LACTOBACILLUS FERMENTUM BACTERIA INHIBITING POST-ACIDIFICATION

Abstract

The present invention relates to a bacterium of the species Lactobacillus fermentum, wherein the bacterium is characterized in that it increases the pH of a fermented milk product comprising the Lactobacillus fermentum during storage after fermentation in comparison to a milk product fermented with the same starter culture not containing the Lactobacillus fermentum. In a related embodiment the present invention provides a bacterium of the species Lactobacillus fermentum characterized in that a fermented milk product comprising the Lactobacillus fermentum maintains a pH above 4.0 when stored for at least 14 days at 25 C., wherein the fermented milk product is obtained by a method comprising incubating a milk with the Lactobacillus fermentum in a concentration of at least 10.sup.7 CFU/g and with a starter culture, fermenting until a pH of 4.6 is reached, shaking the fermented product and cooling. The invention further relates to compositions comprising the bacteria, methods for producing fermented milk products using the bacteria and the products thus obtained.

Claims

1-20. (canceled)

21. A method of producing a fermented milk product, comprising: adding a Lactobacillus fermentum bacterium and a starter culture to milk or to a milk product to obtain a mixture, and fermenting the mixture at a temperature between about 22 C. and about 43 C. until a pH of 4.6 or less 6 is reached, wherein the Lactobacillus fermentum bacterium increases the pH of a fermented milk product comprising it during storage after fermentation in comparison to a milk product fermented with the same starter culture not containing the Lactobacillus fermentum.

22. The method of claim 21, wherein the Lactobacillus fermentum bacterium increases the pH by at least 0.1 as compared to a milk product fermented with the same starter culture not containing the Lactobacillus fermentum.

23. The method of claim 21, wherein the ability of the Lactobacillus fermentum bacterium to increase pH during storage after fermentation is determined after storing a fermented product fermented with a starter culture and the Lactobacillus fermentum at a concentration of at least 10.sup.7 cfu/g for 21 days at 25 C.

24. The method of claim 21, wherein the Lactobacillus fermentum bacterium maintains the pH of a fermented milk product above 4.0 when the fermented milk product is stored for at least 14 days at 25 C., wherein said fermented milk product is obtained by a method comprising inoculating a milk with the Lactobacillus fermentum at a concentration of at least 10.sup.7 CFU/g and with a starter culture, fermenting the inoculated milk product until a pH of 4.6 is reached, shaking the fermented product and cooling the fermented product.

25. The method of claim 21, wherein the method comprises inoculating the milk or milk product with the Lactobacillus fermentum at a concentration of at least 10.sup.7 CFU/g and with the culture, fermenting the inoculated milk product until a pH of 4.6 is reached, shaking the fermented product, and cooling the fermented product, wherein the Lactobacillus fermentum maintains the pH of the fermented milk product above 4.0 when the fermented milk product is stored for at least 14 days at 25 C.

26. The method of claim 21, wherein the method comprises fermenting the mixture (a) such that the concentration of the Lactobacillus fermentum is at least 110.sup.6 cfu/g in the fermented milk product at the termination of fermentation; and/or (b) such that the concentration of the Lactobacillus fermentum bacteria is at least 110.sup.5 cfu/cm.sup.2 on the surface of the fermented milk product.

27. The method of claim 21, further comprising storing the fermented product at a temperature between 7 C. and 25 C.

28. The method of claim 27, wherein the fermented product is stored for a period of at least 14 days and wherein the pH of the fermented milk product is maintained above pH 4.0.

29. The method of claim 21, wherein the starter culture comprise lactic acid bacteria (LAB) which are able to decrease the pH of a milk product during fermentation to a value of pH 4.6 in 10 hours or less.

30. The method of claim 21, wherein the starter culture comprises a mixture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus.

31. The method of claim 21, wherein the Lactobacillus fermentum bacterium is selected from: (a) the Lactobacillus fermentum strain deposited with the German Collection of Microorganisms and Cell Cultures (DSMZ) as DSM32084; (b) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32085; (c) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32086; (d) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32087; (e) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32088; (f) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32089; (g) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32090; (h) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32091; (i) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32096; (j) the Lactobacillus fermentum strain deposited with the DSMZ as DSM22584; or (k) a mutant strain obtained from of (a) to (j), wherein the mutant strain increases the pH of a fermented milk product comprising it during storage after fermentation as compared to a milk product fermented with the same starter culture not containing the Lactobacillus fermentum.

32. The method of claim 21, wherein the method further comprises adding to the milk or to the milk product at least one further bacterium selected from: (a) Lactobacillus rhamnosus bacterium of strain CHCC15860 as deposited with the DSMZ under accession No. DSM32092; (b) Lactobacillus rhamnosus bacterium of strain CHCC5366 as deposited with the DSMZ under accession No. DSM23035; (c) Lactobacillus rhamnosus bacterium of strain CHCC12697 as deposited with the DSMZ under accession No. DSM24616; (d) Lactobacillus paracasei bacterium of strain CHCC12777 as deposited with the DSMZ under accession No. DSM24651; and (e) Lactobacillus paracasei bacterium of strain CHCC14676 as deposited with the DSMZ under accession No. DSM25612.

33. A fermented milk product obtained by the method of claim 21.

34. A method of producing a food, feed, or pharmaceutical product, comprising adding a fermented milk product according to claim 33 to a food, feed, or pharmaceutical product.

35. A food, feed, or pharmaceutical product obtained by a method of claim 34.

36. A Lactobacillus fermentum bacterium, wherein the Lactobacillus fermentum bacterium increases the pH of a fermented milk product comprising it during storage after fermentation in comparison to a milk product fermented with the same starter culture not containing the Lactobacillus fermentum.

37. The Lactobacillus fermentum bacterium of claim 36, selected from: (a) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32084; (b) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32085; (c) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32086; (d) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32087; (e) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32088; (f) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32089; (g) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32090; (h) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32091; (i) the Lactobacillus fermentum strain deposited with the DSMZ as DSM32096; (j) the Lactobacillus fermentum strain deposited with the DSMZ as DSM22584; or (k) a mutant strain obtained from one of (a) to (j), wherein the mutant strain has the ability to inhibit the growth of the fungus Penicillium solitum deposited with the DSMZ under accession number DSM32093 or the fungus Penicillium brevicompactum deposited with the DSMZ under accession number DSM32094 by at least 50%.

38. A composition comprising a Lactobacillus fermentum bacterium according to claim 36, optionally further comprising a cryoprotective compound.

39. The composition of claim 38, wherein the composition further comprises at least one further bacterium selected from: (a) Lactobacillus rhamnosus bacterium of strain CHCC15860 as deposited with the DSMZ under accession No. DSM32092; (b) Lactobacillus rhamnosus bacterium of strain CHCC5366 as deposited with the DSMZ under accession No. DSM23035; (c) Lactobacillus rhamnosus bacterium of strain CHCC12697 as deposited with the DSMZ under accession No. DSM24616; (d) Lactobacillus paracasei bacterium of strain CHCC12777 as deposited with the DSMZ under accession No. DSM24651; and (e) Lactobacillus paracasei bacterium of strain CHCC14676 as deposited with the DSMZ under accession No. DSM25612.

40. The composition according to claim 39, wherein the composition is a solid frozen or freeze dried starter culture comprising lactic acid bacteria at a concentration of at least 10.sup.9 colony forming units per g of frozen material.

41. A food, feed, or pharmaceutical product comprising a Lactobacillus fermentum bacterium according to claim 36, further comprising one or more of: (a) one or more further bacterium selected from one or more of the following genera Lactococcus spp., Streptococcus spp., Lactobacillus spp., Leuconostoc spp., Pseudoleuconostoc spp., Pediococcus spp., Brevibacterium spp. and Enterococcus spp.; (b) Lactobacillus rhamnosus bacterium of strain CHCC15860 deposited with the DSMZ under accession No. DSM32092; (c) Lactobacillus rhamnosus bacterium of strain CHCC5366 deposited with the DSMZ under accession No. DSM23035; (d) Lactobacillus rhamnosus bacterium of strain CHCC12697 deposited with the DSMZ under accession No. DSM24616; (e) Lactobacillus paracasei bacterium of strain CHCC12777 deposited with the DSMZ under accession No. DSM24651; and (f) Lactobacillus paracasei bacterium of strain CHCC14676 deposited with the DSMZ under accession No. DSM25612.

Description

DESCRIPTION OF THE FIGURES

[0092] FIG. 1: pH development in fermented milk products over time when stored at (A) 71 C. or (B) 251 C. for 21 days. The products are fermented with starter culture only (Reference, ) or starter culture in combination with FreshQ4 (), Holdbac YM-C Plus (o) or Lb. fermentum CHCC14591 ().

[0093] FIG. 2: pH development in fermented milk products over time when stored at (A) 71 C. and (B) 251 C. for 28 days. The products are fermented with starter culture only, Reference, or starter culture in combination with FreshQ4, Holdbac YM-C Plus or Lb. fermentum strains.

[0094] FIG. 3: Acetaldehyde levels after storage at 71 C. for 14 days in fermented milk products fermented with starter culture alone (Reference), or starter cultures in combination with Lb. fermentum strains. LOD: Limit of detection. LOQ: Limit of quantification.

[0095] FIG. 4: Acetaldehyde levels after storage at 71 C. for 14 days in fermented milk products fermented with starter culture alone (Reference), or starter cultures in combination with Lb. fermentum CHCC14591. LOD: Limit of detection. LOQ: Limit of quantification.

[0096] FIG. 5: Acidification curves of four commercial starter cultures, FD-DVS YF-L812, F-DVS YF-L901, F-DVS YoFlex Mild 2.0 and F-DVS CH-1, grown in milk (1% fat and 4.5% protein) at 43 C.

[0097] FIG. 6: Post-acidification curves of yoghurt fermented with one of four commercial starter cultures, FD-DVS YF-L812, F-DVS YF-L901, F-DVS YoFlex Mild 2.0 and F-DVS CH-1 after storage at 6 C. for up to 43 days.

[0098] FIG. 7: Acetaldehyde levels after storage at 71 C. for 14 days in fermented milk products fermented with starter culture, FD DVS YF-L812 or F-DVS CH-1, alone (Reference), or starter cultures in combination one of the nine Lb. fermentum strains. LOD: Limit of detection. LOQ: Limit of quantification.

EXAMPLE 1

[0099] Analysis of the Effect of Lb. fermentum CHCC14591 on Post-Acidification

[0100] Reduced-fat (1.5% w/v) homogenized milk was heat-treated at 901 C. for 20 min and cooled immediately. A commercial starter culture (F-DVS Mild 2.0) was inoculated at 0.02% (v/w), and the inoculated milk was distributed into 200 ml bottles. One bottle was inoculated with Lb. fermentum CHCC14591 in total concentration of 210.sup.7 CFU/g, two bottles were inoculated with two commercial bioprotective cultures (FreshQ4 and Holdbac YM-C Plus) in recommended dosages (100 U/T and 20 DCU/100 L for FreshQ4 and Holdbac YM-C Plus, respectively), and one bottle was used as a reference and only inoculated with the starter culture. All bottles were incubated in a water bath at 431 C. and fermented at these conditions until pH of 4.600.1 was reached. After fermentation, the bottles were vigorously shaken to break the coagulum and cooled on ice.

[0101] To monitor the effect on post acidification, the four fermented milk samples (starter-only, FreshQ4, Holdbac YM-C Plus and Lb. fermentum CHCC14591) were stored at 71 C. and 251 C. for 21 days and pH was measured on day 1, 7, 14 and 21.

[0102] The effect is illustrated in FIG. 1, showing that addition of Lb. fermentum CHCC14591 during milk fermentation result in lower post acidification compared to the use of the starter culture alone and particularly compared to the use of the two commercial bioprotective cultures both contributing to post-acidification.

EXAMPLE 2

[0103] Analysis of the Effect of Ten Lb. fermentum Strains on Post-Acidification

[0104] Reduced-fat (1.5% w/v) homogenized milk was heat-treated at 901 C. for 20 min and cooled immediately. A commercial starter culture (F-DVS YF-L901) was inoculated at 0.02% (v/w), and the inoculated milk was distributed into 200 ml bottles. Ten bottles were inoculated with the Lb. fermentum strains in concentrations of 110.sup.7 CFU/g and one bottle was used as a reference and only inoculated with the starter culture. All bottles were incubated in a water bath at 431 C. and fermented at these conditions until pH of 4.600.1 was reached. After fermentation, the bottles were vigorously shaken to break the coagulum and cooled on ice.

[0105] The tested Lb. fermentum strains were: Lb. fermentum CHCC12798, Lb. fermentum CHCC12797, Lb. fermentum CHCC14591, Lb. fermentum CHCC14588, Lb. fermentum CHCC15844, Lb. fermentum CHCC15865, Lb. fermentum CHCC15847, Lb. fermentum CHCC15848, Lb. fermentum CHCC15926, and Lb. fermentum CHCC2008.

[0106] To monitor the effect on post acidification, the eleven fermented milk samples (starter culture alone and starter culture in combination with the ten Lb. fermentum strains) were stored at 71 C. and 251 C. for 28 days and pH was measured on day 1, 7, 14, 21 and 28.

[0107] The effects on post-acidification are illustrated in FIG. 2 and show that each of the Lb. fermentum strains Lb. fermentum CHCC12798, Lb. fermentum CHCC12797, Lb. fermentum CHCC14591, Lb. fermentum CHCC14588, Lb. fermentum CHCC15844, Lb. fermentum CHCC15865, Lb. fermentum CHCC15847, Lb. fermentum CHCC15848, Lb. fermentum CHCC15926, and Lb. fermentum CHCC2008 does not contribute to post-acidification or even reduce post-acidification compared to reference yoghurt.

[0108] These findings were unexpected and are highly significant, as prior art antifungal food-grade bacteria were observed to increase the post-acidification effects caused by the starter culture.

EXAMPLE 3

[0109] Effect of the Ten Lb. fermentum Strains on Acetaldehyde Content

[0110] Ten Lb. fermentum strains were tested for their ability to lower acetaldehyde content.

[0111] Reduced-fat (1.5% w/v) homogenized milk was heat-treated at 901 C. for 20 min and cooled immediately. A commercial starter culture (F-DVS YF-L901 Yo-Flex) was inoculated at 0.02% (v/w), and the inoculated milk was distributed into 200 ml bottles. Ten bottles were inoculated with the Lb. fermentum strains in concentrations of 110.sup.7 CFU/g and one bottle was used as a reference and only inoculated with the starter culture. All bottles were incubated in a water bath at 431 C. and fermented at these conditions until pH of 4.600.1 was reached. After fermentation, the bottles were vigorously shaken to break the coagulum and cooled on ice. The bottles were stored at 71 C. for 14 days.

[0112] On day 14 samples were analyzed for acetaldehyde by static head space gas chromatography (HSGC), a sensitive method for analyzing volatiles in complex matrices. The setup consisted of a Static Head Space sampler connected to Gas Chromatograph with Flame Ionization Detector (FID). For that purpose the following equipment was used:

[0113] HS-autosampler: HS40XI, TurboMatrix 110, Perkin Elmer. [0114] HS-software: HSControl v.2.00, Perkin Elmer. [0115] GC: Autosystem XL, Perkin Elmer. [0116] GC-software: Turbochrom navigator, Perkin Elmer. [0117] Column: HP-FFAP 25 m0.20 mm0.33 i, Agilent Technologies

[0118] Standards of known concentration were used to determine response factors (calibration), controls were used to control that the used response factors were stable within an analytical series as well as in-between series and over time (months). Concentration of volatiles (ppm) in samples and controls was determined using response factors coming from standards. Samples were prepared by adding 200 l of 4N H.sub.2SO.sub.4 to 1 g yoghurt sample and immediately analyzed by HSGC.

[0119] The results are illustrated in FIG. 3 and show that each of the strains Lb. fermentum CHCC12798, Lb. fermentum CHCC12797, Lb. fermentum CHCC14591, Lb. fermentum CHCC14588, Lb. fermentum CHCC15844, Lb. fermentum CHCC15865, Lb. fermentum CHCC15847, Lb. fermentum CHCC15848, Lb. fermentum CHCC15926, and Lb. fermentum CHCC2008 has the ability to reduce the concentration of acetaldehyde produced by a starter culture during fermentation in a fermented milk product.

EXAMPLE 4

[0120] Effect of One Lb. fermentum Strain on Acetaldehyde Content

[0121] One Lb. fermentum strain was tested for the ability to lower acetaldehyde content.

[0122] Reduced-fat (1.5% w/v) homogenized milk was heat-treated at 901 C. for 20 min and cooled immediately. A commercial starter culture (F-DVS Mild 2.0) was inoculated at 0.02% (v/w), and the inoculated milk was distributed into two 200 ml bottles. One bottle was inoculated with the Lb. fermentum strains in concentrations of 110.sup.7 CFU/g and one bottle was used as a reference and only inoculated with the starter culture. Both bottles were incubated in a water bath at 431 C. and fermented at these conditions until pH of 4.600.1 was reached. After fermentation, the bottles were vigorously shaken to break the coagulum and cooled on ice. The bottles were stored at 71 C. for 14 days.

[0123] On day 14 samples were analyzed for acetaldehyde by static head space gas chromatography (HSGC), a sensitive method for analyzing volatiles in complex matrices. The setup consisted of a Static Head Space sampler connected to Gas Chromatograph with Flame Ionization Detector (FID). For that purpose the following equipment was used: [0124] HS-autosampler: HS40XI, TurboMatrix 110, Perkin Elmer. [0125] HS-software: HSControl v.2.00, Perkin Elmer. [0126] GC: Autosystem XL, Perkin Elmer. [0127] GC-software: Turbochrom navigator, Perkin Elmer. [0128] Column: HP-FFAP 25 m0.20 mm0.33 i, Agilent Technologies

[0129] Standards of known concentration were used to determine response factors (calibration), controls were used to control that the used response factors were stable within an analytical series as well as in-between series and over time (months). Concentration of volatiles (ppm) in samples and controls was determined using response factors coming from standards. Samples were prepared by adding 200 l of 4N H.sub.2SO.sub.4 to 1 g yoghurt sample and immediately analyzed by HSGC.

[0130] The results are illustrated in FIG. 4 and show that Lb. fermentum CHCC14591 has the ability to reduce the concentration of acetaldehyde produced by a starter culture during fermentation in a fermented milk product.

EXAMPLE 5

[0131] Functional Analysis of Commercial Starter Starter Cultures

[0132] The three commercial starter cultures included herein were chosen based on their different acidification profiles. Three were frozen, F-DVS CH-1, F-DVS YoFlex Mild 2.0 and F-DVS YF-L901, and one was freeze dried, FD-DVS YF-L812. To test the difference in acidification profiles, semi fat milk was standardized to 1% fat and 4.5% protein with skim milk powder and heat-treated at 851 C. for 30 min and cooled immediately. One of four different commercial starter cultures (F-DVS CH-1, F-DVS YoFlex Mild 2.0, F-DVS YF-L901 or FD-DVS YF-L812) was inoculated at 0.02% (v/w), and the inoculated milk was distributed into 200 ml bottles. The bottles were incubated in a water bath at 431 C. and fermented under these conditions until pH 4.5 was reached. The pH was measured continually throughout the fermentation. Subsequently, the bottles were stored at 6 C. for 43 for days and pH was measured with intervals of 7 days to determine the level of post-acidification.

[0133] The acidification profiles of the three commercial starter cultures, F-DVS CH-1, F-DVS YoFlex Mild 2.0, F-DVS YF-L901 and FD-DVS YF-L812, are shown in FIG. 5. F-DVS CH-1 showed fast fermentation time reaching pH 4.55 in 4.87 hours. F-DVS YoFlex Mild 2.0 showed intermediate fermentation time reaching pH 4.55 in 5.29 hours. FD-DVS YF-L812 and F-DVS YF-L901 showed slower fermentation reaching pH 4.55 in 6.45 and 5.87 hours, respectively. Post-acidification profiles showed very low levels of post-acidification for FD-DVS YF-L812 and F-DVS YoFlex Mild 2.0 (pH=0.12 and pH=0.11 after storage at 6 C. for 43 days, respectively), intermediate levels of post-acidification for F-DVS YF-L901 (pH=0.26 after storage at 6 C. for 43 days and high degree of post-acidification for F-DVS CH-1 (pH=0.55 after storage at 6 C. for 43 days) (FIG. 6).

EXAMPLE 6

[0134] Effect of the Nine Lb. fermentum Strains on Acetaldehyde Content When Fermented With Two Different Starter Cultures

[0135] Nine Lb. fermentum strains were tested for their ability to lower acetaldehyde content.

[0136] Reduced-fat (1.5% w/v) homogenized milk was heat-treated at 901 C. for 20 min and cooled immediately. Milk was inoculated with one of two commercial starter cultures (F-DVS CH-1 or FD-DVS YF-L812) at 0.02% (v/w), and the inoculated milk was distributed into 200 ml bottles. Nine bottles were inoculated with the Lb. fermentum strains in concentrations of 110.sup.7 CFU/g and one bottle inoculated with each starter culture was used as a reference and only inoculated with the starter culture. All bottles were incubated in a water bath at 431 C. and fermented at these conditions until pH of 4.550.1 was reached. After fermentation, the bottles were vigorously shaken to break the coagulum and cooled on ice. The bottles were stored at 71 C. for 14 days.

[0137] The tested Lb. fermentum strains were: Lb. fermentum CHCC12798, Lb. fermentum CHCC12797, Lb. fermentum CHCC14591, Lb. fermentum CHCC14588, Lb. fermentum CHCC15844, Lb. fermentum CHCC15865, Lb. fermentum CHCC15847, Lb. fermentum CHCC15926, and Lb. fermentum CHCC2008.

[0138] On day 14 samples were analyzed for acetaldehyde by static head space gas chromatography (HSGC), a sensitive method for analyzing volatiles in complex matrices. The setup consisted of a Static Head Space sampler connected to Gas Chromatograph with Flame Ionization Detector (FID). For that purpose the following equipment was used: [0139] HS-autosampler: HS40XI, TurboMatrix 110, Perkin Elmer. [0140] HS-software: HSControl v.2.00, Perkin Elmer. [0141] GC: Autosystem XL, Perkin Elmer. [0142] GC-software: Turbochrom navigator, Perkin Elmer. [0143] Column: HP-FFAP 25 m0.20 mm0.33 ui, Agilent Technologies

[0144] Standards of known concentration were used to determine response factors (calibration), controls were used to control that the used response factors were stable within an analytical series as well as in-between series and over time (months). Concentration of volatiles (ppm) in samples and controls was determined using response factors coming from standards. Samples were prepared by adding 200 l of 4N H.sub.2SO.sub.4 to 1 g yoghurt sample and immediately analyzed by HSGC.

[0145] The results are illustrated in FIG. 7 and show that each of the strains Lb. fermentum CHCC12798, Lb. fermentum CHCC12797, Lb. fermentum CHCC14591, Lb. fermentum CHCC14588, Lb. fermentum CHCC15844, Lb. fermentum CHCC15865, Lb. fermentum CHCC15847, Lb. fermentum CHCC15926, and Lb. fermentum CHCC2008 has the ability to reduce the concentration of acetaldehyde produced by a starter culture during fermentation in a fermented milk product.

REFERENCES

[0146] 1. Tamime, A. Y., and H. C. Deeth. 1980. Yoghurt: technology and biochemistry. J. Food Prot. 43:939-977. [0147] 2. A. C. S. D. Chaves, M. Fernandez, A. L. S. Lerayer, I. Mierau, M. Kleerebezem, and J. Hugenholtz, 2002, Metabolic Engineering of Acetaldehyde Production by Streptococcus thermophilus [0148] 3. Lees, G. J., and G. R. Jago. 1976. Formation of acetaldehyde from threonine by lactic acid bacteria. J. Dairy Res. 43:75-83. [0149] 4. Kurmann, J. A. (1982) Die Obersauerung der Joghurtgallerte, ein haufig auftretender und zu wenig beachteter Produktionsfehler, dessen Entstehung und Bekampfung. Dt. Molkerei-Zeitung 103, 690-698.

[0150] Deposits and Expert Solution

[0151] The applicant requests that a sample of the deposited micro-organisms stated below may only be made available to an expert, until the date on which the patent is granted.

[0152] The Lactobacillus fermentum strain CHCC12798 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32084.

[0153] The Lactobacillus fermentum strain CHCC12797 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32085.

[0154] The Lactobacillus fermentum strain CHCC14591 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32086.

[0155] The Lactobacillus fermentum strain CHCC14588 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32087.

[0156] The Lactobacillus fermentum strain CHCC15844 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32088.

[0157] The Lactobacillus fermentum strain CHCC15865 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32089.

[0158] The Lactobacillus fermentum strain CHCC15847 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32090.

[0159] The Lactobacillus fermentum strain CHCC15848 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32091.

[0160] The Lactobacillus fermentum strain CHCC15926 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32096.

[0161] The Lactobacillus fermentum strain CHCC2008 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 22584.

[0162] The Lactobacillus rhamnosus strain CHCC15860 was deposited at German Collection of Microorganisms and Cell Cultures (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH; DSMZ), Inhoffenstr. 7B, D-38124 Braunschweig deposited on Jul. 16, 2015 under the accession No.: 32092.

[0163] The deposits were made according to the Budapest treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure.