EP2094829A2 - Procédé de préparation de cultures bactériennes congelées utilisées dans les domaines alimentaire, diététique, nutraceutique et pharmaceutique - Google Patents
Procédé de préparation de cultures bactériennes congelées utilisées dans les domaines alimentaire, diététique, nutraceutique et pharmaceutiqueInfo
- Publication number
- EP2094829A2 EP2094829A2 EP07825662A EP07825662A EP2094829A2 EP 2094829 A2 EP2094829 A2 EP 2094829A2 EP 07825662 A EP07825662 A EP 07825662A EP 07825662 A EP07825662 A EP 07825662A EP 2094829 A2 EP2094829 A2 EP 2094829A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- lactobacillus
- bifidobacterium
- ssp
- culture
- frozen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
- 238000000034 method Methods 0.000 title claims abstract description 43
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 17
- 235000013305 food Nutrition 0.000 title claims abstract description 17
- 235000005911 diet Nutrition 0.000 title abstract description 8
- 230000000378 dietary effect Effects 0.000 title abstract description 8
- 238000007710 freezing Methods 0.000 claims abstract description 25
- 230000008014 freezing Effects 0.000 claims abstract description 25
- 238000011109 contamination Methods 0.000 claims abstract description 11
- 238000004806 packaging method and process Methods 0.000 claims abstract description 9
- 241000194017 Streptococcus Species 0.000 claims description 27
- 239000007858 starting material Substances 0.000 claims description 24
- 241000186660 Lactobacillus Species 0.000 claims description 22
- 229940039696 lactobacillus Drugs 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 22
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 19
- 241000186000 Bifidobacterium Species 0.000 claims description 18
- 238000009629 microbiological culture Methods 0.000 claims description 17
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 9
- 229910052757 nitrogen Inorganic materials 0.000 claims description 9
- 244000057717 Streptococcus lactis Species 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 7
- 244000199866 Lactobacillus casei Species 0.000 claims description 6
- 241000186606 Lactobacillus gasseri Species 0.000 claims description 6
- 241000186840 Lactobacillus fermentum Species 0.000 claims description 5
- 241000894007 species Species 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 241000186673 Lactobacillus delbrueckii Species 0.000 claims description 4
- 241000191940 Staphylococcus Species 0.000 claims description 4
- 235000014897 Streptococcus lactis Nutrition 0.000 claims description 4
- 235000013365 dairy product Nutrition 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 235000013958 Lactobacillus casei Nutrition 0.000 claims description 3
- 241000186684 Lactobacillus pentosus Species 0.000 claims description 3
- 241000194036 Lactococcus Species 0.000 claims description 3
- 229940017800 lactobacillus casei Drugs 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 241000186018 Bifidobacterium adolescentis Species 0.000 claims description 2
- 241000186011 Bifidobacterium catenulatum Species 0.000 claims description 2
- 241001608472 Bifidobacterium longum Species 0.000 claims description 2
- 241001134772 Bifidobacterium pseudocatenulatum Species 0.000 claims description 2
- 241000194033 Enterococcus Species 0.000 claims description 2
- 240000001046 Lactobacillus acidophilus Species 0.000 claims description 2
- 241000192132 Leuconostoc Species 0.000 claims description 2
- 241000192001 Pediococcus Species 0.000 claims description 2
- 241000186429 Propionibacterium Species 0.000 claims description 2
- 241000235070 Saccharomyces Species 0.000 claims description 2
- 241000191973 Staphylococcus xylosus Species 0.000 claims description 2
- 241000194020 Streptococcus thermophilus Species 0.000 claims description 2
- 235000013351 cheese Nutrition 0.000 claims description 2
- 241001134770 Bifidobacterium animalis Species 0.000 claims 2
- 229940118852 bifidobacterium animalis Drugs 0.000 claims 2
- 241000894006 Bacteria Species 0.000 claims 1
- 241000186016 Bifidobacterium bifidum Species 0.000 claims 1
- 241000186012 Bifidobacterium breve Species 0.000 claims 1
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 claims 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims 1
- 241000218588 Lactobacillus rhamnosus Species 0.000 claims 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 1
- 235000013334 alcoholic beverage Nutrition 0.000 claims 1
- 229940002008 bifidobacterium bifidum Drugs 0.000 claims 1
- 229940004120 bifidobacterium infantis Drugs 0.000 claims 1
- 229940009291 bifidobacterium longum Drugs 0.000 claims 1
- 235000008429 bread Nutrition 0.000 claims 1
- 235000015140 cultured milk Nutrition 0.000 claims 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims 1
- 229940012969 lactobacillus fermentum Drugs 0.000 claims 1
- 229940072205 lactobacillus plantarum Drugs 0.000 claims 1
- 239000006041 probiotic Substances 0.000 claims 1
- 230000000529 probiotic effect Effects 0.000 claims 1
- 235000018291 probiotics Nutrition 0.000 claims 1
- 235000013580 sausages Nutrition 0.000 claims 1
- 235000013618 yogurt Nutrition 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 10
- 241000166315 Vulcaniibacterium thermophilum Species 0.000 description 23
- 239000002028 Biomass Substances 0.000 description 11
- 230000000813 microbial effect Effects 0.000 description 11
- 239000000047 product Substances 0.000 description 7
- 239000002577 cryoprotective agent Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 5
- 239000003344 environmental pollutant Substances 0.000 description 5
- 231100000719 pollutant Toxicity 0.000 description 5
- 240000001929 Lactobacillus brevis Species 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 238000005325 percolation Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 241000194041 Lactococcus lactis subsp. lactis Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 239000004411 aluminium Substances 0.000 description 1
- 229940093740 amino acid and derivative Drugs 0.000 description 1
- RZOBLYBZQXQGFY-UHFFFAOYSA-N ammonium lactate Chemical class [NH4+].CC(O)C([O-])=O RZOBLYBZQXQGFY-UHFFFAOYSA-N 0.000 description 1
- 235000019286 ammonium lactate Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical class [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000002826 coolant Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000008150 cryoprotective solution Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000000832 lactitol Substances 0.000 description 1
- VQHSOMBJVWLPSR-JVCRWLNRSA-N lactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-JVCRWLNRSA-N 0.000 description 1
- 235000010448 lactitol Nutrition 0.000 description 1
- 229960003451 lactitol Drugs 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920000570 polyether Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- -1 polyethylenes Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000012602 primary packaging material Substances 0.000 description 1
- ULWHHBHJGPPBCO-UHFFFAOYSA-N propane-1,1-diol Chemical compound CCC(O)O ULWHHBHJGPPBCO-UHFFFAOYSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 230000002623 sporogenic effect Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000011514 vinification Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/02—Making cheese curd
- A23C19/032—Making cheese curd characterised by the use of specific microorganisms, or enzymes of microbial origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
Definitions
- the present invention relates to a method for preparing frozen bacterial cultures for use in the food, dietary, nutraceutics and pharmaceutical fields.
- Said method includes the packaging of the bacterial culture under aseptic conditions, followed by the freezing process, thus avoiding all those handlings which are a potential contamination source.
- starter cultures which are added to the starting raw materials in order to obtain the desired end products in the dairy-, baked products-, fermented sausages- and vinification sectors .
- Said starter cultures are selected grafts, consisting of known microbial strains (or mixtures thereof) , with well defined biochemical properties and which therefore allow to obtain repeatable and enduring results.
- One of the physical forms in which starter cultures are marketed is the frozen form, which shows the advantage of combining a good shelf life of the same with an absolutely acceptable cost, if compared with the one of cultures in a freeze-dried form.
- the freezing process is carried out by subjecting the bacterial culture to a quick falling of the temperature of the product itself (for example up to temperatures ⁇ -40 0 C) , followed by the packaging of the same .
- the most currently used methods foresee the rapid freezing of the same usually through a contact with refrigerated surfaces or through air flows at temperatures generally between -40 and -50 0 C, as well as through the direct use of a coolant, such as liquid nitrogen or carbon dioxide at the solid state, which is contacted with the food.
- a coolant such as liquid nitrogen or carbon dioxide
- the freezing process is, at present, generally performed through direct percolation in liquid nitrogen, with a consequent immediate change of the existing water from the liquid form to the solid one.
- a disadvantage resulting from such process is that the starter culture, once frozen as above described, must then be stored at very low temperatures, which are on average between -70 and -55°C, with the obvious need of making use of particularly exacting and expensive cooling environments and devices.
- a further substantial drawback connected with the freezing processes commonly used is due to the fact that, during the execution of such processes, there is a direct, more or less extensive, contact between the freezing agent, or the device, and the product intended for the freezing, with consequent possible microbial contamination phenomena of different origin and nature. Said type of contamination persists in the frozen end product also after its packaging, although this can occur in controlled hygienic conditions.
- liquid cultures In the specific case of liquid cultures, a possible contamination source can result from the contacting surfaces of the freezing plant itself and the air existing therein.
- the liquid nitrogen allows the preservation of the existing biological pollutants, if any, in a frozen state but viable and capable of reproducing themselves when permitted by the environmental conditions.
- the presence of biological pollutants in a frozen starter culture intended for dairy applications is particularly harmful, as the inoculum temperature of the milk in the boiler and its following processing is on average suitable to the growth of most of said pollutants .
- the sanitary situation of the food or the product for a food use is particularly serious, as the spores are generally able to withstand most of the heat treatments to which foods, before their consumption, are generally subjected. Therefore, there remains thus the .need of being able to provide frozen bacterial cultures for the use in ⁇ food, dietary, nutraceutics and pharmaceutical fields which are free of each kind of contamination.
- bacterial cultures must be free of contaminations due to the plants and means used for the freezing (especially when the freezing operation is carried out by percolation in liquid nitrogen) .
- the object of the present invention is to provide an adequate answer to the need above pointed out .
- an object of the present invention is a method for preparing frozen bacterial cultures for the use in food, dietary, nutraceutics and pharmaceutical fields which are free of contaminations, having the features described in the appended independent claim.
- Another object of the present invention is a bacterial culture which can be obtained with the above method, having the features described in the appended independent claim.
- Another aspect of the present invention is the use of said bacterial culture in a food, dietary, nutraceutics and pharmaceutical fields, having the features described in the appended independent claim.
- Preferred embodiments of the present invention are re- ported in the appended dependent claims .
- the method subject of the present invention, relates to the preparation of a bacterial culture including: a) at least a first step wherein the starting culture is packaged under completely sterile conditions; b) at least a second step wherein said culture resulting from a) is subjected to freezing.
- the first step a) is performed on the product under complete sterile conditions by using known equipments and modes and usually employed in the sector. For example, one works in a sterile environment at a temperature on average between 1°C and 20 0 C, preferably from 3°C to 1O 0 C, and having an air quality at least equal to the class 100, obtained through opportune filtering systems of the HEPA (High Efficiency Particulate Air) type.
- HEPA High Efficiency Particulate Air
- the environment ⁇ in which the packaging takes place is kept in overpressure conditions of sterile air (with a quality at least of class 100) equal to at least 1.01 atmospheres.
- the method according to the present invention is characterized in that the frozen microbial culture obtained with said method is substantially free of pollutants .
- the method allows to eliminate/reduce the contamina- tion sources in a production line of frozen microbial cultures; in particular, in the food and pharmaceutical industries .
- said process for use in the food field is a starter micro- bial culture in a liquid form.
- Said starter culture in a liquid form is prepared, for example, by adopting the following steps: - development of the microbial biomass in an industrial fermenter using a proper fermentation medium and opportune development conditions of the microbial bio- mass according to the microorganism to be reproduced;
- Cryoprotectants are substances commonly used in the freezing sector. Said substances are usually carefully selected, as a function of the microorganism of interest, among physiologically compatible substances capa- ble of protecting the microbial cells from the osmotic and mechanical stresses caused by the freezing process.
- said substances are selected from the group including; amino acids and derivatives thereof; monosaccharides and disacch.arid.es; polymers presenting alcoholic functions within their structure; polyethers; other substances known to a skilled in the art.
- cryoprotectants are selected from the group including: sodium glutamate, mannitol, lactitol, sucrose, fructose, starch, glycerol, propanediol, polyvinyl pyrrolidone (PVP) , dimethyl sulfoxide
- DMSO dimethyl methacrylate
- PEG polyethylene glycol
- Said cryoprotectants are additioned to the water volume used for re-suspending the washed microbial biomass in a quantity on average between 10% and 65% (weight/volume or volume/volume) based on the total weight of the water; preferably, from 25% to 50% (weight/volume or volume/volume) .
- the used quantity of the cryoprotectant is, however, varying as a function of the physical-chemical features of the cryoprotectant and the biomass cell con- centration after its re-suspension in the cryoprotec- tive solution.
- the starter culture in a liquid form includes at least a microorganism physiologically compatible with the human and/or animal organism.
- said at least one microorganism is se- lected from the group including the following genera: Lactobacillus, Bifidobacterium, Lactococcus, Leu- conostoc, Pediococcus, Streptococcus, Bacillus, Propi- onibacterium, Saccharomyces, Enterococcus, Staphylo- coccus .
- Lactobacillus are preferred the species: L. pentosus, L. plantarum, L. casei, L. casei ssp. paracasei , L. casei ssp. rhamnosus, L. acidophilus, L. delbrueckii ssp. bulgaricus, L. fermen- turn, L. gasseri.
- B. longum B. breve
- B. lactis B. adolescentis
- B. pseudocatenulatum B. catenulatum
- L. lactis L. lactis ssp. lactis.
- Streptococcus are preferred the species: S. thermophilus.
- the starter culture includes a mixture of two or more bacterial strains selected among those above mentioned. It is understood to mean that the freezing method according to the present invention is, in any case, applicable to any type of starter microbial culture in a liquid form, independently from the type of process with which the same is prepared.
- Starter cultures in liquid form once ready for being frozen, are packaged (preferably under a laminar flow of sterile air at a temperature between 1 and 10 0 C) , in sterile primary packaging materials, preferably selected from: polyethylenes , aluminium and polyesters and/or opportune multilayered combinations thereof .
- the same are preferably tightly sealed under the same aseptic conditions above described.
- the following freezing (relative to the step b) above described) is carried out using known techniques in the art of the refrigeration; preferably, through the use of liquid nitrogen or an opportune refrigerator system at a temperature ⁇ -18 0 C for a sufficient time to obtain the freezing of the water existing in the packaged product .
- the freezing time of a total volume between 10 1 and 100 1 is on average between 5 seconds and 2 minutes .
- a starter culture in liquid form resulting from the step a) is frozen by using liquid nitrogen at -200 0 C for times between 3 seconds and 1 minute.
- the viability of the frozen product obtained with the method of the present invention is equivalent to the one typical of the traditional frozen starter cultures.
- the Applicant has found that, in the starter liquid cultures in which the filtration and the washing of the microbial biomass have been carried out, there can be found a sensible decrease, or a total ab- sence, of the lactic acid organic salts (such as ammonium lactates and/or calcium lactates, usual metabolites resulting from the fermentation and development steps of the biomass within the fermenter) .
- This working mode allows to obtain a stability increase of the culture itself after the freezing.
- frozen starter cultures according to the method of the present invention have shown to have an excellent shelf life at temperatures between -24 0 C and -18 0 C, with consequent much more limited storage costs.
- refrigerator devices at lower temperatures usually between, in the general use routine procedure, -70 0 C and -55 0 C) are not needed anymore, while, on the contrary, they are still needed in case of traditional frozen cultures .
- the shelf-life of a starter culture frozen according to the method of the present invention has been evaluated based on the maintenance, over time, of the ability of exact volumes of said culture, inoculated in a proper medium, for example milk, of lowering the pH until precise values after the same time intervals.
- the shelf-life of said frozen starter culture, stored at a temperature between -24 0 C and -18 0 C, has proved to be > 4 months, preferably between 4 and 12 months; more preferably, > 6 months.
- the frozen starter culture obtained with the method of the present invention, is used as starter in the food sector, for example in the dairy industry; dietary sector, for example in the supplements; nutraceu- tic and pharmaceutical sectors.
- the cheese production process to be followed with the frozen starter culture obtained with the method of the present invention is equal to the one which is followed with the traditional frozen cultures .
- Products which originate therefrom have the same organoleptic characteristics, but show high guarantees from a sanitary point of view.
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ITMI20062287 ITMI20062287A1 (it) | 2006-11-28 | 2006-11-28 | Metodo per la preparazi0ne di colture batteriche congelate per utilizzo in campo alimentare dietetico nutraceutico e farmaceutico |
PCT/IB2007/003485 WO2008065491A2 (fr) | 2006-11-28 | 2007-11-12 | Procédé de préparation de cultures bactériennes congelées utilisées dans les domaines alimentaire, diététique, nutraceutique et pharmaceutique |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2094829A2 true EP2094829A2 (fr) | 2009-09-02 |
Family
ID=39468315
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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EP07825662A Ceased EP2094829A2 (fr) | 2006-11-28 | 2007-11-12 | Procédé de préparation de cultures bactériennes congelées utilisées dans les domaines alimentaire, diététique, nutraceutique et pharmaceutique |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP2094829A2 (fr) |
IT (1) | ITMI20062287A1 (fr) |
WO (1) | WO2008065491A2 (fr) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102883627B (zh) | 2010-04-27 | 2015-06-10 | 科·汉森有限公司 | 将酵母接种到果汁中的方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3891773A (en) * | 1971-02-26 | 1975-06-24 | Us Agriculture | Culture of sour dough bacteria |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3592740A (en) * | 1968-03-28 | 1971-07-13 | Niles Lab Inc | Production of cell culture concentrates |
US4304868A (en) * | 1977-09-06 | 1981-12-08 | Microlife Technics, Inc. | Process for the treatment of meat with compositions including Micrococcus varians and a lactic acid producing bacteria |
GB8323094D0 (en) * | 1983-08-26 | 1983-09-28 | Franks F | Preservation of cells |
FR2746109B1 (fr) * | 1996-03-12 | 1998-04-17 | Rhone Poulenc Rorer Sa | Milieu pour la conservation de materiel biologique |
ATE417093T1 (de) * | 1998-08-26 | 2008-12-15 | Chr Hansen As | Flüssige starterkulturen mit verbesserter lagerstabilität und ihre verwendung |
EP1441027A1 (fr) * | 2003-01-22 | 2004-07-28 | Chr. Hansen A/S | Bactéries productrices d'acide lactique congelées pour conservation stable |
GB0610330D0 (en) * | 2006-05-24 | 2006-07-05 | Cab Internat | Method for preservation of micro-organisms |
-
2006
- 2006-11-28 IT ITMI20062287 patent/ITMI20062287A1/it unknown
-
2007
- 2007-11-12 WO PCT/IB2007/003485 patent/WO2008065491A2/fr active Application Filing
- 2007-11-12 EP EP07825662A patent/EP2094829A2/fr not_active Ceased
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3891773A (en) * | 1971-02-26 | 1975-06-24 | Us Agriculture | Culture of sour dough bacteria |
Also Published As
Publication number | Publication date |
---|---|
ITMI20062287A1 (it) | 2008-05-29 |
WO2008065491A3 (fr) | 2008-10-23 |
WO2008065491A2 (fr) | 2008-06-05 |
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