Lactic acid bacteria for heat-treated food products stored at ambient temperature

Abstract

A method for producing an ambient storage food product, the method comprising: providing a food product having a pH of 3.4 to 4.4, heat-treating the food product to obtain a heat-treated food product, aseptically adding one or more ambient storage lactic acid bacteria strains to the heat-treated food product to obtain an ambient storage food product, and storing the ambient storage food product at ambient temperature for a period of time, wherein the ambient storage lactic acid bacteria strain is selected from the group consisting of the following strains: (i) wherein the strain is capable of maintaining viability at the end of 20 days at a temperature of 25°C; and (ii) wherein the pH decreases by at most 0.8 units during the storage period; and (iii) wherein the strain is selected from the group consisting of strains of the genus Oenococcus and mutants thereof.

Description

Technical Field

[0001] The present invention relates to lactic acid bacteria suitable for addition to heat-treated food products having a pH of 3.4 to 4.4 to be stored at ambient temperature. Background Art

[0002] In recent years, fermented dairy products (such as yogurt) that can be stored, transported, processed and eaten for several months under non-refrigerated conditions (that is, at ambient temperature) have been widely used. This type of yogurt allows consumers to carry yogurt with them over a period of time, without the need to refrigerate it as many beverages may be refrigerated, so this type of yogurt provides consumers with significant convenience advantages. In order to obtain this long-term shelf life at ambient temperature, yogurt has been heat-treated after the fermentation process is completed to kill or at least suppress the further growth of a large amount of lactic acid bacteria used in the fermentation process. The further growth of lactic acid bacteria in the live bacteria and starter culture can cause continuous fermentation and cause post-acidification. Heat treatment can be, for example, a pasteurization process or an ultra-high temperature (UHT) process. This type of yogurt is sometimes referred to as pasteurized yogurt (Post Pasteurization Yogurt) or ambient temperature yogurt (Ambient Yogurt).

[0003] Pasteurized yogurt products contain no or only a small amount of active lactic acid bacteria. However, it is desirable that pasteurized yogurt products contain lactic acid bacteria and / or probiotics to provide consumers with the various benefits of such bacteria, such as health and dietary supplement benefits. Of course, adding live bacteria to pasteurized yogurt products to be stored at ambient temperature introduces a technical challenge: for example, due to a decrease in pH caused by an increase in lactic acid concentration due to fermentation, the bacteria will multiply to the point where the yogurt will spoil. In the prior art, this technical challenge has been addressed in many different ways. For example, bacterial cultures in pasteurized yogurt products have been added to the pasteurized yogurt products in the form of spores. Additionally, bacterial cultures for addition to pasteurized yogurt products have been added in the form of powdered, dried, freeze-dried, coated, or encapsulated cultures. Furthermore, bacterial cultures for addition to pasteurized yogurt products have been inactivated, for example, by radiation, microwave treatment, antibiotics, mild pasteurization, chemical agents (inhibitors), or by adjusting pH, water activity, or temperature.

[0004] WO2009 / 116864 discloses a dairy product containing probiotic spores, wherein the dairy product can be stored at non-refrigerated temperature for an extended period of time.

[0005] WO2004 / 069156 discloses a food product containing probiotics, wherein the probiotics have been inactivated by irradiation, microwave treatment, antibiotics, mild pasteurization and chemical agents (inhibitors).

[0006] EP1289380B1 discloses a food product, such as a dairy product, containing inactive Lactobacillus bacteria. Lactobacillus can be inactivated by, for example, mild heat treatment, pH adjustment or water activity adjustment.

[0007] EP1514553B1 discloses a double-coated lactic acid bacteria powder having a high survival rate in the human body, wherein the lactic acid bacteria have been double-coated with protein and polysaccharide.

[0008] CN101323850 discloses a method for producing microcapsules of Lactobacillus helveticus in a microencapsulated form having strong heat resistance.

[0009] EP0555618B1 discloses a dietary product containing freeze-dried lactic acid bacteria.

[0010] CN102492643 discloses a Lactobacillus rhamnosus strain GRX19 and its use in a starter culture for producing a fermented dairy product containing live Lactobacillus bacteria. The fermented dairy product is heat-treated, for example, at 70°C-75°C for 15-20 seconds, to which Lactobacillus strains are resistant because a small fraction of bacteria (e.g., 10exp7 CFU / mL) survives the heat treatment. After the heat treatment, the heat-treated product is aseptically packed into containers and stored at room temperature for, for example, 30 days.

[0011] WO2015 / 169928 discloses a liquid dairy composition suitable for preparing a foamed dairy product, wherein the composition is shelf-stable under ambient storage conditions, has a pH of 3.8 to 4.4, and comprises fermented milk, up to 0.12% hydrolyzed whey protein, up to 5% fat and up to 1% high methyl ester pectin.

[0012] US20100009034 discloses a method for preparing a fermented milk beverage that maintains a high viable cell count at ambient temperature, the method comprising: performing milk fermentation using a conventional lactic acid bacteria starter culture, diluting, mixing and sterilizing, and adding Lactobacillus rhamnosus ATCC 53103 to the mixed milk beverage under aseptic conditions.

[0013] US20100015285 discloses a method for preparing a directly acidified milk beverage that maintains a high viable cell count at ambient temperature, the method comprising: performing direct acidification by adjusting the pH to 4.0-4.5 to obtain an acidified milk beverage, sterilizing, and adding Lactobacillus rhamnosus ATCC 53103 together with 0.01%-0.3% growth promoting factors (e.g., carbohydrates) to the mixed milk beverage under aseptic conditions.

[0014] WO2017 / 194650 discloses a method for producing an ambient storage food product, the method comprising: providing a food product with a pH of 3.4 to 4.4, heat-treating the food product to obtain a heat-treated food product, and aseptically adding one or more ambient storage lactic acid bacteria strains to the heat-treated food product, wherein the lactic acid bacteria strains are selected from the group consisting of Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum and Lactobacillus delbrueckii subsp. bulgaricus and their mutants and variants.

[0015] Further development of improved pasteurized yogurt products containing active lactic acid bacteria is needed. Summary of the Invention

[0016] The present invention relates to a method for producing an ambient storage food product, wherein the food product has been prepared by the following steps: fermentation of a lactic acid bacteria starter culture, heat treatment to inactivate the lactic acid bacteria so as to prevent or substantially prevent post-acidification, and adding a strain of the genus Oenococcus and its derived mutants to the heat-treated food product.

[0017] In one aspect, the present invention relates to a method for producing an ambient storage food product, the method comprising: providing a food product having a pH of 3.4 to 4.4, heat-treating the food product to reduce the bacteria level to no more than 1×10 exp02 CFU / g to obtain a heat-treated food product, aseptically adding one or more ambient storage lactic acid bacteria strains to the heat-treated food product in a total amount of at least 1.0×10 exp03 CFU / g to obtain an ambient storage food product, and storing the ambient storage food product at ambient temperature for a period of time.

[0018] wherein the environmentally stored lactic acid bacteria strain is selected from the group consisting of the following strains,

[0019] (i) wherein the strain is capable of maintaining viability in an amount of at least 1.0×10 exp0 3 CFU / g when added in an amount of 1.0×10 exp0 7 CFU / g to a fermented milk test product in the form of a yogurt obtained by fermentation at a temperature of 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii bulgaricus to a pH of 4.3 and having been heat-treated at 75°C for 30 seconds at the end of a storage period of 120 days at 25°C of the test product, and

[0020] (ii) wherein the pH of the test product decreases by at most 0.8 units during the storage period, and

[0021] (iii) wherein the strain is selected from the group consisting of strains of the genus Oenococcus and mutants thereof.

[0022] The present invention is based on the unexpected experimental discovery that strains of bacteria from the genus Oenococcus can maintain a certain level of viability for at least 120 days when added to a yogurt product stored at ambient temperature without any significant decrease in pH levels. This is a surprising finding because, in a milk matrix, lactic acid bacteria typically either grow and have a reduced pH, or die, as they grow on available carbohydrate sources while simultaneously lowering the pH until the pH reaches a level at which the bacteria cannot survive.

[0023] In another aspect, the present invention relates to an ambient storage food product comprising an ambient storage lactic acid bacteria strain, wherein the product has a pH of 3.4 to 4.4, wherein the product contains at least 1.0×10 exp0 3 CFU / g of the strain, wherein the ambient storage food product is stored at ambient temperature for a period of time, and

[0024] wherein the lactic acid bacteria strain is selected from the group consisting of the following strains,

[0025] (i) wherein the strain is capable of maintaining viability in an amount of at least 1.0×10 exp0 3 CFU / g when added in an amount of 1.0×10 exp0 7 CFU / g to a fermented milk test product in the form of a yogurt obtained by fermentation at a temperature of 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to a pH of 4.3 and which has been heat-treated at 75°C for 30 seconds, at the end of a storage period of 120 days at 25°C of the test product, and

[0026] (ii) wherein the pH of the test product decreases by at most 0.8 units during the storage period, and

[0027] (iii) wherein the strain is selected from the group consisting of strains of the genus Oenococcus and mutants thereof. DETAILED DESCRIPTION

[0028] Methods for storing food products in production environments

[0029] The present invention relates to a method for producing an ambient storage food product, the method comprising: providing a food product having a pH of 3.4 to 4.4, heat-treating the food product to reduce the bacteria level to no more than 1×10 exp02 CFU / g to obtain a heat-treated food product, aseptically adding one or more ambient storage lactic acid bacteria strains to the heat-treated food product in a total amount of at least 1.0×10 exp03 CFU / g to obtain an ambient storage food product, and storing the ambient storage food product at ambient temperature for a period of time, wherein the ambient storage lactic acid bacteria strain is selected from the group consisting of:

[0030] (i) wherein the strain is capable of maintaining viability in an amount of at least 1.0×10 exp0 CFU / g at the end of a storage period of 120 days at 25°C of a fermented milk test product in the form of a yoghurt obtained by fermentation at 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii bulgaricus to a pH of 4.3 and having been heat-treated at 75°C for 30 seconds when added in an amount of 1.0×10 exp07 CFU / g to the test product, and

[0031] (ii) wherein the pH of the test product decreases by at most 0.8 units during the storage period, and

[0032] (iii) wherein the strain is selected from the group consisting of strains of the genus Oenococcus and mutants thereof.

[0033] In a preferred embodiment, the environmental storage strain is capable of maintaining viability in an amount of at least 1.0×10 exp03 CFU / g, preferably at least 5.0×10 exp03 CFU / g, more preferably at least 1.0×10 exp04 CFU / g, more preferably at least 5.0×10 exp04 CFU / g, and most preferably at least 1.0×10 exp05 CFU / g at the end of the storage period.

[0034] Preferably, the pH decreases during the storage period by at most 0.7, preferably 0.6, preferably 0.5, preferably 0.4, preferably 0.3, most preferably 0.2.

[0035] In a preferred embodiment of the present invention, when added to the test product in an amount of 1.0×10 exp07 CFU / g, the strain increases to an amount of at least 5.0×10 exp07 CFU / g, preferably 7.5×10 exp07 CFU / g, most preferably 1.0×10 exp08 CFU / g.

[0036] Preferably, the increase in cell mass occurs within 45 days, preferably within 40 days, preferably within 35 days, preferably within 30 days, preferably within 25 days, preferably within 20 days, and most preferably within 15 days of adding the strain to the test product. Preferably, the cell mass reaches a maximum within 45 days, preferably within 40 days, preferably within 35 days, preferably within 30 days, preferably within 25 days, preferably within 20 days, and most preferably within 15 days of adding the strain to the test product.

[0037] In a particular embodiment of the present invention, the ambient stored food product is stored at ambient temperature for a period of at least 1 day, preferably at least 2 days, more preferably at least 3 days, more preferably at least 4 days, more preferably at least 5 days, more preferably at least 6 days, more preferably at least 7 days, more preferably at least 8 days, more preferably at least 9 days, and most preferably at least 10 days.

[0038] In a specific embodiment of the present invention, the Oenococcus strain is selected from the group consisting of an Oenococcus oeni strain, an Oenococcus kitaharae strain, an Oenococcus sicerae strain, and mutants thereof. In a specific embodiment of the present invention, the Oenococcus strain is selected from the group consisting of an Oenococcus oeni strain and mutants thereof.

[0039] In a specific embodiment of the present invention, the Oenococcus strain of the present invention is selected from the group consisting of the Oenococcus oeni strain with a deposition number of DSM 33144, the Oenococcus oeni strain with a deposition number of DSM 33145, the Oenococcus oeni strain with a deposition number of DSM 33146, the Oenococcus oeni strain with a deposition number of DSM 33147, the Oenococcus oeni strain with a deposition number of DSM 14498, the Oenococcus oeni strain with a deposition number of DSM 15568, the Oenococcus oeni strain with a deposition number of DSM 15569, the Oenococcus oeni strain with a deposition number of DSM 15570, the Oenococcus oeni strain with a deposition number of DSM 15571, and mutants thereof.

[0040] In a specific embodiment of the present invention, the Oenococcus strain of the present invention is selected from the group consisting of the Oenococcus oeni strain with a deposition number of DSM 33144, the Oenococcus oeni strain with a deposition number of DSM 33145, the Oenococcus oeni strain with a deposition number of DSM 33146, the Oenococcus oeni strain with a deposition number of DSM 33147 and the Oenococcus oeni strain with a deposition number of DSM 14498 and mutants thereof.

[0041] In a specific embodiment of the present invention, the Oenococcus strain of the present invention is a citrate-negative strain. The term "citrate-negative" is used in the context of the present invention to characterize a strain that, when placed in a culture medium containing a predetermined amount of citric acid, is only capable of degrading up to 80% of the citric acid. In particular, when placed in a culture medium containing a predetermined amount of citric acid, the Oenococcus strain of the present invention is only capable of degrading up to 70%, preferably up to 60%, more preferably up to 50%, more preferably up to 40%, more preferably up to 30%, more preferably up to 20%, more preferably up to 15%, and most preferably up to 10% of the citric acid.

[0042] Citric acid-negative Oenococcus strains are described in WO2004 / 113488, which is incorporated herein by reference. In a specific embodiment of the present invention, the citric acid-negative Oenococcus strain of the present invention is selected from the group consisting of the Oenococcus oeni strain with a deposit number of DSM 15568, the Oenococcus oeni strain with a deposit number of DSM 15569, the Oenococcus oeni strain with a deposit number of DSM 15570, and the Oenococcus oeni strain with a deposit number of DSM 15571, and mutants thereof.

[0043] In a particular embodiment of the present invention, the Oenococcus strain of the present invention is sucrose positive.

[0044] In a particular embodiment of the present invention, the Oenococcus strain of the present invention is glucose positive.

[0045] In a particular embodiment of the present invention, the Oenococcus strain of the present invention is galactose positive.

[0046] In a particular embodiment of the present invention, the Oenococcus strain of the present invention is fructose positive.

[0047] In a particular embodiment of the present invention, the Oenococcus strain of the present invention is lactose deficient.

[0048] The ability of the Oenococcus strains of the present invention to grow on various carbohydrate sources can be tested using the method described in Example 2.

[0049] In a particular embodiment of the method of the present invention, the food product having a pH of 3.4 to 4.4 is a starter culture fermented dairy product provided by fermenting a milk base with a lactic acid bacteria starter culture to obtain the starter culture fermented dairy product.

[0050] In a particular embodiment of the method of the present invention, the starter culture fermented dairy product has a protein content of more than 5.1% (w / w) on a weight basis.

[0051] In a particular embodiment of the method according to the invention, the starter culture fermentation of the dairy product is not diluted.

[0052] The method of the present invention is described in more detail below, which relates to a method for storing a fermented dairy product in a production environment.

[0053] The starter culture may be any conventional lactic acid bacteria starter culture used for the production of a particular type of fermented dairy product, including single strain cultures and culture mixtures. In a preferred embodiment of the above method of the present invention, the fermentation is carried out to obtain a pH of 3.0 to 5.0, preferably 3.9 to 4.8, more preferably 4.0 to 4.6, most preferably 4.1 to 4.4.

[0054] The starter culture-fermented dairy product is preferably heat-treated at a temperature of 50°C to 90°C, preferably 60°C to 85°C, more preferably 65°C to 82°C, and most preferably 70°C to 80°C to reduce the bacterial level in the starter culture to no more than 1.0×10 exp02 CFU / g fermented milk. The heat treatment is preferably performed for a time of 10 to 180 seconds, preferably 12 to 120 seconds, more preferably 14 to 90 seconds, more preferably 16 to 60 seconds, more preferably 18 to 50 seconds, and most preferably 20 to 40 seconds. Preferably, the bacterial level in the starter culture is reduced to no more than 1.0×10 exp01 CFU / g fermented milk, more preferably 0 CFU / g. For the environment Lactic acid bacteria strains for stored food products

[0055] In one aspect, the present invention relates to a lactic acid bacteria strain for use in an ambient storage food product, wherein the product has a pH of 3.4 to 4.4, wherein the product contains at least 1.0×10 exp0 3 CFU / g of the strain, wherein the ambient storage food product is stored at ambient temperature for a period of time, and

[0056] wherein the lactic acid bacteria strain is selected from the group consisting of the following strains,

[0057] (i) wherein the strain is capable of maintaining viability in an amount of at least 1.0×10 exp0 3 CFU / g when added in an amount of 1.0×10 exp0 7 CFU / g to a fermented milk test product in the form of a yogurt obtained by fermentation at a temperature of 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to a pH of 4.3 and which has been heat-treated at 75°C for 30 seconds, at the end of a storage period of 120 days at 25°C of the test product, and

[0058] (ii) wherein the pH of the test product decreases by at most 0.8 units during the storage period, and

[0059] (iii) wherein the strain is selected from the group consisting of strains of the genus Oenococcus and mutants thereof.

[0060] In a particular embodiment of the strains of the invention, the product is a chemically acidified product.

[0061] In a particular embodiment of the strain of the present invention, the product is a fermented dairy product obtained by fermenting a milk base with a lactic acid bacteria starter culture, wherein the product contains no more than 1×10 exp02 CFU / g of the starter culture and at least 1×10 exp03 CFU / g of the environmental storage lactic acid bacteria strain.

[0062] In a particular embodiment of the strain of the present invention, the product is a fermented dairy product obtained by fermenting a dairy base with a lactic acid bacteria starter culture, wherein the fermented dairy product after fermentation has been subjected to a heat treatment to reduce the bacterial level of the starter culture to no more than 1×10 exp02 CFU / g, and wherein after the heat treatment, the environmental storage strain according to claim 1 has been aseptically added to the heat-treated product in an amount of at least 1.0×10 exp03 CFU / g. Preferably, the environmental storage strain of the present invention has been aseptically added to the heat-treated product in an amount of at least 1.0×10 exp04 CFU / g, more preferably at least 1.0×10 exp05 CFU / g, more preferably at least 1.0×10 exp06 CFU / g, more preferably at least 1.0×10 exp07 CFU / g, and most preferably at least 1.0×10 exp08 CFU / g.

[0063] The Oenococcus strain of the present invention has been described above in conjunction with the method of the present invention, whereby reference is made to the method of the present invention.

[0064] The strains of the present invention can be formulated into a composition comprising one or more strains according to the present invention. Thus, in one embodiment, the strains of the present invention are formulated into a composition comprising a single strain according to the present invention. In another embodiment, the strains are formulated into a composition comprising two or more strains of the present invention. The composition can be in the form of lyophilized or frozen pellets.

[0065] Environmental storage of food products

[0066] In one embodiment, the present invention relates to an ambient storage food product comprising an ambient storage lactic acid bacteria strain, wherein the product has a pH of 3.4 to 4.4, wherein the product contains at least 1.0×10 exp0 3 CFU / g of the strain, wherein the ambient storage food product is stored at ambient temperature for a period of time, and

[0067] wherein the lactic acid bacteria strain is selected from the group consisting of the following strains,

[0068] (i) wherein the strain is capable of maintaining viability in an amount of at least 1.0×10 exp0 3 CFU / g when added in an amount of 1.0×10 exp0 7 CFU / g to a fermented milk test product in the form of a yogurt obtained by fermentation at a temperature of 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to a pH of 4.3 and which has been heat-treated at 75°C for 30 seconds, at the end of a storage period of 120 days at a temperature of 25°C of the test product, and

[0069] (ii) wherein the pH of the test product decreases by at most 0.8 units during the storage period, and

[0070] (iii) wherein the strain is selected from the group consisting of strains of the genus Oenococcus and mutants thereof.

[0071] In one embodiment, the present invention is directed to an ambient stored food product, wherein the product is a chemically acidified product.

[0072] In one embodiment, the present invention relates to an ambient storage food product, wherein the product is a fermented dairy product obtained by fermenting a milk base using a lactic acid bacteria starter culture, wherein the product contains not more than 1×10 exp02 CFU / g of the starter culture and at least 1×10 exp03 CFU / g of the ambient storage lactic acid bacteria strain.

[0073] In one embodiment, the present invention relates to an ambient storage food product, wherein the product is a fermented dairy product obtained by fermenting a dairy base with a lactic acid bacteria starter culture, wherein the fermented dairy product after fermentation has been subjected to a heat treatment to reduce the bacteria level of the starter culture to no more than 1×10 exp02 CFU / g, and wherein after said heat treatment, the ambient storage strain according to claim 1 has been aseptically added to the heat treated product in an amount of at least 1.0×10 exp03 CFU / g.

[0074] In one embodiment, the present invention relates to an ambient stored food product, wherein the Oenococcus strain is selected from the group consisting of Oenococcus oeni, Oenococcus kitamoto, Oenococcus ciliata, and mutants thereof.

[0075] In one embodiment, the present invention relates to an ambient stored food product, wherein the strain is selected from the group consisting of the Oenococcus oeni strain deposited with number DSM 33144, the Oenococcus oeni strain deposited with number DSM 33145, the Oenococcus oeni strain deposited with number DSM 33146, the Oenococcus oeni strain deposited with number DSM 33147, the Oenococcus oeni strain deposited with number DSM 14498, the Oenococcus oeni strain deposited with number DSM 15568, the Oenococcus oeni strain deposited with number DSM 15569, the Oenococcus oeni strain deposited with number DSM 15570 and the Oenococcus oeni strain deposited with number DSM 15571 and mutants thereof.

[0076] In a preferred embodiment of the present invention, the ambient stored food product is selected from the group consisting of fermented dairy products, chemically acidified dairy products, fruit drinks, fermented cereal products, chemically acidified cereal products, soy milk products and any mixtures thereof. Preferably, the ambient stored food product is a fermented dairy product, wherein the milk is mammalian milk.

[0077] Fermented dairy products typically contain protein at a level of 2.0% to 3.5% by weight. Fermented dairy products can also be low-protein products with a protein level of 1.0% to 2.0% by weight. Alternatively, fermented dairy products can be high-protein products with a protein level of more than 3.5% by weight, preferably more than 5.1% by weight. In a particular embodiment of the fermented dairy product of the present invention, the product is a mixture of a fermented dairy product and a cereal product (e.g., an oat product), wherein the cereal product can be a fermented cereal product, such as a fermented oat product.

[0078] In one embodiment of the present invention, the ambient stored food product is a fermented cereal product. The fermented cereal product can be prepared by milling grains of a cereal biological source material to produce cereal flour, which is then fermented. The fermentation of the cereal flour can be carried out using the same lactic acid bacteria (starter culture) as described elsewhere in this application for milk-based fermentation.

[0079] In one embodiment of the present invention, the ambient storage food product is a fruit drink. The fruit drink may further contain, for example, oats, soy, almonds, whey and / or non-fermented milk (e.g., in the form of milk powder). In one embodiment, the fruit drink of the present invention does not contain dairy ingredients, such as milk. In another embodiment of the fruit drink of the present invention, the fruit drink further contains a fermented dairy product.

[0080] In another embodiment of the present invention, the ambient stored food product of the present invention is a chemically acidified product. Acidification can be performed using any acidulant suitable for addition to a food product, such as lactic acid, citric acid, fruit juice, fruit pulp, and fruit complexes. In a specific embodiment, the ambient stored food product is milk acidified with fruit juice.

[0081] In a specific embodiment of the present invention, the ambient-stored food product is a chemically acidified cereal product. The chemically acidified cereal product can be prepared by milling grains of a cereal biological source material to produce a cereal flour, which is then used to produce an aqueous suspension, and then adjusting the pH of the suspension to a desired level. In a specific embodiment, the ambient-stored food product is a cereal product acidified with a fruit beverage.

[0082] Any combination of the above-described elements, aspects, and embodiments in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0083] definition

[0084] The following terms and expressions used in connection with the present invention have the following meanings:

[0085] The expression "heat treatment" refers to any treatment using any temperature, for any period of time and by any means or equipment, which inactivates at least a portion of the bacteria of the starter culture. In this context, the term "inactivation" refers to any cessation, reduction or inhibition of bacterial growth, such as cell lysis.

[0086] The expression "ambient storage" refers to storage at ambient temperature. The expression "ambient temperature" refers to the temperature of the surrounding environment, for example, room temperature. For example, the ambient temperature may be 5°C to 40°C, more particularly 10°C to 35°C, more particularly 15°C to 30°C, and most particularly 18°C ​​to 27°C. The ambient temperature may be controlled, i.e., the temperature is the same throughout the day (24 hours), or uncontrolled, i.e., the temperature varies throughout the day (24 hours).

[0087] The expression "viability" means that the bacteria are able to show growth (form colonies) on MRS agar plates incubated under anaerobic conditions at 30°C for 3 days. MRS agar has the following composition (g / L):

[0088] Peptone: 10.0

[0089] Beef extract: 10.0

[0090] Yeast extract: 5.0

[0091] Glucose: 20.0

[0092] Polysorbate 80:1.0

[0093] Ammonium citrate: 2.0

[0094] Sodium acetate: 5.0

[0095] Magnesium sulfate: 0.1

[0096] Manganese sulfate: 0.05

[0097] Dipotassium hydrogen phosphate: 2.0

[0098] Agar: 15.0

[0099] The expression "ambient storage lactic acid bacteria strain" refers to a lactic acid bacteria strain which, when added to a fermented dairy product, is suitable for ambient storage for a certain period of time.

[0100] The expression "starter culture fermented dairy product" refers to a fermented dairy product containing a starter culture for fermenting milk.

[0101] The expression "heat-treated fermented dairy product" refers to a fermented dairy product that has been subjected to a heat treatment.

[0102] The expression "ambient storage fermented dairy product" refers to a fermented dairy product that is suitable for ambient storage for a certain period of time.

[0103] The expression "lactic acid bacteria" refers to Gram-positive bacteria, microaerophilic or anaerobic, which ferment sugars while producing acids, including lactic acid, acetic acid and propionic acid as the main acids produced. The most industrially useful lactic acid bacteria are found in the order "Lactobacilli", which includes the genera Lactococcus, Streptococcus, Lactobacillus, Leuconostoc, Pseudomonas, Pediococcus, Brevibacterium, Enterococcus and Propionibacterium. These lactic acid bacteria are often used as food cultures, alone or in combination with other lactic acid bacteria.

[0104] Lactic acid bacteria, including bacteria of the genera Lactobacillus and Lactococcus, are commonly supplied to the dairy industry in the form of frozen or freeze-dried cultures for bulk starter propagation, or in the form of so-called "direct-vase starter" (DVS) cultures for direct inoculation into fermentation vessels or tanks for the production of dairy products, such as fermented dairy products or cheese. Such lactic acid bacteria cultures are often referred to as "starter cultures" or "starters."

[0105] The term "milk" is to be understood as the lacteal secretion obtained by milking any mammal, such as a cow, sheep, goat, buffalo or camel. In a preferred embodiment, the milk is bovine milk. The term "milk" also includes protein / fat solutions made from plant materials, such as soy milk.

[0106] The term "milk matrix" may be any raw milk material and / or processed milk material that can be fermented according to the method of the present invention. Thus, useful milk matrices include, but are not limited to, solutions / suspensions of any milk or milk-like product containing protein, such as whole or low-fat milk, skim milk, buttermilk, reconstituted milk powder, condensed milk, milk powder, whey, whey permeate, lactose, mother liquor from lactose crystallization, whey protein concentrate, or cream. Obviously, the milk matrix may be derived from any mammal, for example substantially pure mammalian milk or reconstituted milk powder.

[0107] Prior to fermentation, the milk base may be homogenized and pasteurized according to methods known in the art.

[0108] As used herein, "homogenization" means mixing thoroughly to obtain a soluble suspension or emulsion. If homogenization is performed before fermentation, the milk fat is broken down into smaller sizes so that it no longer separates from the milk. This can be achieved by forcing the milk through small orifices under high pressure.

[0109] As used herein, "pasteurization" refers to treating a milk matrix to reduce or eliminate the presence of living organisms, such as microorganisms. Preferably, pasteurization is achieved by maintaining a specific temperature for a specific period of time. Typically, the specific temperature is achieved by heating. The temperature and duration can be selected to kill or inactivate certain bacteria, such as harmful bacteria. This can be followed by a rapid cooling step.

[0110] "Fermentation" in the method of the present invention refers to the conversion of carbohydrates into alcohols or acids by the action of microorganisms. Preferably, the fermentation in the method of the present invention comprises the conversion of lactose into lactic acid.

[0111] The fermentation processes to be used in the production of dairy products are well known, and those skilled in the art will understand how to select suitable process conditions, such as temperature, oxygen, the amount and characteristics of microorganisms, and process time. Obviously, the fermentation conditions are selected to support the implementation of the present invention, i.e. to obtain dairy products in solid form (e.g., cheese) or liquid form (e.g., fermented dairy products).

[0112] In the context of the present invention, the term "mutant" should be understood as a strain derived from the strain of the present invention by, for example, genetic engineering, radiation and / or chemical treatment, and / or selection, adaptation, screening, etc. Preferably, the mutant is a functionally equivalent mutant, for example, a mutant having substantially the same or improved properties as the parent strain in terms of adaptability to environmental storage. Such a mutant is a part of the present invention. In particular, the term "mutant" refers to a strain obtained by subjecting the strain of the present invention to any conventional mutagenesis treatment or to a spontaneously generated mutant, including treatment with chemical mutagens such as ethyl methanesulfonate (EMS) or N-methyl-N'-nitro-N-nitroguanidine (NTG), or UV light. The mutant may be subjected to several mutagenesis treatments (a single treatment being understood as a mutagenesis step followed by a screening / selection step), but is currently preferably treated no more than 20 times, no more than 10 times, or no more than 5 times. In currently preferred mutants, less than 1%, or less than 0.1%, less than 0.01%, less than 0.001% or even less than 0.0001% of the nucleotides in the bacterial genome are altered (e.g., by substitutions, insertions, deletions or a combination thereof) compared to the parent strain.

[0113] Preferably, the “mutant” of the strain according to the invention has the property that, when added in an amount of 1.0×10 exp07 CFU / g to a fermented milk test product in the form of yogurt obtained by fermentation at a temperature of 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii bulgaricus to a pH of 4.3 and heat-treated at 75°C for 30 seconds, the strain is able to maintain viability in an amount of at least 1.0×10 exp03 CFU / g at the end of a storage period of 120 days at a temperature of 25°C of the test product, and wherein the pH decreases by at most 0.8 units during the storage period.

[0114] Preferably, the "mutant" of the strain of the present invention has less than 25, more preferably less than 10, more preferably less than 9, more preferably less than 8, more preferably less than 7, more preferably less than 6, more preferably less than 5, more preferably less than 4, more preferably less than 3, more preferably less than 2 mutations in the amino acid sequence of one or more proteins of the strain. In this connection, the term "mutation" refers to a mutation selected from the group consisting of substitution, deletion and insertion.

[0115] In the context of describing the present invention (especially in the context of the following claims), unless otherwise stated herein or clearly contradictory to the context, the terms "a (one)" and "an (one)" and "the / said" and similar references should be understood to include singular and plural. Unless otherwise stated, the terms "comprise", "have", "include" and "contain" should be understood as open terms (i.e., meaning "including but not limited to"). Unless otherwise stated herein, the description of numerical ranges herein is only intended to be used as a shorthand method for individually referring to each individual value falling within the range, and each individual value is incorporated into this specification as if it were individually described herein. Unless otherwise stated herein or clearly contradictory to the context, all methods described herein can be performed in any appropriate order. Unless otherwise required, the use of any and all examples or exemplary languages ​​(e.g., "such as") provided herein is only intended to better illustrate the present invention without limiting the scope of the present invention. The language in the specification should not be understood to indicate that any unclaimed element is necessary for the practice of the present invention.

[0116] The expression "fermented dairy product" refers to a food or feed product, wherein the preparation of the food or feed product involves fermentation of a milk base with lactic acid bacteria. As used herein, "fermented dairy product" includes, but is not limited to, products such as thermophilic fermented dairy products (e.g., yogurt), mesophilic fermented dairy products (e.g., sour cream and buttermilk), cheese, and fermented whey.

[0117] The term "thermophile" herein refers to microorganisms that grow best at temperatures above 43°C. The most industrially useful thermophilic bacteria include Streptococcus and Lactobacillus. The term "high-temperature fermentation" herein refers to fermentation at a temperature above about 35°C, such as from about 35°C to about 45°C. The term "high-temperature fermented dairy product" refers to a fermented dairy product prepared by thermophilic fermentation of a thermophilic starter culture, and includes fermented dairy products such as set yogurt, stirred yogurt, and drinking yogurt (e.g., Yakult).

[0118] The term "mesophile" herein refers to microorganisms that grow best at moderate temperatures (15°C-40°C). The most industrially useful mesophilic bacteria include the genera Lactococcus and Leuconostoc. The term "mesophilic fermentation" herein refers to fermentation at a temperature of about 22°C to about 35°C. The term "mesophilic fermented dairy product" refers to a fermented dairy product prepared by mesophilic fermentation of a mesophilic starter culture, and includes fermented dairy products such as buttermilk, yogurt, fermented milk, smetana, sour cream, kefir, and fresh cheeses such as quark, tvarog, and cream cheese.

[0119] The term "cheese" should be understood to encompass any cheese, including hard cheese, semi-hard cheese and soft cheese, such as the following types of cheese: cottage cheese, feta cheese, cheddar cheese, parmesan cheese, mozzarella cheese, emmental cheese, danbo cheese, gouda cheese, edam cheese, feta-type cheese, blue cheese, brine cheese, camembert cheese and brie cheese. Those skilled in the art understand how to convert coagulum into cheese, and methods can be found in the literature, see for example Kosikowski, FV and VV Mistry, "Cheese and Fermented Milk Foods", 1997, 3rd edition, FV Kosikowski, LLC Westport, CT. As used herein, cheeses having a NaCl concentration of less than 1.7% (w / w) are referred to as "low-salt cheeses".

[0120] In the context of the present invention, the term "juice" refers to the liquid naturally contained in fruit, prepared by mechanically pressing or macerating fresh fruit in the absence of heat and solvents. "Juice" may consist of juice from one type of fruit or a mixture of more than one type of fruit.

[0121] In the context of the present invention, the term "fruit drink" refers to a beverage having a fruit juice content of 0% to 29%.

[0122] In the context of the present invention, the term "nectar" refers to a beverage having a juice content of 30% to 99% juice.

[0123] In the context of the present invention, the term "puree" refers to fruit prepared by grinding, pressing and / or straining in the absence of heat and solvents to the consistency of a thick liquid or soft paste. "Puree" is made from 100% fruit, not just the juice of the fruit.

[0124] In the context of the present invention, the term "fruit beverage" refers to a beverage comprising fruit juice, fruit concentrate and / or fruit puree. The term "fruit beverage" encompasses "juice," "fruit drink," and "nectar" as defined herein. A "fruit beverage" may be a beverage containing fruit pulp or a beverage in which the pulp has been removed by an operation such as centrifugation.

[0125] The term "aseptic addition" means that no or minimal amounts of any microorganisms other than environmentally stored lactic acid bacteria are introduced.

[0126] The term "cereal product" refers to any product obtained from cereals or cereal biological sources, including oats, corn, barley, rye, buckwheat, wheat and rice.

[0127] The term "lactose-deficient" is used in the context of the present invention to characterize LAB that have partially or completely lost the ability to use lactose as a source of cell growth or maintenance of cell viability. Such LAB are able to metabolize one or more carbohydrates selected from sucrose, galactose and / or glucose or another fermentable carbohydrate. Since the amounts of these carbohydrates naturally present in milk are insufficient to support the fermentation of lactose-deficient mutants, it is necessary to add these carbohydrates to the milk. Lactose-deficient and partially lactose-deficient LAB can be characterized by white bacterial colonies on a culture medium containing lactose and X-Gal.

[0128] The term "citrate-negative" strain refers to a strain that forms white colonies on Kempler McKay agar medium, while citrate-positive strains form dark blue colonies on the medium, wherein the Kempler McKay medium is defined in the publication "Improved medium for detection of citrate-fermenting Streptococcus lactis subsp. diacetylactis", G.M. Kempler and L.L. McKay, Applied and Environmental Microbiology, April 1980, Vol. 39, No. 4, pp. 926-927. The Kempler McKay medium has the following composition:

[0129] 1% (weight / volume) skim milk

[0130] 0.25% milk protein hydrolyzed peptone

[0131] 0.5% dextrose

[0132] 1.5% agar

[0133] Preparation method:

[0134] At 10 lb / in 2 After sterilizing the culture medium at 45°C for 12 minutes, adjust the temperature at 45°C. Steam (100°C) for 30 minutes two solutions (one containing 10% potassium ferricyanide and one containing 1g ferric citrate and 1g sodium citrate in 40mL of water). Add 10mL of each solution to 1L of agar medium, gently swirl the agar, and pour it off. Dry the plates at 30°C in the dark for 24 hours.

[0135] The expressions "XX×10expYY" and "X.XEYY" both refer to XX×10 YY , these two expressions can be used interchangeably.

[0136] The expression "CFU" means colony forming units.

[0137] Specific items of the present invention

[0138] 1. An ambient storage lactic acid bacteria strain, wherein when added in an amount of 1.0×10 exp07 CFU / g to a fermented milk test product in the form of yogurt, the strain is capable of maintaining viability in an amount of at least 1.0×10 exp03 CFU / g at the end of a storage period of 120 days at a temperature of 25°C, the yogurt being obtained by fermentation at a temperature of 43°C with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii bulgaricus to a pH of 4.3 and having been heat-treated at 75°C for 30 seconds, and wherein the pH decreases by at most 0.8 units during the storage period, and wherein the strain is selected from the group consisting of strains of the genus Coccus and mutants thereof.

[0139] 2. The strain according to embodiment 1, wherein the strain is capable of maintaining viability in an amount of at least 1.0×10 exp03 CFU / g, preferably at least 5.0×10 exp03 CFU / g, more preferably at least 1.0×10 exp04 CFU / g, more preferably at least 5.0×10 exp04 CFU / g, and most preferably at least 1.0×10 exp05 CFU / g at the end of the storage period.

[0140] 3. The strain according to embodiment 1 or 2, wherein the pH decreases by at most 0.7, preferably 0.6, preferably 0.5, preferably 0.4, preferably 0.3, and most preferably 0.2 during the storage period.

[0141] 4. The strain according to any one of the preceding embodiments, wherein when added to the test product in an amount of 1.0×10 exp07 CFU / g, the strain increases to an amount of at least 5.0×10 exp07 CFU / g, preferably 7.5×10 exp07 CFU / g, most preferably 1.0×10 exp08 CFU / g.

[0142] 5. The strain according to embodiment 4, wherein the increase in the amount of cells occurs within 45 days, preferably within 40 days, preferably within 35 days, preferably within 30 days, preferably within 25 days, preferably within 20 days, and most preferably within 15 days of adding the strain to the test product.

[0143] 6. The strain according to embodiment 4 or 5, wherein the amount of cells reaches a maximum within 45 days, preferably within 40 days, preferably within 35 days, preferably within 30 days, preferably within 25 days, preferably within 20 days, and most preferably within 15 days of adding the strain to the test product.

[0144] 7. A composition comprising one or more of the environmentally stored lactic acid bacteria strains according to any one of embodiments 1-6.

[0145] 8. An ambient storage food product having a pH of 3.4 to 4.4, wherein the product contains at least 1.0×10 exp0 3 CFU / g of the ambient storage strain according to embodiment 1.

[0146] 9. The food product of embodiment 8, wherein the product is a chemically acidified product.

[0147] 10. The food product according to embodiment 8, wherein the product is a fermented dairy product obtained by fermenting a milk base using a lactic acid bacteria starter culture, wherein the product contains no more than 1×10 exp02 CFU / g of the starter culture and at least 1×10 exp03 CFU / g of the environmental storage strain according to embodiment 1.

[0148] 11. The food product according to embodiment 8, wherein the product is a fermented dairy product obtained by fermenting a dairy base with a lactic acid bacteria starter culture, wherein the fermented dairy product after fermentation has been subjected to a heat treatment to reduce the bacterial level of the starter culture to no more than 1×10 exp02 CFU / g, and wherein after the heat treatment, the environmental storage strain according to embodiment 1 has been aseptically added to the heat-treated product in an amount of at least 1.0×10 exp03 CFU / g.

[0149] 12. A method for producing an ambient storage food product, the method comprising: providing a food product having a pH of 3.4 to 4.4, heat-treating the food product to reduce the bacteria level to no more than 1×10 exp02 CFU / g to obtain a heat-treated food product, and aseptically adding one or more ambient storage lactic acid bacteria strains according to embodiment 1 in a total amount of at least 1.0×10 exp03 CFU / g to the heat-treated food product to obtain an ambient storage food product.

[0150] 13. A method for producing an ambient storage fermented dairy product, the method comprising: fermenting a milk base with a lactic acid bacteria starter culture to obtain a starter culture fermented dairy product, heat-treating the starter culture fermented dairy product to reduce the bacteria level of the starter culture to no more than 1×10 exp02 CFU / g to obtain a heat-treated fermented dairy product, and aseptically adding one or more ambient storage lactic acid strains according to embodiment 1 in a total amount of at least 1.0×10 exp03 CFU / g to the heat-treated fermented dairy product to obtain an ambient storage fermented dairy product.

[0151] 14. Use of the ambient-stored lactic acid bacteria strain according to embodiment 1 for aseptic addition to a heat-treated food product in a total amount of at least 1.0×10 exp03 CFU / g, wherein the heat-treated food product has a pH of 3.4 to 4.4 and has been heat-treated to reduce the bacterial level of the starter culture to no more than 1×10 exp02 CFU / g.

[0152] 15. A lactic acid bacteria strain, wherein the strain is selected from the group consisting of the Oenococcus oeni strain with a deposit number of DSM 33144, the Oenococcus oeni strain with a deposit number of DSM 33145, the Oenococcus oeni strain with a deposit number of DSM 33146, the Oenococcus oeni strain with a deposit number of DSM 33147, the Oenococcus oeni strain with a deposit number of DSM 14498, and mutants thereof.

[0153] Example

[0154] Example 1: Testing the environmental storage adaptability of six strains of Oenococcus oeni in pasteurized yogurt (PPY)

[0155] Table 1: Milk matrix

[0156]

[0157] Composition of yogurt after terminal pasteurization

[0158] Fat: 2.6%

[0159] Protein: 3.0%

[0160] Carbohydrates: 11.4%

[0161] Starter cultures: Commercially available Starter culture type FD-DVS YF-L904. The starter culture was inoculated into the milk matrix at a level of 500 U / 2500 L milk.

[0162] Environmentally stored strains: Six commercially available Oenococcus oeni strains were tested. Each strain was inoculated into pasteurized yogurt at a concentration of 1 × 10 exp07 cfu / g.

[0163] Test strain 1: DSM33146

[0164] Test strain 2: DSM33147

[0165] Test strain 3: DSM15570

[0166] Test strain 4: DSM33144

[0167] Test strain 5: DSM33145

[0168] Test strain 6: DSM14498

[0169] Steps to produce test products

[0170] 1. Disperse the dry ingredients into the milk

[0171] 2. Let stand at 10℃ for at least 2 hours with gentle stirring

[0172] 3. Heat the milk until it reaches 65°C

[0173] 4. Homogenize at 150 bar

[0174] 5. Heat treatment to 95℃ for 5 minutes

[0175] 6. Cool to 43°C fermentation temperature

[0176] 7. Pump the milk into the fermentation tank

[0177] 8. Inoculation of YoFlex Culture FD-DVS YF-L904

[0178] 9. Ferment until pH reaches 4.30

[0179] 10. Break the curd and stir until a smooth texture is obtained

[0180] 11. Heat treatment at 75°C for 30 seconds

[0181] 12. Post-processing at 2 bar

[0182] 13. Cool to 25℃

[0183] 14. Aseptic filling into 100mL sterile container

[0184] Steps for testing environmentally stored strains 15. Inoculation of strains and cultures

[0185] 16.Store at room temperature of 23°C for 120 days

[0186] pH determination

[0187] Using temperature compensation, calibrate the pH electrode with standard buffer solutions of pH 4.01, pH 7.00, and pH 9.21. The samples are measured at the same temperature, in this case, room temperature (23°C). The displayed measurement must have a stable signal for at least 30 seconds before recording the value. Rinse the electrode with deionized water and carefully wipe it with a soft tissue between samples.

[0188] pH was measured on day +0 and day +1, and then monthly until day +120.

[0189] Cell counting method

[0190] Cell populations were monitored by colony plate counts (cfu / g) on ​​De Man, Rogosa, and Sharpe (MRS) medium adjusted to pH 5.4. Samples were incubated at 30°C for 10 days under anaerobic conditions. On MRS medium, colonies were visible as small, round, light-colored colonies.

[0191] Cell counts were analyzed on day +0 and day +1, and then measured monthly until day +120.

[0192] MRS medium

[0193] MRS agar has the following composition (g / L):

[0194] Peptone: 10.0

[0195] Beef extract: 10.0

[0196] Yeast extract: 5.0

[0197] Glucose: 20.0

[0198] Polysorbate 80:1.0

[0199] Ammonium citrate: 2.0

[0200] Sodium acetate: 5.0

[0201] Magnesium sulfate: 0.1

[0202] Manganese sulfate: 0.05

[0203] Dipotassium hydrogen phosphate: 2.0

[0204] Agar: 15.0

[0205] result

[0206] Table 2: pH

[0207]

[0208]

[0209] Table 3: Cell count

[0210]

[0211] As shown in Table 2, five of the six strains tested had post-acidification levels below 0.10 pH units over 120 days of storage at 25°C, while the sixth strain tested had a post-acidification level below 0.20 pH units. Thus, all six O. oeni strains tested produced only very low levels of post-acidification.

[0212] As shown in Table 3, the cell counts of all six tested strains after 120 days of storage were higher than the cell counts at inoculation on day 0. This is presumably because the O. oeni cultures tested had already experienced some growth during storage. If so, this growth would have resulted in, at most, a very limited decrease in pH.

[0213] Example 2: Testing the Growth of Oenococcus oeni Strains on Different Carbohydrate Sources

[0214] The ability of the following Oenococcus oeni strains of the present invention to grow on galactose, glucose, fructose and lactose was tested using a commercial test called "Api 50CHL medium" from bioMerieux SA:

[0215] Test strain 2: DSM33147

[0216] Test strain 3: DSM15570

[0217] Test strain 5: DSM33145

[0218] Testing Steps Before Moving to API Test Strips

[0219] - Order the strain from a culture collection (from -80°C bank) and streak on GJ5 agar plates

[0220] - Plates were incubated anaerobically at 30°C for 7 days

[0221] - After growth, check the plates for colony purity

[0222] - Then, pipette 2 mL of API CHL medium onto the agar plate - Use a sterile scraper to scrape the colonies off the agar and transfer the concentrated bacterial suspension into more API CHL medium

[0223] Test steps in API test strips

[0224] -Pipette approximately 100 μL of these concentrated solutions into the wells of the API test -Place paraffin oil over the wells to create a barrier and anaerobic environment

[0225] -Incubate the API strips at 25°C

[0226] - Read the results by recording the color change of the wells after 1 and 4 days

[0227] Table 4: Growth on various carbohydrates

[0228]

[0229] Collection and Expert Solutions

[0230] The applicant requests that, before the date of grant of the patent, samples of the deposited microorganisms described below shall be accessible only to experts. In particular, the applicant requests that the accessibility of the deposited microorganisms referred to in Article 33 EPC shall be achieved only by sending samples to independent experts designated by the applicant (Article 32(1) EPC).

[0231] Table 5: Applicant Chr. Hansen A / S deposited with the following depositary institution which has the status of an international depositary authority under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure: Leibniz-Institut DSMZ - German Collection of Microorganisms, Inhofenstrasse 7B, D-38124 Braunschweig, Germany.

[0232] strain Accession number Deposit date Oenococcus oeni DSM 33144 2019.05.28 Oenococcus oeni DSM 33145 2019.05.28 Oenococcus oeni DSM 33146 2019.05.28 Oenococcus oeni DSM 33147 2019.05.28

Claims

1. A method for storing a food product in a production environment, the method comprising: - providing a food product having a pH of 3.4 to 4.4, wherein the food product is a starter culture fermented dairy product; - heat-treating the food product to reduce the bacterial level to no more than 1×10 exp02 CFU / g to obtain a heat-treated food product, - aseptically adding one or more ambient storage lactic acid bacteria strains to said heat-treated food product in a total amount of at least 1.0×10 exp0 3 CFU / g to obtain an ambient storage food product, and - storing said ambient storage food product at ambient temperature for a period of time, The environmental storage lactic acid bacteria strain is a strain of the genus Oenococcus, wherein the strain of the genus Oenococcus is selected from the group consisting of the Oenococcus oeni strain with a deposition number of DSM 33144, the Oenococcus oeni strain with a deposition number of DSM 33145, the Oenococcus oeni strain with a deposition number of DSM 33146, the Oenococcus oeni strain with a deposition number of DSM 33147 and the Oenococcus oeni strain with a deposition number of DSM 15570.

2. The method according to claim 1 , wherein the environmental storage lactic acid bacteria strain, when added in an amount of 1.0×10 exp0 7 CFU / g to a fermented milk test product in the form of yogurt, is capable of maintaining an activity of at least 1.0×10 exp0 3 CFU / g at the end of a storage period of 120 days at 25° C.; the yogurt being obtained by fermentation at a temperature of 43° C. with a starter culture containing Streptococcus thermophilus and Lactobacillus delbrueckii bulgaricus to a pH of 4.3 and having been heat-treated at 75° C. for 30 seconds.

3. The method of claim 2, wherein the pH of the test product decreases by at most 0.8 during the 120-day storage period.

4. The method according to claim 1 or 2, wherein when added to the test product in an amount of 1.0 x 10exp07 CFU / g, the strain increases to an amount of at least 5.0 x 10exp07 CFU / g.

5. The method according to claim 4, wherein the increase in the amount of cells occurs within 45 days of adding the strain to the test product.

6. The method according to claim 4, wherein the amount of cells reaches a maximum within 45 days of adding the strain to the test product.

7. The method of claim 1 or 2, wherein the ambient stored food product is stored at ambient temperature for a period of at least 1 day.

8. The method according to claim 1 or 2, wherein the starter culture fermented dairy product has a protein content exceeding 5.1% by weight.

9. The method according to claim 1 or 2, wherein the starter culture fermented dairy product is not diluted.

10. An ambient stored food product prepared according to the method of any one of claims 1 to 9.

11. An ambient-stored food product comprising an ambient-stored lactic acid bacteria strain, wherein the ambient-stored food product has a pH of 3.4 to 4.4, wherein the ambient-stored food product contains at least 1.0×10 exp03 CFU / g of the strain, wherein the ambient-stored food product is stored at ambient temperature for a period of time, wherein the ambient-stored food product is a starter culture fermented dairy product; and The environmental storage lactic acid bacteria strain is selected from the strain of the genus Oenococcus, wherein the strain of the genus Oenococcus is selected from the group consisting of the Oenococcus oeni strain with a deposition number of DSM 33144, the Oenococcus oeni strain with a deposition number of DSM 33145, the Oenococcus oeni strain with a deposition number of DSM 33146, the Oenococcus oeni strain with a deposition number of DSM 33147 and the Oenococcus oeni strain with a deposition number of DSM 15570.

12. The ambient stored food product of claim 11, wherein the product is a chemically acidified product.

13. The ambient storage food product according to claim 11, wherein the product is a fermented dairy product obtained by fermenting a milk base using a lactic acid bacteria starter culture, wherein the product contains no more than 1×10 exp02 CFU / g of the starter culture and at least 1×10 exp03 CFU / g of the ambient storage lactic acid bacteria strain.

14. The ambient storage food product according to claim 11, wherein the product is a fermented dairy product obtained by fermenting a dairy base with a lactic acid bacteria starter culture, wherein the fermented dairy product after fermentation has been subjected to a heat treatment to reduce the bacteria level of the starter culture to no more than 1×10 exp02 CFU / g, and wherein after the heat treatment, the ambient storage lactic acid bacteria strain is aseptically added to the heat treated product in an amount of at least 1.0×10 exp03 CFU / g.

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