Dry yeast propionate, production process thereof, method of preserving a food or cosmetic product, food or cosmetic product.

Description

1 / 24 “DRY PROPIONATE YEAST, PROCESS FOR PRODUCING THE SAME, METHOD FOR PRESERVING A FOOD OR COSMETIC PRODUCT, FOOD OR COSMETIC PRODUCT” TECHNICAL FIELD OF THE INVENTION

[0001] The present invention relates to a dry fermented product comprising calcium propionate.

[0002] The dry fermented product of the present invention has a very high propionate content and can be suitablely applied as a preservative in, e.g., food and cosmetic products.

[0003] The invention also relates to a process comprising: □ the supply of an aqueous fermentation medium comprising hydrolyzed glucan; □ Inoculation of the fermentation medium with microorganisms that produce propionic acid; □ Incubating the inoculated fermentation medium to produce a fermented product, while simultaneously adding an alkalizing agent to maintain the pH within a predefined range.

[0004] The invention also relates to the use of the dried fermented product in a food or cosmetic product. BACKGROUND OF THE INVENTION

[0005] Preservatives extend the shelf life of baked goods by inhibiting the growth of spore-forming bacteria and molds. Spore-forming bacteria (Bacillus pumilus) can be carried over during the baking process and cause an unpleasant texture and odor in yeast-fermented products such as bread. Molds can contaminate products after baking and then grow on the outer surface. Mold contamination is considered a serious problem among bakers, and the conditions commonly found in baking provide almost ideal conditions for mold growth. Petition 870250089332, dated 10 / 01 / 2025, pp. 104 / 127 2 / 24

[0006] Acidity reduces the chance of spoilage. Most preservatives have acids as their active ingredient. Preservatives can be classified according to the type of acid they contain: mineral, organic, or fatty. Mineral acids (such as sulfuric, hydrochloric, and phosphoric) are the strongest and therefore have the greatest effect on pH. However, they only work by lowering the pH and do not contain any other inhibitors. Organic acids (such as lactic, benzoic, and tartaric) have less effect on pH than mineral acids, but are better preservatives because they have additional inhibitory activity. Fatty acids (such as acetic, sorbic, and propionic) are a specific type of organic acid with even stronger inhibitory activity.

[0007] Calcium propionate is used as a preservative in a wide variety of products, including: bread, other baked goods, processed meat, whey, and other dairy products. Propionates prevent microbes from producing the energy they need, as do benzoates. However, unlike benzoates, propionates do not require an acidic environment.

[0008] Propionate is a commercially valuable carboxylic acid produced by microbial fermentation. Biological production of propionate is limited by the high downstream purification costs.

[0009] Document WO 2010 / 097362 describes a method for manufacturing a mixed solution comprising propionate and acetate, comprising the steps of: - supply of a fermentation product comprising propionate, acetate and carbonate-related compounds, - Addition of acid to the fermentation product to obtain an acidified fermentation product comprising propionate and acetate with a pH in the range of 2.5 to less than 8.

[0010] Document WO 2016 / 146721 describes a process for manufacturing propionate products by fermentation, comprising the following steps: Petition 870250089332, dated 10 / 01 / 2025, pp. 105 / 127 3 / 24 - fermentation of a carbon source selected from sugars and lactate in a fermentation medium by a propionic acid-producing microorganism to provide a first fermentation broth comprising a propionate salt, - recovery of the propionic acid-producing microorganism from the first fermentation broth, - subjecting the first fermentation broth, from which propionic acid-producing microorganisms were recovered, to a water removal step to form a first propionate salt product, - fermentation of a carbon source comprising glycerol with the propionic acid-producing microorganism recovered from the first fermentation broth in the presence of an inorganic alkaline salt to provide a second fermentation broth comprising a propionate salt, - subjecting the second fermentation broth to a purification step comprising at least one precipitation step, to form a second propionate salt product.

[0011] Document WO 2019 / 108064 describes an organic acid preservative system comprising an acetate component and a propionate component, wherein these constitute at least 85% by weight of the total amount of carboxylic acids and carboxylic acid salts in the preservative system and wherein the propionate component constitutes 12 to 50% by weight of said total amount of carboxylic acids and carboxylic acid salts.

[0012] Document CN 115 678 925 describes a method for preparing calcium propionate by fermenting a medium containing 0.1% Na2HPO4 and 0.1% MgSO4.7H2O with Propionibacterium, followed by membrane filtration, reaction of the isolated propionate with calcium hydroxide, evaporation and crystallization.

[0013] Document CN 112 176 005 describes a method for producing calcium propionate, comprising fermenting a fermentation medium with an acid-producing strain while maintaining the pH between 7 and 2.5, Petition 870250089332, dated 10 / 01 / 2025, pp. 106 / 127 4 / 24 sterilization, nanofiltration, and the addition of an aqueous solution of calcium hydroxide. After the reaction is complete, the mixture is concentrated and dried to obtain solid calcium propionate. SUMMARY OF THE INVENTION

[0014] The inventors discovered that a dry propionate ferment with a very high propionate content and a bland taste can be obtained if a carefully balanced combination of alkaline calcium salt and alkaline metal salt is used in the preparation of the ferment.

[0015] The dry fermented product of the present invention comprises, per kg of dry matter: □ 4.7 to 8.1 mol of propionate; □ 1.2 to 3.0 mol of calcium cation; □ 2.4 to 7.8 mol of alkali metal cation selected from sodium, potassium, and combinations thereof; □ 1 to 50 grams of glucose equivalent of hydrolyzed glucan; The calcium cation and propionate are present in the dry fermented product in a molar ratio of 0.2 to 0.6.

[0016] In addition to the intrinsic buffering capacity of the acid salts present, the dry fermentate of the present invention has limited buffering capacity, probably due to the removal of buffering components contained in the biomass, such as proteins. This reduced buffering capacity is advantageous because the fluctuations in buffering capacity observed between production batches are effectively minimized. Likewise, if a pH adjustment is required when the dry fermentate is applied to a food product, less acid or base is needed to achieve it.

[0017] The invention also provides a process for producing a fermented product, said process comprising the following steps: □ provision of an aqueous fermentation medium comprising 30 to 200 g / L of glucose equivalent of hydrolyzed glucan; Petition 870250089332, dated 10 / 01 / 2025, pp. 107 / 127 5 / 24 □ Inoculation of the fermentation medium with a propionic acid-producing microorganism; □ Incubation of the inoculated fermentation medium for at least 20 hours to produce a fermented product, simultaneously adding an alkalizing agent to maintain the pH in the range of 5.0 to 7.5, said alkalizing agent including a water-soluble alkali metal salt and a water-soluble alkali calcium salt; wherein the fermented product at the end of the incubation stage comprises calcium cation and propionate in a molar ratio of 0.3 to 0.8 and alkali metal cation and propionate in a molar ratio of 0.3 to 1.4.

[0018] The fermented product obtained by this process offers the advantage of introducing less bitterness than existing calcium propionate fermented products when used to provide a certain propionate content in a final product. As a result, the present fermented product can be appropriately applied in higher concentrations than existing propionate fermented products, thus achieving better preservation.

[0019] The invention also relates to a method of preserving a food or cosmetic product, said method comprising adding the dry fermented product of the present invention at a concentration of 0.1 to 5% by weight of the food product.

[0020] Another aspect of the invention relates to a food or cosmetic product obtained through the preservation method mentioned above. DETAILED DESCRIPTION OF THE INVENTION

[0021] A first aspect of the invention relates to a dry fermented product having a moisture content of less than 15% by weight and comprising, per kg of dry matter: □ 4.7 to 8.1 mol of propionate; □ 1.2 to 3.0 mol of calcium cation; □ 2.4 to 7.8 mol of alkali metal cation selected from sodium, potassium, and combinations thereof; Petition 870250089332, dated 10 / 01 / 2025, pp. 108 / 127 6 / 24 □ 1 to 50 grams of glucose equivalent of hydrolyzed glucan; The calcium cation and propionate are present in the dry fermented product in a molar ratio of 0.2 to 0.6.

[0022] The term “propionate”, as used herein, encompasses propionic acid as well as its salts. Likewise, the terms “acetate”, “succinate”, and “lactate” also encompass both the acidic and salt forms.

[0023] The term “glucan,” as used herein, refers to a linear or branched polymer consisting of glucose monomeric units. Examples of glucans include amylose, amylopectin, cellulose, and glycogen.

[0024] Hydrolyzed glucan concentrations, expressed as “glucose equivalent,” refer to the total amount of glucose present in the hydrolyzed glucan, including both free glucose and glucose contained in hydrolysis fragments with a degree of polymerization of two or more. The amount of glucose equivalent present in a hydrolyzed glucan can be determined by completely hydrolyzing the hydrolyzed glucan and measuring the glucose content.

[0025] The dry fermented product of the present invention preferably has a moisture content of less than 12% by weight, more preferably from 1 to 10% by weight.

[0026] Propionate is preferably contained in the dry fermentate of the present invention at a concentration of 5.2 to 7.8 mol, more preferably 5.8 to 7.2 mol, per kg of dry matter.

[0027] The dry fermented product preferably contains calcium cations at a concentration of 1.4 to 2.8 mol, more preferably 1.7 to 2.5 mol, per kg of dry matter.

[0028] The hydrolyzed glucan is preferably contained in the dry fermented product at a concentration of 2 to 30 grams, more preferably 3 to 15 grams, of glucose equivalent per kg of dry matter.

[0029] According to a particularly preferred embodiment, the dry ferment contains calcium cation and propionate in a molar ratio of 0.2 to 0.5, most preferably 0.22 to 0.4. Petition 870250089332, dated 10 / 01 / 2025, pp. 109 / 127 7 / 24

[0030] The hydrolyzed glucan in the dry fermented product is preferably selected from hydrolyzed amylose, hydrolyzed amylopectin, hydrolyzed cellulose, hydrolyzed glycogen, and combinations thereof. More preferably, the hydrolyzed glucan comprises hydrolyzed amylose and / or hydrolyzed amylopectin. Both amylose and amylopectin are components of starch.

[0031] The hydrolyzed glucan preferably comprises at least one or more saccharides selected from glucose, maltose, isomaltose, maltotriose, maltotetraose and combinations thereof. According to a particularly preferred embodiment, the dry ferment contains, per kg of dry matter, 0.5 to 10 grams of one or more saccharides selected from glucose, maltose, isomaltose, maltotriose, maltotetraose and combinations thereof, more preferably 0.8 to 5 grams of one or more saccharides selected from glucose, maltose, isomaltose, maltotriose, maltotetraose and combinations thereof.

[0032] According to a preferred embodiment, the dry fermentate of the present invention comprises DNA from a propionic acid-producing microorganism, preferably from a bacterium belonging to the genera Propionibacterium and Acidipropionibacterium, more preferably from a bacterium selected from Acidipropionibacterium acidipropionici, Propionibacterium freudenreichii, Propionibacterium shermanii, Acidipropionibacterium thoenii, Acidipropionibacterium jensenii and combinations thereof. The presence of DNA from a propionic acid-producing microorganism can be suitably determined using a PCR test.

[0033] In a particular preferred embodiment, the dry ferment comprises, per kg of dry matter, 3.0 to 7.44 mol, more preferably 4.0 to 7.0 mol and most preferably 5.3 to 6.5 mol of alkali metal cation selected from sodium, potassium and combinations thereof.

[0034] In another preferred embodiment, the dry fermented product comprises, per kg of dry matter, at least 3.4 mol, more preferably at least 4.0 mol and most preferably at least 5.3 mol of sodium cation. Petition 870250089332, dated 10 / 01 / 2025, pp. 110 / 127 8 / 24

[0035] In addition to propionate, the dry fermentate of the present invention may comprise other salts of organic acids, such as acetate and / or succinate.

[0036] Preferably, the dry ferment contains, per kg of dry matter, 1.5 to 3.0 mol, more preferably 1.7 to 2.8 mol and most preferably 1.9 to 2.6 mol of acetate.

[0037] Succinate is preferably present in the dry fermented product at a concentration of 0.4 to 1.2 mol per kg of dry matter, more preferably 0.5 to 1.1 mol per kg of dry matter and most preferably 0.55 to 1.0 mol per kg of dry matter.

[0038] The combination of propionate, acetate and succinate is preferably contained in the dry fermented product at a concentration of 7.5 to 11.0 mol per kg of dry matter, more preferably 8.2 to 10.5 mol per kg of dry matter and most preferably 8.7 to 10.2 mol per kg of dry matter.

[0039] The dry ferment preferably contains carbonate at a concentration of 0 to 20 mmol per kg of dry matter, more preferably 0 to 10 mmol per kg of dry matter and most preferably 0 to 5 mmol per kg of dry matter.

[0040] Furthermore, in a preferred embodiment, the dry ferment contains calcium cation and alkali metal cation in a molar ratio of 0.25 to 0.62, more preferably 0.30 to 0.55 and most preferably 0.32 to 0.52.

[0041] According to another preferred embodiment, the combination of propionate, acetate, calcium cation and alkali metal cation constitutes 90 to 99% by weight, more preferably 92 to 98% by weight and most preferably 93 to 97% by weight of the dry matter contained in the fermented product.

[0042] According to a particularly preferred embodiment, the dry ferment contains propionate at a concentration X, acetate at a concentration Y, succinate at a concentration Z, calcium cation at a concentration A, sodium cation at a concentration B, and potassium at a concentration C, wherein all said concentrations are expressed in mol / kg of dry matter and wherein 0.8 < Petition 870250089332, dated 10 / 01 / 2025, pp. 111 / 127 9 / 24 (X+Y+2Z) / (2A+B+C) < 1.2, preferably 0.85 < (X+Y+2Z) / (2A+B+C) < 1.15 and most preferably 0.90 < (X+Y+2Z) / (2A+B+C) < 1.12.

[0043] The dry fermented product of the present invention advantageously has a nearly neutral pH. Accordingly, in a preferred embodiment, dispersing 100 grams of fermented product in 1 liter of water at 20 °C produces an aqueous liquid having a pH in the range of 6 to 8.

[0044] The dry fermented product of the present invention is highly soluble in water. Preferably, when 100 grams of the fermented product are dispersed in 1 liter of water at 20 °C, at least 95% by weight, more preferably 98% by weight, and most preferably at least 99% by weight of the fermented product are dissolved.

[0045] The protein content of the dry fermented product preferably ranges from 0 to 30 grams, more preferably from 0 to 10 grams, and most preferably from 0 to 5 grams, per kg of dry matter.

[0046] The dry ferment is preferably a powder, in which at least 80% by weight of the particles have a diameter not exceeding 500 μm. More preferably, the dry ferment is a powder in which at least 80% by weight of the particles have a diameter in the range of 40 to 400 μm. The particle size distribution of the dry ferment can be suitably determined by laser diffraction.

[0047] As mentioned above, the dry fermentate of the present invention offers the advantage that, in addition to the theoretical buffering capacity of the acid salts present in the fermentate, it has a limited actual buffering capacity. The dry fermentate of the present invention preferably has an average dynamic buffering capacity (β) of less than 27 mM / g, more preferably from 12 to 26 mM / g, or more preferably from 16 to 25 mM / g. The static buffering capacity (β) is defined as the mmoles of HCl per gram of dry fermentate required to change the pH of a solution of said dry fermentate (0.2 g / 100 mL of water) by 1. The buffering capacity (β) of a dry fermentate as a function of pH can be determined by titrating an aqueous solution of fermentate (0.2 grams of dry fermentate in 100 mL of water) Petition 870250089332, dated 10 / 01 / 2025, pp. 112 / 127 10 / 24 mL of water) with 0.1 N HCl solution to reduce the pH to 2.5. The average dynamic buffer capacity (β), which takes into account the impact of the titrant volume, can be easily determined by plotting the buffer capacity (β) as a function of pH and applying the mean volume theorem to the titration data of β according to the calculations below. The average dynamic buffer capacity (β) best illustrates the differences in fermented products that affect their functionality. P«2 β =---π----π-· í β(ρΗ)·άρΗ m pH1 — pH2J PH1 where m = sample mass, c = sample concentration, and V = titrant volume.

[0048] Dry ferments containing significant amounts of buffering components other than organic acid salts typically have a higher average dynamic buffering capacity than the dry ferments of the present invention. Likewise, the pKa pH of these dry ferments is usually higher than the pKa pH of the dry ferments of the present invention.

[0049] As mentioned earlier, dry ferment has a desirable flavor profile. In particular, dry ferment has a low bitterness intensity. Preferably, a solution obtained when dry ferment is dissolved in demineralized water at 20 °C to achieve a propionate concentration of 140 mmol / L has a bitterness index of less than 50 on a line scale of 0 to 100. This bitterness index can be measured using conventional sensory methods, such as difference tests with anchored line scales. For example, to compare and evaluate pairs of samples, a rating test can be used with instructions to rate the samples according to bitterness intensity using an anchored line scale with “low” at one end and “high” at the other. Responses can be captured electronically using software such as EyeQuestion®, and analyzed using software such as Petition 870250089332, dated 10 / 01 / 2025, pp. 113 / 127 11 / 24 EyeOpen® software assigns values ​​from 0 to the "low" end of the scale and 100 to the "high" end, and then determines the degree of statistical significance of the values ​​by comparing the jurors' scores. Trained jurors are not required for these difference and ranking tests.

[0050] Furthermore, the flavor profile of the dry fermented product contributes to desirable hedonic scores for food products that include it. In some embodiments, the food product comprising the dry fermented product at a concentration that provides 140 mmol / L of propionate per kg has a hedonic preference score not very different from a food product comprising calcium propionate at the same molar concentration of 140 mmol / L. For hedonic testing, a questionnaire with a nine-point preference scale can be used. The nine points include: Extremely Unpleasant, Very Unpleasant, Moderately Unpleasant, Slightly Unpleasant, Neither Pleasant Nor Unpleasant, Slightly Pleasant, Moderately Pleasant, Very Pleasant, and Extremely Pleasant. A range of attributes that can be evaluated for preference includes overall preference, aroma preference, flavor preference, and aftertaste preference.The jurors' responses can be captured and analyzed electronically using software such as EyeQuestion® and EyeOpenR.

[0051] Another aspect of the invention relates to a process for preparing a fermented product, preferably a dry fermented product according to the present invention, said process comprising the following steps: □ provision of an aqueous fermentation medium comprising 30 to 200 g / L of glucose equivalent of hydrolyzed glucan; □ Inoculation of the fermentation medium with a propionic acid-producing microorganism; □ Incubate the inoculated fermentation medium for at least 20 hours to produce a fermented product, adding an alkalizing agent at the same time. Petition 870250089332, dated 10 / 01 / 2025, pp. 114 / 127 12 / 24 to maintain the pH in the range of 5.0 to 7.5, said alkalizing agent including a water-soluble alkali metal salt and a water-soluble alkali calcium salt; wherein the fermented product at the end of the incubation stage comprises calcium cation and propionate in a molar ratio of 0.2 to 0.8 and alkali metal cation and propionate in a molar ratio of 0.3 to 1.4.

[0052] The aqueous fermentation medium preferably comprises 40 to 180 g / L, more preferably 50 to 150 g / L and most preferably 60 to 120 g / L, of glucose equivalent of hydrolyzed glucan.

[0053] According to a particularly preferred embodiment, the glucan hydrolysate in the aqueous fermentation medium is selected from hydrolyzed amylose, hydrolyzed amylopectin, hydrolyzed cellulose, hydrolyzed glycogen, and combinations thereof. More preferably, the hydrolyzed glucan hydrolysate comprises hydrolyzed amylose and / or hydrolyzed amylopectin. Both amylose and amylopectin are components of starch.

[0054] According to a particularly preferred embodiment, hydrolyzed glucan is obtained by hydrolysis of wheat starch.

[0055] In a particularly preferred embodiment of the invention, the hydrolyzed glucan employed in the present process is a partially hydrolyzed starch and, during incubation, the partially hydrolyzed starch is further hydrolyzed by added enzymes. Preferably, glucose does not constitute more than 70% by weight, more preferably 10 to 60% by weight, of the partially hydrolyzed starch.

[0056] Examples of enzymes that can be added to the fermentation medium to further hydrolyze the partially hydrolyzed starch include glucoamylase, pullulanase, alpha-amylase, beta-amylase and combinations thereof. According to a particularly preferred embodiment, the added enzyme includes glucoamylase.

[0057] Examples of propionic acid-producing microorganisms that can be employed in the present process include Acidipropionibacterium acidipropionici, Acidipropionibacterium freudenreichii, Acidipropionibacterium Petition 870250089332, dated 10 / 01 / 2025, pp. 115 / 127 13 / 24 shermanii, Acidipropionibacterium thoenii, Acidipropionibacterium jensenii and combinations thereof. More preferably, the propionic acid-producing microorganism used is Acidipropionibacterium acidipropionici.

[0058] The incubation stage is preferably carried out under anaerobic conditions.

[0059] The fermentation medium is preferably incubated at a temperature in the range of 20 to 45 °C, more preferably 25 to 42 °C and most preferably 30 to 40 °C.

[0060] The duration of the incubation phase is preferably in the range of 22 to 120 hours, more preferably from 24 to 72 hours.

[0061] To achieve a high yield of propionate, it is advantageous to continue the incubation step until virtually all the hydrolyzed glucan has been digested. Accordingly, in a preferred embodiment, the incubation is continued until the fermentation medium contains no more than 10 g / L, more preferably no more than 5 g / L, more preferably 0.1 to 3 g / L and most preferably 0.3 to 2 g / L, of free glucose.

[0062] The incubation step is preferably continued until the propionate concentration in the fermentation medium has increased to at least 100 mmol / L, more preferably to at least 150 mmol / L and most preferably to 250 to 300 mmol / L.

[0063] The alkalizing agent used to control pH includes (i) a water-soluble alkali metal salt and (ii) a water-soluble alkaline calcium salt. These two different alkalizing agents may be added to the fermentation medium simultaneously or sequentially. Preferably, the alkali metal salt and the calcium salt are added simultaneously.

[0064] The water-soluble alkali metal salt is preferably selected from sodium hydroxide, potassium hydroxide, and combinations thereof.

[0065] The water-soluble alkaline calcium salt is preferably calcium hydroxide. Petition 870250089332, dated 10 / 01 / 2025, pp. 116 / 127 14 / 24

[0066] According to a particularly preferred embodiment of the present process, the alkalizing agent is added to the fermentation medium to maintain the pH in the range of 5.5 to 7.4, more preferably from 5.8 to 7.2.

[0067] At the end of the incubation stage, the fermented product preferably comprises calcium cation and propionate in a molar ratio of 0.4 to 0.7, more preferably 0.45 to 0.68.

[0068] The alkali metal cation and propionate are preferentially present in the fermented product at the end of the incubation stage in a molar ratio of 0.4 to 1.2, more preferably 0.5 to 1.0.

[0069] According to a particularly preferred embodiment of the present invention, the present process contains the additional steps of: □ Removal of undissolved solids from the fermented product by centrifugation or filtration to produce a fermented product with a low solids content; and □ Drying of the fermented product with a low solids content.

[0070] The present process offers the advantage of enabling a surprisingly effective removal of undissolved solids, including biomass, thus producing a fermented product with a low solids content, in which most of the solute consists of propionate. In this way, it is possible to obtain a dry fermented product having a propionate content greater than 50% propionic acid equivalent. This dry fermented product offers the additional advantage of having a low average dynamic buffering capacity.

[0071] In the present process, undissolved solids can be removed by known solid-liquid separation techniques, such as centrifugation, filtration, decantation, and hydrocyclones. More preferably, undissolved solids are removed by centrifugation or filtration. More preferably, undissolved solids are removed by disc stack centrifugation.

[0072] The inventors observed that the undissolved solids removed from the fermented product include a significant amount of calcium salts. As a result, the molar ratio of calcium cation to propionate is substantially reduced by Petition 870250089332, dated 10 / 01 / 2025, pages 117 / 127 15 / 24 removal of undissolved solids. The low-solids ferment obtained after removal of undissolved solids preferably comprises calcium cation and propionate in a molar ratio of 0.2 to 0.5, more preferably 0.2 to 0.4.

[0073] The removal of undissolved solids does not significantly affect the molar ratio of alkali metal cation to propionate. Preferably, the alkali metal cation and propionate are present in the low-solids fermentate in a molar ratio of 0.3 to 1.4, more preferably 0.4 to 1.2, and most preferably 0.5 to 1.0.

[0074] After the removal of undissolved solids, the present process preferably comprises the additional step of concentrating the low-solids fermented product to a dry solids content of at least 10% by weight, more preferably from 12 to 40% by weight and most preferably from 16 to 30% by weight.

[0075] Low solids fermented product or low solids concentrated fermented product is preferably dried to a moisture content of less than 15% by weight, more preferably less than 12% by weight and most preferably from 1 to 10% by weight.

[0076] Drying of low-solids fermented product or low-solids concentrated fermented product is preferably carried out by spray drying, drum drying, freeze-drying or a combination thereof. Spray drying is most preferably carried out.

[0077] According to a particularly preferred embodiment, the present process produces a dry fermented product as described above.

[0078] Another aspect of the invention relates to a method of preserving a food or cosmetic product with the dried ferment, wherein said method comprises incorporating the dried ferment of the present invention at a concentration of 0.1 to 5% by weight of the food or cosmetic product, more preferably 0.2 to 2%. Petition 870250089332, dated 10 / 01 / 2025, pp. 118 / 127 16 / 24

[0079] The present method is particularly suitable for the preservation of food products, especially bakery products and processed meat products. More preferably, the present method comprises adding the dry ferment to a bakery product.

[0080] Yet another aspect of the invention relates to a food or cosmetic product obtained through the present preservation method.

[0081] In a preferred embodiment, the product obtained through the preservation method is a food product, more preferably one selected from among a bakery product and a processed meat product, and most preferably a bakery product.

[0082] The food product of the present invention preferably has a hedonic preference score not significantly lower than that of a food product in which the dry fermented product has been replaced by a mixture of calcium propionate and starch.

[0083] The invention is further illustrated by the following non-limiting examples. EXAMPLES Example 1

[0084] Two dry fermented products containing propionate were prepared from a fermentation medium containing wheat starch syrup as the sole source of fermentable carbohydrate. The composition of the fermentation medium is shown in Table 1. Table 1 Compounds Concentration [g / kg] Glucose equivalent of wheat starch syrup 70 Yeast extract 6.6 Vitamins and minerals 2.1 28% ammonia solution w / v 6.2 Inoculum 1 75.5 Enzyme mixture 2 1.1 Petition 870250089332, dated 10 / 01 / 2025, pp. 119 / 127 17 / 24 Remaining water Contains Acidipropionibacterium acidipropionici It comprises glucoamylase, pullulanase, alpha-amylase and lysophospholipase

[0085] Water, magnesium sulfate, potassium sulfate, potassium dihydrogen phosphate, thiamine hydrochloride, biotin and calcium pantothenate were added to the fermenters and the trace element solution thus obtained was sterilized in situ at 121 °C for 20 minutes.

[0086] Next, the fermenters were cooled to ~40°C and a sterile aqueous dilution in UAT of wheat starch syrup and yeast extract was added to the fermenters. The trace element solution was added aseptically to the fermenters and the flasks containing the solution were subsequently rinsed with sterile demineralized water to ensure the transfer of all trace elements to the fermenter.

[0087] Before inoculation, the pH of the fermentation medium was adjusted to 7.0 using an ammonia solution. The initial glucose concentration was approximately 27 g / L.

[0088] Two different fermentations were carried out. Fermentation 1 was carried out in accordance with the invention. Fermentation A was not in accordance with the invention.

[0089] In Fermentation 1, an ammonia solution and a Ca(OH)2 / NaOH mixture (60:40 w / w) were used to control the pH. The ammonia solution was added during the first 12 hours of fermentation. After the ammonia solution was depleted, it was switched to the Ca(OH)2 / NaOH mixture. Fermentation A was carried out in the same way, except that Ca(OH)2 was used as the sole base to control the pH.

[0090] Fermentations were initiated by adding the inoculum. After inoculation, the enzyme mixture was aseptically added to the fermenter and the flasks were subsequently rinsed with sterile demineralized water to ensure the addition of all enzymes. Petition 870250089332, dated 10 / 01 / 2025, pages 120 / 127 18 / 24

[0091] Fermentations were carried out under sterile conditions at pH 7.0, 35 °C and under constant stirring.

[0092] When free glucose concentrations reached 8 g / L (total glucose of approximately 13 g / L), the addition of base was stopped and fermentation continued until a pH of 5.8 was reached. This procedure minimized carbonate and foam formation during downstream processing, as well as reducing the amount of solids in the final product.

[0093] The fermented products thus obtained were concentrated, sterilized and spray-dried, producing Product 1 (from Fermented Product 1) and Product A (from Fermented Product A).

[0094] The compositions of the spray-dried ferments were analyzed. The results are summarized in Table 2. Table 2 Percentage by weight Product 1 Product A Acetate 7.2 7.4 Lactate 0.8 0.8 Propionate 31.5 30.0 Succinate 5.7 4.3 Calcium 11 18 Sodium 9.4 1.0 Moisture 2.0 2.4 pH at 10% (w / v) dilution 6.3 6.6

[0095] At least 95% by weight of both spray-dried ferments had a particle size not exceeding 300 µm. Carbohydrate analysis of Product 1 showed that it contained: □ Glucose: 0.75 g / kg □ Fructose: 0.1 g / kg Petition 870250089332, dated 10 / 01 / 2025, pages 121 / 127 19 / 24 □ Isomaltose: 2.25 g / kg □ Maltose: 0.35 g / kg Example 2

[0096] Example 1 was repeated to produce Fermented 1. Then, the biomass was removed from the fermented by disc-stack centrifugation and then the clarified fermented was concentrated, sterilized and spray-dried.

[0097] The composition of the spray-dried product (Product 2) The results are summarized in Table 3. Table 3 Percentage by weight: Acetate 14.7%, Lactate 1.7%, Propionate 50.4%, Succinate 8.1%, Calcium 8.3%, Sodium 15%, Moisture 1.7%, pH at 10% (w / v) dilution: 7.8

[0098] At least 95% by weight of the spray-dried fermented product had a particle size not exceeding 300 µm. Example 3

[0099] The composition of the solids removed in Example 2 via disc stack centrifugation was analyzed. The results are presented in Table 4. Table 4 % by weight H2O 65.9 CO32- / HCO3- 10.9 Organic acids 4.5 Petition 870250089332, dated 10 / 01 / 2025, pages 122 / 127 20 / 24 Protein 4.4g, Glucose 1.5g, Ca2+ 2.7g, Remainder 10.1g Comparative Example A

[0100] Example 1 was repeated, but this time NaOH was used instead of the Ca(OH)2 / NaOH mixture to control the pH. It was found that, compared to Product 1 of Example 1, the product obtained (Product B) had a substantially lower propionate content and a much higher succinate content. Example 4

[0101] The fermentation experiments described in Example 1 were repeated. Product 3 was obtained using the same procedure as Product 1 in Example 1. Product C was obtained using the same procedure as Product 1 in Example A.

[0102] Products 3 and C were analyzed. The results of these analyses are summarized in Table 5 along with the analytical results for Products 1 and A. Table 5 Percentage by weight Product 1 Product A Product 3 Product C Acetate 7.2 7.4 7.6 7.4 Lactate 0.8 0.8 1.5 1.6 Propionate 31.5 30 26.9 27 Succinate 5.7 4.3 4.2 4.2 Calcium 11 18 10 18 Sodium 9.4 1 8.6 0.57 Moisture 2 2.4 2.9 2.8 pH at 10% (w / v) dilution 6.3 6.6 5.3 5.5 Petition 870250089332, dated 10 / 01 / 2025, pages 123 / 127 21 / 24

[0103] Each sample was dissolved in water to a concentration of 5% (w / v). Each sample was evaluated by 10 untrained judges. The results are shown in Table 6. This table shows the average scores of the Attribute Classification Test using a line scale anchored with “low” at one end and “high” at the opposite end. Table 6 Solution Intensity Bitterness Intensity Sweetness Intensity Astringent Intensity Powdery Intensity Product 1 32.9 32.25 29.3 30.2 Product A 45.6 26.8 28.4 30.5 Product 3 17.3 31.8 22.0 28.9 Product C 49.8 23.7 34.2 34.1 Example 5

[0104] The spray-dried yeast samples from Example 4 were applied to a typical baked product, bread. The bread recipes are shown in Table 7 (yeasts were added in quantities providing 3.9 grams of propionic acid). Table 7 Grams Bread 1 Bread A Bread 2 Bread B Flour 1000.00 1000.00 1000.00 1000.00 Salt 20.00 20.00 20.00 20.00 Sugar 80.00 80.00 80.00 80.00 Enzymes with preservative action1 5.00 5.00 5.00 5.00 Petition 870250089332, dated 10 / 01 / 2025, pp. 124 / 127 22 / 24 Dough conditioner 2 3.25 3.25 3.25 3.25 Yeast - pressed 120.00 120.00 120.00 120.00 Soybean oil 20.00 20.00 20.00 20.00 60% lactic acid solution 8.90 10.30 9.90 10.90 Water 566.00 566.00 566.00 566.00 Product 1 12.38 Product A 13.00 Product 3 14.50 Product C 14.45 Enzymes with preservative action may include ULTRAFRESH Premium 250, available at Corbion. The pasta conditioner may include PRISTINE Concentrate and PRISTINE Relaxer, available from Corbion. The bread was prepared using the following process: 1. Mix the dry ingredients. 2. Add the soybean oil and lactic acid solution to the mixture of dry ingredients. 3. Cool the water and add it to the mixing container. 4. Add the dry ingredients along with the soybean oil to the chilled water. 5. Add the yeast. 6. Mix for 2 minutes on low speed. 7. Mix for 11 to 13 minutes at medium speed. 8. Cover the dough with a cloth and let it rest for 5 minutes. 9. Cut into 6 portions, shape into balls, and let rest for another 5 minutes. Petition 870250089332, dated 10 / 01 / 2025, pp. 125 / 127 23 / 24 10. Grease a baking pan, line it with parchment paper, and adjust it to the dimensions. 11. Place in the proofing chamber until the dough reaches the mark. 12. Bake at 420°F (216°C) for 23 minutes. 13. Let it cool for 1 hour. 14. Pack and close: Untrained judges evaluated each bread sample on various aspects of preference. The results are shown in Table 8. Table 8 Global Bread Aroma Flavor Aftertaste 1 5.8 6.2 5.9 5.1 A 4.9 5.7 5.0 4.9 3 5.8 6.1 5.9 5.2 C 5.1 5.7 5.1 4.7

[0105] Notes on the test: 1. The test used a questionnaire with a 9-point scale, where 9 = Extremely pleasant, 1 = Extremely unpleasant, and 5 = Neither pleasant nor unpleasant; 2. The samples were tasted in pairs, with Bread Samples 1 and A compared to each other and Bread Samples 2 and B compared to each other. Example 6

[0106] A fermented product was produced in the same manner as Fermented Product 1 of Example 1. The fermented product was divided into two portions. One portion was concentrated, sterilized, and spray-dried, producing Product 4. The other portion was first centrifuged in a disc stack centrifuge and then the Petition 870250089332, dated 10 / 01 / 2025, pp. 126 / 127 24 / 24 supernatant was concentrated, sterilized and spray-dried to produce Product 5.

[0107] Both products were analyzed. The results are shown in Table 9. Table 9 Percentage by weight Product 4 Product 5 Acetate 8.7 13.9 Lactate 0.8 1.2 Propionate 29.4 46.1 Succinate 6.2 10.0 Calcium 11.1 9.6 Sodium 9.3 14.2 Moisture 1.6 1.5 pH at 10% (w / v) dilution 6.4 5.5 Average dynamic buffering capacity 29.7 23.8 pH of maximum buffering capacity Approx. 5.8 4.85 Petition 870250089332, dated 10 / 01 / 2025, page 127 / 127

Claims

1 / 4 CLAIMS 1. Dry fermented product having a moisture content of less than 15% by weight and characterized by comprising, per kg of dry matter: □ 4.7 to 8.1 mol, preferably 5.2 to 7.8 mol and more preferably 5.8 to 7.2 mol of propionate; □ 1.2 to 3.0 mol, preferably 1.4 to 2.8 mol and more preferably 1.7 to 2.5 mol of calcium cation; □ 2.4 to 7.8 mol, preferably 4.0 to 7.0 mol and more preferably 5.3 to 6.5 mol of alkali metal cation selected from sodium, potassium and combinations thereof; □ 1 to 50 grams, preferably 2 to 30 grams, and more preferably 3 to 15 grams of glucose equivalent of hydrolyzed glucan; the calcium cation and propionate being present in the dry fermented product in a molar ratio of 0.2 to 0.

6.

2. Dry fermented product, according to claim 1, characterized in that the fermented product comprises DNA originating from a propionic acid-producing microorganism, preferably from a bacterium belonging to the genus Acidipropionibacterium and more preferably from a bacterium selected from Acidipropionibacterium acidipropionici, Acidipropionibacterium freudenreichii, Acidipropionibacterium shermanii, Acidipropionibacterium thoenii, Acidipropionibacterium jensenii and combinations thereof.

3. Dry fermented product, according to any of the preceding claims, characterized in that the fermented product comprises, per kg of dry matter, at least 3.4 mol, preferably at least 4.0 mol and more preferably at least 5.3 mol of sodium cation.

4. Dry fermented product, according to any of the preceding claims, characterized in that the fermented product comprises, per kg of dry matter, 1.5 to 3.0 mol, preferably 1.7 to 2.8 mol and more preferably 1.9 to 2.6 mol of acetate. Petition 870250089332, dated 10 / 01 / 2025, p. 99 / 127 2 / 4 5. Dry fermented product, according to any of the preceding claims, characterized in that the fermented product comprises, per kg of dry matter, 0.4 to 1.2 mol, preferably 0.5 to 1.1 mol and more preferably 0.55 to 1.0 mol of succinate.

6. Dry fermented product, according to any of the preceding claims, characterized in that the fermented product contains calcium cation and propionate in a molar ratio of 0.25 to 0.45, preferably from 0.28 to 0.

40.

7. Dry fermented product, according to any of the preceding claims, characterized in that the fermented product contains calcium cation and alkali metal cation in a molar ratio of 0.25 to 0.62, preferably from 0.30 to 0.52 and more preferably from 0.32 to 0.

45.

8. Dry fermented product, according to any of the preceding claims, characterized in that the fermented product contains propionate at a concentration X, acetate at a concentration Y, succinate at a concentration Z, calcium cation at a concentration A, sodium cation at a concentration B, and potassium at a concentration C, wherein all said concentrations are expressed in mol / kg of dry matter and wherein 0.8 < (X+Y+2Z) / (2A+B+C) < 1.2, preferably 0.85 < (X+Y+2Z) / (2A+B+C) < 1.15 and more preferably 0.90 < (X+Y+2Z) / (2A+B+C) < 1.

12.

9. Dry fermented product, according to any of the preceding claims, characterized by the dispersion of 100 grams of the fermented product in 1 liter of water, producing an aqueous liquid having a pH in the range of 6 to 8.

10. Dry fermented product, according to any of the preceding claims, characterized in that the dry fermented product has an average dynamic buffering capacity of less than 27 mM / g.

11. Dry fermented product, according to any of the preceding claims, characterized by a solution obtained when the dry fermented product is dissolved in demineralized water at 20 °C to achieve a propionate concentration of 140 mmol / L, having a bitterness index of less than 50 on a line scale of 0 to 100.

12. A process for producing a fermented product, said process characterized by comprising: □ providing an aqueous fermentation medium comprising 30 to 200 g / L of glucose equivalent of hydrolyzed glucan; □ inoculating the fermentation medium with a propionic acid-producing microorganism; □ incubating the inoculated fermentation medium for at least 20 hours to produce a fermented product, adding at the same time an alkalizing agent to maintain the pH in the range of 5.0 to 7.5, said alkalizing agent including a water-soluble alkali metal salt and a water-soluble alkali calcium salt; wherein the fermented product at the end of the incubation step comprises calcium cation and propionate in a molar ratio of 0.3 to 0.8 and alkali metal cation and propionate in a molar ratio of 0.3 to 1.

4.

13. Process according to claim 12, characterized in that the propionic acid-producing microorganism belongs to the genus Acidipropionibacterium.

14. Process, according to any one of claims 12 to 13, characterized in that the water-soluble alkali metal salt is selected from sodium hydroxide, potassium hydroxide and combinations thereof, and in that the water-soluble alkali calcium salt is calcium hydroxide.

15. Process, according to any one of claims 12 to 14, characterized in that the process comprises the additional steps of: □ removing undissolved solids from the fermented product by centrifugation or filtration to produce a fermented product with a low solids content; and □ drying the fermented product with a low solids content. Petition 870250089332, dated 10 / 01 / 2025, pp. 101 / 127 4 / 4 16. Process according to any one of claims 12 to 15, characterized in that the process produces a dry fermented product, as defined in any one of claims 1 to 11.

17. A method for preserving a food or cosmetic product, said method characterized by comprising the incorporation of a dry fermented product, as defined in any one of claims 1 to 11, at a concentration of 0.1 to 5% by weight of the food or cosmetic product.

18. Food or cosmetic product characterized by being obtained through the method as defined in claim 17. Petition 870250089332, dated 10 / 01 / 2025, pp. 102 / 127