WO2015193449A1 - Procédé de production de produit laitier fermenté ayant une concentration réduite en lactose résiduel - Google Patents

Procédé de production de produit laitier fermenté ayant une concentration réduite en lactose résiduel Download PDF

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Publication number
WO2015193449A1
WO2015193449A1 PCT/EP2015/063742 EP2015063742W WO2015193449A1 WO 2015193449 A1 WO2015193449 A1 WO 2015193449A1 EP 2015063742 W EP2015063742 W EP 2015063742W WO 2015193449 A1 WO2015193449 A1 WO 2015193449A1
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Prior art keywords
milk product
lactase
lactic acid
nlu
strain
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PCT/EP2015/063742
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English (en)
Inventor
Nanna Christensen
Kim Ib Soerensen
Gäelle Lettier Buchhorn
Poul Møller HANSEN
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Chr. Hansen A/S
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Publication of WO2015193449A1 publication Critical patent/WO2015193449A1/fr

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
    • A23C9/1206—Lactose hydrolysing enzymes, e.g. lactase, beta-galactosidase
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A23C9/1238—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
    • C12N9/2471—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C2260/00—Particular aspects or types of dairy products
    • A23C2260/05—Concentrated yoghurt products, e.g. labneh, yoghurt cheese, non-dried non-frozen solid or semi-solid yoghurt products other than spreads; Strained yoghurt; Removal of whey from yoghurt
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/21—Streptococcus, lactococcus
    • A23V2400/249—Thermophilus
    • 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
    • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

Definitions

  • the present invention relates to methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of specific lactic acid bacteria and in the presence of lactase.
  • the methods of the present invention provide a fermented milk product with a reduced residual lactose concentration, i.e. a particularly low concentration of lactose.
  • milk primarily obtained from cows, as well as dairy products made from milk, such as cheese, cream, butter, yoghurt or ice cream, contribute significantly to the human diet.
  • Lactose is the main carbohydrate of milk and is usually present in a concentration of about 5%. Lactose is a disaccharide of ⁇ -D galactose and ⁇ -D glucose. In humans lactose can be metabolized only if the disaccharide is hydrolyzed in the duodenum by the enzyme lactase. Hydrolysis of lactose produces the constituent monomers, which are absorbed by humans in the small intestine.
  • Lactose intolerance is a condition characterized by symptoms including abdominal bloating, cramps, flatulence, diarrhea, nausea, borborygmy and vomiting.
  • the condition is caused by the presence of lactose in the colon, where the disaccharide is taken up by bacteria resulting in fermentation processes producing a gas, which causes the abdominal symptoms.
  • the presence of lactose in the colon in turn is caused by the absence of lactase production in the duodenum.
  • the severity of the symptoms of patients suffering from lactose intolerance typically corresponds to the amount of lactose consumed.
  • Lactose intolerance is a well-known condition affecting numerous humans and consequently different approaches have been used to produce low lactose milk or lactose free milk.
  • Methods available for reducing the lactose concentration in milk include hydrolyzing lactose by lactase enzyme, nanofiltration, electrodialysis, ion exchange chromatography and centrifugation.
  • lactase based prior art methods for reducing the lactose concentration in milk always required the use of a very high lactase concentration, such as a concentration of 5000 NLU/I or more.
  • the present invention provides methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism
  • lactase is present in an initial concentration in the range of 40to 1000 NLU/I or in the range of 40 to 2000 NLU/I of the composition comprising the milk and the bacteria.
  • the present inventors surprisingly found that the combination of lactic acid bacteria (subsequently LAB) with a deficiency in glucose metabolism and a low concentration of lactase can be used in these methods to generate a milk product with a very low concentration of lactose (such as a lactose concentration of ⁇ 5mg/g).
  • lactose such as a lactose concentration of ⁇ 5mg/g.
  • the present invention therefore provides methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism
  • the lactase is present in an initial concentration in the range of 40 to 1000 NLU/I or in the range of 40 to 2000 NLU/I of the composition comprising the milk and the bacteria, wherein the lactose concentration of the fermented milk product is ⁇ 5mg/g at the termination of fermentation. Accordingly, the lactose concentration may be significantly below 5mg/g at the termination of fermentation, such as ⁇ 0.5mg/g at the termination of fermentation, or ⁇ 0.1mg/g at the termination of fermentation.
  • the present invention also provides methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism
  • lactase is present in an initial concentration in the range of 55 to 550 NLU/I of the composition comprising the milk and the bacteria, wherein the lactose concentration of the fermented milk product is ⁇ 0.5mg/g at the termination of fermentation.
  • the present invention provides compositions comprising lactic acid bacteria and lactase, wherein the lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism, and wherein the lactic acid bacteria are present in a concentration of > 10 9 CFU per g of composition and the lactase is present in a concentration in the range of 100 to 4500 NLU/g or in the range of 100 to 9000 NLU/g of composition.
  • the present invention provides a starter culture comprising lactic acid bacteria and lactase, wherein the lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism, and wherein the lactic acid bacteria are present in a concentration of > 10 9 CFU per g of starter culture and the lactase is present in a concentration in the range of 100 to 4500 NLU/g or in the range of 100 to 9000 NLU/g of starter culture.
  • the present invention provides fermented milk products comprising lactase and lactic acid bacteria, wherein the lactic acid bacteria comprise a strain having a deficiency in glucose metabolism, and wherein the lactose is present in the fermented milk product in a concentration of ⁇ 0.5mg/g.
  • the fermented milk product can be a yoghurt, a fruit yoghurt, a yoghurt beverage, a skyr, quark, fortified Greek yogurt, strained Greek yogurt, set yogurt, Dahi, Labneh, buttermilk or a cheese.
  • the present invention relates to methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactase is present in an initial concentration in the range of 40to 1000 NLU/I or in the range of 40 to 2000 NLU/I of the composition comprising the milk and the bacteria.
  • milk is broadly used in its common meaning to refer to liquids produced by the mammary glands of animals.
  • the milk may have been processed and the term “milk” includes whole milk, skim milk, fat-free milk, low fat milk, full fat milk and concentrated milk.
  • Fat-free milk is non-fat or skim milk product.
  • Low-fat milk is typically defined as milk that contains from about 0.5% to about 2% fat.
  • Full fat milk often contains 2% fat or more.
  • milk is intended to encompass milk from different mammal sources. Mammal sources of milk include, but are not limited to cow, sheep, goat, buffalo, camel, llama, mare and deer.
  • milk derived from cows is most preferably used as a starting material for the fermentation.
  • milk base is broadly used in the present application to refer to a composition based on milk or milk components which can be used as a medium for growth of and fermentation with LAB.
  • the milk base comprises components derived from milk and any other component that can be used for the purpose of growing or fermenting LAB.
  • lactic acid bacteria or "LAB” is used to refer to food-grade bacteria producing lactic acid as the major metabolic end-product of carbohydrate fermentation. These bacteria are related by their common metabolic and physiological characteristics and are usually Gram positive, low-GC, acid tolerant, non-sporulating, non- resipiring, rod-shaped bacilli or cocci. During the fermentation stage, the consumption of lactose by these bacteria causes the formation of lactic acid, reducing the pH and leading to the formation of a protein coagulum. The lactose found in the fermented product is thus residual lactose, i.e. lactose that was not consumed by the bacteria.
  • lactic acid bacteria encompasses, but is not limited to, bacteria belonging to the genus of Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., Leuconostoc spp., Lactococcus spp., such as Lactobacillus delbrueckii subsp.
  • the fermentation step of the process for manufacturing fermented milk products comprises the addition of a starter culture to milk.
  • starter or “starter culture” as used in the present context refers to a culture of one or more food-grade micro-organisms, in particular lactic acid bacteria, which amongst other cause the acidification of the milk base.
  • the starter cultures may be fresh, frozen or freeze-dried form and may be added in that form or previously dissolved, diluted or mixed with a milk base or other medium components. Freeze dried cultures are frequently regenerated by rehydration before use.
  • the starter can be added in an amount from 0.01 to 3%, preferably from 0.01 and 0.025 % by volume of the total amount of milk.
  • deficiency in glucose metabolism and the term “glucose deficient” are both used in the context of the present invention to characterize LAB which either partially or completely lost the ability to use glucose as a source for cell growth or maintaining cell viability.
  • a respective deficiency in glucose metabolism can for example be caused by a mutation in a gene inhibiting or inactivating expression or activity of the glucokinase protein or the glucose transporter protein.
  • LAB with a deficiency in glucose metabolism can be characterized by increasing the glucose concentration in a culture medium, when grown on lactose as carbohydrate source. The increase of glucose is caused by glucose secretion of the glucose deficient LABs. Increase of glucose concentration in a culture medium can be determined by HPLC analysis, for example using a Dionex CarboPac PA 20 3*150mm column (Thermo Fisher Scientific, product number 060142).
  • bulgaricus mutant LAB strain which cannot take up glucose or galactose present in the medium.
  • the strain can take up lactose, digest lactose and use the glucose obtained from lactose digestion for growth and acid production.
  • the publication of Pool et al., 2006 discloses a further method to obtain LAB with a deficiency in glucose metabolism. Using the general principles described in these publications numerous other LAB with the same or a similar deficiency in glucose metabolism can be generated by one of ordinary skill.
  • the LAB with a deficiency in glucose metabolism may comprise a glucose deficient but galactose-fermenting Streptococcus thermophilus strain.
  • the LAB with a deficiency in glucose metabolism may comprise a Lactobacillus delbrueckii subsp. bulgaricus strain with deficiency in glucose uptake.
  • the starter culture may comprise a glucose deficient but galactose-fermenting Streptococcus thermophilus strain and a Lactobacillus delbrueckii subsp. bulgaricus strain with deficiency in glucose uptake.
  • the starter culture may comprise additional LABs or may consist of a mixture of these strains.
  • the present invention provides methods of producing a fermented milk product comprising a step wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism and bacteria with the ability to metabolize glucose
  • the lactase is present in an initial concentration in the range of 40 to 2000 NLU/I of the composition comprising the milk and the bacteria.
  • a slightly higher concentration of lactase is preferably used for the production of a lactose free fermented product.
  • the mixture comprises Streptococcus thermophilus strain with a deficiency in glucose metabolism and which is galactose- fermenting and a Lactobacillus delbrueckii subsp. bulgaricus strain with the ability to take up and metabolize glucose.
  • the methods of the invention are carried out using a starter culture that consists entirely of LAB with a deficiency in glucose metabolism.
  • glucose deficient LAB in the methods of the invention provides the additional advantage that the fermented milk product thus produced has a sweet taste.
  • Lactase is an enzyme that hydrolyzes lactase and produces glucose and galactose. Lactase is commercially available from many sources and can be prepared according to methods well known in the art. Lactase activity may be determined as units of neutral lactase activity (NLU) . In accordance with the present invention the lactase activity is determined using the method as described in the Food Chemicals Codex (FCC) IV method.
  • NLU neutral lactase activity
  • the methods of the present invention produce a fermented milk product with very low lactose concentration.
  • lactose milk products or lactose free milk products There is no generally accepted definition for low lactose milk products or lactose free milk products.
  • a concentration of less than lg lactose per lOOg of milk or milk product characterizes a low lactose product
  • a concentration of less than lOmg of lactose per lOOg of milk or milk product characterizes a lactose free product.
  • the lactose concentration in low lactose or lactose free products may be considerably higher in other member states.
  • the lactose concentration in milk, milk base or a fermented milk product can be determined by any of a number of known methods for determining lactose concentration in respective liquids.
  • the lactose concentration is determined using HPLC.
  • HPLC can be carried out with different devices and columns.
  • the HPLC can be carried out using a Dionex CarboPac PA 20 3*150mm column (Thermo Fisher Scientific, product number060142).
  • the present invention provides methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism
  • lactase is present in an initial concentration in the range of 40 to 1000 NLU/I of the composition comprising the milk and the bacteria, wherein the lactose concentration of the fermented milk product is ⁇ 5mg/g at the termination of fermentation, or ⁇ 0.5mg/g at the termination of fermentation, or ⁇ 0.1mg/g at the termination of fermentation.
  • the method of producing a fermented milk product uses the lactase in the fermentation in a low initial concentration, such as in the range of 50 to 800 NLU/I, or in the range of 55 to 550 NLU/I, or in the range of 60 to 250 NLU/I .
  • a low initial concentration such as in the range of 50 to 800 NLU/I, or in the range of 55 to 550 NLU/I, or in the range of 60 to 250 NLU/I .
  • the method of producing a fermented milk product use lactase in an initial concentration that is optimized to achieve very high reduction of lactose concentration in the product at the end of fermentation.
  • the initial concentration of lactase can be in the range of 250 to 1600 NLU/I, or in the range of 350 to 1500 NLU/I, or in the range of 450 to 1500 NLU/I. This approach is directed to achieving a lactose-free product using as little lactase as possible.
  • the present invention provides therefore methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • the lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism comprise (i) a glucose deficient but galactose-fermenting Streptococcus thermophilus strain and (ii) a Lactobacillus delbrueckii subsp. bulgaricus strain with a deficiency in glucose uptake or a Lactobacillus delbrueckii subsp.
  • the lactase is present in an initial concentration in the range of 350 to 600 NLU/I or in the range of 350 to 1600 NLU/I of the composition comprising the milk and the bacteria, wherein the lactose concentration of the fermented milk product is ⁇ 0.5mg/g at the termination of fermentation.
  • the lactase is preferably present in an initial concentration in the range of 350 to 600 NLU/I of the composition comprising the milk and the bacteria.
  • the lactase is preferably present in an initial concentration in the range of 350 to 1600 NLU/I of the composition comprising the milk and the bacteria.
  • the present invention provides therefore methods of producing a fermented milk product comprising a step, wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • the lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism comprise a glucose deficient but galactose-fermenting Streptococcus thermophilus strain and a Lactobacillus delbrueckii subsp. bulgaricus strain with a deficiency in glucose uptake or a Lactobacillus delbrueckii subsp. bulgaricus strain with the ability to take up and metabolize glucose, and
  • the lactase is present in an initial concentration in the range of 450 to 550 NLU/I or in the range of 450 to 1500 NLU/I of the composition comprising the milk and the bacteria, wherein the lactose concentration of the fermented milk product is ⁇ 0.1mg/g at the termination of fermentation.
  • the lactase is preferably present in an initial concentration in the range of 450 to 550 NLU/I of the composition comprising the milk and the bacteria.
  • the method uses a Lactobacillus delbrueckii subsp.
  • the lactase is preferably present in an initial concentration in the range of 450 to 1500 NLU/I of the composition comprising the milk and the bacteria.
  • the methods of the present invention can be used to generate any known fermented milk product, including yoghurt, a fruit yoghurt, a yoghurt beverage, a skyr, quark, fortified Greek yogurt, strained Greek yogurt, set yogurt, Dahi, Labneh, buttermilk or a cheese. According to the most preferred embodiment of the invention the methods are used for producing yoghurt type fermented milk products.
  • yoghurt is used in the context of the present invention to refer to fermented milk products obtained by fermenting milk with a starter culture comprising Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus.
  • a starter culture comprising Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus.
  • Numerous different types of yoghurt are known in the art and encompassed by the present invention, including fruit yoghurt, yoghurt beverages, skyr, set yogurt, and Greek style yoghurt characterized by a high protein level, including fortified Greek yogurt, strained Greek yogurt.
  • the present invention provides compositions comprising lactic acid bacteria and lactase, wherein the lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism, and wherein the lactic acid bacteria are present in a concentration of 10 9 CFU per g of composition and the lactase is present in a concentration in the range of 100 to 4500 NLU/g or in the range of 100 to 9000 NLU/g of the composition.
  • compositions may comprise one or several additional components, including a buffer, residual components of fermentation media, stabilizers, cryoprotective agents, etc.
  • compositions are starter cultures, which comprise lactic acid bacteria and lactase, wherein the lactic acid bacteria comprise bacteria with a deficiency in glucose metabolism, and wherein the lactic acid bacteria are present in a concentration of 10 9 CFU per g of starter culture and the lactase is present in a concentration in the range of 100 to 4500 NLU/g or in the range of 100 to 9000 NLU/g of the starter culture.
  • the lactase is preferably present in a concentration in the range of 300 to 2500 NLU/g of composition or starter culture or in the range of 500 to 2500 NLU/g of composition or starter culture or in the range of 300 to 5000 NLU/g of composition or starter culture or in the range of 500 to 5000 NLU/g of composition or starter culture.
  • the starter culture may comprise lactic acid bacteria with a deficiency in glucose metabolism, including a galactose-fermenting Streptococcus thermophilus strain, a Lactobacillus delbrueckii subsp. bulgaricus strain, a galactose-fermenting Lactococcus lactis strain and a galactose-fermenting Leuconostoc strain.
  • the present invention provides fermented milk products comprising lactase and lactic acid bacteria, wherein the lactic acid bacteria comprise a strain having a deficiency in glucose metabolism, and wherein the lactose is present in the fermented milk product in a concentration of less than 0.5mg/g.
  • the fermented milk products may comprise a glucose deficient but galactose- fermenting Streptococcus thermophilus strain and/or a Lactobacillus delbrueckii subsp. bulgaricus strain with deficiency in glucose uptake.
  • the fermented milk product is a yoghurt, a fruit yoghurt, a yoghurt beverage, a skyr, quark, fortified Greek yogurt, strained Greek yogurt, set yogurt, Dahi, Labneh, buttermilk or a cheese.
  • the present invention provides a Streptococcus thermophilus strain, wherein the strain is:
  • the deposited strain (DSM 28889) was found to provide a particularly advantageous combination of high glucose secretion and high production of compounds improving the texture of the fermented food product.
  • the glucose secretion can easily be determined by one of ordinary skill.
  • the production of compounds improving the texture of the fermented food products is particularly advantageous for fermented products such as yoghurt.
  • the present invention also provides methods of producing a fermented food product which methods comprise a step, wherein a milk product is fermented using a starter culture comprising a Streptococcus thermophilus strain, wherein the strain is:
  • the deposited strain may also be used for the methods of producing a fermented milk product in the presence of lactase. Accordingly, the present invention also provides methods of producing a fermented food product which methods comprise a step wherein milk is fermented in the presence of lactic acid bacteria and in the presence of lactase,
  • lactic acid bacteria comprise a Streptococcus thermophilus strain, which strain is:
  • the present invention is directed to fermented food products, which comprise a Streptococcus thermophilus strain, wherein the strain is:
  • the fermented food products can be a yoghurt, a fruit yoghurt, a yoghurt beverage, a skyr, quark, fortified Greek yogurt, strained Greek yogurt, set yogurt, Dahi, Labneh, buttermilk or a cheese.
  • Figure 1 shows the correlation between lactose levels before and after fermentation in trials 1 and 2 as described below.
  • CHCC10019 was deposited for WO2007147890 with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on April 4, 2007 under the accession no. DSM 19252.
  • CHCC16159 was deposited for WO2013160413with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on 2012-09-06 under the accession no. DSM 26420.
  • Streptococcus thermophilus CHCC15757 was deposited for WO2013160413 with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on 2012-04-03 under the accession no. DSM 25850.
  • Streptococcus thermophilus CHCC16731 was deposited for WO2013160413 with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on 2012-04-03 under the accession no. DSM 28889.
  • Streptococcus thermophilus CHCC16404 was deposited for WO2013160413 with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on 2012-12-12 under the accession no. DSM 26722.
  • CHCC14994 was deposited for WO2013160413 with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on 2012-04-03 under the accession no. DSM 25838.
  • CHCC11976 was deposited for WO2011026863 with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on 2009-09-08 under the accession no. DSM 22934.
  • Lactobacillus bulgaricus strain CHCC4351 was deposited with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig on April 4, 2007 under the accession no. DSM 22586.
  • Streptococcus thermophilus CHCC16731 (2deoxyglucose resistant, hyper-lactose fermenting and glucose secreting mutant of CHCC11976).
  • Streptococcus thermophilus CHCC16404 (2-deoxyglucose resistant, hyper-lactose fer- meriting and glucose secreting mutant of CHCC15757).
  • Lactobacillus delbrueckli subsp. bulgaricus CHCC16159 (2-deoxyglucose resistant mutant of CHCC10019).
  • Lactobacillus delbrueckli subsp. bulgaricus CHCC16159 was replaced by Lactobacillus delbrueckli subsp. bulgaricus strain CHCC4351.
  • Ingredients Mix of commercial milk (1.5% fat + 0.5% fat) + skim milk powder to reach the required fat and protein level
  • Skimmed milk powder Aria Foods, lot 990214
  • Lactose Product: Lactose monohydrate, lot 500449
  • Lactose concentration (mg/g) was determined using HPLC and a Dionex CarboPac PA 20 3*150mm column (Thermo Fisher Scientific, product number 060142).
  • EXAMPLE 1 Lactose concentration in milk base with different lactose starting concentration fermented with a starter culture containingbacteria with a deficiency in glucose metabolism
  • Milk base 1 and 2 were fermented with a Frozen Direct Vat Set culture(F-DVS) starter culture containing glucose deficient LABs comprising the strains CHCC15757, CHCC16159, CHCC16404, andCHCC16731. Before fermentation the lactose concentration in milk base 1 and 2 was adjusted by addition of lactose to the level shown in Table 1.
  • F-DVS Frozen Direct Vat Set culture
  • Table 1 Results of Trials 1 and 2 showing start concentration of lactose (mg/g) and residual concentration of lactose (mg/g) after fermentation
  • the starter culture was inoculated at 0.024% in 3 liter scale, and samples were fermented at 43°C. Acidification was monitored with a pH-Meter 1120 (Mettler-Toledo AG, 52120653), and fermentation terminated at pH 4.55.
  • the fermented milk products of trial 3 were stirred in a standardized way, pressurized and cooled (2bar, 25°C), before storing at 6 °C. Samples of trial 3 were taken for HPLC analysis on day 1. Residual lactose (mg/g) levels are shown in Table 2. Table 2: HPLC data from trial 3, residual lactose levels (mg/g) after fermentation with starter culture containing glucose deficient strains as described above with and without addition of lactase (0-1000 NLU/II).
  • the data in Table 2 show that the residual level of lactose decreases with an increase of lactase.
  • the level of added lactase is at preciselyLactose Free" level, when added in a concentration of about 500 NLU/I or slightly more than 500 NLU/I to a milk base fermented with the starter culture containing bacteria with a deficiency in glucose metabolism as described above under the conditions of this example.
  • EXAMPLE 3 Lactose concentration in milk base fermented with a starter culture containing glucose deficient LAB in the presence of 500 NLU/I lactase compared to milk base fermented with YoFlex®Mild 1.0 in the presence of 5000 NLU/I lactase.
  • Table 3 The residual level of lactose after fermentation using F-DVS YoFlex® Mild 1.0 in the presence of 5000 NLU/I lactase and starter culture containing bacteria with a deficiency in glucose metabolism as described above in the presence of 500 NLU/I
  • the starter cultures containing glucose deficient LABs reach similar levels of residual lactose in the end fermented milk product in trial 3, by using 1/10 of the concentration of lactase as YoFlex® Mild 1.0 in trial 4. Fermentation using F-DVS YoFlex® Mild 1.0 and 500 NLU/I does not provide a product with a residual lactose concentration that can be characterized as lactose free.
  • EXAMPLE 4 Lactose concentration in milk base fermented with a starter consisting of glucose deficient Streptococcus thermophilus strains and a Lactobacillus bulgari- cus strain with the ability to take up and metabolize lactose and in the presence of different concentrations of lactase
  • lactase (0, 1000, 1500 or 2000 NLU/I) were added to milk base 2 and the same was fermented with a starter consisting of Streptococcus thermophilus strains with a deficiency in glucose metabolism and a Lactobacillus bulgaricus strain with the ability to take up and metabolize glucose (CHCC4351).
  • the starter culture was inoculated at 0.02% in 3 liter scale, and samples were fermented at 43°C. Acidification was monitored with a pH-Meter 1120 (Mettler-Toledo AG, 52120653). Fermentation terminated at pH 4.55.
  • the fermented milk products of trial 5 were stirred in a standardized way, pressurized and cooled (2bar, 25°C), before storing at 6 °C. Samples of trial 3 were taken for HPLC analysis on day 1. Residual lactose (mg/g) levels are shown in Table 4.
  • Table 5 HPLC data from trial 5, residual lactose levels (mg/g) before and after fermentation with starter culture containing Streptococcus thermophilus strains with a deficiency in glucose metabolism and a Lactobacillus bulgaricus strain with the ability to take up and metabolize glucose with different amounts of lactase (1000-2000 NLU/I).
  • Streptococcus thermophilus CHCC16731 was deposited with DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, D-38124 Braunschweig, on 2014- 06-04 under the accession no. DSM 28889.

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Abstract

La présente invention concerne des procédés de production de produit laitier fermenté, comprenant une étape dans laquelle le lait est fermenté en présence de bactéries d'acide lactique et en présence de lactase, les bactéries d'acide lactique comprenant des bactéries ayant une carence en métabolisme du glucose, et la lactase étant présente à une concentration initiale dans la plage de 40 à 2000 NLU/I de la composition comprenant le lait et les bactéries. Les procédés de la présente invention sont particulièrement appropriés pour produire un produit laitier fermenté ayant une faible concentration en lactose, telle qu'une concentration en lactose ≤ 5 mg/g à la fin de la fermentation, ou ≤ 0,5 mg/g à la fin de la fermentation, ou ≤ 0,1 mg/g à la fin de la fermentation.
PCT/EP2015/063742 2014-06-19 2015-06-18 Procédé de production de produit laitier fermenté ayant une concentration réduite en lactose résiduel WO2015193449A1 (fr)

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WO2017103051A1 (fr) * 2015-12-18 2017-06-22 Chr. Hansen A/S Bactérie lactique pour préparer des produits alimentaires fermentés ayant un goût sucré naturel augmenté et une texture élevée
WO2017109532A1 (fr) * 2015-12-24 2017-06-29 Compagnie Gervais Danone Utilisation d'une lactase pour améliorer la préparation d'un produit laitier fermenté à souches
WO2017112883A1 (fr) * 2015-12-24 2017-06-29 The Dannon Company, Inc. Utilisation d'une lactase dans la préparation d'un produit laitier fermenté à souches
CN107136214A (zh) * 2017-06-05 2017-09-08 光明乳业股份有限公司 一种零乳糖发酵乳及其制备方法
EP3228194A1 (fr) * 2016-04-07 2017-10-11 DSM IP Assets B.V. Produit de lait fermenté avec du diacétyle préparée à l'aide de lactase
WO2018130630A1 (fr) * 2017-01-13 2018-07-19 Chr. Hansen A/S Procédé de production d'un produit laitier fermenté
WO2018177835A1 (fr) 2017-03-28 2018-10-04 Chr. Hansen A/S Composition de bactéries lactiques pour préparer des produits alimentaires fermentés ayant une saveur et un goût sucré naturels accrus
KR20190013181A (ko) * 2017-07-31 2019-02-11 롯데푸드 주식회사 유당 무함유 발효유 및 이의 제조방법
WO2019065651A1 (fr) * 2017-09-27 2019-04-04 株式会社明治 Lait fermenté, et procédé de fabrication de celui-ci
WO2019219973A1 (fr) * 2018-05-18 2019-11-21 Arla Foods Amba Ingrédient laitier fermenté
WO2020008250A1 (fr) * 2018-07-05 2020-01-09 Synbio Tech Inc. Culture de yaourt contenant un mélange de souches de bactéries d'acide lactique, et produit fermenté préparé à l'aide d'une telle culture de yaourt et utilisation de ce produit fermenté
CN112592868A (zh) * 2020-12-30 2021-04-02 杭州娃哈哈科技有限公司 一种用于制备低乳糖或零乳糖酸奶的乳酸菌直投式发酵剂
CN112625971A (zh) * 2020-12-30 2021-04-09 杭州娃哈哈科技有限公司 一株可大量消耗乳糖且兼具耐酸、耐胆盐和抗氧化性的德氏乳杆菌及其应用
US20210259266A1 (en) * 2018-06-20 2021-08-26 Chr. Hansen A/S A method for producing a cheese with reduced amount of galactose
CN115191484A (zh) * 2021-12-29 2022-10-18 苏州福赛思生物科技有限公司 一种含有益生元的低乳糖酸奶及制备方法
WO2023110858A1 (fr) 2021-12-17 2023-06-22 Chr. Hansen A/S Procédé de production d'un produit laitier fermenté
US11690384B2 (en) 2017-08-28 2023-07-04 Chr. Hansen A/S Streptococcus thermophtlus (ST) cell to make e.g. mozzarella cheese

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US11540527B2 (en) 2015-12-18 2023-01-03 Chr. Hansen A/S Lactic acid bacteria for preparing fermented food products with increased natural sweetness and high texture
WO2017103051A1 (fr) * 2015-12-18 2017-06-22 Chr. Hansen A/S Bactérie lactique pour préparer des produits alimentaires fermentés ayant un goût sucré naturel augmenté et une texture élevée
WO2017109532A1 (fr) * 2015-12-24 2017-06-29 Compagnie Gervais Danone Utilisation d'une lactase pour améliorer la préparation d'un produit laitier fermenté à souches
WO2017112883A1 (fr) * 2015-12-24 2017-06-29 The Dannon Company, Inc. Utilisation d'une lactase dans la préparation d'un produit laitier fermenté à souches
RU2730030C2 (ru) * 2015-12-24 2020-08-14 Компани Жерве Данон Применение лактазы для повышения качества приготовления сцеженного ферментированного молочного продукта
EP3228194A1 (fr) * 2016-04-07 2017-10-11 DSM IP Assets B.V. Produit de lait fermenté avec du diacétyle préparée à l'aide de lactase
EP3228194B1 (fr) 2016-04-07 2021-03-17 DSM IP Assets B.V. Produit de lait fermenté avec du diacétyle préparée à l'aide de lactase
EP3804526A1 (fr) * 2016-04-07 2021-04-14 DSM IP Assets B.V. Produit laitier fermenté avec diacétyle produit à l'aide de lactase
EP3821712A1 (fr) * 2017-01-13 2021-05-19 Chr. Hansen A/S Procédé de production d'un produit laiter fermenté
WO2018130630A1 (fr) * 2017-01-13 2018-07-19 Chr. Hansen A/S Procédé de production d'un produit laitier fermenté
JP2020515265A (ja) * 2017-03-28 2020-05-28 セーホーエル．ハンセン アクティーゼルスカブ 増強された天然の甘味およびフレーバーを有する発酵食品を調製するための乳酸菌組成物
EP3599877B1 (fr) 2017-03-28 2022-07-06 Chr. Hansen A/S Composition de bactéries d'acide lactique pour préparer des produits alimentaires fermentés à sucrosité naturelle et saveur accrues
JP7383482B2 (ja) 2017-03-28 2023-11-20 セーホーエル．ハンセン アクティーゼルスカブ 増強された天然の甘味およびフレーバーを有する発酵食品を調製するための乳酸菌組成物
WO2018177835A1 (fr) 2017-03-28 2018-10-04 Chr. Hansen A/S Composition de bactéries lactiques pour préparer des produits alimentaires fermentés ayant une saveur et un goût sucré naturels accrus
CN107136214B (zh) * 2017-06-05 2024-01-12 光明乳业股份有限公司 一种零乳糖发酵乳及其制备方法
CN107136214A (zh) * 2017-06-05 2017-09-08 光明乳业股份有限公司 一种零乳糖发酵乳及其制备方法
KR20190013181A (ko) * 2017-07-31 2019-02-11 롯데푸드 주식회사 유당 무함유 발효유 및 이의 제조방법
KR101983687B1 (ko) 2017-07-31 2019-05-29 롯데푸드 주식회사 유당 무함유 발효유 및 이의 제조방법
US11690384B2 (en) 2017-08-28 2023-07-04 Chr. Hansen A/S Streptococcus thermophtlus (ST) cell to make e.g. mozzarella cheese
WO2019065651A1 (fr) * 2017-09-27 2019-04-04 株式会社明治 Lait fermenté, et procédé de fabrication de celui-ci
WO2019219973A1 (fr) * 2018-05-18 2019-11-21 Arla Foods Amba Ingrédient laitier fermenté
US20210259266A1 (en) * 2018-06-20 2021-08-26 Chr. Hansen A/S A method for producing a cheese with reduced amount of galactose
US12035726B2 (en) * 2018-06-20 2024-07-16 Chr. Hansen A/S Method for producing a cheese with reduced amount of galactose
WO2020008250A1 (fr) * 2018-07-05 2020-01-09 Synbio Tech Inc. Culture de yaourt contenant un mélange de souches de bactéries d'acide lactique, et produit fermenté préparé à l'aide d'une telle culture de yaourt et utilisation de ce produit fermenté
CN112625971A (zh) * 2020-12-30 2021-04-09 杭州娃哈哈科技有限公司 一株可大量消耗乳糖且兼具耐酸、耐胆盐和抗氧化性的德氏乳杆菌及其应用
CN112592868A (zh) * 2020-12-30 2021-04-02 杭州娃哈哈科技有限公司 一种用于制备低乳糖或零乳糖酸奶的乳酸菌直投式发酵剂
WO2023110858A1 (fr) 2021-12-17 2023-06-22 Chr. Hansen A/S Procédé de production d'un produit laitier fermenté
CN115191484A (zh) * 2021-12-29 2022-10-18 苏州福赛思生物科技有限公司 一种含有益生元的低乳糖酸奶及制备方法

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