FIG. 1 shows representative images from these tests at time 0 (i.e. immediately after water addition) and after 60 minutes in the water.
  • the darker spots show germinated spores, the lighter spots show non-germinated spores.
  • the images in group A represent spores that had been germinated using a nutrient-germ inant composition and heated during the incubation period at 42°C according to a preferred composition and preferred method of the invention.
  • the images in group B represent spores that had been germinated using a nutrient-germ inant composition and heated during the incubation period at 65°C according to a preferred composition and preferred method of the invention.
  • the images in group C represent spores that had been germinated using a nutrient-germ inant composition and heated during the incubation period at 75°C according to a preferred composition and preferred method of the invention.
  • the images in group D represent spores that had been germinated using a nutrient-germ inant composition and heated during the incubation period at 85°C according to a preferred composition and preferred method of the invention.
  • the images in group E represent represents spores that had been germinated using a nutrient-germ inant composition and heated during the incubation period at 100°C according to a preferred composition and preferred method of the invention.
  • EXAMPLE 2 A probiotic blend of Bacillus spores (B. subtilis and B. licheniformis) at a final concentration of approx. 4 x 10 6 CFU/mL and a nutrient germinant composition, according to a preferred embodiment of the invention, comprising L-alanine (0.21 g), monosodium phosphate (0.14g), and disodium phosphate (0.5g), were added to approx. 8 fluid ounces of tap water at 85°C.
  • a tea bag containing the indicated variety of tea was steeped in the water for approx. 3 minutes, as recommended by the tea manufacturer. After steeping, the tea bags were removed while the spores remained in the mixture to be ingested as a probiotic dose.
  • FIG. 2 shows representative images from these tests after 60 minutes in the water.
  • the darker spots show germinated spores, the lighter/brighter spots show non-germinated spores.
  • the top images represent spores germinated in steeped black tea (LiptonTM) while the bottom images represent spores germinated in steeped green tea (Bigelow®).
  • compositions and methods of the invention may be used with other types of food or beverage products, such as soups, gravies, or hot chocolate. Most preferably, such food and beverage products are liquids or fluids and are consumed after heating, but other types of food and beverage products may also be used.

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Abstract

La présente invention concerne une composition et un procédé d'activation de spores probiotiques dans des produits alimentaires et des boissons, tels que du thé infusé, du café, des soupes et des sauces. Une composition d'agent de germination nutritive comprend un ou plusieurs acides L-aminés, éventuellement un ou plusieurs tampons pour maintenir le pH de la composition, lorsqu'elle est ajoutée à de l'eau, dans une plage d'environ 6 à 8, éventuellement du D-glucose, du D-fructose, ou à la fois du D-glucose et du D-fructose, et éventuellement un osmoprotecteur. La composition d'agent de germination nutritive, une ou plusieurs espèces de spores de Bacillus et un produit alimentaire ou une boisson peuvent être pré-mélangés dans n'importe quelle combinaison. De l'eau est ajoutée au mélange et chauffée dans une plage de température de 42 à 100 °C pour faire germer les spores probiotiques avant d'être consommées.
EP18873222.6A 2017-11-03 2018-11-02 Composition et procédé pour des composés germinatifs dans des produits alimentaires et des boissons probiotiques pour la consommation humaine Pending EP3703721A4 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201762581324P 2017-11-03 2017-11-03
US201762596424P 2017-12-08 2017-12-08
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US12097226B2 (en) 2016-04-05 2024-09-24 Nch Corporation System and method for using a single-serve nutrient spore composition for small scale farm applications
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WO2021159068A1 (fr) 2020-02-06 2021-08-12 Nch Corportion Composition et méthode d'utilisation des composés germinatifs dans les probiotiques pour réduire l'inflammation chez les êtres humains
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BR112020008712A2 (pt) 2020-11-03
CA3079993A1 (fr) 2019-05-09
PH12020550820A1 (en) 2021-07-05
WO2019090065A1 (fr) 2019-05-09
CO2020006844A2 (es) 2020-08-21
MX2020004473A (es) 2020-08-03
MY199858A (en) 2023-11-25
CN111295197A (zh) 2020-06-16
CA3079993C (fr) 2022-09-20
NZ763867A (en) 2025-06-27
WO2019090065A8 (fr) 2019-06-06

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