Process for the preparation of dry particles comprising viable microorganisms and dry particles thus obtained

Abstract

The present invention relates to a process for the preparation of dry particles comprising viable microorganisms trapped in an alginate matrix, comprising: a) providing a suspension including an alginate salt and viable microorganisms; b) forming in the gelation bath one or more solid elongated elements comprising viable microorganisms trapped in an alginate matrix by dispensing a continuous or intermittent trickle-type flow of the suspension in a gelation bath including at least one gelling agent; c) separating the solid elongated elements from the gelation bath so as to obtain one or more wet solid elongated elements; d) drying the wet solid elongated elements so as to obtain one or more dried solid elongated elements; and e) grinding the dried solid elongated elements so as to obtain dry particles comprising viable microorganisms trapped in an alginate matrix. The dry particles obtained have a mechanical compressive strength greater than or equal to 500 g and less than 3200 g. The present invention also relates to the use of dry particles in an aid for the preparation of doughs for baked food products, an aid for the preparation of dough for baked food products, the related production process, the use of such an aid in the preparation of dough for baked food products, a dough for baked food products and a process for the preparation of a baked food product. Fig. 3

Description

[0001] Process for the preparation of dry particles comprising viable microorganisms and dry particles thus obtained

[0002] Field of the invention

[0003] The present invention concerns a process for the preparation of dry particles comprising viable microorganisms trapped in an alginate matrix, as well as the dry particles incorporating viable microorganisms thus obtained.

[0004] The present invention also relates to the use of such dry particles in an aid for the preparation of dough for baked food products, an aid for the preparation of dough for baked food products, the related production process, and the use of such an aid in the preparation of doughs for baked food products.

[0005] Finally, the present invention relates to a dough comprising the aforementioned aid and a process for the preparation of a baked food product using the aforementioned dough.

[0006] Background of the invention

[0007] Encapsulation processes of microorganisms, e.g. probiotic microorganisms, in polysaccharide microparticles are known in various technical fields, including nutrition, food, pharmaceuticals, cosmetics, livestock and agriculture.

[0008] The main function of the encapsulation of microorganisms within the microparticles is to protect the microorganisms from the external environment and / or from substances that are toxic to the microorganisms in that environment, e.g. during storage or during use. For example, in the nutritional or pharmaceutical fields, the encapsulation of probiotic microorganisms protects them during transit through the gastric tract where they are exposed to a very low pH and the activity of enzymes and bile salts.

[0009] Conversely, in the food industry, such as in the case of so-called baking aids, the encapsulation of microorganisms, e.g. lactic acid bacteria, i.e. viable, metabolically active bacteria belonging to the lactic acid culture group, protects microorganisms from a loss of viability both during storage and during dough preparation when the aids are mixed with water and food flours. A second, equally important function of the encapsulation of microorganisms is to release them in as controlled a manner as possible into the desired environment, e.g. the human intestinal tract in the case of probiotics, or in as rapid and controlled a manner as possible in doughs during the preparation of baked food products.

[0010] Related art

[0011] Document KR20160051902A discloses compositions including lactic acid bacteria encapsulated in alginate beads.

[0012] Alginate beads are produced by mixing lactic acid bacteria with an aqueous solution of alginate and adding this solution drop by drop to a calcium chloride solution to harden the beads. The resulting beads are made dry by freeze-drying.

[0013] The document highlights that the use of freeze-drying as a method of drying capsules containing lactic acid bacteria affects the viability of the bacteria, being associated with a survival rate of only 60-70% after drying. To overcome this drawback and increase the survival rate of the bacteria, various cryoprotectants are used, e.g. soy flour, sugars, amino acids, peptides, gelatin, glycerol, sugar alcohols, whey, alginic acid, ascorbic acid, yeast extract, skim milk, trehalose, garlic extract.

[0014] The Applicant has observed that freeze-drying is a complex and expensive drying technique, as well as rather energy inefficient. The process requires subjecting the material to be dried to very low temperatures under high vacuum conditions for several days. These and other critical issues make this technique difficult to scale on an industrial level.

[0015] Freeze-drying also has a strong impact on the viability of encapsulated microorganisms. To address this problem, KR20160051902A relies on the use of cryoprotectant additives which, however, as acknowledged by the same document, change the taste, texture and / or production costs of the food in which the disclosed lactic acid bacteria compositions are used, which may make them unsuitable for the preparation of many foods.

[0016] It is also noted that the document does not focus on the structural, organoleptic and nutritional characteristics of the finished products obtained from compositions including the particles, characteristics that are, in the Applicant’s opinion, particularly important when the lactic acid bacteria are used in the preparation of baked food products with the intention of reproducing the effects of fermentation promoted by natural sourdough.

[0017] To address these issues, international patent application WO 2024 / 009249 A1 , in the name of the same Applicant, discloses a process for microencapsulating viable lactic acid bacteria in a casing including at least one gelling biopolymer in order to obtain particles that can be used in the preparation of a food additive for baked food products.

[0018] In particular, the process disclosed by application WO 2024 / 009249 A1 comprises: a) providing a solution including a gelling biopolymer, preferably sodium alginate, and lactic acid bacteria; b) feeding drops of said solution of gellable biopolymer and viable lactic acid bacteria into a gelation bath including at least one gelling agent and allowing these drops to gel, and c) drying the gelled droplets by air drying, obtaining particles containing viable lactic acid bacteria belonging to at least one species, said particles having an average size D50 comprised between 350 pm and 550 pm, preferably between 390 pm and 490 pm, more preferably between 400 pm and 460 pm, and a mechanical compressive strength, measured as detailed in Example 4 of the present document, comprised between 3200 g and 9000 g, preferably between 3400 g and 6000 g, more preferably between 3500 g and 5700 g.

[0019] In accordance with this document, the above dimensions and mechanical strength values are an optimal compromise to give the particles the ability to remain intact and preserve the lactic acid bacteria in the absence of mechanical stress and in a dry environment, and to open quickly when exposed to the mechanical stresses typical of dough preparation, whether applied manually or mechanically by kneading machines, and to the wet environment of the dough. The Applicant noted that scaling up this process industrially presents problematic aspects due to the specific production equipment that must be used, as well as the way in which these particles are obtained. In fact, the latter are obtained by mechanically micronizing the solution including the gelling biopolymer and lactic acid bacteria into very small droplets that are then dropped into the gelation bath where they solidify to form substantially spheroidal microparticles.

[0020] The mechanical micronization of the solution, however, is a very slow operation that severely limits the production capacity of the process as a whole. The slowness of the micronization step of the solution including the gelling biopolymer and lactic acid bacteria also inevitably results in very different dwell times of the droplets in the gelation bath between the droplets dispensed first and those dispensed last, with a possible undesirable variation in the mechanical characteristics of the substantially spheroidal microparticles obtained.

[0021] Finally, the Applicant observed that, irrespective of the dwell time in the gelation bath, the microparticles are in any case in some cases excessively rigid, thus making the release of viable lactic acid bacteria more difficult than would be desirable, e.g. in doughs for baked products.

[0022] European patent application EP 3 735 840 A1 discloses a process for the preparation of very dense alginate gel beads comprising an active ingredient, alginate gel beads obtainable by this process, a food product comprising these alginate gel beads and the use of the beads as a food additive.

[0023] In particular, the process disclosed by application EP 3 735 840 A1 comprises the steps of: a. mixing a dry alginate salt with a liquid, wherein the liquid is selected from the group consisting of: (i) an aqueous solution, (ii) an oil-in-water emulsion and (iii) an aqueous suspension, in a ratio of 1 :5 to 1 :0.4 w / w of dry alginate salt to liquid, b. adding an active ingredient to the mixture obtained in step (a), and c. adding a salt of a water-soluble bivalent or trivalent cation, thus obtaining alginate gel beads, or alternatively, a. mixing a dry alginate salt with a liquid, wherein the liquid is selected from the group consisting of: (i) an aqueous solution, (ii) an oil-in-water emulsion and (iii) an aqueous suspension, in a ratio of 1 :5 to 1 :0.4 w / w of dry alginate salt to liquid, wherein the liquid also comprises an active ingredient, and b. adding a salt of a water-soluble bivalent or trivalent cation, thus obtaining alginate gel beads.

[0024] In particular, the active ingredient may be a microorganism and, more precisely, a bacterium of the genus Rhizobium.

[0025] The Applicant noted that this document discloses a type of alginate having a viscosity greater than 90 mPa s when dissolved in a 1 % by weight aqueous solution at a temperature of 20°C, a type that is ill-suited for the rapid release of microorganisms, in particular microorganisms that are viable under the processing conditions adopted in the production of a baked food product.

[0026] In particular, the patent application EP 3 735 840 A1 does not provide any example of alginate gel beads incorporating viable microorganisms, nor does it provide any teachings on how to release these in a rapid and controlled manner into the desired environment, in particular by using viable microorganisms useful in food and cosmetics such as lactic acid bacteria.

[0027] Summary of the invention

[0028] In view of the aforementioned state of the art, the Applicant has set itself the goal of providing dry particles containing viable microorganisms that are capable of both preserving the viability of microorganisms as well as releasing them rapidly during use.

[0029] The Applicant has also set itself the goal of providing a process for the preparation of dry particles comprising viable microorganisms that is very simple and inexpensive and can be efficiently industrially scaled up. In light of these objectives, the Applicant has perceived that by means of appropriate dosage methods of a suspension comprising an alginate salt and viable microorganisms in a gelation bath, it is advantageously possible to obtain dry particles having both optimal viability protection properties of the microorganisms during production, and subsequent storage and use, and breakage resistance properties that at the same time consent rapid and controlled release of the microorganisms into the desired medium or environment.

[0030] The present invention therefore relates, in a first aspect thereof, to a process for the preparation of dry particles comprising viable microorganisms trapped in an alginate matrix.

[0031] Preferably, the process involves a) providing a suspension including an alginate salt and viable microorganisms.

[0032] Preferably, the process comprises b) forming in the gelation bath one or more solid elongated elements comprising viable microorganisms trapped in an alginate matrix by dispensing a continuous or intermittent trickle-type flow of said suspension in a gelation bath including at least one gelling agent.

[0033] Preferably, the process comprises c) separating said one or more solid elongated elements from the gelation bath.

[0034] Preferably, the process comprises d) drying said one or more wet solid elongated elements so as to obtain one or more dried solid elongated elements.

[0035] Preferably, the process comprises e) grinding said one or more dried solid elongated elements to obtain dry particles comprising viable microorganisms trapped in an alginate matrix.

[0036] Preferably, said dry particles have a mechanical compressive strength, measured as indicated in the present description, greater than or equal to 500 g and less than 3200 g.

[0037] In accordance with a second aspect thereof, and within the scope of achieving the objectives outlined above, the invention further relates to dry particles comprising viable microorganisms trapped in an alginate matrix, wherein said dry particles have a mechanical compressive strength, measured as set forth in the present description, greater than or equal to 500 g and less than 3200 g.

[0038] Preferably, said dry particles are particles obtainable by the process as disclosed in the present description.

[0039] In accordance with a third aspect thereof, and within the scope of achieving the objectives outlined above, the invention further relates to the use of dry particles containing viable microorganisms trapped in an alginate matrix, as disclosed herein, in an aid in the preparation of doughs for baked food products.

[0040] In accordance with a fourth aspect of the invention and within the scope of achieving the objectives outlined above, the invention further relates to an aid for the preparation of doughs for baked food products, comprising a food-grade carrier for baked food products and dry particles containing live lactic acid bacteria belonging to at least one species as disclosed herein.

[0041] Preferably, said dry particles have a mechanical compressive strength, measured as indicated in the present description, greater than or equal to 500 g and less than 3200 g.

[0042] Preferably, said dry particles are dispersed in said food-grade carrier.

[0043] Preferably, said dry particles have an average size D50 comprised between 50 pm and 5000 pm.

[0044] In the present description and subsequent claims, the expression “average size D50” used with reference to particles means the percentile diameter relative to 50% of the particle size distribution. D50 is defined as the value below which the diameter of 50% of the particles in the size distribution falls.

[0045] In accordance with a fifth aspect thereof, and within the scope of achieving the objectives outlined above, the invention further relates to the use of an aid as disclosed herein in the preparation of doughs for baked food products.

[0046] In accordance with a sixth aspect of the invention and within the scope of achieving the objectives outlined above, the invention further relates to a process for producing an aid for the preparation of baked food products, comprising dispersing dry particles containing live lactic acid bacteria belonging to at least one species as disclosed in the present description in a quantity of a food-grade carrier for baked food products.

[0047] In accordance with a seventh aspect thereof, and within the scope of achieving the objectives outlined above, the invention further relates to a dough for baked food products comprising dry particles containing live lactic acid bacteria belonging to at least one species as disclosed herein, water and optionally further ingredients.

[0048] In accordance with an eighth aspect thereof, and within the scope of achieving the objectives outlined above, the invention further relates to a process for preparing a baked food product comprising forming a dough by mixing dry particles containing live lactic acid bacteria belonging to at least one species as disclosed in the present description with water and, optionally, further ingredients such as flour.

[0049] Preferably, the process of preparing a baked food product comprises dividing the dough into portions.

[0050] Preferably, the process of preparing a baked food product comprises giving the dough or portions of dough a desired shape.

[0051] Preferably, the process of preparing a baked food product comprises baking the dough or portions of dough in an oven.

[0052] The Applicant has found that the process for the preparation of dry particles comprising viable microorganisms according to the invention is less complex, less costly, faster to implement and more easily scalable than known microencapsulation processes such as those disclosed by KR20160051902A and WO 2024 / 009249 A1 comprising the formation of droplets of the solution comprising the gelling biopolymer and lactic acid bacteria.

[0053] In fact, the process for the preparation of dry particles of the invention does not require for its implementation the use of dedicated and relatively complex apparatuses, but only structural elements, such as containers possibly equipped with agitators, and piping, low-cost and readily available components on the market, capable of enabling the continuous or intermittent trickle-type flow of the alginate suspension and viable microorganisms in the gelation bath.

[0054] The Applicant has also experimentally found that by this simple method of dispensing the suspension including an alginate salt and viable microorganisms in the gelation bath, it is advantageously possible to very quickly and efficiently obtain the aforementioned solid elongates from the gelation of the alginate with the consequent formation of an alginate matrix within which the viable microorganisms are trapped.

[0055] The rapidity of formation of these solid elongated elements makes it possible to obtain an alginate matrix having mechanical characteristics, primarily a mechanical compressive strength, which are much more uniform than those obtainable by the relatively slow processes disclosed by KR20160051902A and WO 2024 / 009249 A1 , which envisage a rather slow operation of mechanical droplet formation of the solution including the gelling biopolymer and lactic acid bacteria, and a different dwell time of the spheroidal microparticles in the gelation bath, with timescales that may be particularly long for the first microparticles dispensed.

[0056] This results, after drying and grinding of the solid elongated elements, in more uniform mechanical characteristics of the dry particles obtained by the process of the invention both within each production batch and between different production batches.

[0057] The Applicant has also experimentally found that the dry particle preparation process of the invention achieves a significant increase in production yield of the wet product understood as the weight of the wet gelled matrix obtained from the suspension of alginate and viable microorganisms with respect to the weight of the initial suspension.

[0058] In particular, the Applicant observed that in the case of the process of the invention that yield can reach values of 90-95% by weight of the weight of the initial suspension, whereas it is limited to values of 80-85% in the case of the process disclosed by WO 2024 / 009249 A1 . This results in an advantageous increase in the production yield of dry particles, understood as the weight of the dry particles obtained with respect to the weight of the initial suspension, a yield that is in the range of 10-15% with respect to values that at most reach 5% in the case of the process disclosed by WO 2024 / 009249 A1 .

[0059] The Applicant has experimentally found that the viability of the microorganisms trapped in an alginate matrix of the dry particles obtained by the process of the invention remains substantially unaltered for long periods of time (e.g., approximately 10 months).

[0060] The Applicant has also experimentally found that the viability of lactic acid bacteria that can be used in cosmetics, nutrition or baked food products is maintained by storing the dry particles and / or the products containing them at room temperature, such as the aforementioned aid for the preparation of doughs for baked food products, demonstrating the high stability of the dry particles and of the aid of the invention.

[0061] A further advantage of the process for the preparation of dry particles comprising viable microorganisms according to the invention is that it is possible to proceed without with anti-aggregating agents, such as silicon dioxide (E551 ), typically used in the microorganism encapsulation process disclosed by WO 2024 / 009249 A1 to reduce the aggregation of substantially spheroidal microparticles and the associated formation of lumps.

[0062] With regard to the specific application of the dry particles of the invention to the specific field of baked food products, the Applicant has experimentally found that baked food products obtained by employing such dry particles, preferably formulated in a suitable processing aid, exhibit organoleptic, rheological and structural or textural properties similar to those of products obtained from sourdough.

[0063] Without wishing to bind itself here to any interpretative theory, the Applicant considers that the positive effect of improving the organoleptic, rheological and structural or textural properties of baked food products is to be attributed to a combination of improved viability characteristics of the microorganisms, in this case lactic acid bacteria, and the mechanical resistance of the dry particles, which allow rapid and complete release of the live lactic acid bacteria, which are thus able to better carry out their metabolic functions within the food doughs.

[0064] In particular, the Applicant considers that the specific mechanical strength characteristics of the dry particles obtained according to the process of the invention, which are reduced compared to those disclosed by the prior art mentioned herein, facilitate their better dispersion and homogeneous distribution in the doughs during mixing.

[0065] Detailed description of embodiments of the invention

[0066] The present invention can be presented in one or more of its aspects or one or more of the preferred characteristics reported below, which can be combined with one another as preferred according to the application requirements.

[0067] In the context of the present description and subsequent claims, “gelling” means a cross-linking reaction of the alginate salt. As known to a person skilled in the art, an alginate gel can be obtained from an alginate salt by ionic cross-linking with bivalent or trivalent cations, which will be discussed below.

[0068] In the context of this description and in subsequent claims, “baked food product” means any savoury or sweet product obtained by total or partial baking of a leavened and / or fermented dough obtained by mixing food flour with water and any additives or aids. By way of non-limiting example, this term may designate products such as bread, pizza, focaccia, biscuits, crackers, rusks, brioches, cakes and baked desserts, rustic, taralli.

[0069] Within the context of the present description and following claims, all the numerical magnitudes indicating quantities, parameters, percentages, and so on are to be considered preceded in every circumstance by the term “about” unless indicated otherwise. Furthermore, all ranges of numerical quantities are to be understood as including extremes, unless otherwise indicated, and include all possible combinations of maximum and minimum numerical values and all possible intermediate ranges, in addition to those specifically indicated below. Preferably, said step a) of providing a suspension including an alginate salt and viable microorganisms, e.g. viable lactic acid bacteria, comprises forming an aqueous suspension of the alginate salt, adding the viable microorganisms and mixing until homogenization.

[0070] Preferably, viable microorganisms can be supplied as fresh concentrated biomass in aqueous suspension, as fresh concentrated biomass stored in frozen form or as biomass in freeze-dried form, which can be stored at room temperature, refrigerated or frozen.

[0071] When viable microorganisms are supplied as fresh or freeze-dried biomass stored in frozen form, a thawing step of the microorganisms is envisaged.

[0072] Preferably, the suspension including an alginate salt and viable microorganisms is an aqueous suspension.

[0073] As explained above, the polymer matrix of the particles of the invention is obtained by gelling an alginate salt.

[0074] Preferably, the alginate salt is sodium alginate.

[0075] Sodium alginate is a water-soluble polysaccharide isolated from the cell wall of algae, composed of two monomeric units: mannuronic acid and guluronic acid. Having a total negative charge, when exposed to multivalent cations such as calcium ions, the carboxyl groups on the guluronic acid monomers interact with the multivalent cations to form a cross-linked hydrogel. Advantageously, polymerization can take place at room temperature and no heating step is required, which could have a negative impact on the viability of the microorganisms.

[0076] Preferably, sodium alginate has a viscosity between 5 and 50 mPa s, more preferably between 8 and 40 mPa s, and even more preferably between 10 and 20 mPa s, when dissolved in a 1 % by weight aqueous solution at a temperature of 20°C.

[0077] An alginate with the above viscosity values is referred to as a low-viscosity alginate. A low-viscosity alginate exhibits high solubility, i.e. it is possible to dissolve relatively high concentrations of alginate in the suspension including an alginate salt and viable microorganisms while maintaining the viscosity of the resulting solution suitable for processing to form particles.

[0078] Preferably, a concentration of said alginate salt, preferably of said sodium alginate, in said suspension is between 1 % and 8% by weight, more preferably between 1 % and 6% by weight, and even more preferably between 2% and 5% by weight, of the total weight of the suspension.

[0079] Preferably, the concentration of said alginate salt, preferably of said sodium alginate, in the suspension is greater than or equal to 1 % by weight, more preferably greater than or equal to 2% by weight, even more preferably greater than or equal to 3% by weight, even more preferably greater than or equal to 4% by weight, even more preferably greater than or equal to 5% by weight, even more preferably greater than or equal to 6% by weight, even more preferably greater than or equal to 7% by weight, of the total weight of the suspension.

[0080] Preferably, the concentration of said alginate salt, preferably of said sodium alginate, in the suspension is less than or equal to 8% by weight, even more preferably less than or equal to 7% by weight, even more preferably less than or equal to 6% by weight, even more preferably less than or equal to 5% by weight, even more preferably less than or equal to 4% by weight, even more preferably less than or equal to 3% by weight, even more preferably less than or equal to 2% by weight, of the total weight of the suspension.

[0081] Preferably, the suspension of alginate salt, preferably of said sodium alginate, has a viscosity greater than or equal to 20 mPa s and less than or equal to 650 mPa s, more preferably greater than or equal to 30 mPa s and less than or equal to 200 mPa s, and even more preferably greater than or equal to 40 mPa s and less than or equal to 100 mPa s.

[0082] Advantageously, the above-mentioned preferred concentrations of said alginate salt in the suspension including an alginate salt and viable microorganisms ensure that the alginate matrix of the dry particles within which the microorganisms, e.g. live lactic acid bacteria, are trapped is sufficiently resistant, thus offering the microorganisms greater protection and chances of survival.

[0083] Experiments carried out by the Applicant have shown that the use of alginate, in particular sodium alginate, having the above-mentioned viscosity and concentration values in the suspension with the viable microorganisms results in dry particles having a size and mechanical properties within the ranges described and claimed below.

[0084] In preferred embodiments of the invention, the suspension may include non-salified alginic acid in addition to the alginate salt.

[0085] Preferably, a concentration of unsalted alginic acid is between 10% and 60% by weight, more preferably between 15% and 45% by weight, and even more preferably between 20% and 30% by weight of the total weight of the alginate salt in the suspension.

[0086] The Applicant has experimentally observed that a beneficial effect on the viability of microorganisms can be achieved by using appropriate amounts of non-salified alginic acid.

[0087] Without wishing to bind itself to any interpretative theory, the Applicant believes that this beneficial effect is related to a reduction in the sodium content of the alginate matrix of the particles.

[0088] Preferably, the dry particles of the invention have a compressive strength, measured as set forth in the present description, greater than or equal to 1500 g and less than or equal to 3000 g, more preferably greater than or equal to 1800 g and less than or equal to 2500 g.

[0089] In this way, it is advantageously possible to have adequate mechanical strength characteristics of the dry particles, which are necessary for the protection of microorganisms, combined with easy disintegration and breakage of the particles at the time of use.

[0090] In the present description and in subsequent claims, “dry particles” means particles with a moisture content less than or equal to 20% by weight. Preferably, a concentration of viable microorganisms in said suspension is between 0.5% and 4% by weight, more preferably between 0.5% and 3% by weight, and even more preferably between 1 .0% and 2.5% by weight of the total weight of the suspension.

[0091] Thus, it is advantageously possible to have an optimal amount of viable microorganisms in the dry particles obtained by the process of the invention.

[0092] Preferably, said suspension has a pH between 4 and 7, more preferably between 4.5 and 6, and even more preferably between 5 and 5.5.

[0093] In this way, effective gelling of the alginate is advantageously achieved.

[0094] Preferably, preparing said suspension comprises keeping said suspension at a temperature between 15°C and 35°C, more preferably between 18°C and 30°C, and even more preferably between 20°C and 25°C.

[0095] In this way, effective gelling of the alginate is advantageously achieved. Excessively high temperatures can also lead to a weakening of the gel from a mechanical point of view.

[0096] In preferred embodiments and depending on the field of application, these viable microorganisms may be one or more of: live lactic acid bacteria belonging to at least one species, preferably from the family of Lactobacillaceae, Streptococcaceaeae, Enterococcaceae, Carnobacteriaceae, microorganisms of the genus Rhizobium, microorganisms of the genus Bifidobacterium, yeasts such as Saccharomyces ssp, non-Saccharomyces ssp, Kluyveromyces spp, sporigenic microorganisms such as Bacillus spp, Weizmannia spp, new generation probiotics, such as Akkermansia spp, Faecalibacterium spp, Clostridium spp. Bacteroides spp, Roseburia spp, non- toxigenic molds of the genus Aspergillus, mycorrhizal fungi, fungi of the genus Trichoderma.

[0097] In preferred embodiments, particularly applicable to the field of food and cosmetics, said viable microorganisms comprise lactic acid bacteria belonging to at least one species. In preferred embodiments, particularly applicable to the field of baked goods, said viable microorganisms are live lactic acid bacteria that can be isolated from natural sourdough or flour.

[0098] More preferably, live lactic acid bacteria can be isolated from traditional sourdough starter, also known in the industry as traditional sourdough starter or type 1 .

[0099] The Applicant has found that baked products made with an aid including live lactic acid bacteria that can be isolated from traditional sourdough starter have recognisable aromas, fragrances and taste that are attributable, in the Applicant's opinion, to the release by the live lactic acid bacteria of a broad spectrum of characteristic volatile compounds such as acids, alcohols, aldehydes, ketones.

[0100] The Applicant further noted that the presence of live lactic acid bacteria in the dry particles of the invention promotes the production, during the fermentation of the dough, of lipolytic enzymes capable of breaking down triglycerides and other lipids releasing glycerol, fatty acids and other molecules having a favourable emulsifying power which contributes to improving the quality and structure of the dough.

[0101] The Applicant has also found that, due to metabolic mechanisms that have not yet been fully clarified, the baked goods made from dry particles according to the invention, e.g. formulated in a suitable aid, are advantageously subject to a slowing down of the cooling process and loss of freshness, and keep longer than baked goods made without the use of the dry particles covered by the invention.

[0102] In particular, the Applicant has found that the shelf life of baked products obtained using the dry particles of the invention, evaluated in terms of the speed of starch retrogradation, is 30% or more longer than the shelf life of similar baked products obtained without the dry particles of the invention.

[0103] Preferably, the live lactic acid bacteria contained in the dry particles belong to one or more lactic acid bacteria species selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, Lactiplantibacillus plantarum, Companilactobacillus alimentarius, Levilactobacillus brevis, Lentilactobacillus buchneri, Latilactobacillus curvatus, Lactobacillus delbrueckii, Limosilactobacillus fermentum, Limosilactobacillus frumenti, Levilactobacillus hammesii, Lactobacillus helveticus, Lentilactobacillus hilgardii, Limosilactobacillus panis, Companilactobacillus paralimentarius, Lacticaseibacillus paracasei, Lactiplantibacillus paraplantarum, Lactiplantibacillus pentosus, Limosilactobacillus pontis, Limosilactobacillus reuteri, Latilactobacillus sakei, Lactococcus lactis, Leuconostoc citreum, Leuconostoc mesenteroides, Pediococcus pentosaceus, Weissella cibaria, and Weissella confusa.

[0104] More preferably, the live lactic acid bacteria contained in the dry particles belong to one or more species of lactic acid bacteria selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae and Lactiplantibacillus plantarum.

[0105] In preferred embodiments of the invention, the dry particles contain a combination of live lactic acid bacteria belonging to one or more species of lactic acid bacteria selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, and Lactiplantibacillus plantarum.

[0106] In other preferred embodiments of the invention, the aid comprises a combination of dry particles containing, each, live lactic acid bacteria belonging to one or more species of lactic acid bacteria selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, and Lactiplantibacillus plantarum.

[0107] In this case, the aid preferably comprises a first dry particle fraction containing live lactic acid bacteria belonging to the species Fructilactobacillus sanfranciscensis.

[0108] Preferably, the aid also comprises a second dry particle fraction containing live lactic acid bacteria belonging to the species Furfurilactobacillus rossiae.

[0109] Preferably, the aid also comprises a third dry particle fraction containing live lactic acid bacteria belonging to the species Lactiplantibacillus plantarum.

[0110] The live lactic acid bacteria belonging to the species Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae and Lactiplantibacillus plantarum are part of the microbiota of many traditional type 1 sourdoughs. In particular, Fructilactobacillus sanfranciscensis and Lactiplantibacillus plantarum are considered key species of sourdough, as their presence is reported in approximately 50% of sourdoughs (Ganzle et al. 2016,

[0001] ). Furfurilactobacillus rossiae is also widely distributed in sourdoughs, especially in those produced in Italy (Settanni et al. 2005, [2]).

[0111] Fructilactobacillus sanfranciscensis is an obligate heterofermentative bacterial species that metabolises maltose efficiently and is therefore very well suited to the sourdough environment. Fructilactobacillus sanfranciscensis produces heterohexopolysaccharides (EPS) that positively influence the volume, texture and general rheological characteristics of baked food products. Fructilactobacillus sanfranciscensis also produces metabolites such as phenylactic acid (PLA) that can inhibit fungal growth, and contributes to the definition of the flavour of baked food products through the metabolism of maltose and amino acids. Other technologically relevant characteristics of strains of this species may be the increased bioavailability of minerals and bioactive compounds, as well as the ability to degrade gluten (Zhang et al. 2019, [3]).

[0112] Lactiplantibacillus plantarum is a facultative heterofermentative species capable of producing antifungal compounds that can improve the shelf life of baked goods (Ventimiglia et al. 2015, [4]). In sourdough, Lactiplantibacillus plantarum is important for acid generation: it ferments hexoses to lactic acid via the homolactic pathway, and degrades pentoses via the pentose phosphate pathway, producing lactic acid and acetic acid. Some Lactiplantibacillus plantarum strains also possess extracellular protease activity that can have a positive impact on the rheological and sensory characteristics of products.

[0113] Furfurilactobacillus rossiae is an obligate heterofermentative bacterial species that is also highly suited to the sourdough environment. Strains belonging to this species show interesting technological properties, such as acidifying capacity and peptidase activities (Di Cagno et al. 2007, [5]).

[0114] The Applicant observed that the aforementioned lactobacillus species, and in particular Fructilactobacillus sanfranciscensis and Lactiplantibacillus plantarum, are particularly efficient in releasing a broad spectrum and high quantities of volatile compounds that positively impact on the aromatic qualities of bakery products. In preferred embodiments, particularly applicable to the field of baked food products, said suspension may include deactivated yeast in addition to live lactic acid bacteria.

[0115] In the context of the present description and subsequent claims, the term “deactivated yeast” is intended to refer to a yeast that has undergone a deactivation treatment (e.g. a heat treatment) that has resulted in the death of the cell and the blocking of its fermentation activity.

[0116] Preferably, the deactivated yeast is generally, but not necessarily, obtained from yeasts of the genus Saccharomyces (Saccharomyces cerevisiae and related species). As such, it is rich in amino acids, vitamins and other micro- and macronutrients. These include glutathione, a tripeptide with antioxidant properties, and beta glucans present in the cell wall of yeasts.

[0117] Depending on how it was obtained, the deactivated yeast may also contain yeast walls.

[0118] Preferably, a concentration of deactivated yeast in said suspension is between 1 % and 8% by weight, more preferably between 1 % and 6% by weight, and even more preferably between 2% and 5% by weight, of the total weight of the suspension.

[0119] The Applicant has experimentally observed that the use of deactivated yeast, which is also incorporated into the alginate matrix of the dry particles, has a beneficial effect on the viability and stability of the microorganisms, particularly when these are lactic acid bacteria.

[0120] In particular, the Applicant has experimentally observed that the deactivated yeast acts positively both on the survival of microorganisms throughout the process of incorporation into the alginate matrix, particularly during the drying step, and on maintaining the viability of the microorganisms during storage of the particles with an advantageous extension of their shelf life.

[0121] In addition, deactivated yeast advantageously plays a positive role in the preparation of baked goods as its addition to doughs increases the extensibility of the gluten mesh. In relation to the possible and advantageous use of deactivated yeast, the Applicant has experimentally observed that the process for the preparation of the dry particles of the invention overcomes the technological production difficulties related to the poor solubility of deactivated yeast in water, which makes the production of spheroidal microparticles including this ingredient very difficult when applying the known processes described in document KR20160051902A or WO 2024 / 009249 A1.

[0122] Preferably, a) preparing said suspension comprises keeping the suspension in agitation, preferably for between 1 h and 24 h, depending on whether or not one wishes to promote the replication of microorganisms in the suspension.

[0123] Thus, in a preferred embodiment, a) providing said suspension comprises keeping the suspension in agitation for between 1 h and 4h, more preferably between 1.5h and 3h, and even more preferably between 2h and 3h in case one does not wish to encourage replication of the microorganisms in the suspension.

[0124] In a further preferred embodiment, a) preparing said suspension comprises keeping the suspension in agitation for between 8h and 24h, more preferably between 10h and 18h, and even more preferably between 12h and 16h if a growth step of the microorganisms in the suspension and production of their associated metabolites is also desired.

[0125] In this way, it is advantageously possible both to achieve adequate solubilization of the bacteria and the alginate salt, and possibly to implement a growth step of the microorganisms present in the suspension with consequent production and accumulation of functional metabolites that can then be trapped in the alginate matrix together with the microorganisms, e.g. live lactic acid bacteria, that produced them.

[0126] This advantageously enhances the technical effects related to the presence of microorganisms and the functional metabolites they produce, such as exopolysaccharides and antioxidant substances, in the final dry particles. In particular, the Applicant has experimentally observed that the beneficial effects related to this biomass growth step are even more pronounced in the presence of deactivated yeast due to the presence of the nutrients contained therein.

[0127] Preferably, b) forming in a gelation bath one or more wet solid elongated elements comprising viable microorganisms trapped in an alginate matrix by dispensing a continuous or intermittent trickle-type flow of said suspension into said gelation bath including at least one gelling agent comprising feeding said suspension into said gelation bath from a storage vessel by gravity or by dispensing at a predetermined pressure from one or more pipelines or dispensing ports.

[0128] In this way, and as explained above, it is advantageously possible to have a process for the production of dry particles that can be implemented in a very simple and low- cost manner using very simple components that are readily available cheaply on the market.

[0129] In the context of the present description and subsequent claims, “dispensing a continuous trickle-type flow” means a method of feeding a suspension of alginate and viable microorganisms into a gelation bath that avoids the fractionation of the suspension into droplets, i.e. maintaining continuity of the liquid flow dispensed. Preferably, this is done by gravity or through the use of piping or dispensing openings of an appropriate diameter.

[0130] In preferred embodiments, these pipes or dispensing openings have a diameter between 1 mm and 3 cm, more preferably between 5 mm and 1.5 cm, and even more preferably between 8 mm and 1 cm.

[0131] Preferably, b) forming one or more wet solid elongated elements comprising viable microorganisms entrapped in an alginate matrix in a gelation bath by dispensing a continuous or intermittent trickle-type of said suspension into the gelation bath comprising at least one gelling agent comprises keeping the gelation bath in agitation.

[0132] Conveniently, this can be done using conventional solution / suspension agitators known in the art. Preferably, the gelling agent can be a salt that originates bivalent cations, e.g. or more of chloride, Ca, Mg, Mn, Zn, Co, Sr, Ba and Cu lactate or gluconate.

[0133] In a particularly preferred embodiment, a calcium salt is used. In an even more preferred embodiment, the calcium salt is calcium chloride, lactate or gluconate.

[0134] In other preferred embodiments, alginate gelling is also possible using trivalent cations, such as Al3+and Fe3+.

[0135] Preferably, a concentration of the salt originating bivalent or trivalent cations in the gelation bath is between 1 % and 12% by weight of the total weight of the gelation bath.

[0136] More preferably, a concentration of calcium chloride in the gelation bath is between 1 % and 8% by weight of the total weight of the gelation bath.

[0137] More preferably, a concentration of calcium lactate in the gelation bath is between 3% and 9% by weight of the total weight of the gelation bath.

[0138] More preferably, a concentration of calcium gluconate in the gelation bath is between 5% and 12% by weight of the total weight of the gelation bath.

[0139] In preferred embodiments, the process for preparing dry particles comprising viable microorganisms trapped in an alginate matrix of the invention further comprises f) maintaining said one or more elongated elements in said suspension for a time between 5 and 30 minutes, more preferably between 10 and 20 minutes, and even more preferably between 12 and 17 minutes.

[0140] Preferably, the suspension of alginate and viable microorganisms is kept in agitation.

[0141] In this way, it is advantageously possible to have more homogeneous incorporation and distribution of the bivalent element, e.g. calcium, in the alginate matrix.

[0142] Preferably, step c) of separating said one or more solid elongated elements from the gelation bath is implemented by feeding the gelation bath containing the solid elongated elements to a sieve that percolates the liquid but retains the wet solid elongated elements. Preferably, the wet solid elongated elements obtained from the separation step from the gelation bath have a moisture content between 80% and 97% by weight, more preferably between 85% and 95% by weight, and even more preferably between 92% and 95% by weight, of the total weight of said wet solid elongated elements.

[0143] Preferably, step d) of drying said one or more wet solid elongated elements is carried out at a temperature between 20°C and 45°C, more preferably between 20°C and 35°C, and even more preferably between 25°C and 30°C.

[0144] Preferably, step d) of drying said one or more wet solid elongated elements is carried out for between 30 min and 16 h, more preferably between 1 h and 8 h, and even more preferably between 2 h and 6 h.

[0145] In this way, it is advantageously possible to minimise possible thermal degradation of the viable microorganisms trapped in the alginate matrix and, at the same time, to have relatively fast drying times.

[0146] In preferred embodiments, step d) of drying said one or more wet solid elongated elements can be carried out in a forced-air circulation oven or fluidised bed dryer.

[0147] Preferably, such a forced air circulation oven comprises one or more support trays for the wet solid elongated elements, an electric coil configured to heat the air in the inner chamber of the oven and a fan configured to move the hot air in the chamber.

[0148] Preferably, such a forced-air circulation oven comprises at the back a discharge configured to release moisture-laden air and a discharge system for exhaust gases.

[0149] In alternative preferred embodiments, said step c) of drying the wet solid elongated elements can be carried out in a vibrational fluidised bed dryer.

[0150] Preferably, such a vibrational fluidised bed dryer comprises a tank containing the bulk mass of wet solid elongated elements set in vibration by special vibrating elements, into which a flow of hot air is conveyed through the bulk mass of wet solid elongated elements as they are moved by the vibration of the tank.

[0151] The use of this method results in particularly gentle and uniform drying of the particles. In further preferred embodiments, said step c) of drying the wet solid elongated elements can be carried out by means of a vacuum drying process. This drying method is based on the drastic reduction of pressure inside a drying chamber in which only wet solid elongated elements are placed, e.g. in suitable trays, thus lowering the boiling point of water. This allows moisture to evaporate from the wet solid elongated elements at lower temperatures than forced-air drying and thus has less negative effects on the viability of the microorganisms and the stability of any other heat-sensitive components. In addition, the process is faster and achieves lower residual moisture levels than conventional methods. A further positive aspect of this type of drying is that heat is transferred mainly by conduction through the heated walls of the chamber and by radiation, rather than by convection as in forced- air drying, allowing more precise temperature control and more even heat distribution.

[0152] In this preferred embodiment, step c) of drying the wet solid elongated elements can be carried out by keeping the drying chamber within which only the wet solid elongated elements are placed at an absolute pressure preferably between 20 and 50 mbar, more preferably between 25 and 45 mbar, and even more preferably between 30 and 40 mbar.

[0153] Preferably, step d) of drying said one or more wet solid elongated elements is carried out until said one or more dried solid elongated elements reach a moisture content between 4% and 18% by weight, more preferably between 6% and 15% by weight, and even more preferably between 8% and 12% by weight, of the total weight of said dry solid elongated elements.

[0154] In this way, it is advantageously possible to maintain a low level of residual water in the dry particles that trap microorganisms, which has a positive effect on their viability.

[0155] Preferably, step e) of grinding said one or more dried solid elongated elements is implemented by exerting mechanical compression on the dried solid elongated elements. This is carried out in order to minimise the mechanical stress on microbial cells that could cause morphological, biochemical and genetic changes, resulting in lethal or sub-lethal damage and consequent loss of viability of the microorganisms.

[0156] Preferably, step e) of grinding said one or more dried solid elongated elements is implemented by means of a grinding apparatus comprising one or more pairs of grinding cylinders.

[0157] Preferably, step e) of grinding said one or more dried solid elongated elements can be implemented in at least two successive steps, a first step of coarse grinding into small, irregularly shaped pieces of a few centimetres in size, and a second step of fine grinding to obtain dry particles of the desired size.

[0158] In some preferred embodiments, it is also possible to have more than two grinding steps, depending on the degree of stress to be placed on the material being crushed in each step and the particle size to be obtained.

[0159] Preferably, the dry particles of the invention, e.g. obtained from step e) of grinding said one or more dried solid elongated elements, have an average size D50 greater than or equal to 50 pm, more preferably greater than or equal to 100 pm, even more preferably greater than or equal to 250 pm, even more preferably greater than or equal to 300 pm, even more preferably greater than or equal to 350 pm and even more preferably greater than or equal to 400 pm.

[0160] Preferably, the dry particles of the invention, for example obtained from the step e) of grinding said one or more dried solid elongated elements, have an average size D50 less than or equal to 5000 pm, more preferably less than or equal to 3000 pm, even more preferably less than or equal to 1500 pm, even more preferably less than or equal to 1000 pm, even more preferably less than or equal to 800 pm, and even more preferably less than or equal to 600 pm.

[0161] Due to this feature, the dry particles of the invention demonstrate more effective protection of the encapsulated microorganisms and a higher disintegration capacity, allowing optimised and more effective release of the microorganisms incorporated into the particles. Purely by way of example, in the case of dough preparation aids for baked food products, the dry particles preferably have an average particle size D50 greater than or equal to 50 pm, more preferably greater than or equal to 100 pm, even more preferably greater than or equal to 250 pm, even more preferably greater than or equal to 300 pm, even more preferably greater than or equal to 350 pm and even more preferably greater than or equal to 400 pm.

[0162] Preferably, the dry particles of the invention in aids for the preparation of doughs for baked food products have an average size D50 less than or equal to 1500 pm, more preferably less than or equal to 1000 pm, even more preferably less than or equal to 800 pm and even more preferably less than or equal to 600 pm.

[0163] Purely by way of example, in the case of probiotics, the dry particles preferably have an average size D50 greater than or equal to 50 pm and less than or equal to 800 pm, preferably greater than or equal to 250 pm and less than or equal to 800 pm, and in the case of mycorrhizal inocula and biocontrol agents used in agriculture, the dry particles preferably have an average size D50 greater than or equal to 500 pm and less than or equal to 3000 pm.

[0164] Preferably, the dry particles of the invention have an average size D10 between 100 pm and 320 pm, more preferably between 230 pm and 300 pm, even more preferably between 260 pm and 280 pm.

[0165] In the context of the present description and subsequent claims, the expression “average size D10” used with reference to the dry particles means the percentile diameter relative to 10% of the size distribution of the dry particles. D10 is defined as the value below which the diameter of 10% of the dry particles of the size distribution falls.

[0166] Preferably, the dry particles of the invention have an average size D90 between 580 pm and 1000 pm, more preferably between 600 pm and 850 pm, even more preferably between 620 pm and 770 pm.

[0167] In the context of the present description and subsequent claims, the expression “average size D90” used with reference to the dry particles means the percentile diameter relative to 90% of the size distribution of the dry particles. D90 is defined as the value below which the diameter of 90% of the dry particles of the size distribution falls.

[0168] Preferably, the dry particles of the invention, e.g. obtained from step e) of grinding said one or more dried solid elongated elements, have a moisture content similar to that indicated above for the dried solid elongated elements, i.e. between 4% and 18% by weight, more preferably between 6% and 15% by weight and, even more preferably, between 8% and 12% by weight, of the total weight of said dry particles.

[0169] Preferably, the dry particles of the invention, for example obtained from step e) of grinding said one or more dried solid elongated elements, have a bulk density at 20°C, evaluated on an uncompacted bulk mass of dry particles, comprised between 0.6 kg / L and 0.8 kg / L, preferably between 0.65 and 0.78 kg / L more preferably between 0.7 and 0.78 kg / L.

[0170] In the case of dough preparation aids for baked food products, the dry particles are preferably dispersed in a food-grade carrier for baked food products.

[0171] Preferably, said food-grade carrier for baked food products comprises one or more among soft wheat flour, durum wheat flour, maize flour, rice flour, spelt flour, oat flour, pea flour, chickpea flour, soy flour, rye flour, millet flour, linseed flour, sesame seed flour, sunflower seed flour, gluten, maize starch, dried sourdough, brewer's yeast, salt, or mixtures thereof.

[0172] Preferably, the aid comprises a combination of particles each containing live lactic acid bacteria belonging to a species selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, and Lactiplantibacillus plantarum.

[0173] Preferably, said aid also includes an effective amount of at least one added enzyme.

[0174] In the context of the present description and in the subsequent claims, “added enzyme” means an enzyme distinct from and additional to enzymes possibly produced exogenously by the metabolism of microorganisms - in particular, live lactic acid bacteria - present in the aid and / or in the dough to which such aid is added. This added enzyme is added to the aid during its production, before the aid is used in a dough.

[0175] In the context of the present description and subsequent claims, “effective amount of enzyme” means the smallest amount that enables the enzyme to exert its characteristic effect.

[0176] Preferably, the aid comprises an amount of said at least one enzyme comprised between 0.01 and 5 g / kg of aid.

[0177] In preferred embodiments, said at least one added enzyme is selected from xylanase, transglutaminase, cellulase, amylase, amyloglucosidase, lipase, phospholipase, asparaginase, oxidase of fungal or bacterial origin or mixtures thereof.

[0178] Preferably, said at least one added enzyme is of the hydrolysing and / or non- proteolytic type.

[0179] More preferably, said at least one added enzyme comprises a xylanase enzyme capable of hydrolysing arabinoxylans, thus producing xylose.

[0180] Arabinoxylans are branched side-chain polysaccharides built from pentose sugars - arabinose and xylose - located in the cell walls of the starchy endosperm, in bran tissues, as well as in the husk of different types of grains. By breaking the arabinoxylan chains and producing xylose, the specifically selected enzyme xylanase increases the availability of soluble, fermentable sugars in the dough.

[0181] This increased availability of sugars has a positive effect on the metabolism of live xylose-fermenting lactic acid bacteria, in particular lactic acid bacteria with a heterofermenting metabolism. Compared to the fermentation of glucose and maltose (hexose sugars), the fermentation of pentose sugars such as xylose leads to the formation of acetic acid as well as lactic acid, with positive effects on product quality and shelf life.

[0182] Being able to freely express their metabolic activity also with a competitive advantage over the yeasts used in leavening bread, live lactic acid bacteria give baked food products high quality sensory, nutritional and textural properties comparable to those observed in products made from natural sourdough.

[0183] Preferably, the additional ingredients used in dough for baked food products may include one or more of the following: food flour, salt, yeast, sugar, fat, additives, or baking aids.

[0184] As discussed above, in a preferred embodiment the dry particles of the invention comprise viable microorganisms trapped in a calcium alginate matrix obtained by gelling an alginate salt with a calcium salt, preferably calcium chloride.

[0185] Advantageously and due to the specific production methods of the process according to the invention described above, in this preferred embodiment, it is possible to limit the amount of calcium in the dry particles to very low values while achieving the desired mechanical compressive strength of the dry particles, measured as described herein, of less than 3200 g.

[0186] Preferably, an amount of calcium in the dry particles is between 0.4 and 5% by weight, preferably between 0.7 and 4% by weight, more preferably between 1.0 and 3.0% by weight of the total weight of said dry particles.

[0187] In this way, it is advantageously possible to achieve the desired mechanical compressive strength of the dry particles, measured as indicated in the present description, of less than 3200 g.

[0188] Preferably, the aid described herein finds advantageous use in the preparation of doughs for baked food products by means of a so-called “straight dough” preparation process.

[0189] In the present description and subsequent claims, the terms “straight dough method / process” and “indirect dough method / process” are used in their common meaning in the field of baked food preparation.

[0190] In particular, “straight dough method / process” refers to a method / process for preparing a dough for baked food products in which the ingredients are added all together or one after the other in sequence and immediately kneaded. On the other hand, “indirect dough method / process” refers to a method / process in which a pre-dough is made with only part of the ingredients, this dough is left to rest for a predetermined time that can range from a few minutes to a few hours, and then the remaining ingredients are added.

[0191] Brief description of the figures

[0192] In the appended drawings:

[0193] • FIG. 1 shows an image of wet solid elongated elements in filament form obtained by a preferred embodiment of the process according to the invention;

[0194] • FIG. 2 shows an image of dried solid elongated elements in filament form obtained by a preferred embodiment of the process according to the invention;

[0195] • FIG. 3 shows an image of dry particles according to the invention obtained after grinding the dried solid elongated elements in filament form of FIG. 2;

[0196] • FIGS. 4a, 4b and 4c show as many SEM images (increasing magnifications of 32X, 1000X and 4000X respectively) of the dry particles of FIG. 3;

[0197] • FIG. 5 shows a diagram of the sensory analysis to which loaves obtained using dry particles according to the invention incorporating live lactic acid bacteria (F. ross / ae), dry particles according to the prior art incorporating the same live lactic acid bacteria and reference loaves obtained without using particles incorporating live lactic acid bacteria were subjected.

[0198] Further features of the invention will be illustrated in the following examples.

[0199] EXAMPLE 1 - Preparation of dry particles comprising viable microorganisms trapped in an alginate matrix (Invention)

[0200] Preparation of wet solid elongated elements

[0201] In accordance with the process of the invention, wet solid elongated elements in filament form of a sodium alginate gel gelled with calcium chloride were prepared as illustrated below.

[0202] In a first step, 1 litre of a suspension of sodium alginate, live lactic acid bacteria and deactivated yeast was prepared by the following sequence of operations: dissolving 40 grams of sodium alginate with a viscosity of 10 to 50 mPa s (measured with a Brookfield DV plus viscometer, with SPINDLE LV-01 cylindrical probe) and 40 grams of deactivated yeast in 900 grams of sterile deionised water (final concentration of alginate and deactivated yeast in the solution 4% w / w); leaving the suspension in agitation at room temperature (25°C) for one and a half hours to allow the alginate and deactivated yeast to solubilise; adding to the suspension of alginate and deactivated yeast, 20 g of freeze- dried biomass of live lactic acid bacteria, specifically a strain of the species Furfurilactobacillus rossiae (final concentration in the suspension 2% w / w); leaving the alginate suspension, deactivated yeast and live lactic acid bacteria under stirring at room temperature (25°C) for 15 minutes to allow homogeneous dispersion of the bacterial cells in the suspension.

[0203] A suspension with a pH of 5.8 was obtained.

[0204] Table 1 below shows the quantities of components used.

[0205] Table 1 - Composition of alginate suspension, live lactic acid bacteria and deactivated yeast

[0206] Instead, the gelation bath was prepared using calcium chloride as the gelling agent by dissolving the latter in sterile deionised water in the quantities specified in Table 2 below. Table 2 - Composition of the gelation bath

[0207] In order to gel the suspension of alginate, deactivated yeast and live lactic acid bacteria, the suspension was transferred into a pressurised container (applied pressure 0.2-0.5 bar) connected to a 5 mm diameter tube through which the suspension was dispensed by means of a continuous trickle-type flow into the gelation bath. This trickle-type flow dispensing was carried out by dropping the suspension without interrupting the flow into the gelation bath.

[0208] The gelation bath was kept in agitation at room temperature (25°C) for 20-30 min to allow the alginate to cross-link. A total of 950 g of wet solid elongated elements in filament form were obtained, which were then washed with deionised water and placed lying in a tray.

[0209] The moisture content of the alginate filaments, live lactic acid bacteria and deactivated yeast was about 90 g / 100 g (90% by weight of the total weight of the solid elongated elements).

[0210] FIG. 1 illustrates the wet solid elongated elements in filament form obtained by the gelation process described above.

[0211] Drying of the wet solid elongated elements

[0212] The drying of the wet solid elongated elements was conducted in a dryer consisting of a cylindrically shaped steel drying chamber fitted with a hermetic door allowing a trolley on which the trays containing the wet filaments of alginate, live lactic acid bacteria and deactivated yeast were placed. Previously heated and dehumidified air was introduced into the drying chamber (air temperature in the chamber approx. 30-32°C and maximum relative humidity 5%). The trolley inside the drying chamber was kept in rotation about an axis of rotation perpendicular to the tray support surface for the duration of the drying step to ensure uniform drying of all the material distributed in the trays. The filaments were dried for 12 hours to obtain 97g of dried solid elongated elements having a moisture content of 10% by weight of the total weight of the dried solid elongated elements.

[0213] FIG. 2 illustrates dried solid elongated elements in filament form obtained by the drying process described above. elements

[0214] The dried filaments of alginate, live lactic acid bacteria and deactivated yeast were subjected to grinding.

[0215] In particular, an initial coarse grinding (pre-grinding) was carried out in which the filaments were passed between two granite grinding cylinders placed at an adjustable distance, resulting in dry particles with a diameter of approximately 5000 pm.

[0216] Subsequently, the dry particles obtained in the pre-grinding step were further grinded by passing them several times through two granite grinding cylinders (refining step), until the desired size was obtained (average size D50 comprised between 50 pm and 2000 pm).

[0217] Dry particles were thus obtained, comprising live lactic acid bacteria and deactivated yeast trapped in an alginate matrix, the characteristics of which will be described below.

[0218] FIG. 3 illustrates dry particles of alginate, live lactic acid bacteria and deactivated yeast obtained after grinding.

[0219] The dry particles thus obtained were subjected to some analyses using a scanning electron microscope in order to characterise their morphology. Specifically, the dry particles were deposited on a copper grid coated with a carbon film (400 mesh) and observed by SEM (Nova 600i, FEI Italy). The images were collected using a voltage of 10.0 kV and current of 0.54 A. The samples were inspected at different magnifications to appreciate the surface characteristics of the dry particles.

[0220] FIGS. 4a, 4b and 4c show some SEM images of the dry particles obtained as described above at increasing magnifications. In particular, FIG. 4a shows the dry particles at 32X magnification, FIG. 4b shows the dry particles at 1000X magnification and FIG. 4c shows the dry particles at 4000X magnification.

[0221] As can be seen from FIG. 4a, the larger dry particles constituting the sample have an irregular shape and very pronounced angles; in the Applicant’s opinion, this morphology of the dry particles is quite characteristic and can be attributed to the grinding process the filaments undergo after drying.

[0222] As can be seen from FIGS. 4b and 4c, there are smooth and porous areas on the surface of the dry particles (FIG. 4b) and in the cavities of the porous areas, smaller dry particles with an ovoid, slightly elongated shape are present (FIG. 4c). In the Applicant’s opinion, these smaller dry particles can be traced back to live lactic acid bacteria trapped in the alginate matrix (F. ross / ae which is characterised by a small stick-shaped morphology).

[0223] In the Applicant’s opinion, the portion of smooth surface area represents the intact calcium alginate coating beneath which the bacterial cells are embedded.

[0224] EXAMPLE 2 - Preparation of dry particles comprising viable microorganisms trapped in an alginate matrix (Invention)

[0225] In accordance with the preparation methods described in Example 1 above, dry particles were prepared incorporating live lactic acid bacteria of the species Furfurilactobacillus rossiae (RO) but without the presence of deactivated yeast.

[0226] Dry particles similar in every respect to those in Example 1 were obtained.

[0227] EXAMPLE 3 - Production of dry alginate microcapsules incorporating viable microorganisms (Comparative) In accordance with the process disclosed in Example 1 of international patent application WO 2024 / 009249 A1 , dry microcapsules incorporating freeze-dried lactic acid bacteria belonging to the species Furfurilactobacillus rossiae (RO) were prepared.

[0228] For droplet micronization, a final suspension of alginate and live lactic acid bacteria was prepared, with the composition shown in Table 3 below.

[0229] In this case, deactivated yeast could not be used due to the clogging of the dispensing nozzles of the alginate suspension and live lactic acid bacteria.

[0230] Table 3 - Composition of gelling biopolymer solution and live lactic acid bacteria for subsequent droplet micronization

[0231] The resulting suspension of gelling biopolymer and live lactic acid bacteria had a pH of 5.8.

[0232] This suspension was then sieved with an 80 pm sieve to remove any unwanted aggregates and then extruded into droplets using a vibrating nozzle with an outlet diameter of 350 pm (500 Hz, 2750 mV, 150 mBar), dropping the droplets into a gelation bath containing divalent ions from calcium chloride and polyethylene glycol of an average molecular weight of 1500 (optional ingredient), in the quantities specified in Table 4.

[0233] Table 4 - Composition of the gelation bath

[0234] The droplets were left in the gelation bath, resulting in gelled droplets with an average size comprised between 800 pm and 1000 pm.

[0235] The resulting gelled drops were washed with a 0.85% NaCI solution and then treated with SiO2 anti-caking agent.

[0236] Finally, the gelled drops were dried in a forced circulation drying oven by setting an air temperature of 30°C, until a relative humidity of the microcapsules of 12% to 15.5% was reached, resulting in dry microcapsules containing Furfurilactobacillus rossiae.

[0237] EXAMPLE 4 - Compression test of dry microcapsules containing live lactic acid bacteria

[0238] The compressive strength of the dry particles obtained according to Examples 1 and 2 of the invention and, respectively, according to Comparative Example 3, was measured using a texture analyser (Perten TVT 6700 Texture Analyzer, with compression probe No. 670159 with a diameter of 21 mm) according to the following process.

[0239] A volume of sample (approximately 6 g of sample) was placed in a cylindrical stainless steel container with an internal diameter of 40 mm and a height of 20 mm.

[0240] The compression probe was positioned 5 mm above the maximum level of the sample.

[0241] The vertical descent speed of the probe was set to 1 .7 mm / s and a minimum force of 5g was set for the start of the measurement. The downward movement of the probe was stopped upon reaching 40% of the initial height measured at the beginning of the measurement.

[0242] For each type of sample, particles or dry microcapsules, a force-time curve was obtained and maximum force peaks calculated. For each sample of microcapsules, the compressive strength was then evaluated as the peak force recorded in these temporal force trends.

[0243] The results of the compressive strength measurement, as an average of 3 measurements, are shown in Table 5.

[0244] Table 5 - Compression test

[0245] Compressive strength tests carried out show that dry particles obtained by grinding dried solid elongated elements according to the invention (Examples 1 and 2) exhibit lower mechanical strength, which facilitates the opening of the alginate matrix during use of the dry particles, for example during dough preparation steps for baked food products, with more effective release and distribution of the live lactic acid bacteria incorporated in the dry particles.

[0246] Tests have also shown an unexpected further decrease in the mechanical compressive strength of the dry particles containing deactivated yeast with respect to both the dry microcapsules of the prior art and the dry particles without deactivated yeast according to the invention, which further decrease in mechanical compressive strength could result from a further unexpected positive effect of the deactivated yeast on the breakability of the product.

[0247] EXAMPLE 5 - Characterization of dry particles containing live lactic acid bacteria

[0248] The dry particles obtained according to Examples 1 and 2 of the invention and, respectively, according to Comparative Example 3, were characterised by assessing their size distribution and the amount of calcium, two parameters that the Applicant considers significant for characterising the performance of the dry particles in use.

[0249] Size distribution

[0250] The size distribution of the samples was carried out using the ISO 13320-1 :2020 method. The results of these analyses are shown in Table 6 where:

[0251] - D10 is the value below which the 10% diameter of the size distribution of dry particles / microcapsules falls;

[0252] - D50 is the value below which the diameter of 50% of the size distribution of the dry particles / microcapsules falls; and

[0253] - D90 is the value below which the diameter of 90% of the size distribution of dry particles / microcapsules falls.

[0254] Table 6. - Results of laser particle size analysis The dry particles according to the invention examined in this particle size analysis test are distributed in larger size intervals than the dry particles obtained from conventional capsules. In this respect, it should be noted that the size of the dry particles according to the invention can be advantageously altered according to the use requirements by adjusting the parameters of the grinding step (e.g. by bringing the refining rolls closer together or moving them farther apart, or by repeated passes between the rolls), which can lead to different sizes in relation to the different scope of application of the dry particles.

[0255] Ca content

[0256] The calcium concentrations in the dry particles of Examples 1 and 2 according to the invention and in the dry microcapsules of Example 3 according to the prior art were assessed by ICP (plasma emission spectrometry) using the AOAC test method 2011 .14 “Calcium, Copper, Iron, Magnesium, Manganese, Potassium, Phosphorus, Sodium, and Zinc in Fortified Food Products. Microwave Digestion and Inductively Coupled Plasma-Optical Emission Spectrometry”.

[0257] The results obtained are reported in Table 7.

[0258] Table 7 - Calcium content of dry particles / microcapsules

[0259] From the above data, it can be seen that the dry particles according to the invention contain significantly less calcium than the dry microcapsules according to the prior art. Without wishing to bind itself herein to any interpretative theory, the Applicant considers that the aforementioned reduced amount of calcium in the dry particles according to the invention has an important effect in reducing the compressive strength of the dry particles themselves due to the creation of a less dense and compact matrix of alginate which more easily releases the microorganisms trapped in that matrix during use of the dry particles.

[0260] In this respect, the Applicant considers that the process for producing dry particles according to the invention has a strong positive impact on the control of the calcium concentration present in the alginate matrix of the dry particles both between dry particles of the same production batch and between dry particles of different production batches.

[0261] In fact, the formation of the solid elongated elements within the process for preparing dry particles according to the invention is very rapid and results in the dwell time of the solid elongated elements in the gelation bath being very similar between different parts of the solid elongated elements or between different solid elongated elements.

[0262] Conversely, and as explained above, the known gelling process involves the formation of microspheres that have very different dwell times in the gelation bath depending on whether the microspheres were produced at the beginning or end of the production batch.

[0263] As a result, microspheres obtained by the much slower known gelling process not only have a higher calcium content in absolute terms, but are also inhomogeneous within the batch with obvious negative consequences on the performance of the microspheres in use.

[0264] Example 6 - Evaluation of the viability of the microorganisms incorporated into the dry particles

[0265] The viability of live lactic acid bacteria incorporated into the dry particles obtained according to Examples 1 and 2 of the invention and, respectively, encapsulated into the dry microspheres according to the comparative Example 3, was measured by means of viable plate counts at time TO and time T3 (3 months) in sample aliquots maintained under stress conditions i.e. at a temperature of 30°C and uncontrolled humidity. The following samples were analysed:

[0266] - dry particles according to the invention containing live lactic acid bacteria and deactivated yeast embedded in an alginate matrix (Example 1 ),

[0267] - dry particles according to the invention containing live lactic acid bacteria embedded in an alginate matrix without deactivated yeast (Example 2),

[0268] - dry alginate microspheres incorporating live lactic acid bacteria obtained by spherification without deactivated yeast (Comparative Example 3). The method and results of counting Furfurilactobacillus rossiae are reported below:

[0269] - sample preparation: first suspension in phosphate solution (4.45 g / l Na2HPO4 x 2H2O+ 1.5 g / l KH2PO4), subsequent dilutions in maximum recovery diluent,

[0270] - culture medium: unacidified MRS agar, - incubation: 30°C for 72 h.

[0271] The results obtained are reported in Table 8 below.

[0272] Table 8 - Count of F. rossiae at TO and T3. Alog = Log cfu / g T3 - Log cfu / g TO From the data shown in the table, it can be seen that at time TO there was a viable cell count two orders of magnitude higher in the dry particles according to the invention (Examples 1 and 2) than in the dry microcapsules obtained in accordance with the prior art (Comparative Example 3).

[0273] In this regard, but without wishing to bind itself herein to any interpretative theory, the Applicant believes that the increased breakability of the dry particles according to the invention may play a positive role in making microorganisms incorporated in the alginate matrix more readily available with an ease of opening of the dry particles resulting in increased performance of the dry particles in the matrices or substrates of use.

[0274] Furthermore, the presence of deactivated yeast in the dry particles according to Example 1 of the invention effectively contributed to maintaining the viability of the microorganisms trapped in the alginate matrix over time, with a cfu count as much as three orders of magnitude higher than that of the dry microcapsules obtained in accordance with the prior art.

[0275] Example 7 - Evaluation of productivity and production yield of dry parti cl es / m i crocapsu I es

[0276] The productivity and production yields of the dry particle production processes described in Examples 1 and 2 according to the invention and, respectively, in the comparative Example 3 according to the prior art, were evaluated by means of production trials of pilot batches of the order of about 12 kg in the manner illustrated in the aforementioned Examples.

[0277] In the tests performed according to Comparative Example 3, droplet micronization of the microorganism suspension in alginate was carried out using nozzles of different diameter or number.

[0278] From the tests performed, it was found that the process according to the invention, which does not involve any micronization into droplets of the suspension of microorganisms and alginate, achieves an hourly output of up to about 50 kg / h of processed suspension, while the process according to the prior art achieved a variable maximum hourly output between about 1.5 kg / h and about 7 kg / h, depending on whether a single nozzle or a plate with multiple nozzles is used.

[0279] The results of the tests performed are shown in Table 9 below.

[0280] Table 9 - Productivity comparison between the process according to the invention and according to the prior art

[0281] Key: LD=deactivated yeast

[0282] The dry yield was calculated as a % of final dry product to wet product weight

[0283] From the data shown in the table, it can be seen that the process according to the invention, in its variants with or without deactivated yeast (the latter basically not dispensable with the nozzles of the process according to the prior art, which tend to clog after a short time), achieves both a significantly higher hourly output and dry yield than can be achieved with the process of the prior art.

[0284] EXAMPLE 8 (invention) - Baking tests

[0285] By means of the process described in Example 1 above, dry particles were produced using a single species of live lactic acid bacteria (F. rossiae) both with deactivated yeast, Example 1A, and without deactivated yeast, Example 2A.

[0286] Baking tests were then carried out in comparison with a traditional dough without the use of F. rossiae, Comparative Example 4.

[0287] In particular, four doughs were prepared, referred to as doughs 1A and 2A (invention), 3 (comparative) and 4 (reference) using a straight dough method, mixing the ingredients in a single step. These ingredients were as follows:

[0288] - Dough 1 A: 3000g flour, 39g dry yeast and 30g aid containing dry particles incorporating F. rossiae with deactivated yeast obtained according to Example 1 ,

[0289] - Dough 2A: 3000g flour, 39g dry yeast and 30g aid containing dry particles incorporating F. rossiae without deactivated yeast obtained according to Example 2,

[0290] - Dough 3: 3000g flour, 39g dry yeast and 30g aid microcapsules incorporating F. rossiae obtained in accordance with Comparative Example 3,

[0291] - Dough 4: 3000g flour and 39g dry yeast (reference).

[0292] The aid of doughs 1 A, 2A and 3 had the following composition: 0.03% dry particles incorporating F. rossiae and 0.035% of the enzyme Xylanase (Panzea®, Novozyme).

[0293] For each dough 1800 ml water and all the above ingredients, except salt, were mixed in a spiral mixer (VMI BERTO ITALIA SRL model 80 MAG 2V) at speed 1 for 5 minutes, followed by 8 minutes at speed 2. As soon as the dough became smooth, the remaining water (450 g) and salt were added. The dough was then left to rest at 26°C for 45 minutes.

[0294] After this step, each dough was divided into 100 g pieces that were left to rise for 60 minutes at 28°C and 80% RH and then baked for 18 minutes in a bridge oven at 210°C. Finally, the loaves were cooled to room temperature before being evaluated with sensory analysis.

[0295] A total of 40 loaves were produced for each dough. The bread obtained with these recipes and this process is called “Zoccoletto”, a type of bread traditionally characterised by large cavities in the crumb and a very crisp, golden and fragrant crust.

[0296] The breads obtained in the baking tests were evaluated on the basis of colour, texture, aroma and taste by a panel of six judges trained to recognise bread quality parameters. For each bread sample, the judges assigned a score on a 10-point hedonic scale taking into account sensory characteristics of the different breads such as the colour of the crust, the presence of surface fractures, the increase in volume in width and length compared to the original dough, the alveolation of the crumb, the fragrance, the toasted aroma of the crust, the lactic notes, the acidity, the overall flavour, the sweetness, the softness of the crumb during chewing, and the crunchiness of the crust.

[0297] The results of the sensory analysis are shown in Table 10 and FIG. 5. For each sensory parameter and each bread, the table shows the average of the scores awarded by each judge.

[0298] Table 10 - Sensory analysis | DOUGH 4 | 4.3 | 3.3 | 3 | 4 | 3 | 3.5 | 1.5 | 3 | 3.3 | 2.6 | 3.5 | 2.9 |

[0299] From the data in the table and Fig. 5, it is inferred that the loaves obtained using the dry particles according to the invention (Examples 1 and 2) exhibit better sensory characteristics than breads obtained using the dry particles according to the prior art (Comparative Example 3) and significantly better sensory characteristics than breads obtained from the reference dough without lactic acid bacteria.

[0300] Furthermore, in all the parameters tested, albeit with different intensities, the loaves obtained using the dry particles according to the invention with deactivated yeast presented higher scores than the loaves obtained from a dough with dry particles without deactivated yeast.

[0301] Naturally, those skilled in the art may make modifications and variants to the abovedescribed invention with the purpose of meeting specific and contingent application needs, variants and modifications in any case falling within the scope of protection as defined by the successive claims.

[0302] Thus, for example, the process of the invention allows the preparation of dry particles comprising viable microorganisms trapped in an alginate matrix that may find useful and advantageous use in a variety of fields of application.

[0303] This is due to the versatility of the process and the ability of dry particles to significantly improve the stability and effectiveness of microorganisms trapped in an alginate matrix in various applications. This results in higher quality products, longer shelf life and potential reduction of production costs in various industries.

[0304] Purely, by way of example, the possible areas of use for the dry particles of the invention are shown in Table 11 below.

[0305] Table 11 - Potential industrial applications of the dry particles of the invention

Claims

CLAIMS1. Process for the preparation of particles comprising viable microorganisms trapped in an alginate matrix, comprising: a) providing a suspension including an alginate salt, preferably sodium alginate, having a viscosity between 5 and 50 mPa s, more preferably between 8 and 40 mPa s, more preferably between 10 and 20 mPa s, when dissolved in a 1 % by weight aqueous solution at a temperature of 20°C, and viable microorganisms; b) forming in the gelation bath one or more wet solid elongated elements comprising viable microorganisms trapped in an alginate matrix by dispensing a continuous or intermittent trickle-type flow of said suspension in a gelation bath including at least one gelling agent, c) separating said one or more wet solid elongated elements from the gelation bath; d) drying said one or more wet solid elongated elements so as to obtain one or more dried solid elongated elements; e) grinding said one or more dried solid elongated elements so as to obtain dry particles comprising viable microorganisms trapped in an alginate matrix, said dry particles having a mechanical compressive strength, measured as set forth in the present description, greater than or equal to 500 g and less than 3200 g, preferably greater than or equal to 1500 g and less than or equal to 3000 g, more preferably greater than or equal to 1800 g and less than or equal to 2500 g.

2. Process according to claim 1 , wherein a concentration of said alginate salt, preferably of said sodium alginate, in said suspension is comprised between 1 % and 8% by weight, preferably between 1 % and 6% by weight, more preferably between 2% and 5% by weight of the total weight of the suspension.

3. Process according to claim 1 or 2, wherein a concentration of viable microorganisms in said suspension is comprised between 0.5% and 4% by weight, preferably between 0.5% and 3% by weight, more preferably between 1.0% and 2.5% by weight of the total weight of the suspension.

4. Process according to any one of the preceding claims, wherein said suspension has a pH between 4 and 7, preferably between 4.5 and 6, more preferably between 5 and 5.5.

5. Process according to any one of the preceding claims, wherein a) providing said suspension comprises keeping said suspension at a temperature comprised between 15°C and 35°C, preferably between 18°C and 30°C, more preferably between 20°C and 25°C.

6. Process according to any one of the preceding claims, wherein said viable microorganisms comprise live lactic acid bacteria belonging to at least one species and wherein said suspension preferably includes deactivated yeast.

7. Process according to claim 6, wherein a concentration of said deactivated yeast in said suspension is comprised between 1 % and 8% by weight, preferably between 1 % and 6% by weight, more preferably between 2% and 5% by weight, of the total weight of the suspension.

8. Process according to any one of the preceding claims, wherein a) providing said suspension comprises keeping the suspension under stirring preferably for a time comprised between 1 h and 24h.

9. Process according to any one of the preceding claims, wherein b) forming in the gelation bath one or more wet solid elongated elements comprising viable microorganisms trapped in an alginate matrix by dispensing a continuous or intermittent trickle-type flow of said suspension into said gelation bath including at least one gelling agent comprising feeding said suspension into said gelation bath from a storage vessel by gravity or by dispensing at a predetermined pressure from one or more pipelines or dispensing ports.

10. Process according to any one of the preceding claims, further comprising f) maintaining said one or more elongated elements in said suspension, preferably kept under stirring, for a time comprised between 5 and 30 minutes, preferably between 10 and 20 minutes, more preferably between 12 and 17 minutes.

11. Process according to any one of the preceding claims, wherein said wet solid elongated elements have a moisture content comprised between 80% and 97% by weight, preferably between 85% and 95% by weight, more preferably, between 92% and 95% by weight, of the total weight of said wet solid elongated elements.

12. Process according to any one of the preceding claims, wherein d) drying said one or more wet solid elongated elements is carried out at a temperature preferably comprised between 20°C and 45°C, preferably between 20°C and 35°C and, more preferably, between 25°C and 30°C, until said one or more dried solid elongated elements achieve a moisture content comprised between 4% and 18% by weight, preferably between 6% and 15% by weight, more preferably between 8% and 12% by weight, of the total weight of said dried solid elongated elements.

13. Process according to any one of the preceding claims, wherein e) grinding said one or more dried solid elongated elements is carried out by means of a grinding apparatus comprising one or more pairs of grinding cylinders.

14. Dry particles comprising viable microorganisms trapped in an alginate matrix, wherein said dry particles have a mechanical compressive strength, measured as set forth in the present description, greater than or equal to 500 g and less than 3200 g, preferably greater than or equal to 1500 g and less than or equal to 3000 g, more preferably greater than or equal to 1800 g and less than or equal to 2500 g.

15. Dry particles according to claim 14, wherein said dry particles have an average size D50 comprised between 50 pm and 5000 pm, preferably between 250 pm and 5000 pm, more preferably between 300 pm and 3000 pm, even more preferably between 350 pm and 1000 pm, even more preferably between 400 pm and 600 pm and a moisture content comprised between 4% and 18% by weight, preferably between 6% and 15% by weight, more preferably between 8% and 12% by weight of the total weight of said dry particles.

16. Dry particles according to claim 14 or 15, wherein said dry particles have an apparent density, evaluated on an uncompacted bulk mass of dry particles, comprised between 0.6 kg / L and 0.8 g / ml, preferably between 0.65 and 0.78 kg / L, more preferably between 0.7 and 0.78 kg / L.

17. Dry particles according to any one of claims 14-16, wherein said viable microorganisms are one or more of: live lactic acid bacteria belonging to at least one species, preferably of the family of Lactobacillaceae, Streptococcaceae, Enterococcaceae, Carnobacteriaceae, microrganismi del genere Rhizobium, microorganisms of the genus Bifidobacterium, yeasts such as Saccharomyces ssp, non-Saccharomyces ssp, Kluyveromyces spp, spore-forming microorganisms, such as Bacillus spp, Weizmannia spp, new generation probiotics, such as Akkermansia spp, Faecalibacterium spp, Clostridium spp. Bacteroides spp, Roseburia spp., non- toxigenic molds of the genus Aspergillus, mycorrhizal fungi, fungi of the genus Trichoderma.

18. Dry particles according to claim 17, wherein said microorganisms are live lactic acid bacteria belonging to one or more species selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, Lactiplantibacillus plantarum, Companilactobacillus alimentarius, Levilactobacillus brevis, Lentilactobacillus buchneri, Latilactobacillus curvatus, Lactobacillus delbrueckii, Limosilactobacillus fermentum, Limosilactobacillus frumenti, Levilactobacillus hammesii, Lactobacillus helveticus, Lentilactobacillus hilgardii, Limosilactobacillus panis, Companilactobacillus paralimentarius, Lacticaseibacillus paracasei, Lactiplantibacillus paraplantarum, Lactiplantibacillus pentosus, Limosilactobacillus pontis, Limosilactobacillus reuteri, Latilactobacillus sakei, Lactococcus lactis, Leuconostoc citreum, Leuconostoc mesenteroides, Pediococcus pentosaceus, Weissella cibaria, and Weissella confusa, preferably selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, and Lactiplantibacillus plantarum.

19. Dry particles according to claim 17 or 18, wherein said viable microorganisms comprise one or more live lactic acid bacteria belonging to at least one species and wherein said particles preferably include deactivated yeast.

20. Dry particles according to any one of claims 18 or 19, wherein the particles contain a combination of live lactic acid bacteria belonging to the species Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, andLactiplantibacillus plantarum.

21. Dry particles according to any one of claims 14-20, comprising viable microorganisms trapped in a calcium alginate matrix, wherein an amount of calcium in the dry particles is comprised between 0.4% and 5% by weight, preferably between 0.7% and 4% by weight, more preferably between 1.0% and 3.0% by weight of the total weight of said dry particles.

22. Use of dry particles according to any of claims 14-21 in an aid for the preparation of doughs for baked food products.

23. Aid for the preparation of doughs for baked food products, comprising a foodgrade carrier for baked food products and dry particles according to any one of claims 14-21 containing live lactic acid bacteria belonging to at least one species according to any one of claims 18-21 , wherein said dry particles are dispersed in said food-grade carrier and have an average size D50 comprised between 50 pm and 1500 pm, preferably between 250 pm and 1500 pm, more preferably between 300 pm and 1000 pm, even more preferably between 350 pm and 800 pm and, even more preferably, between 400 pm and 600 pm.

24. Aid according to claim 23, wherein said food-grade carrier for baked food products comprises one or more of soft wheat flour, durum wheat flour, maize flour, rice flour, spelt flour, oat flour, pea flour, chickpea flour, soy flour, rye flour, millet flour, linseed flour, sesame seed flour, sunflower seed flour, gluten, maize starch, dried sourdough, baker’s yeast, salt, or mixtures thereof.

25. Aid according to any one of claims 23 or 24, wherein the aid comprises a combination of dry particles containing, each, live lactic acid bacteria belonging to a species selected from Fructilactobacillus sanfranciscensis, Furfurilactobacillus rossiae, and Lactiplantibacillus plantarum.

26. Aid according to any one of claims 23-25, further comprising an effective amount of at least one added enzyme, preferably of the hydrolysing and / or non-proteolytic type, said effective amount being preferably comprised between 0.01 and 5 g / kg of aid.

27. Aid according to claim 26, wherein said at least one added enzyme is selected from xylanase, transglutaminase, cellulase, amylase, amyloglucosidase, lipase, phospholipase, asparaginase, oxidase of fungal or bacterial origin, or mixtures thereof.

28. Use of an aid according to any one of claims 23-27, in the preparation of doughs for baked food products by means of a straight dough preparation process.

29. Process for producing an aid for the preparation of baked food products comprising dispersing dry particles according to any one of claims 14-21 containing live lactic acid bacteria belonging to at least one species according to any one of claims 18-21 in an amount of a food-grade carrier for baked food products.

30. Dough for baked food products, comprising dry particles according to any one of claims 14-21 containing live lactic acid bacteria belonging to at least one species according to any one of claims 18-21 , water and optionally further ingredients.31 . Process for the preparation of a baked product, preferably a straight dough process, comprising:- forming a dough by mixing dry particles according to any one of claims 14-21 containing live lactic acid bacteria belonging to at least one species according to any one of claims 18-21 with water and optionally further ingredients;- optionally, dividing the dough into portions;- optionally, giving the dough or portions of dough a desired shape; and- baking the dough or portions of dough in an oven.

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