METHOD FOR THE PREPARATION OF NATURALLY THICKENED AND STABILIZED FERMENTED MILK PRODUCTS AND PRODUCTS PRODUCED THEREBY

Abstract: A method for the preparation of naturally stabilized, thick bodied, fermented milk products by fermentation is described. Mixed cultures of milk fermenting, non-slime, lactic acid producing bacteria and slime producing Strepto­ coccus lactis, Streptococcus cremoris or mixtures thereof having the thickening characteristics in milk of Streptococcus cremoris NRRL-B-12,361, 12,362 or 12,363 are used, prefer­ ably in addition with a diacetyl producing bacterium for flavor. The fermented milk products are thick bodied without any ropiness or sliminess and are stable to separation of whey from curd upon storage at refrigeration temperatures, with little or no added stabilizing agents such as gums and starches or thickening agents such as added non-fat milk solids. The preferred product is a thick bodied buttermilk.

Description: METHOD FOR THE PREPARATION OF NATURALLY THICKENED AND STABILIZED FERMENTED MILK PRODUCTS AND PRODUCTS PRODUCED THEREBY BACKGROUND OF THE INVENTION The present invention relates to naturally stabilized, thick bodied fermented milk products which are produced by fermentation without the need for added stabilizing or thickening agents. In particular the present invention relates to a method which uses mixed cultures of milk fermenting, non-slime, lactic acid producing bacteria and' slime producing Streptococcus Zactis, Strepto coccu8 cremcris or mixtures thereof having the thickening and stabilizing characteristics of Streptococcus cremoris NRRL-B-12,361, 12,362 or 12,363 for the fermentations. Prior Art Dairy products, such as yogurt and buttermilk, prepared by fermentation have a characteristic thick lumpy texture which is not uniformly cohesive and smooth when stirred and also these products have a tendency to "whey off" upon standing by the separation or shrinking of the curd which exudes the whey. Various additive stabilizing agents, such as plant gums, starches and cellulose derivatives, are used commercially in an attempt to improve the texture by making the products smoother and to improve the storage stability of the fermented milk products. U.S. Patent Nos. 3,080,236; 3,235,387; 3,269,842; 3,539,363; 3,932,680 and 3,969,534 describe the use of various added stabilizers and are representative of this prior art practice which is widely used commercially. Such stabilizing agents add to the cost of the products and in addition introduce a sometimes unwanted ingredient into the fermented milk product. Generally non-fat milk solids are added to the fermented milk products prior to the fermentation in order to thicken them. Stabilizing agents are generally not added after the fermentation because microbial contamination will occur and the product will spoil faster. U.S. Patent No. 4,243,684 shows the use of *ropy" cultures of¯lactic acid producingbactena in place of non-ropy cultures for making cheese. Also ropy cultures are used in the Scandinavian countries to make ropy products. None of these products have the texture of the hereinafter described products of the present invention. Objects It is therefore an object of the present invention to provide novel naturally thick bodied and stabilized, fermented milk products. It is further an object of the present invention to provide such thick bodied, fermented milk products which are relatively inexpensive to manufacture and which have a storage stability to wheying off which'is at least equivalent to products which are produced with added stabilizing agents. Additionally, it is an object,of the present invention to provide thick bodied, fermented milk products which have a unique silky' smooth or "velvet" texture without any ropiness or sliminess by'mesons of -the use of selected slime producing strains of Strepto- coccus ikrdis, Streptococcus cremorts or mixtures thereof ad- mixed with lactic acid producing bacteria. These and other objects will become increasingly apparent by reference to the following description. Description of the Invention The present invention relates to a thick bodied fermented milk product which comprises.- milk with a pH between about 4.2 and 4.7 which has been fermented with a mixed culture containing a milk fermenting, non-slime, lactic acid producing bacteria and slime producing Streptococcus tcrctis, Streptococcus cremoris or mixtures thereof having the thickening and stabilizing characteristics of Streptococcus smor*s NRRL-B-12,361, 12,362 or 12,363 to produce the thick bodied milk product without need for addition of non-fat milk solids and without resulting in a slimy or topy product, wherein the product is stable to separation of whey from curd upon storage at refrigeration temperatures without added stabilizing agents. rurther the present invention relates to the method of producing a thick bodied fermented milk product which comprises: providing in milk a mixed culture containing milk fermenting, non-slime, lactic acid producing bacteria which reduce the pH of the milk and slime producing Strepto coccus Zactis, Streptococcus cremoris or mixtures thereof having the thickening and stabilizing characteristics of Strepto coccus crempris NRRL-B-12,361, 12,362 or 12,363; and fermenting the milk with the culture to produce the thick bodied milk product without need for addition of non-fat milk solids and without resulting in a slimy or ropy product, wherein the product is stable to separation of whey from curd upon storage at refrigeration temperatures without added stabilizing agents. Streptococcus cremorss NRRL,B-12,361 (which is pre ferried) is available along with NRRL-B-12,362 and fRRL-B- 12,363 to anyone who requests it by number from the United States Department of Agriculture, Northern Regional Research Center, 1815 North University Street, Peoria, Illinois 61604. Other related strains having the thickening characteristics of NRRL-B-12,361 can be selected from available cultures and tested by fermentation in milk, Thus Streptococcus cremoris NRRL-B-12,362 and Streptococcus cremoris NRRL-B-12, 363 were also found to have this characteristic. It is contemplated that genetic manipula tion of any strain of Streptococcus cremor*s or Streptococcus Zactis by recombinant or mutagenic techniques could produce strains having the same essential fermentation characteristics of NARL-B-12,361, 12,362 or.12,363 and it is intended that these be included in the present invention. The fermentation characteristics of strains NRRL-B-12,361, 12,362 and 12,363 are particularly unique in that they have the ability to produce a silky smooth velvet" texture in fermented milk in combination with non Slime, lactic acid producing bacteria, without any ropiness or sliminess. The term "ropiness" is defined as meaning the tendency of the milk to agglomerate into long' strands as a result of the fermentation. The term "slimi- ness" refers to a product which is tacky such that the tongue tends to stick to the palate upon tasting An example of "sliminess" is the slick feel of the white of a raw egg. It is a characteristic of a few strains of Streptococcus lactis and Streptococcus cremoris that in pure culture they produce fermented milk products which, are ropy and slimy, characteristics which are undesirable. In the present invention, when these selected slime producing strains of Streptococcus Zactis and/or Streptococcus cremoris are mixed in specific proportions with other traditionally used nonslime, lactic acid bacteria, ropy or slimy characteristics contributed by the slime producing Streptococcus Zactis and/or Streptococcus cremoris are overcome. The result is a uniformly smooth, velvety, non-slimy product. The mixed culture contains the traditional milk fermenting, non-slime lactic acid producing bacteria. Nonslime producing Streptococcus Zactis, Streptococcus cremoris, Streptococcus thermophiZus, LactoDaciZZus butgaricus, Lactobacillus acidophilus, LaCtoba1us lactisj including particularly NRRL-B-5628 or mixtures thereof can be used with flavor Cdiacetyl) producing bacteria such as Streptococcus lactis sub species diacetyZactis and Leucooostoc cremoris. Many other species of non-slime, streptococcus and Lactob=ZZus commonly used in milk fermentations to generate lactic acid can be used. In order to avoid the problem of sliminess or ropiness and to produce the thick .body, it is necessary to include the milk fermenting, non-slime, lactic acid producing bacteria in a volume ratio of between about 1:9 and 4=1 bacterial cells per part by volume of the slime producing Streptococcus lactis, Streptbcoccus cremoris or mixtures thereof. The cells to be mixed are generally at the same concentration level per, unit volume. Within this range, variations in product, body can occur depending upon the specific strain of slime producing Streptococcus Zactis and/or Streptococcus cremoris selected. The slime producing bacteria for the clutures can be grown in milk or other suitable media to a level of at least about 108 cells per milliliter of growth medium. They can be used directly or they can be concentrated above the level of growth by various conventional means such as by centrifugation or ultrafiltration by dialysis of the growth medium to a higher cell count level, particularly above about 1010 cells per milliliter up to about 1012 cells per milliliter as is known to those skilled in the art. Cell counts are based upon the number of chains or colony forming units (CFU). The slime producing cultures and the non-slime producing cultures can be added to milk as individual single strain preparations (concentrated or non-concentrated) or the slime producing cultures (concentrated or non-concentrated) can be mixed with non-slime producing cultures (concentrated or non-concentrated and then added directly to milk. Also, milk can be inoculated with single or mixed strain slime producing culture (concentrated or non-concentrated) and milk can be separately inoculated ,with single or mixed strain non-slime producing culture (concentrated or non-concentrated). Both inoculated milks are fermented at 10-32 C and then the resultant ropy body buttermilk can be blended with the resultant normal body buttermilk to produce a non-ropy, non-slimy, naturally stabilized, thick bodied buttermilk. When concentrates of slime producing and non-slime producing cultures are prepared and stored-for any length of time prior to preparation of the fermented milk product, freezing stabilizing agents can be added for freezing or the concentrate can be lyophilized'in a manner conventionally known to the prior'art. U.S. Pat. No. 3,420,742 illustrates a preferred method for the preparation of the bacterial concentrates which are frozen for storage with a freezing stabilizing agent such as glycerol or the like. The bacterial concentrate can be prepared from culture grown in a medium inoculated with a certain ratio of slime producing to non-slime producing Streptococcus Zactis strains, Streptococcus cremoris strains or mixtures thereof or the individual slime producing and non-slime producing strains can be prepared separately and then blended together in the desired ratio of slime producing Streptococcus cremoria and/or Streptococcus lactLs to non-slime, lactic acid producing bacteria along with Leuconostoc cremoris and Streptococcus Zactis subspecies diacetyZactwis flavor producing bacteria prior to freezing. Many different freezing stabilizing agents which maintÅain viability of the bacteria upon storage are known to those skilled in the art. Inoculated milk is fermented with the mixed culture at temperatures between about 10 to 320C, pre ferably 240C. The fentation is generally conducted for between about 12 to 18 hours. The milk usually contains less than nine percent (9%) milk solids; however, between 8 and 14 percent solids can be present. The resulting thick bodied, fermented milk products can be marketed with the bacteria in a viable form in a cooled product to restrict continuing 'fermentation. Alternatively, the product can be marketed with the bacteria killed such as by pasteurization, after the fermentation. Such products are more stable to storage at ambient room temperatures of 15 to 210C. They can be mixed with fruits or other flavorings. Specific Description The following examples demonstrate the preparation of the fermented milk products of the present invention utilizing the mixed cultures of a lactic acid producing bacterium and Streptococcus cremoris NRRL-B-12,361, B-12,362, and B-l2,363. Examples 1-4 illustrate the growth of the bacteria, the preparation of the bacterial culture mixtures and the preparation of the fermented milk products using the bacterial cultures. EXAMPLE 1 A growth medium comparable to that described in U.S. Reissue Patent Nos. 28,276 and 28,488 was prepared, heat-treated and cooled to 240C. The medium was inoculated at the rate of 0.89 volume percent with a 10 to 12 hour milk culture inoculum consisting of 40 volume percent of NRRL-B-12,361 and 60 volume percent of Streptococcus Zactis and/or Streptococcus cremoris non-slime producing mixed strains. An attempt was made to determine the exact composition of the mixed non-slime producing culture since it made no difference. The inoculated medium was held at 240C with constant neutralization at pH 5.8 until carbohydrate was depleted. The bacteria were removed from the spent medium by centrifugation to about 50 x io9 cru per ml and were blended with 10 weight percent of a concentrate of Leuconostoc cremoris containing about 1 x 109 cfu per ml. Ten weight percent of sterile glycerol was added and the resultant thoroughly mixed concentrate was packaged in appropriate containers and was quick-frozen and stored at -34 0C. After storage, the concentrated culture was removed from freezer and placed into 320Cwaterbath to thaw. Skim milk containing 9% solids-not-fat was heat treated at 1800F for 30 minutes and cooled to 240C. The milk was inoculated at rate of 0.567 ml of the thawed concentrated culture per 3785 ml milk. Control milk was inoculated with a commercial concentrate of non-slime producing strains of Streptococcus Uctis and/or Streptococcus cremoris at same rate. The inoculated milk was incubated at 240C for 16 hours, chilled to 100C, thoroughly mixed to break curd and evaluated organoleptically. The control buttermilk inoculated with non-slime producing strains of Streptococcus Lactic and/or Streptococcus cremoris had thin body characteristic of buttermilk made i:h unfortified, nonstabilized skim milk. The test butermilk inoculated with mixture of slime and non-slime producing strains of Streptococcus lactis and/or Streptococcus cremoris had a thick silky smooth "velvet" texture which was not ropy or slimy. The body was comparable in mouth feel (but not in viscosity by Zahn cup measurement per Example 4) to buttermilk made with skim milk fortified with 1% solids-not-fat and ino culated with usual, non-slime producing strains. EXAMPLE 2 BUTTERMILK MADE WITH VARIOUS RATIOS OF SLIME PRODUCING NRRL-B-12,361 to NON-SLIME PRODUC ING STREPTOCOCCUS LACTIS g STREPTOCOCCUS CREMORIS Skim milk (containing 9% solids-not-fat by test) was heat-treated at 1800F for 30 minutes and cooled to 240C. Buttermilk was made by inoculating the milk with various mixtures of slime producing Streptococcus cremoris NRRL-B12,361 and non-slime producing Streptococcus Zactis and/or Streptococcus cremoris strains which contained about 1 x cfu per ml at rate of 1 ml of overnight culture (sterile 11% NFDM incubated at 240C for 16 hours) per 100 ml of milk. The inoculated milk was incubated 16 hours at 240C, chilled to l00C, thoroughly mixed to break curd and evaluated organoleptically. The results are shown in Table I. TABLE I Slime Producing Non-Slime Producing S. cremoris Mixed S. Zactis and/or Texture of NRRL-B-12,361 S. Cremoris strains Buttermilk 908 (volume) 10% (volume) Ropy, "snotty" defect 80% 20% Ropy, wsnottyS defect 75% 25% Too viscous 50% 50% Thick, not viscous, not ropy, not slimy 40% 60% Thick, not viscous, - not ropy, not slimy 30% 70% Not as thick as above, but thicker than control 20% 80% Comparable to above 0 100% Thin body character is tic of unfortified, non-stabilized buttermilk. The results in Table I show that an inoculum consisting of anything more than about 80% by volume of slime producing Streptococcus cremoris NRRL-B-12, 361 results in a product which is characterized by the prior art as being "snotty" or ropy. An inoculum containing less than 40% by volume of NRRL-B-12, 361 resulted in a fermented product which was slightly thicker than regular buttermilk in texture. Thus the thick velvety, non-slimy texture was obtained only in a narrow ratio range of NRRL-B-12, 361 to non-slime forming Streptococcus cremoris and/or Streptococcue lactis which will vary depending upon specific slime producing strains used as can be seen from Example 3. ExAMPLE 3 BUTTERMILK MADE WITH VARIOUS RATIOS OF SLIME PRODUCING S1BOCOCCUS CRE:BJORIS NRRL-B-12,362 TO NON-SLIME PRODUCING STREPTOCOCCUS LACTIS AND/OR STREPTOCOCCUS CREMORIS Buttermilk was made as in Example 2 above and results are shown in Table II. TABLE II Slime Producing Non-slime Producing S. cremoris Mixed S. Zactis and/or Texture of NP-3-l2,362 S. cremoris ButtenniLlc 45% (volume) 55% (volume) Ropy, "snotty" defect 33.3% 66.6% Too viscous, not ropy 20% 80% Thick, not viscous, not ropy, not slimy 0% 100% Thin body character istic of unfortified, non-stabilized buttermilk The results in Table II show that an inoculum consisting of anything more than about 45% by volume of slime producing Streptococcus cremoris NRRL-B-12,362 results in a product which is characterized by the prior art as being "snotty" or ropy. EXAMPLE 4 Buttermilk was made from unfortified skim milk treated as in Example 2. The skim milk was inoculated with different ratios of slime producing Streptococcus camoris strains to non-slime producing mixed Streptococcus Zactis and Streptococcus cremoris strains. (Inoculuin consisted of 16 hours at 24 C milk cultures at rate of 1 ml per 100 ml of milk.) Inoculated skim was incubated for 16 hours at 240C and viscosity of resultant buttermilk was determined with Zahn Viscosimeter cups No.-002 and -004 (CSC Scientific Company Chicago, Illinois 60623, CSC No. 027134). Viscosity is expressed as Centipoises. After viscosity determination, buttermilk was chilled to 10 0C and evaluated organoleptically. The results are shown in Table III. TABLE III Inoculum volume Organoleptic percent t ' Centi- Evaluation of Slime Producing Slime Non-Slime poises Buttermilk Culture Number Producer Producer ' at 25'or Texture NRRI-B-12,361 100 0 725 Ropy, too mucoid B-12,361 75 25 600 Ropy, too mucoid B-12,361 50 50 445 Thick, not ropy B-12,361 25 75 285 Thinner than above B-12,361 0 , 100 50 Thin body characteristic of unfortified, non stabilized buttermilk NRRL-B-12,362 100 0 800 Ropy, too mucoid B-12,362 75 25 650 Ropy, too mucoid B-12,362 50 50 500 Thick, slightly ropy?? B-12,362 25 75 425 Thick, not ropy B-12,362 0 100 50 Thin body characteristic of unfortified, non stabilized, buttermilk NRRL-B-12,363 100 0 300 Ropy1 mucoid B-12,363 75 25 225 Thick, not ropy B-12,363 50 50 60 Thinner than above B-12,363 25 25 50 Thin body. B-12,363 0 100 50 Thin body. . The fermented milk products preferably have a Zahn cup viscosity between about 60 and 500 centipoises; however, mouth feel is the preferred method of determining the desirable thick texture of the products of the present invention. Unexpectedly the buttermilk of the present invention have the same mouth feel when made from skim milk as buttermilk made from 1 to 2 percent by weight cream added to milk and fermented with conventional cultures. The present invention also allows the elimination of a portion of the conventionally added milk solids in sour cream. Lactobacillus lactis NRRL-B-5628 is available to anyone who requests it.by number from the United States Department of Agriculture, Northern Regional Research Center, 1815 North University Street, Peoria, Illinois 61604, U.S.A.

Claims: WE CLAIM: -1 A thick bodied liquid, fermented milk product which comprises: milk with a pH between about 4.2 and 4.7 which has been fermented with a mixed culture containing a milk fermenting, non-slime, lactic acid producing bacteria and slime producing Streptococcus Zactis , Streptococcus cremoris or mixtures thereof to produce the thick bodied milk product without need for added milk solids and without resulting in a slimy or ropy product, wherein the product is stable to separation of whey from curd upon storage at refrigeration temperatures without added stabilizing agents, said slime producing Streptococcus Zactis or Streptococcus cremoris alone further being capable of producing a Zahn cup viscosity of about 300 centipoises or above when incubated in milk for 16 hours at 240C. -2 The product of Claim 1 wherein the mixed culture in additionlcontains Leuconostoc cremoris , Streptococcus Zactis subspecies diacetyZactis or mixtures thereof to impart a diacetyl flavor to the product and the product is a thick bodied buttermilk or sour cream. -3 The product of Claim 1 which has been fermented at between about 100c to 320C. -4 The product of Claim 1 which the slime producing Streptococcus cremoriaS essentially the milk fermentation characteristics of NRRL-B-12361. -5 The'product of Claim-l wherein the slime 'producing Streptococcus cremoris is NRRL-B-l2,361 -6 The product of Claim 1 wherein the slime producingstreptococcus crernorts ' is NRRL-B-12,362. -7 The product of Claim 1 wherein the slime producing Streptococcus cremoris is NRRL-3-12,363. -8 The product of Claim 1 wherein the fermentation has been completed in 12 to 18 hours at 240C. -9 The product of Claim 1 which is mixed with additional flavoring. -10- The product of Claim 9 wherein the flavoring is a fruit flavor. -11 The product of Claim 1 wherein the bacteria after the fermentation are viable in the product cooled so as to retard continuing fermentation. -12 The product of Claim 1 wherein the'bacteria are rendered non-viable after the fermentation. -13 The product of Claim 1 wherein the non-slime, lactic acid producing bacteria are non-slime producing strains ofStreptococcus'lactis , Streptococcus cremoris Streptococcus thermophilus , Lactobacittus butgaricus , Lactobacilius acidophiZug LactobaciZZus Lactic including LactobaciZZus Zactis NRRL-B-5628 or mixtures thereof. The method of producing a thick bodied liquid fermented milk product which comprises: (a) providing in milk a mixed culture containing milk fermenting, non-slime, lactic acid producing bacteria which reduce the pH of the milk and a slime producing Streptococcus Zactts , Streptococcus cremoris or mixtures thereof, said slime producing Streptococcus Zactis or Streptococcus cremoris alone further being capable of producing a Zahn cup viscosity of about 300 centipoises or above when incubated in milk for 16 hours at 240C; and (b) fermenting the milk with the culture to produce the thick bodied milk product without need for addition of non-fat milk solids and without resulting in a slimy or ropy product, wherein the product is stable to separation of whey from curd upon storage at refrigeration temperatures without'added stabilizing agents.

Inventors: SHAH, RAMESH B., VEDAMUTHU, EBENEZER R.
Applicants: MICROLIFE TECHNICS, INC.
Application year: 1982
Main classification: A23C9
IPC codes: A23C13, A23C9