Patent Description:
Raw milk received for cheese production, especially in an industrial cheese plant, has to be stored until it can be used for cheese production, mainly due to bottlenecks in the cheese plant. During the storage, wherein the milk is kept cold, the mineral balance of the milk is displaced, minerals are lost, and it therefore loses some of its original ability to coagulate and undergo syneresis, two very important properties in cheese making (Lane, C. , Sousa, M. , and McSweeney, P.

In order to restore these properties to the milk, especially milk to be used for soft cheese such as camembert, the milk normally undergoes a processing, a so-called "cold-maturation" step, where the purpose is to prepare the milk for cheese making.

Cold maturation consists of physical and biological maturation that aims at obtaining five objectives, generally believed to make the milk more suitable for cheese making (Pernoud S. and Mayer H.

In cold maturation the milk normally undergoes a mild heat treatment (thermization, e.g. <NUM> for <NUM> seconds) or pasteurization (e.g. <NUM> for <NUM> seconds) to remove psycrotroph bacteria such as Listeria species. CaCl2 may be added and the milk is kept at <NUM>-<NUM> for <NUM> to <NUM> hrs. to restore the calcium balance of the milk (physical maturation).

Following physical and biological maturation, the milk is normally pasteurized (e.g., <NUM> for <NUM> seconds) to kill and lyse the culture used for the biological maturation thereby releasing bacterial enzymes that may assist in ripening.

Subsequent to the cold maturation the milk may undergo pasteurization and warm maturation.

During the warm maturation, the milk is most often inoculated with lactic acid bacteria and kept at coagulation temperature for <NUM>-<NUM> mins.

In soft cheese production, this warm maturation step is never less than <NUM> minutes and can go up to <NUM> minutes. In Literature, we can find description of different soft cheese processes where a warm maturation is systematically mentioned. However the duration is not always given. [Goudédranch; Camier-Caudron, Gassi, Schuck. (<NUM>), M. Leclercq-Perlat (<NUM>).

As with the cold maturation, the purpose of the warm maturation comprise a number of objectives generally believed to make the milk more suitable for cheese making, including Biological maturation: Lower the pH of the milk from about pH <NUM> to a level suitable for renneting (normally in the range of pH <NUM> - <NUM> for e.g. Camembert). Normally, lactic acid bacteria cultures that acidify milk well are used to obtain biological maturation and reducing the red-ox potential to favor the growth of strains inhibited by oxygen.

However the warm maturation has significant drawbacks including, but not limited to:.

Being able to bypass the warm maturation step is therefore highly desired as it would remove the investments and risks associated with that step in the cheese production process. Previous attempts to avoid warm maturation in the production of soft cheese have included the use of fast acidifying cultures combined with a physical inactivation by e.g. salt and/or cold. However, this approach is not applicable in the production of surface ripened cheeses such as e.g. brie or camembert type cheese due to the necessity to keep a temperature allowing surface ripening cultures to grow (around <NUM>) which allow also lactic cultures to continue to acidify and create excessive post-acidification.

Present invention allows bypassing warm maturation in the production of soft cheese by providing a specific blend of bacterial strains directly inoculated in high concentration and working in concert with a specifically developed coagulant.

The present invention relates to a method according to claim <NUM>.

The method may further comprise the following steps:.

The slow or fast acidifying bacterial culture may be a culture of lactic acid bacteria, such as a culture of one or more strains, selected from the group consisting of Lactococcus spp. , Streptococcus spp. , Lactobacillus spp. , Leuconostoc spp. , Pseudoleuconostoc spp. , Pediococcus spp. , Brevibacterium spp. , Enterococcus spp. and Propionibacterium spp.

In yet a related aspect, the invention relates to a method wherein the bacterial culture is a culture of one or more strains selected from the group consisting of Streptocccus spp. or mutants or variants of any of these strains.

In yet a related aspect, the invention relates to a method wherein the one or more coagulants are one or more chymosins, such as e.g. a chymosin with a bovine or camel origin.

In yet a related aspect, the invention relates to a method wherein the one or more coagulants is a blend of two or more coagulants.

In yet a related aspect, the invention relates to a method wherein the one or more coagulants is a blend of two or more different chymosins, e.g. a bovine derived and a camel derived chymosin.

In yet a related aspect, the invention relates to a method wherein the one or more coagulants of the blend of coagulant exhibit a C/P ratio which is at least three times higher, such as e.g. <NUM>, <NUM>, <NUM>, <NUM>, or <NUM> times higher than the C/P ratio of bovine derived chymosin such as e.g. ChyMax®.

In yet a related aspect, the invention relates to a method wherein the coagulant is added within 10mins, 5mins or <NUM> mins after step a).

In yet a related aspect, the invention relates to a method as described above wherein the cheese is a brie type or a camembert type cheese.

In yet a related aspect, the invention relates to a method as described above wherein the fat on dry matter content in the cheese is from <NUM>% to <NUM>% such as e.g. around <NUM>%.

In yet a related aspect, the invention relates to a method as described above wherein the salt level is from <NUM>% to <NUM>%, such as e.g. between <NUM> and <NUM>%, such as e.g. <NUM>% on dry matter content in the cheese is from <NUM>% to <NUM>% such as e.g. around <NUM>%.

In yet a related aspect, the invention relates to a method as described above wherein the dry matter content is from <NUM>% to <NUM>%, such as e.g. <NUM>% to <NUM>% such as e.g. <NUM>%.

In yet a related aspect, the invention relates to a method as described above wherein one or more of the bacterial cultures and/or the coagulant is added as a concentrated liquid solution.

In yet a related aspect, the invention relates to a method as described above wherein one or more of the bacterial cultures and/or the coagulant is added as a Direct Vat Set (DVS) formulation.

In yet a related aspect, the invention relates to a method as described above wherein the coagulant is added in a final concentration of <NUM>-to <NUM> such as e.g. <NUM> to <NUM> such as e.g. 4000IMCU's per <NUM> milk.

In yet a related aspect, the invention relates to a method as described above wherein the milk prior to step a) is free or substantially free of microbial cultures.

In yet a related aspect, the invention relates to a method as described above wherein the milk has not been subject to warm maturation prior to step a).

In yet a related aspect, the invention relates to a method as described above wherein the milk is cow's milk.

The method may comprise further cheese making steps. Such steps are known to the person skilled in the art.

In the present context, the term "milk" refers to the lacteal secretion obtained by milking any mammal, such as cows, sheep, goats, buffaloes or camels. In a preferred embodiment, the milk is cow's milk, and especially raw cow's milk. However, it should be understood that the term milk also comprises compositions comprising milk, and milk compositions that have been treated, e.g. chemically, enzymatically, and/or mechanically.

In the context of the present invention, "microorganism" may include any bacterium, or fungus being able to ferment the milk substrate. Lactic acid bacteria and in particular Streptococcus thermophilus ssp. and Lactococcus ssp. are preferred microorganisms.

The microorganisms used for most fermented milk products are selected from the group of bacteria generally referred to as lactic acid bacteria. As used herein, the term "lactic acid bacterium" designates a gram-positive, microaerophilic or anaerobic bacterium, which ferments sugars with the production of acids including lactic acid as the predominantly produced acid, acetic acid and propionic acid. The industrially most useful lactic acid bacteria are found within the order "Lactobacillales" which includes Lactococcus spp. , Streptococcus spp. , Lactobacillus spp. , Leuconostoc spp. , Pseudoleuconostoc spp. , Pediococcus spp. , Brevibacterium spp. , Enterococcus spp. and Propionibacterium spp. Additionally, lactic acid producing bacteria belonging to the group of the strict anaerobic bacteria, bifidobacteria, i.e. Bifidobacterium spp. , are generally included in the group of lactic acid bacteria. These are frequently used as food cultures alone or in combination with other lactic acid bacteria.

As used herein, the term "culture" refers to any sample or item that contains one or more microorganisms. "Pure cultures" are cultures in which the organisms present are only of one strain of a particular genus and species. This is in contrast to "mixed cultures," which are cultures in which more than one genus and/or species of microorganism are present. In some embodiments of the present invention, pure cultures find use, but normally a culture as used in present invention contains more than one strain.

In the context of the present invention, "laboratory milk" is a reconstituted skim milk (RSM) with <NUM>% dry-matter on a weight basis that has been subjected to temperatures of <NUM> for <NUM> minutes before use.

In the context of the present invention, a "slow acidifying" bacterial culture (or a culture which acidifies the milk slowly) is either a mesophlic culture which has a maximum rate of acidification of <NUM> pH units per hour at <NUM> degrees C when inoculated at a quantity of <NUM>^<NUM> cfu (10E6 colony forming units) per ml laboratory milk (For the sake of completeness, if the culture consists of more than one strain, the culture as a whole should have the max rate of acidification of <NUM> UpH per hour at <NUM> degrees C as when inoculated <NUM>^<NUM> cfu/ml milk as defined above) or a thermophilic culture which is not able to decrease pH more than <NUM> pH unit in <NUM> incubation in a Lab milk when inoculated at quantity of <NUM>^<NUM> CFU - this definition for thermophilic cultures is preferred to maximum rate of acidification due to better characterization of fast and slow culture in the context of this invention.

In the context of the present invention, any other culture than a slow acidifying culture may be defined as a "fast acidifying" bacterial culture.

In the context of present invention the culture of bacteria which are mesophilic slow culture, lowers the pH less than <NUM> pH units (such as less than <NUM> pH units, less than <NUM> pH units, or less than <NUM> pH units) per hour at <NUM> degrees C, when inoculated at a quantity of 10E6 cfu (colony forming units) per ml laboratory milk.

In the context of present invention the culture of bacteria which are thermophilic slow culture lowers the pH less than <NUM> pH Unit within <NUM> incubation in a Lab milk when inoculated at quantity of <NUM>^<NUM> CFU.

In the context of present invention the culture of bacteria which are mesophilic fast culture, lowers the pH more than or equal to <NUM> pH units per hour at <NUM> degrees C, when inoculated at a quantity of 10E6 cfu (colony forming units) per ml laboratory milk.

In the context of present invention the culture of bacteria which are thermophilic fast culture lowers the pH more than <NUM> pH Unit within <NUM> incubation in a Lab milk when inoculated at quantity of <NUM>^<NUM> CFU.

By the term "practically simultaneously" is understood within <NUM> to <NUM> mins before or after one or more actions, such as e.g. <NUM> to <NUM> mins before or after.

By the term "warm maturation" is understood holding the milk at a temperature between <NUM> and <NUM>, such as <NUM> to <NUM>, such as e.g. <NUM> to <NUM>, such as e.g. <NUM> in the presence of lactic bacteria and without the addition of coagulants for at least <NUM> minutes.

"Soft cheese" is defined as any Rennet coagulated cheese that contains about <NUM> - <NUM> % moisture on a non-fat solids basis and is produced without scalding and pressing. Hence the preferred examples of Soft Cheese encompass brie, camembert, roquefort, etc..

In the present context, the term "mutant" should be understood as a strain derived, or a strain which can be derived, from a strain of the invention (or the mother strain) by means of e.g. genetic engineering, radiation and/or chemical treatment. It is preferred that the mutant is a functionally equivalent mutant, e.g. a mutant that has substantially the same, or improved, properties (e.g. regarding acidification speed) as the mother strain. Such a mutant is a part of the present invention. Especially, the term "mutant" refers to a strain obtained by subjecting a strain of the invention to any conventionally used mutagenization treatment including treatment with a chemical mutagen such as ethane methane sulphonate (EMS) or N-methyl-N'-nitro-N-nitroguanidine (NTG), UV light, or to a spontaneously occurring mutant. A mutant may have been subjected to several mutagenization treatments (a single treatment should be understood one mutagenization step followed by a screening/selection step), but it is presently preferred that no more than <NUM>, or no more than <NUM>, or no more than <NUM>, treatments (or screening/selection steps) are carried out. In a presently preferred mutant, less than <NUM>%, or less than <NUM>% or even less than <NUM>% of the nucleotides in the bacterial genome have been shifted with another nucleotide, or deleted, compared to the mother strain.

In the present context, the term "variant" should be understood as a strain or enzymes which is functionally equivalent or superior to a strain or enzyme of the invention, e.g. having substantially the same, or improved, properties e.g. regarding acidification speed or coagulation specificity). Such variants, which may be identified using appropriate screening techniques, are a part of the present invention.

The use of the terms "a" and "an" and "the" and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising", "having", "including" and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to,") unless otherwise noted.

The milk used is prepared from milk powders without added whey proteins.

Milk is standardized to <NUM>/L of Proteins and <NUM>/L of fat. pH is about <NUM> - <NUM>.

<NUM> of <NUM>/L-CaCl2 /<NUM> of milk is then added.

Milk is cooled at <NUM>
Milk is kept <NUM> for physical maturation at <NUM>.

Milk is then pasteurized at <NUM>/<NUM>.

Milk is incubated <NUM> minutes for Warm maturation. pH at the end of warm maturation is <NUM> and temperature <NUM>.

Coagulant addition is then performed: Hannilase is used with a dosage of 4000IMCU/<NUM> of milk. Clotting time is <NUM>. Curd is kept for hardening time during <NUM>. Total coagulation time is therefore <NUM>.

The curd is then cut into cubes of 15X15X15mm.

Three stirrings are made: one <NUM> after cutting (<NUM> movements) and one <NUM> minutes later (<NUM> movements) and one <NUM> later (<NUM> movements).

One hour and <NUM> after coagulant addition, the curd is transferred to <NUM> diameter round molds.

The curd is then drained in molds for <NUM> additional hours. The cheeses will be turned after <NUM> and after <NUM> and after <NUM>. Temperature will decrease during drainage to reach <NUM> after <NUM> and <NUM> at the end of drainage.

The cheeses are then removed from the forms. pH is <NUM> and temperature is <NUM>. Dry matter of the cheese at this step is <NUM>% and Fat on Dry Matter is <NUM>%.

The cheeses are then salted in brine (<NUM>/L density, pH5. <NUM> and temperature <NUM>) for <NUM>.

The cheeses will then be dried during <NUM> at <NUM> and <NUM>% relative humidity (RH) atmosphere.

Then the cheese will be ripened at <NUM> for <NUM> days with <NUM>%RH and turned at D+<NUM>.

Then ripening temperature will decrease from <NUM> to <NUM> in <NUM> days.

The cheeses will then be packed into composite paper (OPP) at D+<NUM>. pH at packaging is <NUM>, salt level <NUM>%, fat level <NUM>%, dry Mater <NUM>%.

Tasting session is performed after <NUM> days.

Coagulant addition is then performed: ChyMax® M is used with a dosage of 4000IMCU/<NUM> of milk. Clotting time is <NUM>. curd is kept for hardening time during <NUM>. Total coagulation time is therefore <NUM>.

Two stirrings are made: one <NUM> after coagulant addition (<NUM> movements) and one <NUM> minutes later (<NUM> movements).

One hour after coagulant addition, the curd is transferred to <NUM> diameter round molds.

The curd is then drained in molds for <NUM> additional hours. The cheeses will be turned after <NUM> and after <NUM>. Temperature will decrease during drainage to reach <NUM> after <NUM> and <NUM> at the end of drainage.

The cheeses will then be packed into composite paper at D+<NUM>. pH at packaging is <NUM>, salt level <NUM>%, fat level <NUM>%, dry matter <NUM>%.

Milk is standardized to <NUM>/L of Proteins and <NUM>/L of fat.

<NUM> of Aroma forming culture F-DVS SDMB7/<NUM> is added to the milk.

Coagulant addition is then performed: Naturen® is used with a dosage of <NUM> IMCU/<NUM> of milk. Clotting time is <NUM>. curd is kept for hardening time during <NUM>. Total coagulation time is therefore <NUM>.

One stirring is made <NUM> after cutting (<NUM> movements).

The cheeses are then removed from the forms. pH is <NUM> and temperature is <NUM>. Dry matter of the cheese at this step is <NUM>%, Fat on Dry Matter is <NUM>%, NFHumidity is <NUM>% Ca/NFS is <NUM>%.

The cheeses are then salted in brine (<NUM>/L density, pH4. <NUM> and temperature <NUM>) for <NUM>.

The cheeses will then be dried during <NUM> at <NUM> and <NUM>% relative Humidity atmosphere. Then the cheese will be ripened at <NUM> for <NUM> days with <NUM>%RH.

<NUM> of Secondary culture F-DVS STI06/<NUM> is added to the milk.

<NUM> of Secondary culture F-DVS SSC100 /<NUM> is added to the milk.

<NUM> of Aroma forming culture F-DVS SDMB7/<NUM> is added to the milk Coagulant addition is then performed: ChyMax M® is used with a dosage of <NUM> IMCU/<NUM> of milk. Clotting time is <NUM>. curd is kept for hardening time during <NUM>. Total coagulation time is therefore <NUM>.

One steering is made <NUM> after cutting (<NUM> movements)
One hour and <NUM> after coagulant addition, the curd is transferred to <NUM> diameter round molds.

The cheeses are then removed from the forms after 24hours. pH is <NUM> and temperature is <NUM>. Dry matter of the cheese at this step is <NUM>%, Fat on Dry Matter is <NUM>%, NFHumidity is <NUM>% Ca/NFS is <NUM>%.

Claim 1:
A method for the production of soft-cheese, said method comprising
a) adding to the milk a slow acidifying bacterial culture and adding to the milk a fast acidifying bacterial culture,
b) adding to the milk one or more coagulants,
c) incubating the milk, and
d) using the treated milk to produce the soft-cheese, wherein the soft-cheese is surface ripened, and
wherein:
(i) steps a) and b) are done simultaneously or practically simultaneously within 15mins regardless of order;
(ii) the pH drops at least <NUM> units within <NUM> hours after performing step a) and b);
(iii) the slow acidifying bacterial culture is:
- a mesophilic slow culture which lowers the pH less than <NUM> pH units (such as less than <NUM> pH units, less than <NUM> pH units, or less than <NUM> pH units) per hour at <NUM> degrees C, when inoculated at a quantity of 10E6 cfu (colony forming units) per ml laboratory milk; and/or
- a thermophilic slow culture which lowers the pH less than <NUM> pH Unit within <NUM> incubation in a laboratory milk when inoculated at quantity of <NUM>^<NUM> CFU at <NUM>; and
(iv) the fast acidifying bacterial culture is:
- a mesophilic fast culture which lowers the pH more than or equal to <NUM> pH units per hour at <NUM> degrees C, when inoculated at a quantity of 10E6 cfu (colony forming units) per ml laboratory milk; and/or
- a thermophilic fast culture which lowers the pH more than <NUM> pH Unit within <NUM> incubation in a laboratory milk when inoculated at quantity of <NUM>^<NUM> CFU per ml laboratory milk at <NUM>;
wherein a laboratory milk is a reconstituted skim milk with <NUM>% dry-matter on a weight basis that has been subjected to temperatures of <NUM> for <NUM> minutes before use.