WO2024115523A1 - Procédé de préparation d'une composition d'additif solide congelée ou lyophilisée - Google Patents

Procédé de préparation d'une composition d'additif solide congelée ou lyophilisée Download PDF

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WO2024115523A1
WO2024115523A1 PCT/EP2023/083437 EP2023083437W WO2024115523A1 WO 2024115523 A1 WO2024115523 A1 WO 2024115523A1 EP 2023083437 W EP2023083437 W EP 2023083437W WO 2024115523 A1 WO2024115523 A1 WO 2024115523A1
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equal
freeze
additive composition
less
solid additive
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PCT/EP2023/083437
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English (en)
Inventor
Bernadette Theresia MOREL
Pieter Vonk
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Dsm Ip Assets B.V.
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Publication of WO2024115523A1 publication Critical patent/WO2024115523A1/fr

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    • 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
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

Definitions

  • the invention relates to a process for preparing a frozen or freeze-dried solid additive composition, a solid additive composition obtained or obtainable therewith, a kit of parts comprising such solid additive composition, a process of producing a cultured food product using such solid additive composition or such kit of parts, and the use of such solid additive composition or such kit of parts in a process for making a food or feed product.
  • Lactic acid bacteria are used world-wide in the dairy industry to produce a variety of fermented dairy products such as cheese, yoghurts, sour cream, kefir, butter and koumiss. Selected strains of lactic acid bacteria initiating and carrying out the desired fermentations are essential in the manufacture of the above products. They are often referred to as starter cultures.
  • Milk can be inoculated without intermediate transfer and/or propagation with so-called direct vat set (DVS) or direct-to-vat inoculation (DVI) starter cultures.
  • DVDS direct vat set
  • DVI direct-to-vat inoculation
  • Such starter cultures are generally available from commercial manufacturers in frozen or lyophilized/freeze-dried form. They can comprise only a single lactic acid bacterium species, but can also be mixed cultures comprising two or more different lactic acid bacterium species.
  • cryoprotectants In order to maintain high cell counts and/or improved stability of liquid, frozen or lyophilized/freeze- dried starter cultures, cryoprotectants can be employed.
  • W02009/056979 describes a starter composition made by adding appropriate amounts of powdered forms of the sources of formate and purine to a lyophilized (i.e., freeze dried) cell culture. It also describes a starter composition made by blending a frozen bacterial pellet with a frozen solution containing the sources of formate and purine.
  • US 2005/0069862 describes that measures that have to be provided by an effective cryo protective agent are very much different depending on whether the cryoprotective agent is designed to protect liquid or frozen cultures, and consequently, additives, which are effective cryoprotective agents of liquid cultures, may not be effective for frozen cultures.
  • US 2005/0069862 states that one example of such an additive is Na-formate.
  • US 2005/0069862 refers to WO 00/39281 and mentions that 3% Na-formate is effective to increase the storage stability of liquid lactic acid bacterial starter culture concentrates. However, as illustrated in Example 15 of US 2005/0069862, 3% Na-formate decrease the storage stability of frozen lactic acid bacterial starter culture.
  • WO2012/076665 describes a process for making a starter culture composition comprising a microorganism, a cryoprotectant and at least one stimulating additive, the process comprising the steps of: a) culturing a microorganism in a culture medium; and b) collecting the microorganism from the culture medium; and c) adding a cryoprotectant to the microorganism obtained in step b); and d) freezing the microorganisms obtained in step c); and e) adding at least one stimulating additive to microorganisms obtained in step d).
  • combinations of frozen culture pellets and frozen sodium formate pellets are described and also combinations of freeze-dried culture pellets and freeze-dried sodium formate pellets are described.
  • a cryoprotectant solution containing (in wt%): maltodextrin (28,6%), sorbitol (10,3%), calcium ascorbate (11 ,4%), glutamate (2,9%) and water (46,8%) is added to 65.84 g of the culture concentrate directly after centrifugation and before freezing or freeze-drying to create the culture pellets.
  • the sodium formate pellets contain 40% (w/w) sodium formate, 2% (w/w) Arabic gum with the remainder being water.
  • the invention accordingly provides a process for preparing a frozen or freeze-dried solid additive composition, comprising or consisting of the following steps: a) preparing an aqueous mixture comprising or consisting of (I) formic acid or a salt or ester thereof and (ii) a crystallization inhibitor; b) preparing a frozen solid additive composition by freezing the aqueous mixture of a); and c) optionally preparing a freeze-dried solid additive composition by freeze-drying the frozen solid additive composition of b).
  • the above process can preferably produce the formic acid or a salt or ester thereof in the frozen or freeze-dried solid additive composition existing partly or wholly in an amorphous state.
  • the prepared frozen or freeze- dried solid additive composition can be helpful to prepare a stable starter culture as illustrated in detail below.
  • the process according to the invention can allow for a faster freeze-drying process, which is economically attractive.
  • the invention further provides a solid additive composition, preferably frozen or freeze- dried, preferably in the form of a pellet, a granule, a tablet, a powder or another solid particle, comprising or consisting of:
  • solid additive composition preferably does not comprise any type of lactic acid bacteria, present, individually, in a viable cell count density of equal to or more than 1 x 10 6 cfu/gram; and which solid additive composition preferably is obtained or obtainable by a process as described above.
  • the invention also provides a kit of parts comprising:
  • a second solid composition comprising or consisting of a solid culture composition, preferably frozen or freeze-dried, preferably in the form of a pellet, a granule, a tablet, a powder or another solid particle, wherein the solid culture composition comprises at least one type of lactic acid bacteria in an individual viable cell count density of equal to or more than 1 x 10 6 cfu/gram.
  • the above kit of parts is a starter culture.
  • such starter culture may advantageously have an improved stability.
  • the invention provides a method for producing a cultured food product, the method comprising the steps of:
  • the cultured food product is a fermented milk product.
  • the invention provides a use of the solid additive composition described above or the kit of parts described above in a process for making a food or feed product.
  • This food or feed product is preferably a fermented milk product.
  • Figure 1 illustrates a comparison of freeze drying (I) pellets with a composition, based on dry matter, of 100% w/w sodium-formate (“100% NAF”); (II) pellets with a composition, based on dry matter, of 98 % w/w sodium formate and 2 % w/w maltodextrin (“NAF-MD (2%)”); and (ill) pellets with a composition of 96 % w/w sodium formate and 4 % w/w maltodextrin (“ NAF-MD (4%)”). It is illustrated that the NAF-MD (2%) composition and the NAF-MD (4%) composition allow for a faster freeze-drying process, which is economically attractive.
  • FIG. 2 illustrates a comparison of freeze drying (I) pellets with a composition, based on dry matter, of 100% w/w sodium-formate (“NaF 100%”, the line at the top); (II) pellets with a composition, based on dry matter, of 98 % w/w sodium formate and 2 % w/w sucrose (“NaF 98% + Sucrose 2%”, the second line from the top); (ill) pellets with a composition, based on dry matter, of 98 % w/w sodium formate and 2 % w/w maltodextrin (“NaF 98% + Maltodextrin 2%”, the third line from the top); and (iv) pellets with a composition, based on dry matter, of 80 % w/w sodium formate and 20 % w/w maltodextrin (“ NaF 80% + Maltodextrin 20%”, the bottom dots).
  • the compound in principle includes all enantiomers, diastereomers and cis/trans isomers of that compound that may be used in the particular aspect of the invention; in particular when referring to such as compound, it includes the natural isomer(s).
  • milk is intended to encompass milks from mammals and plant sources or mixtures thereof.
  • the milk is from a mammal source.
  • Mammals sources of milk include, but are not limited to cow, sheep, goat, buffalo, camel, llama, horse or reindeer.
  • the milk is from a mammal selected from the group consisting of cow, sheep, goat, buffalo, camel, llama, horse and deer, and combinations thereof.
  • Plant sources of milk include, but are not limited to, milk extracted from soy bean, pea, peanut, barley, rice, oat, quinoa, almond, cashew, coconut, hazelnut, hemp, sesame seed and sunflower seed. Bovine milk is preferred.
  • milk refers to not only whole milk, but also skim milk or any liquid component derived thereof or reconstituted milk.
  • milk base refers to a base composition, comprising milk or milk ingredients, or derived from milk or milk ingredients.
  • the milk can be from an animal, preferably mammal, source, from a plant source, or a mixture thereof.
  • the milk base can be used as a raw material for the fermentation to produce a fermented milk product.
  • the milk base may for example comprise or consist of skimmed or non-skimmed milk, or reconstituted milk.
  • the milk base may be concentrated or in the form of powder, or may be reconstituted from such.
  • reconstituted milk is herein understood liquid milk obtained by adding liquid, such as water, to a skim milk powder, skim milk concentrate, whole milk powder or whole milk concentrate.
  • the milk base may or may not have been subjected to a thermal processing operation which is at least as efficient as pasteurization.
  • the milk base is from a bovine source.
  • the terms “fermented milk product”, “fermented dairy product” and “acidified milk product” are used interchangeably and are intended to refer to products which are obtained by the multiplication of lactic acid bacteria in a milk base leading to a milk coagulum.
  • the particular characteristics of the various fermented milk products depend upon various factors, such as the composition of milk base, the incubation temperature, the composition of the lactic acid bacteria and/or presence of further non-lactic acid microorganisms.
  • fermented milk products manufactured herein include, for instance, various types of yoghurt (including for example set yoghurt, low fat yoghurt, non-fat yoghurt), kefir, dahi, ymer, buttermilk, butterfat, sour cream and sour whipped cream as well as fresh cheeses such as quark and cottage cheese.
  • Petit Suisse or Mozarella is yet another example of a fermented dairy product.
  • the fermented milk product is a yoghurt.
  • the fermented milk product produced in the current invention can be a stirred yoghurt or a set yoghurt.
  • the fermented milk product is a set yoghurt.
  • yoghurt and “yogurt” are used interchangeably herein.
  • the term “yoghurt” refers to products comprising or obtained by means of lactic acid bacteria that include at least Streptococcus salivarius thermophilus and Lactobacillus delbruekii subsp. bulgaricus, but may also, optionally, include further microorganisms such as Lactobacillus delbruekii subsp. lactis, Bifidobacterium animalis subsp. lactis, Lactococcus lactis, Lactobacillus acidophilus and Lactobacillus casei, or any microorganism derived therefrom.
  • Such lactic acid strains other than Streptococcus salivarius thermophilus and Lactobacillus delbruekii subsp. bulgaricus can give the finished product various properties, such as the property of promoting the equilibrium of the gut microbiota.
  • the term "yoghurt” encompasses set yoghurt, stirred yoghurt, drinking yoghurt, heat treated yoghurt and yoghurt-like products. More preferably, the term "yoghurt” encompasses, but is not limited to, yoghurt as defined according to French and European regulations, e.g. coagulated dairy products obtained by lactic acid fermentation by means of specific thermophilic lactic acid bacteria only (i.e.
  • Yoghurts may optionally contain added dairy raw materials (e.g. cream) or other ingredients such as sugar or sweetening agents, one or more flavouring(s), fruit, cereals, or nutritional substances, especially vitamins, minerals and fibers.
  • dairy raw materials e.g. cream
  • other ingredients such as sugar or sweetening agents, one or more flavouring(s), fruit, cereals, or nutritional substances, especially vitamins, minerals and fibers.
  • Such yoghurt advantageously meets the specifications for fermented milks and yoghurts of the AFNOR NF 04-600 standard and/orthe codex StanA- lla-1975 standard.
  • the product In order to satisfy the AFNOR NF 04-600 standard, the product must not have been heated after fermentation and the dairy raw materials must represent a minimum of 70% (m/m) of the finished product.
  • starter or “starter culture” as used herein refers to a culture of one or more food-grade micro-organisms, more preferably a culture comprising lactic acid bacteria, which are responsible for the acidification of the milk base. Starter cultures may be fresh (liquid), frozen or freeze-dried. Freeze dried cultures need to be regenerated before use. For the production of a yoghurt, the starter culture (i.e.
  • the total weight of all lactic acid bacterial combined can for example be added in an amount from 0.001 to 10% by weight, suitably in an amount of 0.01 to 3% by weight, of the total amount of milk base.
  • dosages in the lower part of the range can be used such as from 0.006% by weight of the total amount of milk base.
  • lactic acid bacteria As used herein, the term "lactic acid bacteria”, “LAB”, “lactic acid bacterial strains” and “lactic bacteria” are used interchangeably and refer to food-grade bacteria producing lactic acid as the major metabolic end-product of carbohydrate fermentation. These bacteria are related by their common metabolic and physiological characteristics and are usually Gram positive, low-GC, acid tolerant, non- sporulating, non-respiring, rod-shaped bacilli or cocci. During the fermentation stage, the consumption of lactose by these bacteria causes the formation of lactic acid, reducing the pH and leading to the formation of a protein coagulum. These bacteria are thus responsible for the acidification of milk and for the texture of the dairy product.
  • lactic acid bacteria encompasses, but is not limited to, bacteria belonging to the genus of Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., Lactococcus spp., such as Lactobacillus delbruekii subsp. bulgaricus, Streptococcus salivarius thermophilus, Lactobacillus lactis, Bifidobacterium animalis, Lactococcus lactis, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus acidophilus and Bifidobacterium breve.
  • a strain is a genetic variant or subtype of a microorganism, in this case a subtype or variant of a lactic acid bacteria.
  • a crystallization inhibitor is herein preferably understood a compound that can stabilize an amorphous state, preferably the amorphous state of formic acid or a salt or ester thereof. Such a crystallization inhibitor can suitably prevent crystallization or re-crystallization. Most preferably the crystallization inhibitor is maltodextrin.
  • the invention provides a process for preparing a frozen or freeze-dried solid additive composition, comprising or consisting of the following steps: a) preparing an aqueous mixture comprising or consisting of (i) formic acid or a salt or ester thereof and
  • the aqueous mixture of step a) does not comprise any type of lactic acid bacteria, individually, in any individual viable cell count density of equal to or more than 1 x 10 6 cfu/gram. More preferably the aqueous mixture of step a) does not comprise any type of lactic acid bacteria, individually, in any individual viable cell count density of equal to or more than 1 x 10 4 cfu/gram, before the freezing and/or freeze-drying of the mixture.
  • the aqueous mixture of step a) comprises a total amount of lactic acid bacteria of equal to or less than 1 x 10 10 cfu/gram, more preferably of equal to or less than 1 x 10 8 cfu/gram, still more preferably of equal to or less than 1 x 10 6 cfu/gram, even more preferably of equal to or less than 1 x 10 5 cfu/gram, yet more preferably of equal to or less than 1 x 10 4 cfu/gram, and yet even more preferably of equal to or less than 1 x 10 3 cfu/gram.
  • the aqueous mixture of step a) comprises a total amount of lactic acid bacteria of equal to or less than 1 x 10 2 cfu/gram, preferably equal to or less than 1 x 10 1 cfu/gram.
  • the aqueous mixture of step a) comprises no lactic acid bacteria.
  • the frozen or freeze-dried solid additive composition does not comprise any type of lactic acid bacteria in any individual viable cell count density of equal to or more than 1 x 10 6 cfu/gram, preferably of equal to or more than 1 x 10 4 cfu/gram.
  • the formic acid or a salt or ester thereof in the frozen or freeze-dried solid additive composition exists partly or wholly in an amorphous state.
  • the aqueous mixture of step a) comprises or consists of:
  • the freezing in step b) can be carried out in any manner known by the person skilled in the art to be suitable therefore.
  • the frozen solid additive composition is prepared by freezing the aqueous mixture to a temperature below -1 °C, more preferably to a temperature equal to or less than -10°C, even more preferably to a temperature equal to or less than -20°C.
  • the frozen solid additive composition is prepared by freezing the aqueous mixture to a temperature equal to or less than -50°C, even more preferably to a temperature equal to or less than -80°C.
  • the frozen solid additive composition is prepared by freezing the aqueous mixture to a temperature equal to or less than -100°C, even more preferably to a temperature equal to or less than -180°C. Most preferably the freezing is carried out with the help of liquid nitrogen (also referred to as cryogenic liquid nitrogen). More preferably step b) comprises dripping or spraying of the aqueous mixture into liquid nitrogen. If so desired the frozen solid additive prepared in step b) can subsequently be milled. That is, step b) may suitably further comprise milling of the frozen solid additive composition.
  • Step b) may comprise preparing a frozen solid additive composition by freezing the aqueous mixture of a), wherein the frozen solid additive composition may have the form of a pellet, a granule, a tablet, a powder or another solid particle. More preferably step b) comprises preparing a frozen solid additive composition by freezing the aqueous mixture of a), into the form of a pellet or a granule.
  • the frozen solid additive composition is preferably freeze-dried until an Aw of equal to or less than 0.2, preferably an Aw of equal to or less than 0.1 is reached.
  • the process comprises step c), comprising or consisting of preparing a freeze-dried solid additive composition by freeze-drying the frozen solid additive composition of b).
  • the frozen solid additive composition is preferably freeze-dried until an Aw of equal to or less than 0.2, preferably an Aw of equal to or less than 0.1 is reached. That is, more preferably the process is a process wherein the mixture is freeze-dried until an Aw of less than 0.2, preferably an Aw of less than 0.1 , is reached.
  • freeze drying can comprise one or more steps and the freeze-drying pressure and temperature can vary. Freeze-drying may further be carried out by for example plate freeze-drying or radiation freeze-drying.
  • the pressure during freeze-drying preferably lies in the range from equal to or more than 0.10 mbar, more preferably equal to or more than 0.20 mbar to equal to or less than 3.0 mbar, more preferably equal to or less than 2.0 mbar.
  • the temperature during the freeze-drying preferably lies in the range from equal to or more than 5°C, more preferably equal to or more than 10°C and still more preferably equal to or more than 20°C to equal to or less than 90°C, more preferably equal to or less than 80°C and most preferably equal to or less than 70°C.
  • the frozen particles can be submitted to plate freeze-drying at 0.50 mbar during primary drying and 0.18 mbar during secondary drying while applying a shelf heating temperature up to 50°C.
  • the time needed for freeze-drying can advantageously be reduced.
  • the drying process preferably takes in the range from equal to or more than 25 to equal to or less than 60 hours. In comparison with conventional processes, drying time can be reduced by using the process according to the invention.
  • the invention also provides a preferred improved freeze-drying process comprising radiation freeze-drying using a pressure in the range from 0.35 mbar to 0.45 mbar, preferably 0.40 mbar, during primary drying and a pressure in the range from 0.05 mbar to 0.15 mbar, preferably 0.10 mbar, during secondary drying, in combination with a shelf heating temperature up to 50°C, preferably in the range from 5°C to 50°C.
  • the frozen and/or freeze-dried solid additive composition is produced in the form of solid particles, preferably in the form of pellets or granules, preferably having a particle size distribution with a mean particle size in the range from preferably equal to or more than 0.5 mm, more preferably equal to or more than 1 mm, still more preferably equal to or more than 2 mm, and most preferably equal to or more than 3 mm to preferably equal to or less than 15 mm, more preferably equal to or less than 10 mm, still more preferably equal to or less than 8 mm and most preferably equal to or less than 6 mm.
  • the pellets or granules have a particle size distribution wherein the mean particle size ranges from 1 mm to 8 mm, preferably from 2 to 6 mm.
  • Particle size distributions and their mean particle size can be determined by sieving (such as for example described in ASTM D6913-04(2009)), size selective particle filtering and/or by particle size analyzers (PSAs) based on laser diffraction (LD).
  • the crystallization inhibitor is chosen from the group consisting of mono-saccharides, disaccharides and polysaccharides.
  • Examples of possible crystallization inhibitors include glucose, fructose, galactose, mannose, maltose, sucrose, lactose, starch, cyclodextrins and maltodextrin. More preferably the crystallization inhibitor is sucrose or maltodextrin.
  • the crystallization inhibitor comprises or consists of sucrose, carboxymethyl cellulose, maltodextrin or a combination of two or more of these.
  • the crystallization inhibitor preferably does not comprise or consist of maltodextrin.
  • the crystallization inhibitor comprises or consists of maltodextrin.
  • the crystallization inhibitor preferably does not comprise or consist of sucrose.
  • the crystallization inhibitor comprises or consists of sucrose.
  • the crystallization inhibitor is preferably present in an amount in the range from equal to or more than 0.05 % w/w to equal to or less than 40 % w/w, more preferably in the range from equal to or more than 0.5 % w/w to equal to or less than 20 % w/w, and most preferably in the range from equal to or more than 1% w/w to equal to or less than 14 % w/w of crystallization inhibitor, based on the total weight of the crystallization inhibitor and the formic acid or salt or ester thereof together.
  • the solid additive composition does not comprise purine or inosine.
  • the solid additive composition does not comprise yeast extract.
  • the solid additive does not comprise Arabic gum or any other type of gum.
  • the pellets further may or may not comprise hydrocolloid molecules such as for example sodium alginate, Cellulose gum, Gellan gum, Guar gum, Locust bean gum, Pectin, Xanthan gum.
  • the pellets do not comprise hydrocolloid molecules such as for example sodium alginate, Cellulose gum, Gellan gum, Guar gum, Locust bean gum, Pectin, Xanthan gum.
  • the pellets do comprise hydrocolloid molecules such as for example sodium alginate, Cellulose gum, Gellan gum, Guar gum, Locust bean gum, Pectin, Xanthan gum.
  • the additive is the formic acid or a salt or ester thereof.
  • the formic acid or a salt or ester thereof can be present as the formic acid itself.
  • the formic acid or a salt or ester thereof is present is a salt.
  • Preferred examples of a formic acid or a salt or ester thereof include sodium formate, potassium formate, ammonium formate, calcium formate, ethyl formate and methyl formate. More preferably the formic acid or a salt or ester thereof is a formate salt. Most preferably the formic acid or a salt or ester thereof is present in the form of ammonium formate, potassium formate or sodium formate. Sodium formate is especially preferred.
  • the formic acid or salt or ester thereof is preferably at least partly present in an amorphous state.
  • at least 10% w/w, more preferably at least 20% w/w, even more preferably at least 30% w/w, still more preferably at least 40% w/w and even still more preferably at least 50% w/w of the formic acid or salt or ester thereof is present in an amorphous state.
  • More preferably at least 60% w/w, even more preferably at least 70% w/w, still even more preferably at least 80% w/w, and yet more preferably at least 90% w/w of the formic acid or salt or ester thereof is present in an amorphous state.
  • Most preferably at least 95% w/w of the formic acid or salt or ester thereof is present in an amorphous state.
  • at least 99% w/w of the formic acid or salt or ester thereof is present in an amorphous state.
  • Amorphous content (amorphicity) in materials can be determined for example by Differential Scanning Calorimetry (DSC), Modulated DSC® (MDSC®), Continuous Relative Humidity Perfusion Microcalorimetry (cRHp), Dynamic Vapor Sorption (DVS), and Solution Calorimetry (SolCal).
  • DSC Differential Scanning Calorimetry
  • MDSC Modulated DSC®
  • CRRHp Continuous Relative Humidity Perfusion Microcalorimetry
  • DVDS Dynamic Vapor Sorption
  • SolCal Solution Calorimetry
  • an additive composition a composition comprising at least one compound that can suitably function as an additive in a starter culture comprising one or more selected strains of lactic acid bacteria.
  • the additive is the formic acid or a salt or ester thereof.
  • Formic acid or a salt thereof can be suitable as an additive for starter cultures, as described for example in WO2012/076665.
  • a solid additive composition is herein understood a composition in a solid form, in contrast to a composition in a liquid form.
  • a frozen form is herewith also understood to be a solid form.
  • the solid additive composition is a solid additive composition in a frozen or freeze-dried form.
  • the solid additive composition is a solid additive composition in a freeze- dried form.
  • the, preferably frozen or freeze-dried, solid additive composition comprises a water activity (Aw) of less than 0.2, more preferably an Aw of less than 0.1 .
  • the solid additive composition does not comprise any type of lactic acid bacteria present, individually, in a viable cell count density of equal to or more than 1 x 10 6 cfu/gram. More preferably the solid additive composition does not comprise any micro-organisms in a viable cell count density of equal to or more than 1 x 10 6 cfu/gram.
  • the solid additive composition is a lactic acid bacteria (LAB) depleted or LAB free composition, comprising, of each potentially present type of LAB individually, a viable cell count density of equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • LAB lactic acid bacteria
  • any lactic acid bacteria will have very limited activity or even be inactive in any process for preparing a cultured food product.
  • the solid additive composition is a microorganism depleted or microorganism free composition, comprising, of each potentially present type of microorganism individually, a viable cell count density of equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • any microorganism will have very limited activity or even be inactive in any process for preparing a cultured food product.
  • the solid additive composition preferably comprises less than 1 ,10 6 cfu/g, preferably less than 10 3 cfu/g, of each of Streptococcus spp, Leuconostoc spp, Lactococcus spp and/or Lactobacillus Leuconostoc spp.
  • the invention also provides a solid additive composition, in the form of a pellet, a granule, a tablet or a powder, comprising a mixture of:
  • composition comprises of any type of LAB present, individually, a viable cell count density of equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • the solid additive composition is a composition comprising a total amount of lactic acid bacteria (LAB) that is equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • LAB lactic acid bacteria
  • the solid additive composition is a composition comprising a total amount of microorganisms that is equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • cfu/g means "colony forming units per gram”. This is also referred to as the viable cell count density.
  • viable cell count density The determination of viable cell count density of the lactic acid bacterial strains described or claimed herein is known to the skilled person.
  • the viable cell count density (expressed in cfu/g or in cfu/ml) of Streptococcus thermophilus is preferably determined using Streptococcus Thermophilus Isolation Agar or STA.
  • This agar has the following composition: Casein enzymic hydrolysate 10.0 g/l; Yeast extract 5.0 g/l; Sucrose 10.0 g/l; Dipotassium phosphate 2.0 g/l; Agar 15.0 g/l; Final pH 6.8 +/- 0.2 at 25°C.
  • streptococci and lactobacilli can be together enumerated on the same STA plate; streptococci are counted as smooth, round colonies and lactobacilli are counted as fluffy colonies.
  • viable cell count density expressed in cfu/g or in cfu/ml
  • lactobacilli especially of Lactobacillus helveticus is preferably determined using tryptone glucose meat extract agar for example as detailed by Galesloot, T. E., F. Hassing, and J.
  • the solid additive composition is suitably present in the form of a pellet, a granule, a tablet, a powder or another solid particle. More preferably the solid additive composition is present as a pellet or a granule.
  • the pellets are preferably pellets manufactured by:
  • the pellets or granules have a particle size distribution wherein the mean particle size ranges from preferably equal to or more than 0.5 mm, more preferably equal to or more than 1 mm, still more preferably equal to or more than 2 mm, and most preferably equal to or more than 3 mm to preferably equal to or less than 15 mm, more preferably equal to or less than 10 mm, still more preferably equal to or less than 8 mm and most preferably equal to or less than 6 mm.
  • the pellets or granules have a particle size distribution wherein the mean particle size ranges from 1 mm to 8 mm, preferably from 2 to 6 mm.
  • Particle size distributions and their mean particle size can be determined by sieving (such as for example described in ASTM D6913-04(2009)), size selective particle filtering and/or by particle size analyzers (PSAs) based on laser diffraction.
  • The, preferably freeze-dried, solid additive composition is preferably uncoated. That is, the, preferably freeze-dried, solid additive composition is preferably not coated.
  • WO2012/076665 describes that in one embodiment of the therein claimed process, the freeze- dried microorganism and/or the stimulating additive added to the freeze-dried microorganism and/or the freeze-dried starter culture composition is coated. Without wishing to be bound by any kind of theory, it is believed that such a coating could disadvantageously reduce the working of the additive, i.e. the working of the formic acid or a salt or ester thereof. It is therefore preferred that the solid additive composition is uncoated. To create optimum contact, most preferably, neither the first solid composition nor the second solid composition is coated.
  • the present invention therefore also provides a kit of parts comprising:
  • a solid culture composition preferably in the form of a pellet, a granule, a tablet, a powder or another solid particle, wherein the solid culture composition comprises at least one type of lactic acid bacteria in an individual viable cell count density of equal to or more than 1 x 10 6 cfu/gram.
  • the kit of parts, or starter culture composition is a kit of parts, respectively starter culture composition, wherein the solid additive composition and/or the solid culture composition is in a frozen or freeze-dried form.
  • the kit of parts, or starter culture composition is a kit of parts, respectively starter culture composition, wherein: - the solid additive composition is present in the form of solid additive composition pellets, granules or tablets; and
  • the solid culture composition is present in the form of solid culture composition pellets, granules or tablets;
  • the solid additive composition pellets, granules or tablets are distinctive, different and/or separate from the solid culture composition pellets, granules or tablets.
  • the solid additive composition pellets, granules or tablets are not the same as the solid culture composition pellets, granules or tablets.
  • the solid additive composition comprises of any type of LAB present, individually, a viable cell count density of equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • a viable cell count density of equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most
  • the solid additive composition comprises a total amount of lactic acid bacteria (LAB) that is equal to or less than 1 x 10 6 cfu/gram, more preferably equal to or less than 1 x 10 5 cfu/gram, still more preferably equal to or less than 1 x 10 4 cfu/gram, even more preferably equal to or less than 1 x 10 3 cfu/gram, still even more preferably equal to or less than 1 x 10 2 cfu/gram and most preferably equal to or less than 1 x 10 1 cfu/gram.
  • LAB lactic acid bacteria
  • the solid culture composition preferably comprises a total amount of lactic acid bacteria (LAB) that is equal to or more than 1 x 10 6 cfu/gram, more preferably equal to or more than 1 x 10 5 cfu/gram, still more preferably equal to or more than 1 x 10 7 cfu/gram, even more preferably equal to or more than 1 x 10 8 cfu/gram, still even more preferably equal to or more than 1 x 10 9 cfu/gram and most preferably equal to or more than 1 x 1O 10 cfu/gram.
  • LAB lactic acid bacteria
  • the solid culture composition comprises of any type of LAB present, individually, a viable cell count density that is equal to or more than 1 x 10 6 cfu/gram, more preferably equal to or more than 1 x 10 5 cfu/gram, still more preferably equal to or more than 1 x 10 7 cfu/gram, even more preferably equal to or more than 1 x 10 8 cfu/gram, still even more preferably equal to or more than 1 x 10 9 cfu/gram and most preferably equal to or more than 1 x 1O 10 cfu/gram.
  • the solid culture composition comprises more than 1 ,10 6 cfu/g, preferably more than 10 9 cfu/g, of one or more of Streptococcus spp, Leuconostoc spp, Lactococcus spp and/or Lactobacillus Leuconostoc spp., bifidobacteria, propioni bacteria, Enterococci.
  • the solid culture composition comprises more than 1.10 6 cfu/g, preferably more than 10 9 cfu/g, of one or more of bacteria belonging to the genus of Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., Lactococcus spp., more preferably one or more of as Lactobacillus delbruekii subsp.
  • the solid culture composition may in addition comprise one or more cryoprotectants. Suitable cryoprotectants include those described in WO2012/076665, herein incorporated by reference. However, the cryoprotectant(s) and the crystallization inhibitors have different functions. Hence, preferably, within one kit of parts or within one starter culture composition, any potential cryoprotectant(s) in the solid culture composition, and the crystallization inhibitor in the solid additive composition, are different compounds. That is, preferably any potential cryoprotectant(s) in the solid culture composition are not the same compound as the crystallization inhibitor in the solid additive composition.
  • the solid culture composition and/or the solid additive composition is frozen or freeze- dried.
  • both the solid culture composition and the solid additive composition are present in the form of frozen or freeze-dried pellets, granules or tablets.
  • kit of parts can suitably be used as a starter culture composition.
  • “Starter culture” is defined herein as a preparation containing microbial cells that is intended for inoculating a medium to be fermented.
  • “Microorganism” as used herein includes, but is not limited to, algae, protozoa, viruses, bacteria (such as lactic acid bacteria, Brevibacterium and Propionibacterium) and fungi (such as the genera Penicillium, Geotrichum, Saccharomyces and Kluyveromyces).
  • the microorganisms may be non-modified (i.e. wild type) or genetically modified. Genetically modified microorganisms may be provided with and without the use of recombinant DNA-technology.
  • the invention advantageously also provides for the use of such a starter culture composition in a process for making a food or feed product.
  • the invention further provides a process of producing a cultured food product, the process comprising the steps of: a) inoculating a precursor material with a starter culture composition prepared according to the process of the invention or a kit of parts as described above. b) culturing the precursor material with the starter culture composition or kit of parts, and c) producing the cultured food product.
  • the cultured food product is a fermented milk product.
  • milks from mammals and plant sources or mixtures thereof are preferred. More preferably, the milk is from a mammal source.
  • Preferred fermented milk products include yoghurt (including for example set yoghurt, low fat yoghurt, non-fat yoghurt), kefir, dahi, ymer, buttermilk, butterfat, sour cream and sour whipped cream as well as fresh cheeses such as quark and cottage cheese.
  • Petit Suisse or Mozarella is yet another example of a fermented milk product.
  • the fermented milk product is a yoghurt.
  • the invention also provides a use of the solid additive composition as described above, or a kit of parts as described above, in a process for making a food or feed product.
  • the solid additive compositions comprising sodium formate formulated with maltodextrin (2 and 4 w/w% on dm) is freeze drying faster than the NaF as such (100%).
  • the overall drying time could therefore advantageously be reduced by more than 15-20%.

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Abstract

L'invention propose un procédé de préparation d'une composition d'additif solide congelée ou lyophilisée, comprenant les étapes suivantes : a) préparation d'un mélange aqueux comprenant ou consistant en (i) de l'acide formique ou un sel ou ester de celui-ci et (ii) un inhibiteur de cristallisation ; b) préparation d'une composition d'additif solide congelée par congélation du mélange aqueux de a) ; et c) éventuellement préparation d'une composition d'solide lyophilisée par lyophilisation de la composition d'additif solide congelée de b). De plus, l'invention concerne une composition d'additif solide ainsi préparée.
PCT/EP2023/083437 2022-11-29 2023-11-28 Procédé de préparation d'une composition d'additif solide congelée ou lyophilisée WO2024115523A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000039281A2 (fr) 1998-08-26 2000-07-06 Chr. Hansen A/S Cultures de levain sous forme liquide faisant preuve d'une stabilite au stockage amelioree et leurs applications
US20050069862A1 (en) 2003-07-02 2005-03-31 Chr. Hansen A/S Use of compounds involved in biosynthesis of nucleic acids as cryoprotective agents
WO2009056979A1 (fr) 2007-10-30 2009-05-07 Danisco A/S Milieu de croissance pour bactéries d'acide lactique
WO2012076665A1 (fr) 2010-12-10 2012-06-14 Dsm Ip Assets B.V. Compositions de ferments
US20220064588A1 (en) * 2018-12-18 2022-03-03 Dsm Ip Assets B.V. Phage resistant lactic acid bacteria

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000039281A2 (fr) 1998-08-26 2000-07-06 Chr. Hansen A/S Cultures de levain sous forme liquide faisant preuve d'une stabilite au stockage amelioree et leurs applications
EP1141233B1 (fr) * 1998-08-26 2008-12-10 Chr. Hansen A/S Cultures starter liquides avec stabilite au stockage amelioree et leurs applications
US20050069862A1 (en) 2003-07-02 2005-03-31 Chr. Hansen A/S Use of compounds involved in biosynthesis of nucleic acids as cryoprotective agents
WO2009056979A1 (fr) 2007-10-30 2009-05-07 Danisco A/S Milieu de croissance pour bactéries d'acide lactique
WO2012076665A1 (fr) 2010-12-10 2012-06-14 Dsm Ip Assets B.V. Compositions de ferments
US10370636B2 (en) * 2010-12-10 2019-08-06 Dsm Ip Assets B.V. Starter culture compositions
US20220064588A1 (en) * 2018-12-18 2022-03-03 Dsm Ip Assets B.V. Phage resistant lactic acid bacteria

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
GALESLOOT, T. E.F. HASSINGJ. STADHOUDERS., NETH. MILK DAIRY, vol. 15, 1961, pages 127 - 150
STÉPHANIE PASSOT ET AL: "Critical water activity and amorphous state for optimal preservation of lyophilised lactic acid bacteria", FOOD CHEMISTRY, vol. 132, no. 4, 1 June 2012 (2012-06-01), pages 1699 - 1705, XP055053098, ISSN: 0308-8146, DOI: 10.1016/j.foodchem.2011.06.012 *

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